Search results for: in silico laboratory

Authors: A. Vinod Kumar, G. Sunny Deol, Rakesh Kumar, B. Chandra

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Application of fiber reinforcement in road construction is gaining some interest in enhancing soil strength. In this paper, the natural Geotextile material obtained from gunny bags was used due to vast local availability material. Construction of flexible pavement on weaker soil such as clay soils are a significant problem in construction as well as in design due to its expansive characteristics. Jute Geotextile (JGT) was used on a foundation layer of flexible pavement on rural roads. This problem will be conquered by increasing the subgrade strength by decreasing sub-base layer thickness by improving their overall pavement strength characteristics which ultimately reduces the cost of construction and leads to economically design. The California Bearing Ratio (CBR), unconfined compressive strength (UCS) and triaxial laboratory tests were conducted on two different soil samples CI and MI. Weaker soil is reinforced with JGT, JGT+Bitumen; JGT+polythene sheet was varied with heights while performing the laboratory tests. Subgrade strength evaluation was investigated by conducting soak CBR test in the laboratory for clayey and silt soils. Laboratory results reveal that reinforced soak CBR value of clayey soil (CI) observed was 10.35%, and silty soil (MI) was 15.6%. This study intends to develop new technique for reinforcing weaker soil with JGT varying parameters for the need of low volume flexible pavements. It was observed that the performance of JGT is inferior when used with bitumen and polyethylene sheets.

Keywords: CBR, Jute geotextile, low volume road, weaker soil

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Authors: Sadaf Moeez, Madiha Khalid, Zoya Khalid, Sania Shaheen, Sumbul Khalid

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Background: Inter-individual variation in response to metformin, which has been considered as a first line therapy for T2DM treatment is considerable. In the current study, it was aimed to investigate the impact of two genetic variants Leu125Phe (rs77474263) and Gly64Asp (rs77630697) in gene SLC47A1 on the clinical efficacy of metformin in T2DM Pakistani patients. Methods: The study included 800 T2DM patients (400 metformin responders and 400 metformin non-responders) along with 400 ethnically matched healthy individuals. The genotypes were determined by allele-specific polymerase chain reaction. In-silico analysis was done to confirm the effect of the two SNPs on the structure of genes. Association was statistically determined using SPSS software. Results: Minor allele frequency for rs77474263 and rs77630697 was 0.13 and 0.12. For SLC47A1 rs77474263 the homozygotes of one mutant allele ‘T’ (CT) of rs77474263 variant were fewer in metformin responders than metformin non-responders (29.2% vs. 35.5 %). Likewise, the efficacy was further reduced (7.2% vs. 4.0 %) in homozygotes of two copies of ‘T’ allele (TT). Remarkably, T2DM cases with two copies of allele ‘C’ (CC) had 2.11 times more probability to respond towards metformin monotherapy. For SLC47A1 rs77630697 the homozygotes of one mutant allele ‘A’ (GA) of rs77630697 variant were fewer in metformin responders than metformin non-responders (33.5% vs. 43.0 %). Likewise, the efficacy was further reduced (8.5% vs. 4.5%) in homozygotes of two copies of ‘A’ allele (AA). Remarkably, T2DM cases with two copies of allele ‘G’ (GG) had 2.41 times more probability to respond towards metformin monotherapy. In-silico analysis revealed that these two variants affect the structure and stability of their corresponding proteins. Conclusion: The present data suggest that SLC47A1 Leu125Phe (rs77474263) and Gly64Asp (rs77630697) polymorphisms were associated with the therapeutic response of metformin in T2DM patients of Pakistan.

Keywords: diabetes, T2DM, SLC47A1, Pakistan, polymorphism

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Authors: Safee Chaudhary, Mahnoor Naseer Gondal, Hira Anees Awan, Abdul Rehman, Ammar Arif, Risham Hussain, Huma Khawar, Zainab Arshad, Muhammad Faizyab Ali Chaudhary, Waleed Ahmed, Muhammad Umer Sultan, Bibi Amina, Salaar Khan, Muhammad Moaz Ahmad, Osama Shiraz Shah, Hadia Hameed, Muhammad Farooq Ahmad Butt, Muhammad Ahmad, Sameer Ahmed, Fayyaz Ahmed, Omer Ishaq, Waqar Nabi, Wim Vanderbauwhede, Bilal Wajid, Huma Shehwana, Muhammad Tariq, Amir Faisal

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Cancer is a manifestation of multifactorial deregulations in biomolecular pathways. These deregulations arise from the complex multi-scale interplay between cellular and extracellular factors. Such multifactorial aberrations at gene, protein, and extracellular scales need to be investigated systematically towards decoding the underlying mechanisms and orchestrating therapeutic interventions for patient treatment. In this work, we propose ‘TISON’, a next-generation web-based multiscale modeling platform for clinical systems oncology. TISON’s unique modeling abstraction allows a seamless coupling of information from biomolecular networks, cell decision circuits, extra-cellular environments, and tissue geometries. The platform can undertake multiscale sensitivity analysis towards in silico biomarker identification and drug evaluation on cellular phenotypes in user-defined tissue geometries. Furthermore, integration of cancer expression databases such as The Cancer Genome Atlas (TCGA) and Human Proteome Atlas (HPA) facilitates in the development of personalized therapeutics. TISON is the next-evolution of multiscale cancer modeling and simulation platforms and provides a ‘zero-code’ model development, simulation, and analysis environment for application in clinical settings.

Keywords: systems oncology, cancer systems biology, cancer therapeutics, personalized therapeutics, cancer modelling

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Authors: Zunnu Raen Akhtar, U. Irshad, M. Majid

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Due to the widespread cultivation of transgenic cotton, intense selection pressure resulted in resistant allele in pink bollworm, Pectinophora gossypiella (Gelechiidae: Lepidoptera). A resistant strain of pink bollworm against transgenic cotton has become a challenge to Integrated Resistance Management (IRM) in the World. Laboratory and field studies were conducted to determine the resistant strains of pink bollworm by performing bioassay, extracting the DNA, conducting PCR of both laboratory as well as field collected pink bollworms to observe the developed resistance. In all of the studies, two Bt varieties FH-142 and FH-118 expressing Cry1Ac compared to non-Bt (Control) were tested against pink bollworm. In the laboratory, bioassay results showed that there was no significant mortality difference between Bt and non-Bt varieties. Similar mortality percentage was observed in transgenic and non-transgenic (control) variety. Insects which were survived after bioassay, as well as those collected from the Bt cotton fields, were selected for further molecular studies. DNA extraction followed by PCR was conducted to check the resistant strains in pink bollworm. In field studies, we also observed the population dynamics of pink boll worms on Bt as compared to non-Bt varieties. Laboratory and field studies confirmed that resistant strains occurs in Pakistani Bt cotton fields. Different strategies should be adopted to combat that serious prevailing resistance issues.

