Search results for: hela cell line

Authors: Ahmed M. Haddad, Hadeel S. El-Shaal, Gadallah M. Abu-Elreesh

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Some of filamentous fungi can be used for biodiesel production as they are able to accumulate high amounts of intracellular lipids when grown at stress conditions. Aspergillus fumigatus sp. was isolated from Nile delta soil in Egypt. The fungus was primarily screened for its capacity to accumulate lipids using Nile red staining assay. The fungus could accumulate more than 20% of its biomass as lipids when grown at optimized minimal medium. After lipid extraction, we could use fungal cell debris to remove some heavy metals from contaminated waste water. The fungal cell debris could remove Cd, Cr, and Zn with absorption efficiency of 73%, 83.43%, and 69.39% respectively. In conclusion, the Aspergillus fumigatus isolate may be considered as a promising biodiesel producer, and its biomass waste can be further used for bioremediation of wastewater contaminated with heavy metals.

Keywords: biodiesel, bioremediation, fungi, heavy metals, lipids, oleaginous

Procedia PDF Downloads 219

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: Yongfang Deng, Dunwen Zuo

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An attempt is made here to join 2024 aluminum alloy plate by friction stir joining (FSJ) using different types of tools. Joint surface morphology was observed, and both arc line spacing and flash were measured. Study is carried out on the effect of pin, shoulder and eccentricity of the tool on the surface topography of the joint and the formation of the joint surface topography is analyzed. It is found that, eccentric squeezing action of the tool is the mainly motive power to form arc lines contour and flash structure. Little flash appears in the advancing side but with severe deformation, while the flash in the retreating side is heavy but with soft deformation. The pin of tool has a deep impact on the flash on the advancing side of the joints. Shoulder can widen the arc lines, refine arcs structure, reduce flash in the retreat side, but will increase the flash in the advancing side. Increasing the amount of eccentricity, it has litter effect on the arc line spacing but will destroy the arc lines morphology in the joint surface and promote the formation of filamentous flash structure in the joint.

Keywords: FSJ, surface morphology, tool, joint

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Authors: Z. Kolacinski, L. Szymanski. G. Raniszewski, D. Koza, L. Pietrzak

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Magnetic Bio-Nano-Fluid (BNF) can be composed of a buffer fluid such as plasma and magnetic nanoparticles such as iron, nickel, cobalt and their oxides. However iron is one of the best elements for magnetization by electromagnetic radiation. It can be used as a tool for medical diagnosis and treatment. Radio frequency (RF) radiation is able to heat iron nanoparticles due to magnetic hysteresis. Electromagnetic heating of iron nanoparticles and ferro-fluids BNF can be successfully used for non-invasive thermal ablation of cancer cells. Moreover iron atoms can be carried by carbon nanotubes (CNTs) if iron is used as catalyst for CNTs synthesis. Then CNTs became the iron containers and they screen the iron content against oxidation. We will present a method of CNTs addressing to the required cells. For thermal ablation of cancer cells we use radio frequencies for which the interaction with human body should be limited to minimum. Generally, the application of RF energy fields for medical treatment is justified by deep tissue penetration. The highly iron doped CNTs as the carriers creating magnetic fluid will be presented. An excessive catalyst injection method using electrical furnace and microwave plasma reactor will be presented. This way it is possible to grow the Fe filled CNTs on a moving surface in continuous synthesis process. This also allows producing uniform carpet of the Fe filled CNTs carriers. For the experimental work targeted to cell ablation we used RF generator to measure the increase in temperature for some samples like: solution of Fe2O3 in BNF which can be plasma-like buffer, solutions of pure iron of different concentrations in plasma-like buffer and in buffer used for a cell culture, solutions of carbon nanotubes (MWCNTs) of different concentrations in plasma-like buffer and in buffer used for a cell culture. Then the targeted therapies which can be effective if the carriers are able to distinguish the difference between cancerous and healthy cell’s physiology are considered. We have developed an approach based on ligand-receptor or antibody-antigen interactions for the case of colon cancer.

Keywords: cancer treatment, carbon nano tubes, drag delivery, hyperthermia, iron

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Authors: Jackson Becker

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Transportation produces the largest amount of carbon dioxide emissions in the United States of America. Today, cars are the predominant mode of transportation across the country, and our cities have been reshaped due to them. This was not always the case. Streetcars were seen in almost every American city of the early 1900s. These streetcar systems were viewed as obsolete with the rise of the automobile. With fewer streetcars came lower public transport ridership. Austin, Texas is one of the fastest growing cities in the country, and it used to have an extensive streetcar line. Today, it plans to build a light rail line with less rail mileage than 100 years ago. This research looks at the areas of Austin that are not included in the city’s new transit plan. Transit connectivity is one factor that goes into walkability rates for cities. This study also looks at the correlation between walkability ratings with median household income levels from the 2019 Census. The results showed a correlation between higher income neighborhoods having higher walkability rates, which was influenced by the lack of public transportation options.

Keywords: transportation, walkability, income, austin

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Authors: Nazanin Amanat, Alison Tayler, Steve Whyard

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While chemical insecticides effectively control many insect pests, they also harm many non-target species. Double-stranded RNA (dsRNA) pesticides, in contrast, can be designed to target unique gene sequences and thus act in a species-specific manner. DsRNA insecticides do not, however, work equally well for all insects, and for some species that are considered refractory to dsRNA, a primary factor affecting efficacy is the relative ease by which dsRNA can enter a target cell’s cytoplasm. In this study, we are examining how different structured dsRNAs (linear, hairpin, and paperclip) can enter mosquito and lepidopteran cells, as they represent dsRNA-sensitive and refractory species, respectively. To determine how the dsRNAs enter the cells, we are using chemical inhibitors and RNA interference (RNAi)-mediated knockdown of key proteins associated with different endocytosis processes. Understanding how different dsRNAs enter cells will ultimately help in the design of molecules that overcome refractoriness to RNAi or develop resistance to dsRNA-based insecticides. To date, we have conducted chemical inhibitor experiments on both cell lines and have evidence that linear dsRNAs enter the cells using clathrin-mediated endocytosis, while the paperclip dsRNAs (pcRNAs) can enter both species’ cells in a clathrin-independent manner to induce RNAi. An alternative uptake mechanism for the pcRNAs has been tentatively identified, and the outcomes of our RNAi-mediated knockdown experiments, which should provide corroborative evidence of our initial findings, will be discussed.

