Search results for: fluorescent probe

Authors: Dalma Dojcsák, Csaba Váradi, Flóra Farkas, Tamás Farkas, János Papp, Béla Viskolcz

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Background: Appendicitis is a common acute inflammatory condition in both children and adults, but current laboratory markers such as C-reactive protein (CRP), white blood cell count (WBC), absolute neutrophil count (ANC), and red blood cell count (RNC) lack specificity in detecting appendicitis-related inflammation. N-glycosylation, an asparagine-linked glycosylation process, plays a vital role in cellular interactions, angiogenesis, immune response, and effector functions. Altered N-glycosylation impacts tumor growth and both acute and chronic inflammatory processes. IgG, the second most abundant glycoprotein in serum, shows altered glycosylation patterns during inflammation, suggesting that IgG glycan modifications may serve as potential biomarkers for appendicitis. Specifically, increased levels of agalactosylated IgG glycans are a known feature of various inflammatory conditions, potentially including appendicitis. Identifying pediatric appendicitis remains challenging due to the absence of specific biomarkers, which makes diagnosis reliant on clinical symptoms, imaging such as ultrasound, and nonspecific lab indicators (e.g., CRP, WBC, ANC). In this study, we analyzed the IgG derived N-glycome in pediatric patients with appendicitis compared with healthy controls. Methodology: The N-glycome was analyzed by high-performance liquid-chromatography combined with mass spectrometry. IgG was isolated from serum samples by Protein G column. The IgG derived glycans were released by enzymatic deglycosylation and fluorescent tags were attached to each glycan moiety, which made necessitates the sample clean-up for further reliable quantitation. Overall, 38 controls and 40 serum samples diagnosed with pediatric appendicitis were analyzed by HILIC-MS methods. Multivariate statistical tests were performed with area percentage under the peak data derived from the integrated peaks, which were obtained from the chromatograms. Conclusions: Our results represented the altered N-glycome of IgG in pediatric appendicitis is similar with other observations. The glycosylation pattern reported so far for IgG is characterized by decreased galactosylation and sialylation, and an increase in fucosylation.

Keywords: N-glycosylation, liquid chromatography, mass spectrometry, inflammation, appendicitis, immunoglobulin G

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Authors: H. M. Fayzan Shakir, Khadija Zubair, Tingkai Zhao

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Electromagnetic (EM) waves are being used widely now a days. Cell phone signals, WIFI signals, wireless telecommunications etc everything uses EM waves which then create EM pollution. EM pollution can cause serious effects on both human health and nearby electronic devices. EM waves have electric and magnetic components that disturb the flow of charged particles in both human nervous system and electronic devices. The shielding of both humans and electronic devices are a prime concern today. EM waves can cause headaches, anxiety, suicide and depression, nausea, fatigue and loss of libido in humans and malfunctioning in electronic devices. Polyaniline (PANI) and polypyrrole (PPY) were successfully synthesized using chemical polymerizing using ammonium persulfate and DBSNa as oxidant respectively. Barium ferrites (BaFe) were also prepared using co-precipitation method and calcinated at 10500C for 8h. Nanocomposite thin films with various combinations and compositions of Polyvinylchloride, PANI, PPY and BaFe were prepared. X-ray diffraction technique was first used to confirm the successful fabrication of all nano fillers and particle size analyzer to measure the exact size and scanning electron microscopy is used for the shape. According to Electromagnetic Interference theory, electrical conductivity is the prime property required for the Electromagnetic Interference shielding. 4-probe technique is then used to evaluate DC conductivity of all samples. Samples with high concentration of PPY and PANI exhibit remarkable increased electrical conductivity due to fabrication of interconnected network structure inside the Polyvinylchloride matrix that is also confirmed by SEM analysis. Less than 1% transmission was observed in whole NIR region (700 nm – 2500 nm). Also, less than -80 dB Electromagnetic Interference shielding effectiveness was observed in microwave region (0.1 GHz to 20 GHz).

Keywords: nanocomposites, polymers, EMI shielding, thermal imaging

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Authors: Lynn Mcquarrie, Charlotte Enns

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The goal of this project was to develop a visually-based, signed language phonological awareness training program and to pilot the intervention with signing deaf children (ages 6 -10 years/ grades 1 - 4) who were beginning readers to assess the effects of systematic explicit American Sign Language (ASL) phonological instruction on both ASL vocabulary and English print vocabulary learning. Growing evidence that signing learners utilize visually-based signed language phonological knowledge (homologous to the sound-based phonological level of spoken language processing) when reading underscore the critical need for further research on the innovation of reading instructional practices for visual language learners. Multiple single-case studies using a multiple probe design across content (i.e., sign and print targets incorporating specific ASL phonological parameters – handshapes) was implemented to examine if a functional relationship existed between instruction and acquisition of these skills. The results indicated that for all cases, representing a variety of language abilities, the visually-based phonological teaching approach was exceptionally powerful in helping children to build their sign and print vocabularies. Although intervention/teaching studies have been essential in testing hypotheses about spoken language phonological processes supporting non-deaf children’s reading development, there are no parallel intervention/teaching studies exploring hypotheses about signed language phonological processes in supporting deaf children’s reading development. This study begins to provide the needed evidence to pursue innovative teaching strategies that incorporate the strengths of visual learners.