Keywords: transgenic cotton, resistance, pectinophora gossypiella, , integrated resistance management (IRM), polymerase chain reaction (PCR)

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Authors: M. Patterson, R. Bond, K. Cowan, M. Mulvenna, C. Reid, F. McMahon, P. McGowan, H. Cormican

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This paper outlines the design of a simulator to allow for the optimisation of clinical workflows through a pathology laboratory and to improve the laboratory’s efficiency in the processing, testing, and analysis of specimens. Often pathologists have difficulty in pinpointing and anticipating issues in the clinical workflow until tests are running late or in error. It can be difficult to pinpoint the cause and even more difficult to predict any issues which may arise. For example, they often have no indication of how many samples are going to be delivered to the laboratory that day or at a given hour. If we could model scenarios using past information and known variables, it would be possible for pathology laboratories to initiate resource preparations, e.g. the printing of specimen labels or to activate a sufficient number of technicians. This would expedite the clinical workload, clinical processes and improve the overall efficiency of the laboratory. The simulator design visualises the workflow of the laboratory, i.e. the clinical tests being ordered, the specimens arriving, current tests being performed, results being validated and reports being issued. The simulator depicts the movement of specimens through this process, as well as the number of specimens at each stage. This movement is visualised using an animated flow diagram that is updated in real time. A traffic light colour-coding system will be used to indicate the level of flow through each stage (green for normal flow, orange for slow flow, and red for critical flow). This would allow pathologists to clearly see where there are issues and bottlenecks in the process. Graphs would also be used to indicate the status of specimens at each stage of the process. For example, a graph could show the percentage of specimen tests that are on time, potentially late, running late and in error. Clicking on potentially late samples will display more detailed information about those samples, the tests that still need to be performed on them and their urgency level. This would allow any issues to be resolved quickly. In the case of potentially late samples, this could help to ensure that critically needed results are delivered on time. The simulator will be created as a single-page web application. Various web technologies will be used to create the flow diagram showing the workflow of the laboratory. JavaScript will be used to program the logic, animate the movement of samples through each of the stages and to generate the status graphs in real time. This live information will be extracted from an Oracle database. As well as being used in a real laboratory situation, the simulator could also be used for training purposes. ‘Bots’ would be used to control the flow of specimens through each step of the process. Like existing software agents technology, these bots would be configurable in order to simulate different situations, which may arise in a laboratory such as an emerging epidemic. The bots could then be turned on and off to allow trainees to complete the tasks required at that step of the process, for example validating test results.

Keywords: laboratory-process, optimization, pathology, computer simulation, workflow

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Authors: Nwadioha Samuel Iheanacho, Abah Paul, Odimayo Simidele Michael

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Background: The World Health Assembly through the Global Health Sector Strategy on viral hepatitis calls for the elimination of viral hepatitis as a public health threat by 2030. All hands are on deck to actualize this goal through an effective and active vaccination and monitoring tool. Aim: To combine the Epidemiologic with Laboratory Hepatitis B Virus vaccination monitoring tools. Method: Laboratory results analysis of subjects recruited during the World Hepatitis week from July 2020 to July 2021 was done after obtaining their epidemiologic data on Hepatitis B virus risk factors, in the Medical Microbiology Laboratory of Benue State University Teaching Hospital, Nigeria. Result: A total of 500 subjects comprising males 60.0%(n=300/500) and females 40.0%(n=200/500) were recruited. A fifty-three percent majority was of the age range of 26 to 36 years. Serologic profiles were as follows, 15.0%(n=75/500) HBsAg; 7.0% (n=35/500) HBeAg; 8.0% (n=40/500) Anti-Hbe; 20.0% (n=100/500) Anti-HBc and 38.0% (n=190/500) Anti-HBs. Immune responses to vaccination were as follows, 47.0%(n=235/500) Immune naïve {no serologic marker + normal ALT}; 33%(n=165/500) Immunity by vaccination {Anti-HBs + normal ALT}; 5%(n=25/500) Immunity to previous infection {Anti-HBs, Anti-HBc, +/- Anti-HBe + normal ALT}; 8%(n=40/500) Carriers {HBsAg, Anti-HBc, Anti-HBe +normal ALT} and 7% (35/500) Anti-HBe serum- negative infections {HBsAg, HBeAg, Anti-HBc +elevated ALT}. Conclusion: The present 33.0% immunity by vaccination coverage in Central Nigeria was much lower than the 41.0% national peak in 2013, and a far cry from the global expectation of attainment of a Global Health Sector Strategy on the elimination of viral hepatitis as a public health threat by 2030. Therefore, more creative ideas and collective effort are needed to attain this goal of the World Health Assembly.

Keywords: Hepatitis B, vaccination status, laboratory tools, resource-limited settings

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Authors: Nicolas Spitz, Nicolas Coniglio, Mohamed El Mansori, Alex Montagne, Sabeur Mezghani

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Nowadays buildings' construction is performed by pouring concrete into molds referred to as formworks that are usually prefabricated metallic modules. Defects such as stripping may possibly form during the removal of the formwork if the interfacial bonding between the concrete and the formwork is high. A new pull-off tensile test was developed in our laboratory to simulate small-scale formwork removals. The concrete-to-formwork adherence force was measured on bare and coated formworks with different surface signatures. The used concrete was a mixture largely used on building sites and contains CEM I Portland cement and calcareous filler. The concrete surface appearance and the type of failures at the concrete-formwork interface have been investigated. The originality of this near-to-surface test was to compare the laboratory-measured adherence forces to the on-site observations. Based upon the small-scale laboratory test results, functional formwork specifications with low adherence to concrete was proposed in terms of superficial signature characteristics.