Keywords: dsRNA, RNAi, uptake, insecticides, dipteran, lepidopteran

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Authors: Jordan Chamarande, Lisiane Cunat, Corentine Alauzet, Catherine Cailliez-Grimal

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Gut microbiota (GM) is now considered a new organ mainly due to the microorganism’s specific biochemical interaction with its host. Although mechanisms underlying host-microbiota interactions are not fully described, it is now well-defined that cell surface molecules and structures of the GM play a key role in such relation. The study of surface structures of GM members is also fundamental for their role in the establishment of species in the versatile and competitive environment of the digestive tract and as a potential virulence factor. Among these structures are capsular polysaccharides (CPS), fimbriae, pili and lipopolysaccharides (LPS), all well-described for their central role in microorganism colonization and communication with host epithelium. The health-promoting Parabacteroides distasonis, which is part of the core microbiome, has recently received a lot of attention, showing beneficial properties for its host and as a new potential biotherapeutic product. However, to the best of the authors’ knowledge, the cell surface molecules and structures of P. distasonis that allow its maintain within the GM are not identified. Moreover, although P. distasonis is strongly recognized as intestinal commensal species with benefits for its host, it has also been recognized as an opportunistic pathogen. In this study, we reported gene clusters potentially involved in the synthesis of the capsule, fimbriae-like and pili-like cell surface structures in 26 P. distasonis genomes and applied the new RfbA-Typing classification in order to better understand and characterize the beneficial/pathogenic behaviour related to P. distasonis strains. In context, 2 different types of fimbriae, 3 of pilus and up to 14 capsular polysaccharide loci, have been identified over the 26 genomes studied. Moreover, the addition of data to the rfbA-Type classification modified the outcome by rearranging rfbA genes and adding a fifth group to the classification. In conclusion, the strain variability in terms of external proteinaceous structure could explain the inter-strain differences previously observed in P. distasonis adhesion capacities and its potential pathogenicity.

Keywords: gut microbiota, Parabacteroides distasonis, capsular polysaccharide, fimbriae, pilus, O-antigen, pathogenicity, probiotic, comparative genomics

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Authors: Danni Cheng

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Purpose:Surgicallyinclusive treatment(SIT)isthemajor treatment fororopharyngealsquamouscellcarcinoma (OPSCC) in Eastern countries, while nonsurgical treatments(NSTs) are the priority treatment in Western countries. The preferred treatmentsforOPSCC patients remaindebated. Methods:Atotalof 153 consecutive OPSCC casesdiagnosed between 2009 and 2019inWCH, and 15,400 OPSCC cases from SEER database (2000-2017) were obtained. Clinical characteristics, treatments, and survival outcomes were retrospectively collected. We conductedKaplan-Meier curves univariate and multivariate analysis to compare the prognosis of OPSCC patients in WCH, SEER Asian, and SEER all ethnic population by different treatment modalities,HPVstatus, ages, and TNM stages. Results: The 5-year overall survival rate was 59% in WCH, 64% in the SEER all ethnic and 67% in SEER Asian group. In both univariate and multivariate analysis, SIT was observed as a consistent benefit factor for OPSCC patients in all three populations when classified by genders, tumor stages, and HPV status. Patients who underwent SIT had significantly better survival outcomes than those who received NSTsin WCH, SEER Asian, and SEER all ethnic groups. HPV positive status was the beneficial factor of OPSCC patients in all three groups. Besides, male patients had worse survival outcomes in both WCH and SEER Asian group, whereas male patients had better outcomes in the SEER all ethnic group. Conclusion: In contrast to nowadaysNSTs are the first-line therapiesfor OPSCC, our ten-year real-world data and SEER data indicated that OPSCC patients who underwent SIT had better prognosis than NSTs.

Keywords: OPSCC, survival outcome, SEER, treatment modalities

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Authors: Daniel Aberdam

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Haploinsufficiency of PAX6 in humans is the main cause of congenital aniridia, a rare eye disease characterized by reduced visual acuity. Patients have also progressive disorders including cataract, glaucoma and corneal abnormalities making their condition very challenging to manage. Aniridia-related keratopathy (ARK), caused by a combination of factors including limbal stem-cell deficiency, impaired healing response, abnormal differentiation, and infiltration of conjunctival cells onto the corneal surface, affects up to 95% of patients. It usually begins in the first decade of life resulting in recurrent corneal erosions, sub-epithelial fibrosis with corneal decompensation and opacification. Unfortunately, current treatment options for aniridia patients are currently limited. Although animal models partially recapitulate this disease, there is no in vitro cellular model of AKT needed for drug/therapeutic tools screening and validation. We used genome editing (CRISPR/Cas9 technology) to introduce a nonsense mutation found in patients into one allele of the PAX6 gene into limbal stem cells. Resulting mutated clones, expressing half of the amount of PAX6 protein and thus representative of haploinsufficiency were further characterized. Sequencing analysis showed that no off-target mutations were induced. The mutated cells displayed reduced cell proliferation and cell migration but enhanced cell adhesion. Known PAX6 targets expression was also reduced. Remarkably, addition of soluble recombinant PAX6 protein into the culture medium was sufficient to activate endogenous PAX6 gene and, as a consequence, rescue the phenotype. It strongly suggests that our in vitro model recapitulates well the epithelial defect and becomes a powerful tool to identify drugs that could rescue the corneal defect in patients. Furthermore, we demonstrate that the homeotic transcription factor Pax6 is able to be uptake naturally by recipient cells to function into the nucleus.