Keywords: American sign language phonological awareness, dual language strategies, vocabulary learning, word reading

Procedia PDF Downloads 333

Authors: Roman Matejka, Vaclav Prochazka, Tibor Izak, Jana Stepanovska, Martina Travnickova, Alexander Kromka

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Cell-based impedance spectroscopy is suitable method for electrical monitoring of cell activity especially on substrates that cannot be easily inspected by optical microscope (without fluorescent markers) like decellularized tissues, nano-fibrous scaffold etc. Special sensor for this measurement was developed. This sensor consists of corning glass substrate with gold interdigitated electrodes covered with diamond layer. This diamond layer provides biocompatible non-conductive surface for cells. Also, a special PPFC flow cultivation chamber was developed. This chamber is able to fix sensor in place. The spring contacts are connecting sensor pads with external measuring device. Construction allows real-time live cell imaging. Combining with perfusion system allows medium circulation and generating shear stress stimulation. Experimental evaluation consist of several setups, including pure sensor without any coating and also collagen and fibrin coating was done. The Adipose derived stem cells (ASC) and Human umbilical vein endothelial cells (HUVEC) were seeded onto sensor in cultivation chamber. Then the chamber was installed into microscope system for live-cell imaging. The impedance measurement was utilized by vector impedance analyzer. The measured range was from 10 Hz to 40 kHz. These impedance measurements were correlated with live-cell microscopic imaging and immunofluorescent staining. Data analysis of measured signals showed response to cell adhesion of substrates, their proliferation and also change after shear stress stimulation which are important parameters during cultivation. Further experiments plan to use decellularized tissue as scaffold fixed on sensor. This kind of impedance sensor can provide feedback about cell culture conditions on opaque surfaces and scaffolds that can be used in tissue engineering in development artificial prostheses. This work was supported by the Ministry of Health, grants No. 15-29153A and 15-33018A.

Keywords: bio-impedance measuring, bioreactor, cell cultivation, diamond layer, gold interdigitated electrodes, tissue engineering

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

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

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

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Authors: Ivana Hrebickova, Maria Sevcikova, Romana Slamberova

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The present study was aimed at evaluation of cognitive changes following scheduled neonatal methamphetamine exposure in combination with long-term exposure in adulthood of female Wistar rats. Pregnant mothers were divided into two groups: group with indirect exposure (methamphetamine in dose 5 mg/ml/kg, saline in dose 1 ml/kg) during early lactation period (postnatal day 1–11) - progeny of these mothers were exposed to the effects of methamphetamine or saline indirectly via the breast milk; and the second group with direct exposure – all mothers were left intact for the entire lactation period, while progeny was treated with methamphetamine (5 mg/ml/kg) by injection or the control group, which was received needle pick (shame, not saline) at the same time each day of period of application (postnatal day 1–11). Learning ability and memory consolidation were tested in the Morris Water Maze, which consisted of three types of tests: ‘Place Navigation Test ‘; ‘Probe Test ‘; and ‘Memory Recall Test ‘. Adult female progeny were injected daily, after completion last trial with saline or methamphetamine (1 mg/ml/kg). We compared the effects of indirect/direct neonatal methamphetamine exposure and adult methamphetamine treatment on cognitive function of female rats. Statistical analyses showed that neonatal methamphetamine exposure worsened spatial learning and ability to remember the position of the platform. The present study demonstrated that direct methamphetamine exposure has more significant impact on process of learning and memory than indirect exposure. Analyses of search strategies (thigmotaxis, scanning) used by females during the Place Navigation Test and Memory Recall Test confirm all these results.