Keywords: concrete-formwork adherence, interfacial bonding, skin formwork functionality, small-scale pull-off tensile test

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Authors: Maria Konstadinou, Etienne Alderlieste, Anderson Peccin da Silva, Ben Arntz, Leonard van der Bijl, Wouter Verschueren

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The shear strength and compression properties of stiff Boom clay from Belgium at the depth of about 30 m has been investigated by means of cone penetration and laboratory testing. The latter consisted of index classification, constant rate of strain, direct, simple shear, and unconfined compression tests. The Boom clay samples exhibited strong swelling tendencies. The suction pressure was measured via different procedures and has been compared to the expected in-situ stress. The undrained shear strength and OCR profile determined from CPTs is not compatible with the experimental measurements, which gave significantly lower values. The observed response can be attributed to the presence of pre-existing discontinuities, as shown in microscale CT scans of the samples. The results of this study demonstrate that the microstructure of the clay prior to testing has an impact on the mechanical behaviour and can cause inconsistencies in the comparison of the laboratory test results with in-situ data.

Keywords: boom clay, laboratory testing, overconsolidation ratio, stress-strain response, swelling, undrained shear strength

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Authors: Santina Sahibon, Sivasooriar Sivaneson, Martin Giddy, Nelson Nheu, Siti Sazeelah, Choo Kok Ming, Thuhairah Abdul Rahman, Fatmawati Binti Kamal

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Introduction: Establishing population-based reference intervals (RI) are essential when evaluating laboratory test results and for method verification. Our laboratory initiated an exercise to establish RI for routine coagulation profile as part of the method verification procedure and to determine any differences in RI between three analyzers planned to be used in the laboratory. Methodology: 145 blood samples were collected and analysed for activated partial thromboplastin time (aPTT), prothrombin time (PT), international normalized ratio (INR), and fibrinogen] using three coagulation analysers which were CA104, CA660, and CS-2500 (Sysmex, USA). RI was established at 2.5th and 97.5th percentiles. Results: The RI for aPTT between C104, C660 and CS-2500 are (RI: 20.5-30.2 sec), (RI: 21.5-29.2 sec) and (RI: 22.7-30.3 sec) respectively. The RI for PT were (RI: 7.5-10.3 sec), (RI: 9.2- 11.1 sec) and (RI: 9.8-11.9 sec) for C104, CA660 and CS-2500 respectively. INR had an RI of (RI: 0.87- 1.16), (RI: 0.89-1.10) and (0.90-1.11) respectively on CA104, C660 and CS-2500. Fibrinogen RI was (RI: 2.04-4.62 g/L) and (2.05-4.76 g/L) on the CA660 and CS-2500, respectively. Conclusion: The RI was similar across the analytical platforms for aPTT, INR, and fibrinogen. However, CA104 showed lower RI compared to the other two analysers for PT. This highlights the potential variability in results between instruments that need to be addressed when verifying RI.

Keywords: coagulation, reference interval, APTT, PT, INR, fibrinogen

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Authors: Maznieda Mahjom, Rohaida Ismail, Masita Arip, Mohd Shaiful Azlan, Nor’Ashikin Othman, Hafizah Abdullah, nor Zahrin Hasran, Joshita Jothimanickam, Syaqilah Shawaluddin, Nadia Mohamad, Raheel Nazakat, Tuan Mohd Amin, Mizanurfakhri Ghazali, Rosmanajihah Mat Lazim

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COVID-19 outbreak is particularly detrimental to the mental health of everyone as well as leaving a long devastating crisis in the healthcare sector. Daily increment of COVID-19 cases and close contact, necessitating the testing of a large number of samples, thus increasing the workload and burden to laboratory personnel. This study aims to determine the prevalence of personal-, work- and client-related burnout as well as to measure the concentration of salivary cortisol among laboratory personnel in the main laboratories in Klang Valley, Malaysia. This cross-sectional study was conducted in late 2021 and recruited a total of 404 respondents from three laboratories in Klang Valley, Malaysia. The level of burnout was assessed using Copenhagen Burnout Inventory (CBI) comprising three sub-dimensions of personal-, work- and client-related burnout. The cut-off score of 50% and above indicated possible burnout. Meanwhile, salivary cortisol was measured using a competitive enzyme immunoassay kit (Salimetrics, State College, PA, USA). Normal levels of salivary cortisol concentration in adults are within 0.094 to 1.551 μg/dl (morning) and can be none detected to 0.359 μg/dl (evening). The prevalence of personal-, work- and client-related burnout among laboratory personnel were 36.1%, 17.8% and 7.2% respectively. Meanwhile, the abnormal morning and evening cortisol concentration recorded were 29.5% and 21.8% excluding 6.9%-7.4% missing data. While the IgA level is normal for most of the respondents, which recorded at 95.53%. Laboratory personnel were at risk of suffering burnout during the COVID-19 pandemic. Thus, mental health programs need to be addressed at the department and hospital level by regularly screening healthcare workers and designing an intervention program. It is also vital to improve the coping skills of laboratory personnel by increasing the awareness of good coping skill techniques. The training must be in an innovative way to ensure that the lab personnel can internalise the technique and practise it in real life.

Keywords: burnout, COVID-19, laborotary personnel, salivary cortisol

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Authors: Menna Ewida, Dalal Abou El-Ella, Dina Lasheen, Huessin El-Subbagh

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Introduction: Vascular Endothelial Growth Factor receptor (VEGF) is a signal protein produced by cells that stimulates vasculogenesis to create new blood vessels. VEGF family binds to three trans-membrane tyrosine kinase receptors,Dihydrofolate reductase (DHFR) is an enzyme of crucial importance in medicinal chemistry. DHFR catalyzes the reduction 7,8 dihydro-folate to tetrahydrofolate and intimately couples with thymidylate synthase which is a pivotal enzyme that catalysis the reductive methylation of deoxyuridine monophosphate (dUMP) to deoxythymidine monophosphate (dTMP) utilizing N5,N10-methylene tetrahydrofolate as a cofactor which functions as the source of the methyl group. Purpose: Novel substituted Thiazole agents were designed as DHFR and VEGF-TK inhibitors with increased synergistic activity and decreased side effects. Methods: Five series of compounds were designed with a rational that mimic the pharmacophoric features present in the reported active compounds that target DHFR & VEGFR. These molecules were docked against Methotrexate & Sorafenib as controls. An in silico ADMET study was also performed to validate the bioavailability of the newly designed compounds. The in silico molecular docking & ADMET study were also applied to the non-classical antifolates for comparison. The interaction energy comparable to that of MTX for DHFRI and Sorafenib for VEGF-TKI activity were recorded. Results: Compound 5 exhibited the highest interaction energy when docked against Sorafenib, While Compound 9 showed the highest interaction energy when docked against MTX with the perfect binding mode. Comparable results were also obtained for the ADMET study. Most of the compounds showed absorption within (95-99) zone which varies according to the type of substituents. Conclusions: The Substituted Thiazole Analogues could be a suitable template for antitumor drugs that possess enhanced bioavailability and act as DHFR and VEGF-TK inhibitors.