Keywords: Pax6, crispr/cas9, limbal stem cells, aniridia, gene therapy

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Authors: Mathieu Imbert, Sebastien Comas-Cardona, Emmanuelle Abisset-Chavanne, David Prono

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Short cycle time Resin Transfer Molding (RTM) applications appear to be of great interest for the mass production of automotive or aeronautical lightweight structural parts. During the RTM process, the two components of a resin are mixed on-line and injected into the cavity of a mold where a fibrous preform has been placed. Injection and polymerization occur simultaneously in the preform inducing evolutions of temperature, degree of cure and viscosity that furthermore affect flow and curing. In order to adjust the processing conditions to reduce the cycle time, it is, therefore, essential to understand and quantify the physical mechanisms occurring in the part during injection. In a previous study, a dual-scale simulation tool has been developed to help determining the optimum injection parameters. This tool allows tracking finely the repartition of the resin and the evolution of its properties during reactive injections with on-line mixing. Tows and channels of the fibrous material are considered separately to deal with the consequences of the dual-scale morphology of the continuous fiber textiles. The simulation tool reproduces the unsaturated area at the flow front, generated by the tow/channel difference of permeability. Resin “storage” in the tows after saturation is also taken into account as it may significantly affect the repartition and evolution of the temperature, degree of cure and viscosity in the part during reactive injections. The aim of the current study is, thanks to experiments, to understand and quantify the “storage” evolution in the tows to adjust and validate the numerical tool. The presented study is based on four experimental repeats conducted on three different types of textiles: a unidirectional Non Crimp Fabric (NCF), a triaxial NCF and a satin weave. Model fluids, dyes and image analysis, are used to study quantitatively, the resin flow in the saturated area of the samples. Also, textiles characteristics affecting the resin “storage” evolution in the tows are analyzed. Finally, fully coupled on-line mixing reactive injections are conducted to validate the numerical model.

Keywords: experimental, on-line mixing, high-speed RTM process, dual-scale flow

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Authors: Albina V. Moskvicheva, Gevorg G. Kazarian, Anna R. Valeeva, Marina A. Efimova, Malik N. Mukminov, Eduard A. Shuralev, Rustam Kh. Ravilov, Kamil S. Khaertynov

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Improving the early diagnosis of tuberculosis and solving a number of problems associated with the differential diagnosis of Mycobacterium bovis infection, nonspecific tuberculin reactions caused by sensitization of the body by non-tuberculosis mycobacteria, is urgent. The filtrates and extracts of M. bovis cell surface components are promising antigens with diagnostic potential. The purpose of this study was to isolate and characterize antigenic proteins and determine the dominant M. bovis antigens recognized by the humoral immune system. The mycobacterial cells were homogenized on FastPrep-24. Gel-filtration chromatography was used to fractionate the lysates of cell surface component extracts and proteins isolated from M. bovis culture supernatant. The separated fractions were analyzed using two-dimensional gel electrophoresis followed by determination of antigen serological activity using immunoblot with specific hyperimmune rabbit blood serum. As a result of electrophoretic separation of components by molecular weight, 23 antigen fractions were obtained. Analysis of densitograms showed that the fractions contained two zones of antigens with pronounced serological activity, corresponding to molecular weights of 28 and 21 kDa. The high serological activity of the 28 kDa antigen was established by immunoblot using hyperimmune blood sera. Separated and characterized by M. bovis specific antigen with a molecular weight of 28 kDa was added to the collection of specific marker antigens for M. bovis.

Keywords: antigen, gel-filtration chromatography, immunoblot, Mycobacterium bovis

Procedia PDF Downloads 133

Authors: Yingjeng James Li, Lung-Yu Sung, Huan-Jyun Ciou

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Durability of Membrane Electrode Assembly for Proton Exchange Membrane Fuel Cells was evaluated in both steady state and accelerated decay modes. Steady state mode was carried out at constant current of 800mA / cm2 for 2500 hours using air as cathode feed and pure hydrogen as anode feed. The degradation of the cell voltage was 0.015V after such 2500 hrs operation. The degradation rate was therefore calculated to be 6uV / hr. Accelerated mode was carried out by switching the voltage of the single cell between OCV and 0.2V. The durations held at OCV and 0.2V were 20 and 40 seconds, respectively, meaning one minute per cycle. No obvious change in performance of the MEA was observed after 10000 cycles of such operation.

Keywords: durability, lifetime, membrane electrode assembly, proton exchange membrane fuel cells

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Authors: Kanyarat Sikhao, Nichakorn Khondee

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Adjuvant is commonly mixed with agricultural spray solution during foliar application to improve the performance of microbial-based biological control, including better spreading, absorption, and penetration on a plant leaf. This research aims to replace chemical surfactants in adjuvant by biosurfactants for reducing a negative impact on antagonistic microorganisms and crops. Biosurfactant was produced from Brevibacterium casei NK8 and used as a cell-free broth solution containing a biosurfactant concentration of 3.7 g/L. The studies of microemulsion formation and phase behavior were applied to obtain the suitable composition of biosurfactant-based adjuvant, consisting of cell-free broth (70-80%), coconut oil-based fatty alcohol C12-14 (3) ethoxylate (1-7%), and sodium chloride (8-30%). The suitable formula, achieving Winsor Type III microemulsion (bicontinuous), was 80% of cell-free broth, 7% of fatty alcohol C12-14 (3) ethoxylate, and 8% sodium chloride. This formula reduced the contact angle of water on parafilm from 70 to 31 degrees. The non-phytotoxicity against plant seed of Oryza sativa and Brassica rapa subsp. pekinensis were obtained from biosurfactant-based adjuvant (germination index equal and above 80%), while sodium dodecyl sulfate and tween80 showed phytotoxic effects to these plant seeds. The survival of Bacillus subtilis in biosurfactant-based adjuvant was higher than sodium dodecyl sulfate and tween80. The mixing of biosurfactant and plant-based surfactant could be considered as a viable, safer, and acceptable alternative to chemical adjuvant for sustainable organic farming.