Keywords: methamphetamine, Morris water maze, neonatal exposure, strategies, Wistar rats

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Authors: Nur Farah Ain Zainee, Ahmad Ismail, Nazlina Ibrahim, Asmida Ismail

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The study of genus Halimeda in Malaysia is in the early stage due to less specific study on its taxonomy. Most of the previous research tend to choose other genus such as Caulerpa and Gracilaria because of the potential of being utilized. The identification of Halimeda is complex by the high morphological variation within individual species due to different types of habitat and the changes in composition of seawater. The study was completed to study the diversity and distribution of Halimeda in Malaysia and to identify the morphological and anatomical differences between Halimeda species. The methods which have been used for this study are collection of Halimeda and seawater, preservation of specimen, identification of the specimen including the preparation of the temporary slide and decalcification of the calcium layer by using diluted hydrochloric acid. The specimen were processed in laboratory and kept as herbarium specimen in Algae Herbarium, Universiti Kebangsaan Malaysia. Environmental parameters were tested by using YSI multiparameter probe and the recorded data were temperature, salinity, pH and dissolved oxygen. The nutrient content of seawater such as nitrate and phosphate were analysed by using Hach kit model DR 2000. In the present study, out of 330 herbarium specimen, ten species were identified as Halimeda cuneata, H. discoidea, H. macroloba, H. macrophysa, H. opuntia, H. simulans, H. stuposa, H. taenicola, H. tuna and H. velasquezii. Of these, five species were new record to Malaysia. They are Halimeda cuneata, H. macrophysa, H. stuposa, H. taenicola and H. velasquezii. H. opuntia was found as the most abundance species with wide distribution in Malaysia coastal area. Meanwhile, from the study of their distribution, two localities in which Pulau Balak Balak, Kudat and Pulau Langkawi, Kedah, were noted having high number of Halimeda species. As a conclusion, this study has successfully identified ten species of Halimeda of Malaysia with full description of morphological characteristics that may assist further researcher to differentiate and identify Halimeda.

Keywords: Distribution, diversity, Halimeda, morphological, taxonomy

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Authors: Abdullah Alnutayfat, Alexander Sutin

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One of the most important types of renewable energy resources is wind energy which can be produced by wind turbines. The blades of the wind turbine are exposed to the pressure of the harsh environment, which causes a significant issue for the wind power industry in terms of the maintenance cost and failure of blades. One of the reliable methods for blade inspection is the vibroacoustic structural health monitoring (SHM) method which examines information obtained from the structural vibrations of the blade. However, all vibroacoustic SHM techniques are based on comparing the structural vibration of intact and damaged structures, which places a practical limit on their use. Methods for nonlinear vibroacoustic SHM are more sensitive to damage and cracking and do not need to be compared to data from the intact structure. This paper presents the Vibro-Acoustic Modulation (VAM) method based on the modulation of high-frequency (probe wave) by low-frequency loads (pump wave) produced by the blade rotation. The blade rotation alternates bending stress due to gravity, leading to crack size variations and variations in the blade resonance frequency. This method can be used with the classical SHM vibration method in which the blade is excited by piezoceramic actuator patches bonded to the blade and receives the vibration response from another piezoceramic sensor. The VAM modification of this method analyzes the spectra of the detected signal and their sideband components. We suggest the VAM model as the simple mechanical oscillator, where the parameters of the oscillator (resonance frequency and damping) are varied due to low-frequency blade rotation. This model uses the blade vibration parameters and crack influence on the blade resonance properties from previous research papers to predict the modulation index (MI).

Keywords: wind turbine blades, damaged detection, vibro-acoustic structural health monitoring, vibro-acoustic modulation

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Authors: Ramin Rezaei, Terry Ng, Abdollah Afjeh

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Near wake development of a wind turbine affects the aerodynamic loads on the tower and the wind turbine. Design considerations of both isolated wind turbines and wind farms must include unsteady wake flow conditions under which the turbines must operate. The consequent aerodynamic loads could lead to over design of wind turbines and adversely affect the cost of wind turbines and, in turn, the cost of energy produced by wind turbines. Reducing the weight of turbine rotors is particularly desirable since larger wind turbine rotors can be utilized without significantly increasing the cost of the supporting structure. Larger rotor diameters produce larger swept areas and consequently greater energy production from the wind thereby reducing the levelized cost of wind energy. To understand the development and structure of the near tower wake of a wind turbine, an experimental study was conducted to describe the flow field of the near wake for both upwind and downwind turbines. The study was conducted under controlled environment of a wind tunnel using a scaled model of a turbine. The NREL 5 MW reference wind turbine was used as a baseline design and was modified as necessary to design and build upwind and downwind scaled wind turbine models. This paper presents the results of the wind tunnel study using turbine models to quantify the near wake of upwind and downwind wind turbine configurations for various lengths of tower-to-turbine spacing. The variations of mean velocity and turbulence are measured using a computer-controlled, traversing hot wire probe. Additionally, smoke flow visualizations were conducted to qualitatively study the wake. The results show a more rapid dissipation of the near wake for an upwind configuration. The results can readily be incorporated into low fidelity system level turbine simulation tools to more accurately account for the wake on the aerodynamic loads of a upwind and downwind turbines.