Keywords: anti-tumor agents, DHFR, drug design, molecular modeling, VEGFR-TKIs

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Authors: Mohammad Shamim Hasan Mandal, Phu Minh, Do Tan Khang, Phung Thi Tuyen, Tran Dang Xuan

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Rice (Oryza sativa L.) is one of the major crops of Vietnam which has more than thousands of varieties. Many of the local varieties have greater potentiality but they are in danger of extinct. Rice plant contains many secondary metabolites that are allelopathic to other plants. Seven rice varieties were cultivated in the field condition at Hiroshima University, Japan; stems and leaves from each variety were collected later, they were extracted with methanol, hexane, ethyl acetate, butanol, and water. Total phenolic content and total flavonoid contents were high in ethyl acetate extracts. DPPH antioxidant assay results showed that the ethyl acetate extracts had the higher IC50 value. Therefore, the ethyl acetate extracts were selected for laboratory experimentation through petri dish assay. Results showed that the two-local variety Re nuoc and Nan chon completely inhibited the germination of radish seedlings. Further laboratory bioassay and field experimentation will be conducted to validate the laboratory bioassay findings.

Keywords: allelopathy, bioassay, Oryza sativa, Raphanus sativus

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Authors: Daniel M. Barry

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Since more than half a century ago, attempts have been made to culture cells and embryos outside the body (in vitro or ex vivo). This was done with different culture media and in various “incubators”. In the present study two different culture media were used: a standard TCM-199 culture medium and first trimester amniotic fluid (BAF) collected sterilely from pregnant cows after slaughter. Two different culture conditions were also investigated, the standard laboratory CO2 incubator versus culturing bovine embryos in the vagina of a goat doe. Two experiments were done: Firstly the permeability of different receptacles to CO2 gas was analyzed for possible culture in the vagina. Four-well plates and straws were used to incubate TCM-199 and BAF for a period of 120 h in the presence or absence of 5% CO2 gas. The pH values were measured and recorded every 24 h. In the second experiment pre-compacted morula stage bovine embryos were cultured in the above culture media in sealed 0.25 mL straws in a standard laboratory incubator and in the vagina of a goat doe. Evaluation was done on (1) stage of development and (2) number of blastomeres after 96 h of culture. In the first experiment it was shown that the CO2 gas diffused out of the 4-well plate as well as through the wall of the straws in the absence of CO2 gas, while in the presence of CO2 the pH of both media stabilized between 7.3 and 7.5. This meant that the semen straws were permeable to CO2 gas and could therefore be used as receptacles for culturing early stage bovine embryos. In the second experiment no statistical differences (p>0.05) were found in the number of pre-compacted bovine embryos that developed to the blastocyst stage, or the hatched blastocyst stage, neither for the culture medium used, or the method of culturing in the two incubators. Neither was there any difference (p>0.05) in the number of blastomeres that developed at the blastocyst stage between the two types of incubators. The bovine embryos tended to develop more blastomeres when cultured in BAF than when cultured in TCM-199 in both the standard laboratory incubator and when using the vagina of a goat doe as an incubator.

Keywords: alternative culture, bovine embryos, vagina, bovine amniotic fluid, incubator

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Authors: Asif Bilal, Fouzia Tanvir, Sibtain Ahmad

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Estrogen receptor alpha, also known as estrogen receptor-1, is highly involved in risk of mammary carcinoma. The objectives of this study were to identify non-synonymous SNPs of estrogen receptor and their association with breast cancer and to identify the chemotherapeutic responses of phytochemicals against it via in-silico study design. For this purpose, different online tools. to identify pathogenic SNPs the tools were SIFT, Polyphen, Polyphen-2, fuNTRp, SNAP2, for finding disease associated SNPs the tools SNP&GO, PhD-SNP, PredictSNP, MAPP, SNAP, MetaSNP, PANTHER, and to check protein stability Mu-Pro, I-Mutant, and CONSURF were used. Post-translational modifications (PTMs) were detected by Musitedeep, Protein secondary structure by SOPMA, protein to protein interaction by STRING, molecular docking by PyRx. Seven SNPs having rsIDs (rs760766066, rs779180038, rs956399300, rs773683317, rs397509428, rs755020320, and rs1131692059) showing mutations on I229T, R243C, Y246H, P336R, Q375H, R394S, and R394H, respectively found to be completely deleterious. The PTMs found were 96 times Glycosylation; 30 times Ubiquitination, a single time Acetylation; and no Hydroxylation and Phosphorylation were found. The protein secondary structure consisted of Alpha helix (Hh) is (28%), Extended strand (Ee) is (21%), Beta turn (Tt) is 7.89% and Random coil (Cc) is (44.11%). Protein-protein interaction analysis revealed that it has strong interaction with Myeloperoxidase, Xanthine dehydrogenase, carboxylesterase 1, Glutathione S-transferase Mu 1, and with estrogen receptors. For molecular docking we used Asiaticoside, Ilekudinuside, Robustoflavone, Irinoticane, Withanolides, and 9-amin0-5 as ligands that extract from phytochemicals and docked with this protein. We found that there was great interaction (from -8.6 to -9.7) of these ligands of phytochemicals at ESR1 wild and two mutants (I229T and R394S). It is concluded that these SNPs found in ESR1 are involved in breast cancer and given phytochemicals are highly helpful against breast cancer as chemotherapeutic agents. Further in vitro and in vivo analysis should be performed to conduct these interactions.