Keywords: biosurfactant, microemulsion, bio-adjuvant, antagonistic microorganisms

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Authors: S. Sambath, P. Palanivel

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Research conducted in the last few decades has proven that an inclusion of Distributed Genaration (DG) into distribution systems considerably lowers the level of power losses and the power quality improved. Moreover, the choice of DG is even more attractive since it provides not only benefits in power loss minimisation, but also a wide range of other advantages including environment, economic, power qualities and technical issues. This paper is an intent to quantify and analyse the impact of distributed generation (DG) in Tamil Nadu, India to examine what the benefits of decentralized generation would be for meeting rural loads. We used load flow analysis to simulate and quantify the loss reduction and power quality enhancement by having decentralized generation available line conditions for actual rural feeders in Tamil Nadu, India. Reactive and voltage profile was considered. This helps utilities to better plan their system in rural areas to meet dispersed loads, while optimizing the renewable and decentralised generation sources.

Keywords: distributed generation, distribution system, load flow analysis, optimal location, power quality

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Authors: Mayuva Youngsabanant, Chanikarn Srinark, Supanyika Sengsai, Soratorn Kerdkriangkrai, Nongnuch Gumlungpat, Mayuree Pumipaiboon

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The follicular fluid proteins of healthy medium size follicles (4-6 mm in diameters) and large size follicles (7-8 mm in diameter) of large white pig ovaries were collected by using sterile technique. They were used for testing the effect on primary in vitro cell culture growth of porcine oviductal epithelial cells (pOEC). Porcine oviductal epithelial cells of luteal phase was culture in M199 and added with 10% fetal calf serum 2.2 mg/mL, NaHCO₃, 0.25 mM pyruvate, 15 µg/mL and 50 µg/mL, gentamycin sulfate at high humidified atmosphere with 5% CO₂ in 95% air atmosphere at 37°C for 96 h before testing. The optimized concentration of pFF of two follicle sizes (at concentration of 2, 4, 20, 40, 200, 400, 500, and 600 µg proteins) in culture medium was observed for 24 h using MTT assay. Results were analyzed with a one-way ANOVA in SPSS statistic. Moreover, pOEC was also studied in morphological characteristic on long-term culture. The results of long-term study revealed that pOEC showed 70-80 percentage of healthy morphology on epithelial-like character and contained 30 percentage of an elongated shape (fibroblast-like morphology) at 4 weeks of culture time. MTT assay reviewed an increase in the percentage of viability of pOEC in 2 treated of follicular fluid groups. Two treatment concentration groups were higher than control group (p < 0.05) but not in positive control group. Interestingly, at 200 µg protein of 2 treated follicular fluid groups were reached the highest cell viability which is higher than a positive control and it is significantly different form control group (P < 0.05). These cells are developed and had fibroblast elongate shape which is longer than the cells in control group and positive control group. This report implies that pFF of medium follicle size at 200 µg proteins and large follicle size at 200 and 500 µg proteins could be optimized concentration for using as a supplement in culture medium to promote cell growth and development instead of growth hormone from fetal calf serum. It could be applied in cell biotechnology researches. Acknowledgements: The project was funded by a grant from Silpakorn University Research and Development Institute (SURDI) and Faculty of Science, Silpakorn University, Thailand.

Keywords: in vitro, porcine follicular fluid protein (pFF), porcine oviductal epithelial cells (pOEC), MTT

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Authors: Ayda Youssef, Mohammed Ali Khalaf, Tarek Rafaat

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Background: Langerhans cell histiocytosis (LCH) is a rare multi-systemic disease that shows an abnormal proliferation of these kinds of cells associated with a granular infiltration that affects different structures of the human body, including the lung, liver, spleen, lymph nodes, brain, mucocutaneous, soft tissue (head and neck), and salivary glands. Evaluation of the extent of disease is one of the major predictors of patient outcome. Objectives: To recognize the pathogenesis of Langerhans cell histiocytosis (LCH), describe the radiologic criteria that are suggestive of LCH in different organs rather than the bones and to illustrate the appropriate differential diagnoses for LCH in each of the common extra-osseous sites. Material and methods: A retrospective study was done on 150 biopsy-proven LCH patients from 2007 to 2012. All patients underwent imaging studies, mostly US, CT, and MRI. These patients were reviewed to assess the extra-skeletal manifestations of LCH. Results: In 150 patients with biopsy-proven LCH, There were 33 patients with liver affection, 5 patients with splenic lesions, 55 patients with enlarged lymph nodes, 9 patient with CNS disease and 11 patients with lung involvement. Conclusions: Because of the frequent LCH children and evaluation of the extent of disease is one of the major predictors of patient outcome. Radiologist need to be familiar with its presentation in different organs and regions of body outside the commonest site of affection (bones). A high-index suspicion should be raised a biopsy is recommended in the presence of radiological suspicion. Chemotherapy is the preferred therapeutic modality.

Keywords: langerhans cell histiocytosis, extra-skeletal, pediatrics, radiology

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Authors: Jingjing Huang, Nengwei Li, Guanghua Wei, Jiabin You, Chao Wang, Junliang Zhang

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In this work, molecular dynamics (MD) simulation is applied to analyze the mass transport process in the cathode of proton exchange membrane fuel cell (PEMFC), of which all types of molecules situated in the cathode is considered. a reasonable and effective MD simulation process is provided, and models were built and compared using both Materials Studio and LAMMPS. The mass transport is one of the key issues in the study of proton exchange membrane fuel cells (PEMFCs). In this report, molecular dynamics (MD) simulation is applied to analyze the influence of Nafion ionomer distribution and Pt nano-particle size on mass transport process in the cathode. It is indicated by the diffusion coefficients calculation that a larger quantity of Nafion, as well as a higher equivalent weight (EW) value, will hinder the transport of oxygen. In addition, medium-sized Pt nano-particles (1.5~2nm) are more advantageous in terms of proton transport compared with other particle sizes (0.94~2.55nm) when the center-to-center distance between two Pt nano-particles is around 5 nm. Then mass transport channels are found to be formed between the hydrophobic backbone and the hydrophilic side chains of Nafion ionomer according to the radial distribution function (RDF) curves. And the morphology of these channels affected by the Pt size is believed to influence the transport of hydronium ions and, consequently the performance of PEMFC.