Keywords: hot wire anemometry, near wake, upwind and downwind turbine. Hot wire anemometry, near wake, upwind and downwind turbine

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Authors: Martin C. Eze, Gao Min

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Lithium bis(trifluoromethanesulfonyl)imide (Li-TFSI) dopant is beneficial in improving the properties of 2,2′,7,7′-Tetrakis (N, N-di-p-methoxyphenylamino)-9,9′-spiro-bifluorene (Spiro-OMETAD) transport layer used in perovskite solar cells (PSCs). Properties such as electrical conductivity, band energy mismatch, and refractive index of Spiro-OMETAD layers are believed to play key roles in PSCs performance but only the dependence of electrical conductivity on Li-TFSI doping has been extensively studied. In this work, the effect of Li-TFSI doping level on highest occupied molecular orbital (HOMO) energy, electrical conductivity, and refractive index of Spiro-OMETAD film and PSC performance was demonstrated. The Spiro-OMETAD films were spin-coated at 4000 rpm for 30 seconds from solutions containing 73.4 mM of Spiro-OMeTAD, 23.6 mM of 4-tert-butylpyridine, 7.6 mM of tris(2-(1H-pyrazol-1-yl)-4-tert-butylpyridine) cobalt(III) tri[bis(trifluoromethane) sulfonimide] (FK209) dopant and Li-TFSI dopant varying from 37 to 62 mM in 1 ml of chlorobenzene. From ultraviolet photoelectron spectroscopy (UPS), ellipsometry, and 4-probe studies, the results show that films deposition from Spiro-OMETAD solution doped with 40 mM of Li-TFSI shows the highest electrical conductivity of 6.35×10-6 S/cm, the refractive index of 1.87 at 632.32 nm, HOMO energy of -5.22 eV and the lowest HOMO energy mismatch of 0.21 eV compared to HOMO energy of perovskite layer. The PSCs fabricated show the best power conversion efficiency, open-circuit voltage, and fill factor of 17.10 %, 1.1 V, and 70.12%, respectively, for devices based on Spiro-OMETAD solution doped with 40 mM of Li-TFSI. This study demonstrates that the optimum Spiro-OMETAD/ Li-TFSI doping ratio of 1.84 is the optimum doping level for Spiro-OMETAD layer preparation.

Keywords: electrical conductivity, homo energy mismatch, lithium bis(trifluoromethanesulfonyl)imide, power conversion efficiency, refractive index

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Authors: Victor S. Klimin, Alexey A. Rezvan, Maxim S. Solodovnik, Oleg A. Ageev

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In this paper, the problems of existing methods of profiling and surface modification of nanoscale arsenide-gallium structures are analyzed. The use of a combination of methods of local anodic oxidation and plasma chemical etching to solve this problem is considered. The main features that make this technology one of the promising areas of modification and profiling of near-surface layers of solids are demonstrated. In this paper, we studied the effect of formation stress and etching time on the geometrical parameters of the etched layer and the roughness of the etched surface. Experimental dependences of the thickness of the etched layer on the time and stress of formation were obtained. The surface analysis was carried out using atomic force microscopy methods, the corresponding profilograms were constructed from the obtained images, and the roughness of the etched surface was studied accordingly. It was shown that at high formation voltage, the depth of the etched surface increased, this is due to an increase in the number of active particles (oxygen ions and hydroxyl groups) formed as a result of the decomposition of water molecules in an electric field, during the formation of oxide nanostructures on the surface of gallium arsenide. Oxide layers were used as negative masks for subsequent plasma chemical etching by the STE ICPe68 unit. BCl₃ was chosen as the chlorine-containing gas, which differs from analogs in some parameters for the effect of etching of nanostructures based on gallium arsenide in the low-temperature plasma. The gas mixture of reaction chamber consisted of a buffer gas NAr = 100 cm³/min and a chlorine-containing gas NBCl₃ = 15 cm³/min at a pressure P = 2 Pa. The influence of these methods modes, which are formation voltage and etching time, on the roughness and geometric parameters, and corresponding dependences are demonstrated. Probe nanotechnology was used for surface analysis.