Keywords: breast cancer, ESR1, phytochemicals, molecular docking

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Authors: Richa Dhingra, Monika, Manav Malhotra, Tilak Raj Bhardwaj, Neelima Dhingra

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5-Alpha-reductases (5AR), a membrane bound, NADPH dependent enzyme and convert male hormone testosterone (T) into more potent androgen dihydrotestosterone (DHT). DHT is the required for the development and function of male sex organs, but its overproduction has been found to be associated with physiological conditions like Benign Prostatic Hyperplasia (BPH). Thus the inhibition of 5ARs could be a key target for the treatment of BPH. In present study, 2D and 3D Quantitative Structure Activity Relationship (QSAR) pharmacophore models have been generated for 5AR based on known inhibitory concentration (IC₅₀) values with extensive validations. The four featured 2D pharmacophore based PLS model correlated the topological interactions (–OH group connected with one single bond) (SsOHE-index); semi-empirical (Quadrupole2) and physicochemical descriptors (Mol. wt, Bromines Count, Chlorines Count) with 5AR inhibitory activity, and has the highest correlation coefficient (r² = 0.98, q² =0.84; F = 57.87, pred r² = 0.88). Internal and external validation was carried out using test and proposed set of compounds. The contribution plot of electrostatic field effects and steric interactions generated by 3D-QSAR showed interesting results in terms of internal and external predictability. The well validated 2D Partial Least Squares (PLS) and 3D k-nearest neighbour (kNN) models were used to search novel 5AR inhibitors with different chemical scaffold. To gain more insights into the molecular mechanism of action of these steroidal derivatives, molecular docking and in silico absorption, distribution, metabolism, and excretion (ADME) studies were also performed. Studies have revealed the hydrophobic and hydrogen bonding of the ligand with residues Alanine (ALA) 63A, Threonine (THR) 60A, and Arginine (ARG) 456A of 4AT0 protein at the hinge region. The results of QSAR, molecular docking, in silico ADME studies provide guideline and mechanistic scope for the identification of more potent 5-Alpha-reductase inhibitors (5ARI).

Keywords: 5α-reductase inhibitor, benign prostatic hyperplasia, ligands, molecular docking, QSAR

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Authors: Muhammad Sufyan, Muhammad D. Gogi, Muhammad Arshad, Ahmad Nawaz, Muhammad Usman

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Cotton (Gossypium hirsutum) universally known as silver fiber and is one of the most important cash crop of Pakistan. A wide array of pests constraints cotton production among which sucking insect pests cause serious losses. Mostly new chemistry insecticides used to control a wide variety of insect pests including sucking insect pests. In the present study efficacy of different neonicotinoids was evaluated against sucking insect pests of cotton in the field and in laboratory for red and dusky cotton bug. The experiment was conducted at Entomology Research Station, University of Agriculture Faisalabad, in a Randomized Complete Block Design (RCBD). Field trial was conducted to evaluate the efficacy of Confidence Ultra (Imidacloprid) 70% SL, Confidor (Imidacloprid) 20% SL, Kendo (Lambda cyhalothrin) 24.7 SC, Actara (Thiamethoxam) 25% WG, Forcast (Tebufenozide+ Emamectin benzoate) 8.8 EW and Timer (Emamectin benzoate) 1.9 EC at their recommended doses. The data was collected on per leaf basis of thrips, aphid, jassid and whitefly before 24 hours of spray. The post treatment data was recorded after 24, 48 and 72 hours. The fresh, non-infested and untreated cotton leaves was collected from the field and brought to the laboratory to assess the efficacy of neonicotinoids against red and dusky cotton bug. After data analysis all the insecticides were found effective against sucking pests. Confidence Ultra was highly effective against the aphid, jassid, and whitefly and gave maximum mortality, while showed non-significant results against thrips. In case of aphid plot which was treated with Kando 24.7 SC showed significant mortality after 72 hours of pesticide application. Similar trends were found in laboratory conditions with all these treatments by making different concentrations and had significant impact on dusky cotton bug and red cotton bug population after 24, 48 and 72 hours after application.

Keywords: cotton, laboratory and field conditions, neonicotinoids, sucking insect pests

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Authors: Dawit Assefa, Kassaye Tekie, Gebrie Alebachew, Degefu Beyene, Bikila Alemu, Naji Mohammed, Asnakech Agegnehu, Seble Tsehay, Geremew Tasew

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Background: Implementation of a Laboratory Quality Management System (LQMS) is critical to ensure accurate, reliable, and efficient laboratory testing of antimicrobial resistance (AMR). However, limited LQMS implementation and progress toward accreditation in the AMR surveillance laboratory testing setting exist in Ethiopia. By addressing non-conformances (NCs) and working towards accreditation, microbiology laboratories can improve the quality of their services, increase staff competence, and contribute to mitigate the spread of AMR. Methods: Using standard ISO 15189 horizontal and vertical assessment checklists, certified assessors identified NCs at Hawassa and Jimma Hospital microbiology laboratories. The Ethiopian Public Health Institute AMR mentors and IDDS staff prioritized closing the NCs through the implementation of an intensified mentorship program that included ISO 15189 orientation training, resource allocation, and action plan development. Results: For the two facilities to clear their NCs, an intensified mentorship approach was adopted by providing ISO 15189 orientation training, provision of buffer reagents, controls, standards, and axillary equipment, and facilitating equipment maintenance and calibration. Method verification and competency assessment were also conducted along with the implementation of standard operating procedures and recommended corrective actions. This approach enhanced the laboratory's readiness for accreditation. After addressing their NCs, the two laboratories applied to Ethiopian Accreditation Services for ISO 15189 accreditation. Conclusions: Clearing NCs through the implementation of intensified mentorship was crucial in preparing the two laboratories for accreditation and improving quality laboratory test results. This approach can guide other microbiology laboratories’ accreditation attainment efforts.

Keywords: non-conformance clearance, intensified mentorship, accreditation, ISO 15189

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Authors: Bright Chukwunwike Uzuegbunam, Wojciech Paslawski, Hans Agren, Christer Halldin, Wolfgang Weber, Markus Luster, Thomas Arzberger, Behrooz Hooshyar Yousefi