Keywords: cathode catalytic layer, mass transport, molecular dynamics, proton exchange membrane fuel cell

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Authors: Vrinda Hari Ankolekar, Anne D. Souza, Mamatha Hosapatna

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Introduction: Precise anatomical assessment of femoral neck anteversion (FNA) and the neck shaft angles (NSA) would be essential in diagnosing the pathological conditions involving hip joint and its ligaments. FNA of greater than 20 degrees is considered excessive femoral anteversion, whereas a torsion angle of fewer than 10 degrees is considered femoral retroversion. Excessive femoral torsion is not uncommon and has been associated with certain neurologic and orthopedic conditions. The enlargement and maturation of the hip joint increases at the 20th week of gestation and the NSA ranges from 135- 140◦ at birth. Material and methods: 48 femurs were tagged according to the GA and two photographs for each femur were taken using Nikon digital camera. Each femur was kept on a horizontal hard desk and end on an image of the upper end was taken for the estimation of FNA and a photograph in a perpendicular plane was taken to calculate the NSA. The images were transferred to the computer and were stored in TIFF format. Microsoft Paint software was used to mark the points and Image J software was used to calculate the angles digitally. 1. Calculation of FNA: The midpoint of the femoral head and the neck were marked and a line was drawn joining these two points. The angle made by this line with the horizontal plane was measured as FNA. 2. Calculation of NSA: The midpoint of the femoral head and the neck were marked and a line was drawn joining these two points. A vertical line was drawn passing through the tip of the greater trochanter to the inter-condylar notch. The angle formed by these lines was calculated as NSA. Results: The paired t-test for the inter-observer variability showed no significant difference between the values of two observers. (FNA: t=-1.06 and p=0.31; NSA: t=-0.09 and p=0.9). The FNA ranged from 17.08º to 33.97 º on right and 17.32 º to 45.08 º on left. The NSA ranged from 139.33 º to 124.91 º on right and 143.98 º to 123.8 º on left. Unpaired t-test was applied to compare the mean angles between the second and third trimesters which did not show any statistical significance. This shows that the FNA and NSA of femur did not vary significantly during the third trimester. The FNA and NSA were correlated with the GA using Pearson’s correlation. FNA appeared to increase with the GA (r=0.5) but the increase was not statistically significant. A decrease in the NSA was also noted with the GA (r=-0.3) which was also statistically not significant. Conclusion: The present study evaluates the FNA and NSA of the femur in fetuses and correlates their development with the GA during second and third trimesters. The FNA and NSA did not vary significantly during the third trimester.

Keywords: anteversion, coxa antetorsa, femoral torsion, femur neck shaft angle

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Authors: Maja Ruzic-Baf, Mirjana Radetic-Paic

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The use of computers by children has many positive aspects, including the development of memory, learning methods, problem-solving skills and the feeling of one’s own competence and self-confidence. Playing on line video games can encourage hanging out with peers having similar interests as well as communication; it develops coordination, spatial relations and presentation. On the other hand, the Internet enables quick access to different information and the exchange of experiences. How kids use computers and what the negative effects of this can be depends on various factors. ICT has improved and become easy to get for everyone. In the past 12 years so many video games has been made even to that level that some of them are free to play. Young people, even some adults, had simply start to forget about the real outside world because in that other, digital world, they have found something that makes them feal more worthy as a man. This article present the use of ICT, forms of behavior and addictions to on line video games. The use of computers by children has many positive aspects, including the development of memory, learning methods, problem-solving skills and the feeling of one’s own competence and self-confidence. Playing on line video games can encourage hanging out with peers having similar interests as well as communication; it develops coordination, spatial relations and presentation. On the other hand, the Internet enables quick access to different information and the exchange of experiences. How kids use computers and what the negative effects of this can be depends on various factors. ICT has improved and become easy to get for everyone. In the past 12 years so many video games has been made even to that level that some of them are free to play. Young people, even some adults, had simply start to forget about the real outside world because in that other, digital world, they have found something that makes them feal more worthy as a man. This article present the use of ICT, forms of behavior and addictions to on line video games.

Keywords: addiction to video games, behaviour, ICT, young people

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Authors: G. S. Ayyappan, Srinivas Kota, Jaffer R. C. Sheriff, C. Prakash Chandra Joshua

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In the present scenario of energy crises, energy conservation in the electrical machines is very important in the industries. In order to conserve energy, one needs to monitor the performance of an induction motor on-site and in-situ. The instruments available for this purpose are very meager and very expensive. This paper deals with the design and development of induction motor performance analyser on-line, on-site, and in-situ. The system measures only few electrical input parameters like input voltage, line current, power factor, frequency, powers, and motor shaft speed. These measured data are coupled to name plate details and compute the operating efficiency of induction motor. This system employs the method of computing motor losses with the help of equivalent circuit parameters. The equivalent circuit parameters of the concerned motor are estimated using the developed algorithm at any load conditions and stored in the system memory. The developed instrument is a reliable, accurate, compact, rugged, and cost-effective one. This portable instrument could be used as a handy tool to study the performance of both slip ring and cage induction motors. During the analysis, the data can be stored in SD Memory card and one can perform various analyses like load vs. efficiency, torque vs. speed characteristics, etc. With the help of the developed instrument, one can operate the motor around its Best Operating Point (BOP). Continuous monitoring of the motor efficiency could lead to Life Cycle Assessment (LCA) of motors. LCA helps in taking decisions on motor replacement or retaining or refurbishment.