Keywords: nanostructures, GaAs, plasma chemical etching, modification structures

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Authors: Dionysios Kakaroumpas, Jesseka Farago, Stephen Webber

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Athletes and spectators depend on the tennis ranking system to represent the truest caliber of athletic prowess; a careful look at the current ranking system though, reveals its main weakness: it undermines expectations of fans and players. Our study proposes several key changes to the existing ranking formula that provide a fair and accurate approach to measure player performance. The study proposes a modification of the system to value: participation, continued advancement, and overall achievement. The new ranking formula facilitates closing the trust gap, encouraging competition equality, engaging the fan base, attracting investment, and promoting tennis involvement worldwide. To probe the crux of our main contention we performed week-by-week comparisons between results procured from the current and proposed formulae. After performing this rigorous case-study of top players of each gender, the findings strongly indicated that there is identifiable inflation in the ranks and enhanced the conviction that the current system should be updated. The new system is accompanied by a web-based software package freely available to anyone involved or interested in tennis rankings. The software package is designed to automatically calculate new player rankings based on a responsive, multi-faceted formula that also generates projected point scenarios and provides separate rankings for the three different court surfaces. By taking a critical look at the current tennis ranking system with consideration to the perspective of fans, players, and businesses involved, an upgrade is in order for it to maintain the balance of trust between fans and the evaluation process. In closure, this proposed solution increases fair play competition, eliminates rank inflation, and better engages fans, players, and sponsors by bringing in a new era of professional tennis.

Keywords: measurement and evaluation, rules and regulations, sports management and marketing, tennis ranking system

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Authors: M. C. Muya, L. J. Erasmus

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Twenty four new born male Holstein calves were divided into two treatments groups and used to evaluate the effects of M. elsdenii NCIMB 41125. The first groups were dosed with 50 ml containing 108 CFU/mL of M. elsdenii NCIMB 41125 (Me) and the control calves were not dosed. Within each of the two treatments groups, calves were divided into three treatment groups (Not dosed: 7 d, 14 d and 21 d vs dosed Me 7 d, Me14 and Me21 d (treatments), each groups contained 4 calves within which two calves were euthanized at 24 h and two calves at 72 h. Calves entered the trial until euthanize at whether 24 or 72 H after dosing time. After receiving colostrum for 3 consecutive days after birth, calves were fed whole milk and had free access to a commercial calf starter pellet and fresh water. Fecal grab samples were taken from each calf in duplicate +24 h or +72 h relative to dosing. Immediately after euthanizing, the digestive tract was harvested, and duplicate rumen and colon digesta samples collected for VFA’s determination and DNA extraction for bacteria count using 16s RNA PCR probe technique. Independent two t-test was performed to compare mean volatile fatty acids. Mixed-effects linear regressions were performed to establish relationships between: 1) M. elsdenii and Me, and between VFA’s and Me using SAS (2009). M. elsdenii NCIMB 41125 was detected in the faeces, colon and rumen of dosed calves at both +24H and +72H and ranged from 1.6 x 106 to 4.9 x 109 cfu/ml, indicating its potential to colonize in the digestive tract of calves. There was a strong positive relationship (R²=0.96; P < 0.0001) between M. elsdenii NCIMB 41125 and M. elsdenii population (cfu/ml) in the rumen, suggesting that the increase in M. elsdenii was due to increased M. elsdenii NCIMB 41125. An increase in butyrate was observed from +24 h to +72 h when calves were dosed on both d 7 and 14. Results showed that Me presented a positive relationship with butyrate (P < 0.001, R² = 0.43) and a concomitant negative relationship with acetate (P = 0.017, R² = -0.33). These results suggest that dosing pre-weaned dairy calves with M. elsdenii NCIMB 41125 has the potential to alter ruminal VFA production through increasing proportions of butyrate at the expense of propionate.

Keywords: calves, megasphaera elsdenii, rumen fermentation, bacteria

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Authors: Nikolai N. Bogolubov, Andrey V. Soldatov

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Terahertz radiation occupies a range of frequencies somewhere from 100 GHz to approximately 10 THz, just between microwaves and infrared waves. This range of frequencies holds promise for many useful applications in experimental applied physics and technology. At the same time, reliable, simple techniques for generation, amplification, and modulation of electromagnetic radiation in this range are far from been developed enough to meet the requirements of its practical usage, especially in comparison to the level of technological abilities already achieved for other domains of the electromagnetic spectrum. This situation of relative underdevelopment of this potentially very important range of electromagnetic spectrum is known under the name of the 'terahertz gap.' Among other things, technological progress in the terahertz area has been impeded by the lack of compact, low energy consumption, easily controlled and continuously radiating terahertz radiation sources. Therefore, development of new techniques serving this purpose as well as various devices based on them is of obvious necessity. No doubt, it would be highly advantageous to employ the simplest of suitable physical systems as major critical components in these techniques and devices. The purpose of the present research was to show by means of conventional methods of non-equilibrium statistical mechanics and the theory of open quantum systems, that a thoroughly studied two-level quantum system, also known as an one-electron two-level 'atom', being driven by external classical monochromatic high-frequency (e.g. laser) field, can radiate continuously at much lower (e.g. terahertz) frequency in the fluorescent regime if the transition dipole moment operator of this 'atom' possesses permanent non-equal diagonal matrix elements. This assumption contradicts conventional assumption routinely made in quantum optics that only the non-diagonal matrix elements persist. The conventional assumption is pertinent to natural atoms and molecules and stems from the property of spatial inversion symmetry of their eigenstates. At the same time, such an assumption is justified no more in regard to artificially manufactured quantum systems of reduced dimensionality, such as, for example, quantum dots, which are often nicknamed 'artificial atoms' due to striking similarity of their optical properties to those ones of the real atoms. Possible ways to experimental observation and practical implementation of the predicted effect are discussed too.