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There is a need to develop a PET tracer that will enable to diagnosis and track the progression of Alpha-synucleinopathies (Parkinson’s disease [PD], dementia with Lewy bodies [DLB], multiple system atrophy [MSA]) in living subjects over time. Alpha-synuclein aggregates (a-syn), which are present in all the stages of disease progression, for instance, in PD, are a suitable target for in vivo PET imaging. For this reason, we have developed some promising a-syn tracers based on a disarylbisthiazole (DABTA) scaffold. The precursors are synthesized via a modified Hantzsch thiazole synthesis. The precursors were then radiolabeled via one- or two-step radiofluorination methods. The ligands were initially screened using a combination of molecular dynamics and quantum/molecular mechanics approaches in order to calculate the binding affinity to a-syn (in silico binding experiments). Experimental in vitro binding assays were also performed. The ligands were further screened in other experiments such as log D, in vitro plasma protein binding & plasma stability, biodistribution & brain metabolite analyses in healthy mice. Radiochemical yields were up to 30% - 72% in some cases. Molecular docking revealed possible binding sites in a-syn and also the free energy of binding to those sites (-28.9 - -66.9 kcal/mol), which correlated to the high binding affinity of the DABTAs to a-syn (Ki as low as 0.5 nM) and selectivity (> 100-fold) over Aβ and tau, which usually co-exist with a-synin some pathologies. The log D values range from 2.88 - 2.34, which correlated with free-protein fraction of 0.28% - 0.5%. Biodistribution experiments revealed that the tracers are taken up (5.6 %ID/g - 7.3 %ID/g) in the brain at 5 min (post-injection) p.i., and cleared out (values as low as 0.39 %ID/g were obtained at 120 min p.i. Analyses of the mice brain 20 min p.i. Revealed almost no radiometabolites in the brain in most cases. It can be concluded that in silico study presents a new venue for the rational development of radioligands with suitable features. The results obtained so far are promising and encourage us to further validate the DABTAs in autoradiography, immunohistochemistry, and in vivo imaging in non-human primates and humans.

Keywords: alpha-synuclein aggregates, alpha-synucleinopathies, PET imaging, tracer development

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Authors: Rijnbout-St.James Willem, Lindner Volkhard, Scholand Mary Beth, Ashton M. Tillett, Di Gennaro Michael Jude, Smith Silvia Enrica

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Introduction: Fibrosing lung diseases are characterized by changes in the lung interstitium and are classified based on etiology: 1) environmental/exposure-related, 2) autoimmune-related, 3) sarcoidosis, 4) interstitial pneumonia, and 4) idiopathic. Among interstitial lung diseases (ILD) idiopathic forms, idiopathic pulmonary fibrosis (IPF) is the most severe. Pathogenesis of IPF is characterized by an increased presence of proinflammatory mediators, resulting in alveolar injury, where injury to alveolar epithelium precipitates an increase in collagen deposition, subsequently thickening the alveolar septum and decreasing gas exchange. Identifying biomarkers implicated in the pathogenesis of lung fibrosis is key to developing new therapies and improving the efficacy of existing therapies. The transforming growth factor-beta (TGF-B1), a mediator of tissue repair associated with WNT5A signaling, is partially responsible for fibroblast proliferation in ILD and is the target of Pirfenidone, one of the antifibrotic therapies used for patients with IPF. Canonical TGF-B signaling is mediated by the proteins SMAD 2/3, which are, in turn, indirectly regulated by Collagen Triple Helix Repeat Containing 1 (CTHRC1). In this study, we tested the following hypotheses: 1) CTHRC1 is more elevated in the ILD cohort compared to unaffected controls, and 2) CTHRC1 is differently expressed among ILD types. Material and Methods: CTHRC1 levels were measured by ELISA in 171 plasma samples from the deidentified University of Utah ILD cohort. Data represent a cohort of 131 ILD-affected participants and 40 unaffected controls. CTHRC1 samples were categorized by a pulmonologist based on affectation status and disease subtypes: IPF (n = 45), sarcoidosis (4), nonspecific interstitial pneumonia (16), hypersensitivity pneumonitis (n = 7), interstitial pneumonia (n=13), autoimmune (n = 15), other ILD - a category that includes undifferentiated ILD diagnoses (n = 31), and unaffected controls (n = 40). We conducted a single-factor ANOVA of plasma CTHRC1 levels to test whether CTHRC1 variance among affected and non-affected participants is statistically significantly different. In-silico analysis was performed with Ingenuity Pathway Analysis® to characterize the role of CTHRC1 in the pathway of lung fibrosis. Results: Statistical analyses of CTHRC1 in plasma samples indicate that the average CTHRC1 level is significantly higher in ILD-affected participants than controls, with the autoimmune ILD being higher than other ILD types, thus supporting our hypotheses. In-silico analyses show that CTHRC1 indirectly activates and phosphorylates SMAD3, which in turn cross-regulates TGF-B1. CTHRC1 also may regulate the expression and transcription of TGFB-1 via WNT5A and its regulatory relationship with CTNNB1. Conclusion: In-silico pathway analyses demonstrate that CTHRC1 may be an important biomarker in ILD. Analysis of plasma samples indicates that CTHRC1 expression is positively associated with ILD affectation, with autoimmune ILD having the highest average CTHRC1 values. While characterizing CTHRC1 levels in plasma can help to differentiate among ILD types and predict response to Pirfenidone, the extent to which plasma CTHRC1 level is a function of ILD severity or chronicity is unknown.

Keywords: interstitial lung disease, CTHRC1, idiopathic pulmonary fibrosis, pathway analyses

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Authors: Seyed Abolhasan Naeini, Eisa Aliagahei

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Today, the use of geosynthetic materials in geotechnical activities is increasing significantly. One of the main uses of these materials is to increase the compressive strength of clay reinforced by geotextile layers. In the present study, the effect of clay reinforcement by geotextile layers in increasing the compressive strength of clay has been investigated using modeling in ABAQUS 6.11.3 software. For this purpose, the modified Drager Prager model has been chosen to simulate the stress-strain behavior of soil layers and the linear elastic model for the geotextile layer. Unreinforced samples and reinforced samples are modeled by geotextile layers (1, 2 and 3 geotextile layers) by software. In order to validate the results, an article in the same field was used and the numerical modeling results were calibrated with the laboratory results. Based on the obtained results, the software has a suitable capability for modeling and the results of the numerical model overlap with the laboratory results to a very acceptable extent, by increasing the number of geotextile layers, the error between the results of the laboratory sample and the software model increases. The highest amount of error is related to the sample reinforced with three layers of geotextile and is 7.3%.

Keywords: Abaqus, cap model, clay, geotextile layer, reinforced soil

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Authors: R. Vishnu, K. S. Reddy, Amrendra Kumar

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The present guideline for the construction of flexible pavement in India, IRC 37: 2012 recommends to use viscous grade VG 40 bitumen in both wearing and binder bituminous layers. However, most of the bitumen production plants in India are unable to produce the air-blown VG40 grade bitumen. This requires plant’s air-blowing technique modification, and often the manufactures finds it as uneconomical. In this context, stiffer grade bitumen can be produced if bitumen is modified. Gilsonite, which is naturally occurring asphalt have been found to be used for increasing the stiffness of binders. The present study evaluates the physical, rheological characteristics of Gilsonite modified binders and the performance characteristics of these binders when used in the mix.