Keywords: energy conservation, equivalent circuit parameters, induction motor efficiency, life cycle assessment, motor performance analysis

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Authors: Raju Ranjha, Surbhi Aggarwal

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Background: Inflammation plays an important role in infectious and non-infectious diseases. MiRNA is also reported to play role in inflammation and associated cancers. Chemokine CXCL12 is also known to play role in inflammation and various cancers. CXCL12/CXCR4 chemokine axis was involved in pathogenesis of IBD specially UC. Supplementation of CXCL12 induces homing of dendritic cells to spleen and enhances control of plasmodium parasite in BALB/c mice. We looked at the regulation of CXCL12β by miRNA in UC colitis. Prolonged inflammation of colon in UC patient increases the risk of developing colorectal cancer. We looked at the expression differences of CXCl12β and its targeting miRNA in cancer susceptible area of colon of UC patients. Aim: Aim of this study was to find out the expression regulation of CXCL12β by miRNA in inflammation. Materials and Methods: Biopsy samples and blood samples were collected from UC patients and non-IBD controls. mRNA expression was analyzed using microarray and real-time PCR. CXCL12β targeting miRNA were looked by using online target prediction tools. Expression of CXCL12β in blood samples and cell line supernatant was analyzed using ELISA. miRNA target was validated using dual luciferase assay. Results and conclusion: We found miR-200a regulate the expression of CXCL12β in UC. Expression of CXCL12β was increased in cancer susceptible part of colon and expression of its targeting miRNA was decreased in the same part of colon. miR-200a regulate CXCL12β expression in inflammation and may be an important therapeutic target in inflammation associated cancer.

Keywords: inflammation, miRNA, regulation, CXCL12

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Authors: Achitphol Chookaew, Paramee Thongsukhsai, Patamarerk Engsontia, Narongwit Nakwan, Pritsana Raugrut

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Lung cancer is one of the most common malignancies and primary cause of death due to cancer worldwide. Non-small cell lung cancer (NSCLC) is the main subtype in which majority of patients present with advanced-stage disease. Herein, we analyzed differentially expressed genes to find potential biomarkers for lung cancer diagnosis as well as prognostic markers. We used transcriptome data from our 2 NSCLC patients and public data (GSE81089) composing of 8 NSCLC and 10 normal lung tissues. Differentially expressed genes (DEGs) between NSCLC and normal tissue and between early-stage and late-stage NSCLC were analyzed by the DESeq2. Pairwise correlation was used to find the DEGs with false discovery rate (FDR) adjusted p-value £ 0.05 and |log2 fold change| ³ 4 for NSCLC versus normal and FDR adjusted p-value £ 0.05 with |log2 fold change| ³ 2 for early versus late-stage NSCLC. Bioinformatic tools were used for functional and pathway analysis. Moreover, the top ten genes in each comparison group were verified the expression and survival analysis via GEPIA. We found 150 up-regulated and 45 down-regulated genes in NSCLC compared to normal tissues. Many immnunoglobulin-related genes e.g., IGHV4-4, IGHV5-10-1, IGHV4-31, IGHV4-61, and IGHV1-69D were significantly up-regulated. 22 genes were up-regulated, and five genes were down-regulated in late-stage compared to early-stage NSCLC. The top five DEGs genes were KRT6B, SPRR1A, KRT13, KRT6A and KRT5. Keratin 6B (KRT6B) was the most significantly increased gene in the late-stage NSCLC. From GEPIA analysis, we concluded that IGHV4-31 and IGKV1-9 might be used as diagnostic biomarkers, while KRT6B and KRT6A might be used as prognostic biomarkers. However, further clinical validation is needed.

Keywords: differentially expressed genes, early and late-stages, gene ontology, non-small cell lung cancer transcriptomics

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Authors: Reham H. Soliman, Noha Noufal, Howayda AbdelAal

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Calcium/calmodulin-dependent serine protein kinase (CASK) belongs to the membrane-associated guanylate kinase (MAGUK) family and has been proposed as a mediator of cell-cell adhesion and proliferation, which can contribute to tumorogenesis. CASK has been linked as a good prognostic factor with some tumor subtypes, while considered as a poor prognostic marker in others. To our knowledge, no sufficient evidence of CASK role in colorectal cancer is available. The aim of this study is to evaluate the expression of Calcium/calmodulin-dependent serine protein kinase (CASK) in non-mucinous colorectal adenocarcinoma and adenomatous polyps as precursor lesions and assess its prognostic significance. The study included 42 cases of conventional colorectal adenocarcinoma and 15 biopsies of adenomatous polyps with variable degrees of dysplasia. They were reviewed for clinicopathological prognostic factors and stained by CASK; mouse, monoclonal antibody using heat-induced antigen retrieval immunohistochemical techniques. The results showed that CASK protein was significantly overexpressed (p <0.05) in CRC compared with adenoma samples. The CASK protein was overexpressed in the majority of CRC samples with 85.7% of cases showing moderate to strong expression, while 46.7% of adenomas were positive. CASK overexpression was significantly correlated with both TNM stage and grade of differentiation (p <0.05). There was a significantly higher expression in tumor samples with early stages (I/II) rather than advanced stage (III/IV) and with low grade (59.5%) rather than high grade (40.5%). Another interesting finding was found among the adenomas group, where the stronger intensity of staining was observed in samples with high grade dysplasia (33.3%) than those of lower grades (13.3%). In conclusion, this study shows that there is significant overexpression of CASK protein in CRC as well as in adenomas with high grade dysplasia. This indicates that CASK is involved in the process of carcinogenesis and functions as a potential trigger of the adenoma-carcinoma cascade. CASK was significantly overexpressed in early stage and low-grade tumors rather than tumors with advanced stage and higher histological grades. This suggests that CASK protein is a good prognostic factor. We suggest that CASK affects CRC in two different ways derived from its physiology. CASK as part of MAGUK family can stimulate proliferation and through its cell membrane localization and as a mediator of cell-cell adhesion might contribute in tumor confinement and localization.