Keywords: terahertz gap, two-level atom, resonant fluorescence, quantum dot, resonant fluorescence, two-level atom

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Authors: William E. Bauta, Jennifer Rebeles, Reggie Jacob

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Porphyrins are noteworthy in biomedical science for their cancer tissue accumulation and photophysical properties. The preferential accumulation of some porphyrins in cancerous tissue has been known for many years. This, combined with their characteristic photophysical and photochemical properties, including their strong fluorescence and their ability to generate reactive oxygen species in vivo upon laser irradiation, has led to much research into the application of porphyrins as cancer diagnostic and therapeutic agents. Porphyrins have been used as dyes to detect cancer cells both in vivo and, less commonly, in vitro. In one example, human sputum samples from lung cancer patients and patients without the disease were dissociated and stained with the porphyrin TCPP (5,10,15,20-tetrakis-(4-carboxyphenyl)-porphine). Cells were analyzed by flow cytometry. Cancer samples were identified by their higher TCPP fluorescence intensity relative to the no-cancer controls. However, quantitative analysis of fluorescence in cell suspensions stained with multiple fluorophores requires particles stained with each of the individual fluorophores as controls. Fluorescent control particles must be compatible in size with flow cytometer fluidics and have favorable hydrodynamic properties in suspension. They must also display fluorescence comparable to the cells of interest and be stable upon storage amine-functionalized spherical polystyrene beads in the 5 to 20-micron diameter range that was reacted with TCPP and EDC in aqueous pH six buffer overnight to form amide bonds. Beads were isolated by centrifugation and tested by flow cytometry. The 10-micron amine-functionalized beads displayed the best combination of fluorescence intensity and hydrodynamic properties, such as lack of clumping and remaining in suspension during the experiment. These beads were further optimized by varying the stoichiometry of EDC and TCPP relative to the amine. The reaction was accompanied by the formation of a TCPP-related particulate, which was removed, after bead centrifugation, using a microfiltration process. The resultant TCPP-functionalized beads were compatible with flow cytometry conditions and displayed a fluorescence comparable to that of stained cells, which allowed their use as fluorescence standards. The beads were stable in refrigerated storage in the dark for more than eight months. This work demonstrates the first preparation of porphyrin-functionalized flow cytometry control beads.

Keywords: tetraaryl porphyrin, polystyrene beads, flow cytometry, peptide coupling

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Authors: Jacques M. Berner, Monja Gerber, Gillian L. Maggs-Kolling, Stuart J. Piketh

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The enigmatic melon species, Acanthosicyos horridus Welw. ex Hook. f., locally known as !nara, is endemic to the hyper-arid Namib Desert, where it thrives in sandy dune areas and dry river banks. The Namib Desert is characterized by extreme weather conditions which include high temperatures, very low rainfall, and extremely dry air. Plant and animals that have made the Namib Dessert their home are dependent on non-rainfall water inputs, like fog, dew and water vapor, for survival. Fog is believed to be the most important non-rainfall water input for most of the coastal Namib Desert and is a life line to many Namib plants and animals. It is commonly assumed that the !nara plant is adapted and dependent upon coastal fog events. The !nara plant shares many comparable adaptive features with other organisms that are known to exploit fog as a source of moisture. These include groove-like structures on the stems and the cone-like structures of thorns. These structures are believed to be the driving forces behind directional water flow that allow plants to take advantage of fog events. The !nara-fog interaction was investigated in this study to determine the dependence of !nara on these fog events, as it would illustrate strategies to benefit from non-rainfall water inputs. The direct water uptake capacity of !nara shoots was investigated through absorption tests. Furthermore, the movement and behavior of fluorescent water droplets on a !nara stem were investigated through time-lapse macrophotography. The shoot water potential was measured to investigate the effect of fog on the water status of !nara stems. These tests were used to determine whether the morphology of !nara has evolved to exploit fog as a non-rainfall water input and whether the !nara plant has adapted physiologically in response to fog. Chlorophyll a fluorescence was used to compare the photochemical efficiency of !nara plants on days with fog events to that on non-foggy days. The results indicate that !nara plants do have the ability to take advantage of fog events as commonly believed. However, the !nara plant did not exhibit visible signs of drought stress and this, together with the strong shoot water potential, indicates that these plants are reliant on permanent underground water sources. Chlorophyll a fluorescence data indicated that temperature stress and wind were some of the main abiotic factors influencing the plants’ overall vitality.