Keywords: bitumen, gilsonite, stiffness, laboratory evaluation

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Authors: Hector Barrios-Piña, Georgia García-Arellano, Salvador García-Rodríguez, Gerardo Bocanegra-García, Shashi Kant

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This work shows the methodology of application and the effectiveness of the Flipped Learning technique for Civil Engineering laboratory classes. It was experimented by some of the professors of the Department of Civil Engineering at Tecnológico de Monterrey while teaching their laboratory classes. A total of 28 videos were created. The videos primarily demonstrate instructions of the experimental practices other than the usage of tools and materials. The technique allowed the students to prepare for their classes in advance. A survey was conducted on the participating professors and students (semester of August-December 2019) to quantify the effectiveness of the Flipped Learning technique. The students reported it as an excellent way of improving their learning aptitude, including self-learning whereas, the professors felt it as an efficient technique for optimizing their class session, which also provided an extra slot for class-interaction. A comparison of grades was analyzed between the students of the traditional classes and with Flipped Learning. It did not distinguish the benefits of Flipped Learning. However, the positive responses from the students and the professors provide an impetus for continuing and promoting the Flipped Learning technique in future classes.

Keywords: flipped learning, laboratory classes, civil engineering, competences development

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Authors: Hamid Karimi, Naegar Mahdavi, Hossein Tayefi Nasrabadi

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Spermatozoids production rate and its quality are more important factors in the diagnosis of infertility. Also, spematozids activity have a more important role in fertilization. Some medicinal plants as Asparagus(Asparagus officinalis) has many antioxidant component. Therefore, They can affect testes tissue to production more and high-quality spermatozoids. In this survey, Asparagus root extract is studied on spermatogenesis index in the laboratory mouse testes. Hydro-alcoholic extract of asparagus root is prepared and examined on four group of the mature male mouse. Blank group without extract, group 1,100ml/kg dose, group 2, 200 ml/kg dose and group 3, 300ml/kg dose. Then, mice are euthanized, and testes are removed. Testes are weighted, and paraffinized blocks are prepared. TDI(Tubular Differentiation Index) and SPI(Spermiation Index) are studied on histological sections by light microscope. This study results were showed that TDI and SPI in treatments groups with 200 and 300 ml/kg dose had significant enhancement (P<0.05). Consequently, Extract of Asparagus root can enhance spermatozoid production and, therefore, cause improve fertility in male laboratory mice.

Keywords: histology, spermatozoid, ASP [aragus, testes

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Authors: Sarah K. Taylor, Miratun M. Saharuddin, Zabri A. Talib

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Cybercrime is on the rise, and yet many Law Enforcement Agencies (LEAs) in Malaysia have no Digital Forensics Laboratory (DFL) to assist them in the attrition and analysis of digital evidence. From the estimated number of 30 LEAs in Malaysia, sadly, only eight of them owned a DFL. All of the DFLs are concentrated in the capital of Malaysia and none at the state level. LEAs are still depending on the national DFL (CyberSecurity Malaysia) even for simple and straightforward cases. A survey was conducted among LEAs in Malaysia owning a DFL to understand their history of establishing the DFL, the challenges that they faced and the significance of the DFL to their case investigation. The results showed that the while some LEAs faced no challenge in establishing a DFL, some of them took seven to 10 years to do so. The reason was due to the difficulty in convincing their management because of the high costs involved. The results also revealed that with the establishment of a DFL, LEAs were better able to get faster forensic result and to meet agency’s timeline expectation. It is also found that LEAs were also able to get more meaningful forensic results on cases that require niche expertise, compared to sending off cases to the national DFL. Other than that, cases are getting more complex, and hence, a continuous stream of budget for equipment and training is inevitable. The result derived from the study is hoped to be used by other LEAs in justifying to their management the benefits of establishing an in-house DFL.

Keywords: digital evidence, digital forensics, digital forensics laboratory, law enforcement agency

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Authors: Vahid Ayan, Joshua R Omer, Alireza Khavandi, Mukesh C Limbachiya

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In Iran, utilization of reclaimed asphalt pavement (RAP) aggregate has become a common practice in pavement rehabilitation during the last ten years. Such developments in highway engineering have necessitated several studies to clarify the technical and environmental feasibility of other alternative materials in road rehabilitation and maintenance. The use of recycled concrete aggregates (RCA) in asphalt pavements is one of the major goals of municipality of Tehran. Nevertheless little research has been done to examine the potential benefits of local RCA. The objective of this study is laboratory investigation of incorporating RCA into RAP for use in unbound subbase application. Laboratory investigation showed that 50%RCA+50%RAP is both technically and economically appropriate for subbase use.

Keywords: Roads & highways, Sustainability, Recycling & reuse of materials

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Authors: Bartosz Kedra, Robert Malkowski

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This paper provides description and presentation of laboratory test unit built basing on 150 kVA power frequency converter and Simulink RealTime platform. Assumptions, based on criteria which load and generator types may be simulated using discussed device, are presented, as well as control algorithm structure. As laboratory setup contains transformer with thyristor controlled tap changer, a wider scope of setup capabilities is presented. Information about used communication interface, data maintenance, and storage solution as well as used Simulink real-time features is presented. List and description of all measurements are provided. Potential of laboratory setup modifications is evaluated. For purposes of Rapid Control Prototyping, a dedicated environment was used Simulink RealTime. Therefore, load model Functional Unit Controller is based on a PC computer with I/O cards and Simulink RealTime software. Simulink RealTime was used to create real-time applications directly from Simulink models. In the next step, applications were loaded on a target computer connected to physical devices that provided opportunity to perform Hardware in the Loop (HIL) tests, as well as the mentioned Rapid Control Prototyping process. With Simulink RealTime, Simulink models were extended with I/O cards driver blocks that made automatic generation of real-time applications and performing interactive or automated runs on a dedicated target computer equipped with a real-time kernel, multicore CPU, and I/O cards possible. Results of performed laboratory tests are presented. Different load configurations are described and experimental results are presented. This includes simulation of under frequency load shedding, frequency and voltage dependent characteristics of groups of load units, time characteristics of group of different load units in a chosen area and arbitrary active and reactive power regulation basing on defined schedule.