Keywords: CASK, colorectal cancer, overexpression, prognosis

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Authors: Eva Filová, Jana Daňková, Věra Sovková, Matej Daniel

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Titanium alloys are biocompatible metals that are widely used in clinical practice as load bearing implants. The chemical modification may influence cell adhesion, proliferation, and differentiation as well as stiffness of the material. The aim of the study was to evaluate the adhesion, growth and differentiation of pig mesenchymal stem cells on the novel beta titanium alloy Ti36Nb6Ta compared to standard medical titanium alloy Ti6Al4V. Discs of Ti36Nb6Ta and Ti6Al4V alloy were sterilized by ethanol, put in 48-well plates, and seeded by pig mesenchymal stem cells at the density of 60×103/cm2 and cultured in Minimum essential medium (Sigma) supplemented with 10% fetal bovine serum and penicillin/streptomycin. Cell viability was evaluated using MTS assay (CellTiter 96® AQueous One Solution Cell Proliferation Assay;Promega), cell proliferation using Quant-iT™ ds DNA Assay Kit (Life Technologies). Cells were stained immunohistochemically using monoclonal antibody beta-actin, and secondary antibody conjugated with AlexaFluor®488 and subsequently the spread area of cells was measured. Cell differentiation was evaluated by alkaline phosphatase assay using p-nitrophenyl phosphate (pNPP) as a substrate; the reaction was stopped by NaOH, and the absorbance was measured at 405 nm. Osteocalcin, specific bone marker was stained immunohistochemically and subsequently visualized using confocal microscopy; the fluorescence intensity was analyzed and quantified. Moreover, gene expression of osteogenic markers osteocalcin and type I collagen was evaluated by real-time reverse transcription-PCR (qRT-PCR). For statistical evaluation, One-way ANOVA followed by Student-Newman-Keuls Method was used. For qRT-PCR, the nonparametric Kruskal-Wallis Test and Dunn's Multiple Comparison Test were used. The absorbance in MTS assay was significantly higher on titanium alloy Ti6Al4V compared to beta titanium alloy Ti36Nb6Ta on days 7 and 14. Mesenchymal stem cells were well spread on both alloys, but no difference in spread area was found. No differences in alkaline phosphatase assay, fluorescence intensity of osteocalcin as well as the expression of type I collagen, and osteocalcin genes were observed. Higher expression of type I collagen compared to osteocalcin was observed for cells on both alloys. Both beta titanium alloy Ti36Nb6Ta and titanium alloy Ti6Al4V Ti36Nb6Ta supported mesenchymal stem cellsˈ adhesion, proliferation and osteogenic differentiation. Novel beta titanium alloys Ti36Nb6Ta is a promising material for bone implantation. The project was supported by the Czech Science Foundation: grant No. 16-14758S, the Grant Agency of the Charles University, grant No. 1246314 and by the Ministry of Education, Youth and Sports NPU I: LO1309.

Keywords: beta titanium, cell growth, mesenchymal stem cells, titanium alloy, implant

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Authors: Yi-Chiang Huang, Hsu-Feng Lee, Yu-Chao Tseng, Wen-Yao Huang

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Proton exchange membranes as a key component in fuel cells have been widely studying over the past few decades. As proton exchange, membranes should have some main characteristics, such as good mechanical properties, low oxidative stability and high proton conductivity. In this work, trifluoromethyl groups had been introduced on polymer backbone and phenyl side chain which can provide densely located sulfonic acid group substitution and also promotes solubility, thermal and oxidative stability. Herein, a series of novel sulfonated aromatic hydrocarbon polyelectrolytes was synthesized by polycondensation of 4,4''''-difluoro-3,3''''- bis(trifluoromethyl)-2'',3''-bis(3-(trifluoromethyl)phenyl)-1,1':4',1'':4'',1''':4''',1''''-quinquephenyl with 2'',3''',5'',6''-tetraphenyl-[1,1':4',1'': 4'',1''':4''',1''''-quinquephenyl]-4,4''''-diol and post-sulfonated was through chlorosulfonic acid to given sulfonated polymers (SFC3-X) possessing ion exchange capacities ranging from 1.93, 1.91 and 2.53 mmol/g. ¹H NMR and FT-IR spectroscopy were applied to confirm the structure and composition of sulfonated polymers. The membranes exhibited considerably dimension stability (10-27.8% in length change; 24-56.5% in thickness change) and excellent oxidative stability (weight remain higher than 97%). The mechanical properties of membranes demonstrated good tensile strength on account of the high rigidity multi-phenylated backbone. Young's modulus were ranged 0.65-0.77GPa which is much larger than that of Nafion 211 (0.10GPa). Proton conductivities of membranes ranged from 130 to 240 mS/cm at 80 °C under fully humidified which were comparable or higher than that of Nafion 211 (150 mS/cm). The morphology of membranes was investigated by transmission electron microscopy which demonstrated a clear hydrophilic/hydrophobic phase separation with spherical ionic clusters in the size range of 5-20 nm. The SFC3-1.97 single fuel cell performance demonstrates the maximum power density at 1.08W/cm², and Nafion 211 was 1.24W/cm² as a reference in this work. The result indicated that SFC3-X are good candidates for proton exchange membranes in fuel cell applications. Fuel cell of other membranes is under testing.