Keywords: Acanthosicyos horridus, chlorophyll a fluorescence, fog, foliar absorption, !nara

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Authors: Chien Y. Lin, Jung Y. Huang, Leu-Wei Lo

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A growing body of data reveals that the membrane cholesterol molecules can alter the signaling pathways of living cells. However, the understanding about how membrane cholesterol modulates receptor proteins is still lacking. Single-molecule tracking can effectively probe into the microscopic environments and thermal fluctuations of receptor proteins in a living cell. In this study we applies single-molecule optical tracking on ligand-induced dimerization process of EGFRs in the plasma membranes of two cancer cell lines (HeLa and A431) and one normal endothelial cell line (MCF12A). We tracked individual EGFR and dual receptors, diffusing in a correlated manner in the plasma membranes of live cells. We developed an energetic model by integrating the generalized Langevin equation with the Cahn-Hilliard equation to help extracting important information from single-molecule trajectories. From the study, we discovered that ligand-bound EGFRs move from non-raft areas into lipid raft domains. This ligand-induced motion is a common behavior in both cancer and normal cells. By manipulating the total amount of membrane cholesterol with methyl-β-cyclodextrin and the local concentration of membrane cholesterol with nystatin, we further found that the amount of cholesterol can affect the stability of EGFR dimers. The EGFR dimers in the plasma membrane of normal cells are more sensitive to the local concentration changes of cholesterol than EGFR dimers in the cancer cells. Our method successfully captures dynamic interactions of receptors at the single-molecule level and provides insight into the functional architecture of both the diffusing EGFR molecules and their local cellular environment.

Keywords: membrane proteins, single-molecule tracking, Cahn-Hilliard equation, EGFR dimers

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Authors: Jiahui Song

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The use of electric fields with high intensity (~ 100kV/cm or higher) and ultra short pulse durations (nanosecond range) has been a recent development. Most of the studies of electroporation have ignored possible thermal effects because of the small duration of the applied voltage pulses. However, it has been predicted membrane temperature gradients ranging from 0.2×109 to 109 K/m. This research focuses on thermal effects that drive for electroporative enhancements, even though the actual temperature values might not have changed appreciably from their equilibrium levels. The dynamics of pore formation with the application of an externally applied electric field is studied on the basis of molecular dynamics (MD) simulations using the GROMACS package. MD simulations of a lipid layer with constant electric field strength of 0.5 V/nm at 25 °C and 47 °C are implemented to simulate the appropriate thermal effects. The GROMACS provides the force fields for the lipid membranes, which is taken to comprise of dipalmitoyl-phosphatidyl-choline (DPPC) molecules. The water model mimicks the aqueous environment surrounding the membrane. Velocities of water and membrane molecules are generated randomly at each simulation run according to a Maxwellian distribution. The high background electric field is typically used in MD simulations to probe electroporation. It serves as an accelerated test of the pore formation process since low electric fields would take inordinately long simulation time. MD simulation shows no pore is formed in a 1-ns snapshot for a DPPC membrane set at a temperature of 25°C after a 0.5 V/nm electric field is applied. A nano-sized pore is clearly seen in a 0.75-ns snapshot on the same geometry, but with the membrane surfaces kept at temperatures of 47°C. And the pore increases at 1 ns. The MD simulation results suggest the possibility that the increase in temperature can result in different degrees of electrically stimulated bio-effects. The results points to the role of thermal effects in facilitating and accelerating the electroporation process.

Keywords: high-intensity, ultra-short, electroporation, thermal effects, molecular dynamics

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Authors: Ahmed El Naggar, Homyan Saleh

Abstract:

Dams are crucial for flood control, water supply, and the creation of hydroelectric power. Every dam has a water conveyance system, such as a spillway, providing the safe discharge of catastrophic floods when necessary. Spillway design has historically been investigated in laboratory research owing to the absence of suitable full-scale flow monitoring equipment and safety problems. Prototype measurements of aerated flows are urgently needed to quantify projected scale effects and provide missing validation data for design guidelines and numerical simulations. In this work, an image-based investigation of free-surface flows on a tiered spillway was undertaken at the laboratory (fixed camera installation) and prototype size (drone video) (drone footage) (drone footage). The drone videos were generated using data from citizen science. Analyses permitted the measurement of the free-surface aeration inception point, air-water surface velocities, fluctuations, and residual energy at the chute's downstream end from a remote site. The prototype observations offered full-scale proof of concept, while laboratory results were efficiently confirmed against invasive phase-detection probe data. This paper stresses the efficacy of image-based analyses at prototype spillways. It highlights how citizen science data may enable academics better understand real-world air-water flow dynamics and offers a framework for a small collection of long-missing prototype data.