Keywords: MATLAB, power converter, Simulink Real-Time, thyristor-controlled tap changer

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Authors: Suganya Chandrababu, Dhundy Bastola

Abstract:

Like in the gut, the subgingival microbiota plays a crucial role in oral hygiene, health, and cariogenic diseases. Human activities like diet, antibiotics, and periodontal treatments alter the bacterial communities, metabolism, and functions in the oral cavity, leading to a dysbiotic state and changes in the plaques of orthodontic patients. Fixed periodontal appliances hinder oral hygiene and cause changes in the dental plaques influencing the subgingival microbiota. However, the microbial species’ diversity and complexity pose a great challenge in understanding the taxa’s community distribution patterns and their role in oral health. In this research, we analyze the subgingival microbial samples from individuals with fixed dental appliances (metal/clear) using an in silico approach. We employ exploratory hypothesis-driven multivariate and regression analysis to shed light on the microbial community and its functional fluctuations due to dental appliances used and identify risks associated with complex disease phenotypes. Our findings confirm the changes in oral microbiota composition due to the presence and type of fixed orthodontal devices. We identified seven main periodontic pathogens, including Bacteroidetes, Actinobacteria, Proteobacteria, Fusobacteria, and Firmicutes, whose abundances were significantly altered due to the presence and type of fixed appliances used. In the case of metal braces, the abundances of Bacteroidetes, Proteobacteria, Fusobacteria, Candidatus saccharibacteria, and Spirochaetes significantly increased, while the abundance of Firmicutes and Actinobacteria decreased. However, in individuals With clear braces, the abundance of Bacteroidetes and Candidatus saccharibacteria increased. The highest abundance value (P-value=0.004 < 0.05) was observed with Bacteroidetes in individuals with the metal appliance, which is associated with gingivitis, periodontitis, endodontic infections, and odontogenic abscesses. Overall, the bacterial abundances decrease with clear type and increase with metal type of braces. Regression analysis further validated the multivariate analysis of variance (MANOVA) results, supporting the hypothesis that the presence and type of the fixed oral appliances significantly alter the bacterial abundance and composition.

Keywords: oral microbiota, statistical analysis, fixed or-thodontal appliances, bacterial abundance, multivariate analysis, regression analysis

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Authors: R. Daïra, V. Chalvidan

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We present the speckle interferometry method to determine the deformation of a piece. This method of holographic imaging using a CCD camera for simultaneous digital recording of two states object and reference. The reconstruction is obtained numerically. This latest method has the advantage of being simpler than the methods currently available, and it does not suffer the holographic configuration faults online. Furthermore, it is entirely digital and avoids heavy analysis after recording the hologram. This work was carried out in the laboratory HOLO 3 (optical metrology laboratory in Saint Louis, France) and it consists in controlling qualitatively and quantitatively the deformation of object by using a camera CCD connected to a computer equipped with software of Fringe Analysis.

Keywords: speckle, nondestructive testing, interferometry, image processing

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Authors: Godfrey Muiya Mukala

Abstract:

This research looks at an array of fields in Sub-Saharan Africa comprising agriculture, food enterprises, medicine, organisms genetically modified, microbiology, and nanotechnology that can be gained from biotechnological research and development. Findings into dangerous organisms, mainly bacterial germs, rickettsia, fungi, parasites, or organisms that are genetically engineered, have immensely posed questions attributed to the biological danger they bring forth to human beings and the environment because of their uncertainties. In addition, the recurrence of previously managed diseases or the inception of new diseases are connected to biosafety challenges, especially in rural set-ups in low and middle-income countries. Notably, biotechnology laboratories are required to adopt biosafety measures to protect their workforce, community, environment, and ecosystem from unforeseen materials and organisms. Sensitization and inclusion of educational frameworks for laboratory workers are essential to acquiring a solid knowledge of harmful biological agents. This is in addition to human pathogenicity, susceptibility, and epidemiology to the biological data used in research and development. This article reviews and analyzes research intending to identify the proper implementation of universally accepted practices in laboratory safety and biological hazards. This research identifies ideal microbiological methods, adequate containment equipment, sufficient resources, safety barriers, specific training, and education of the laboratory workforce to decrease and contain biological hazards. Subsequently, knowledge of standardized microbiological techniques and processes, in addition to the employment of containment facilities, protective barriers, and equipment, is far-reaching in preventing occupational infections. Similarly, reduction of risks and prevention may be attained by training, education, and research on biohazards, pathogenicity, and epidemiology of the relevant microorganisms. In this technique, medical professionals in rural setups may adopt the knowledge acquired from the past to project possible concerns in the future.

Keywords: sub-saharan africa, biotechnology, laboratory, infections, health

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Authors: Tukur Daiyabu Abdulkadir, Wan Ishak Wan Ismail, Muhammad Saufi Mohd Kassim

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Seed singulation and accuracy in seed spacing are the major challenges associated with the adoption of mechanical seeder for system of rice intensification. In this research the metering system of a pneumatic planter was modified and automated for increase precision to meet the demand of system of rice intensification SRI. The chain and sprocket mechanism of a conventional vacuum planter were now replaced with an electro mechanical system made up of a set of servo motors, limit switch, micro controller and a wheel divided into 10 equal angles. The circumference of the planter wheel was determined based on which seed spacing was computed and mapped to the angles of the metering wheel. A program was then written and uploaded to arduino micro controller and it automatically turns the seed plates for seeding upon covering the required distance. The servo motor was calibrated with the aid of labVIEW. The machine was then calibrated using a grease belt and varying the servo rpm through voltage variation between 37 rpm to 47 rpm until an optimum value of 40 rpm was obtained with a forward speed of 5 kilometers per hour. A pressure of 1.5 kpa was found to be optimum under which no skip or double was recorded. Precision in spacing (coefficient of variation), miss index, multiple index, doubles and skips were investigated. No skip or double was recorded both at laboratory and field levels. The operational parameters under consideration were both evaluated at laboratory and field. Even though there was little variation between the laboratory and field values of precision in spacing, multiple index and miss index, the different is not significant as both laboratory and field values fall within the acceptable range.

Keywords: automation, calibration, pneumatic seed planter, system of rice intensification

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