Keywords: fuel cells, polyelectrolyte, proton exchange membrane, sulfonated polymers

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Authors: Asma Alhuqail, Nagwa Aref

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Programmed cell death resembles a real nature active defense in Datura metel against TMV after three days of virus infection. Physiological plant response was assessed for asymptomatic healthy and symptomatic infected detached leaves. The results indicated H2O2 and Chlorophyll-a as the most potential parameters. Chlorophyll-a was considered the only significant predictor variant for the H2O2 dependent variant with a P value of 0.001 and R-square of 0.900. The plant immune response was measured within three days of virus infection using the cutoff value of H2O2 (61.095 lmol/100 mg) and (63.201 units) for the tail moment in the Comet Assay. Their percentage changes were 255.12% and 522.40% respectively which reflects the stress of virus infection in the plant. Moreover, H2O2 showed 100% specificity and sensitivity in the symptomatic infected group using the receiver-operating characteristic (ROC). All tested parameters in the symptomatic infected group had significant correlations with twenty-five positive and thirty-one negative correlations where the P value was <0.05 and 0.01. Chlorophyll-a parameter had a crucial role of highly significant correlation between total protein and salicylic acid. Contrarily, this correlation with tail moment unit was (r = _0.930, P <0.01) where the P value was < 0.01. The strongest significant negative correlation was between Chlorophyll-a and H2O2 at P < 0.01, while moderate negative significant correlation was seen for Chlorophyll-b where the P value < 0.05. The present study discloses the secret of the three days of rapid transient production of activated oxygen species (AOS) that was enough for having potential quantitative physiological parameters for defensive plant response toward the virus.

Keywords: programmed cell death, plant–adaptive immune response, hydrogen peroxide (H2O2), physiological parameters

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Authors: Meihua Fang, Yipan Guo, Tao Fei, Pengyu Tian

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Space plasma caused spacecraft surface charging is the major space environment hazard. Particle in cell (PIC) method can be used to simulate the interaction between space plasma and spacecraft. It was proved that surface charging level of spacecraft in Jupiter’s orbits was high for its’ electron-heavy plasma environment. In this paper, Jovian plasma environment is modeled and surface charging analysis is carried out by PIC based software Spacecraft Plasma Interaction System (SPIS). The results show that the spacecraft charging potentials exceed 1000V at 2Rj, 15Rj and 25Rj polar orbits in the dark side at worst case plasma model. Furthermore, the simulation results indicate that the large Jovian magnetic field increases the surface charging level for secondary electron gyration.

Keywords: Jupiter, PIC, space plasma, surface charging

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Authors: Abdull Halim Abdull, Nasiman Sapari, Mohammad Haikal Asyraf Bin Anuar

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Although soil thermal properties are required in many areas to improve oil recovery, they are seldom measured on a routine basis. Reasons for this are unclear, but may be related to a lack of suitable instrumentation and entropy theory. We integrate single probe thermal gradient for the radial conduction of a short-duration heat pulse away from a single electrode source, and compared it with the theory for an instantaneously heated line source. By measuring the temperature response at a short distance from the line source, and applying short-duration heat-pulse theory, we can extract all the entropy properties, the thermal diffusivity, heat capacity, and conductivity, from a single heat-pulse measurement. Results of initial experiments carried out on air-dry sand and clay materials indicate that this heat-pulse method yields soil thermal properties that compare well with thermal properties measured by single electrode.

Keywords: entropy, single probe thermal gradient, soil thermal, probe heat

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Authors: Suharni Mohamad Suharni Mohamad, Nurul Izzati Hamzan Nurul Izzati Hamzan, Norhayu Abdul Rahman Norhayu Abdul Rahman, Siti Suraiya Md Noor Siti Suraiya Md Noor

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Human papillomavirus (HPV) is an important risk factor for development of oral cancer. HPV16 is the most common type found in HPV-positive squamous cell carcinoma. In the present study, we established a real-time loop-mediated isothermal amplification (real-time LAMP) for detection of HPV16. A set of six primers was specially designed to recognize eight distinct sequences of HPV16-E6. Detection and quantification was achieved by real-time monitoring using a real-time turbidimeter based on threshold time required for turbidity in the LAMP reaction. LAMP reagents (MgSO4, dNTPs, Bst polymerase concentrations) and various incubation times and temperatures were optimized. The sensitivity was determined using 10-fold serial dilutions of HPV16 standard strain. The specificity of was evaluated using other HPV genotypes. The optimized method was established with specifically designed primers by real-time detection in approximately 30 min at 65°C. The limit of detection of HPV16 using the LAMP assay was 10 pg/ml that could be detected in 30 min. The LAMP assay was 10 times more sensitive than the conventional PCR in detecting HPV16. No cross-reactivity with other HPV genotypes was observed. This quantitative real-time LAMP assay may improve diagnostic potential for the detection and quantification of HPV16 in clinical samples and epidemiological studies due to its rapidity, simplicity, high sensitivity and specificity. This assay will be further evaluated with HPV DNAs of saliva from patients with oral squamous cell carcinoma. Acknowledgement: This study was financially supported by the ScienceFund Grant, Ministry of Science, Technology and Innovation (305/PPSG/6113219).

Keywords: Oral Squamous Cell Carcinoma (OSCC), Human Papillomavirus 16 (HPV16), Loop-Mediated Isothermal Amplification (LAMP), rapid detection

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Authors: Shuo Mu, Guangzhi Jiang, Jinsa Chen

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The analysis of Indel for RNA sequencing of clinical samples is easily affected by sequencing experiment errors and software selection. In order to improve the efficiency and accuracy of analysis, we developed an automatic reporting system for Indel recognition and annotation based on image snapshot of transcriptome reads alignment. This system includes sequence local-assembly and realignment, target point snapshot, and image-based recognition processes. We integrated high-confidence Indel dataset from several known databases as a training set to improve the accuracy of image processing and added a bioinformatical processing module to annotate and filter Indel artifacts. Subsequently, the system will automatically generate data, including data quality levels and images results report. Sanger sequencing verification of the reference Indel mutation of cell line NA12878 showed that the process can achieve 83% sensitivity and 96% specificity. Analysis of the collected clinical samples showed that the interpretation accuracy of the process was equivalent to that of manual inspection, and the processing efficiency showed a significant improvement. This work shows the feasibility of accurate Indel analysis of clinical next-generation sequencing (NGS) transcriptome. This result may be useful for RNA study for clinical samples with microsatellite instability in immunotherapy in the future.

Keywords: automatic reporting, indel, next-generation sequencing, NGS, transcriptome

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