Keywords: remote sensing, aerated flows, large dams, proof of concept, dam spillways, air-water flows, prototype operation, remote sensing, inception point, optical flow, turbulence, residual energy

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Authors: Win Zaw Hein, Khin Sandar Linn, Su Su Yi Mon, Yoshitaka Goto

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The Earth ionosphere is located at the altitude of about 70 km to several 100 km from the ground, and it is composed of ions and electrons called plasma. In the ionosphere, these plasma makes delay in GPS (Global Positioning System) signals and reflect in radio waves. The delay along the signal path from the satellite to the receiver is directly proportional to the total electron content (TEC) of plasma, and this delay is the largest error factor in satellite positioning and navigation. Sounding observation from the top and bottom of the ionosphere was popular to investigate such ionospheric plasma for a long time. Recently, continuous monitoring of the TEC using networks of GNSS (Global Navigation Satellite System) observation stations, which are basically built for land survey, has been conducted in several countries. However, in these stations, multi-frequency support receivers are installed to estimate the effect of plasma delay using their frequency dependence and the cost of multi-frequency support receivers are much higher than single frequency support GPS receiver. In this research, single frequency GPS receiver was used instead of expensive multi-frequency GNSS receivers to measure the ionospheric plasma variation such as vertical TEC distribution. In this measurement, single-frequency support ublox GPS receiver was used to probe ionospheric TEC. The location of observation was assigned at Mandalay Technological University in Myanmar. In the method, the ionospheric TEC distribution is represented by polynomial functions for latitude and longitude, and parameters of the functions are determined by least-squares fitting on pseudorange data obtained at a known location under an assumption of thin layer ionosphere. The validity of the method was evaluated by measurements obtained by the Japanese GNSS observation network called GEONET. The performance of measurement results using single-frequency of GPS receiver was compared with the results by dual-frequency measurement.

Keywords: ionosphere, global positioning system, GPS, ionospheric delay, total electron content, TEC

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Authors: Ozlem Oral, Seval Kilic, Ozlem Yedier, Serap Dokmeci, Devrim Gozuacik

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Autophagy is a degradation pathway, activating under stress conditions. It digests macromolecules, such as abnormal proteins and long-lived organelles by engulfing them and by subsequent delivery of the cargo to lysosomes. The members of the phospholipid-dependent serine/threonine kinases, involved in many signaling pathways, which are necessary for the regulation of cellular metabolic activation. Previous studies implicate that, serine/threonine kinases have crucial roles in the mechanism of many diseases depend on the activated and/or inactivated signaling pathway. Data indicates, the signaling pathways activated by serine/threonine kinases are also involved in activation of autophagy mechanism. However, the information about the effect of serine/threonine kinases on autophagy mechanism and the roles of these effects in disease formation is limited. In this study, we investigated the effect of activated serine/threonine kinases on autophagic pathway. We performed a commonly used autophagy technique, GFP-LC3 dot formation and by using microscopy analyses, we evaluated promotion and/or inhibition of autophagy in serine/threonine kinase-overexpressed fibroblasts as well as cancer cells. In addition, we carried out confocal microscopy analyses and examined autophagic flux by utilizing the differential pH sensitivities of RFP and GFP in mRFP-GFP-LC3 probe. Based on the shRNA-library based screening, we identified autophagy-related proteins affected by serine/threonine kinases. We further studied the involvement of serine/threonine kinases on the molecular mechanism of newly identified autophagy proteins and found that, autophagic pathway is indirectly controlled by serine/threonine kinases via specific autophagic proteins. Our data indicate the molecular connection between two critical cellular mechanisms, which have important roles in the formation of many disease pathologies, particularly cancer. This project is supported by TUBITAK-1001-Scientific and Technological Research Projects Funding Program, Project No: 114Z836.

Keywords: autophagy, GFP-LC3 dot formation assay, serine/threonine kinases, shRNA-library screening

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Authors: Varun A Baheti, Sanjay Kashyap, Kamanio Chattopadhyay, Praveen Kumar, Aloke Paul

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Ni–Sn system finds applications in the microelectronics industry, especially with respect to flip–chip or direct chip, attach technology. Here the region of interest is under bump metallization (UBM), and solder bump (Sn) interface due to the formation of brittle intermetallic phases there. Understanding the growth of these phases at UBM/Sn interface is important, as in many cases it controls the electro–mechanical properties of the product. Cu and Ni are the commonly used UBM materials. Cu is used for good bonding because of fast reaction with solder and Ni often acts as a diffusion barrier layer due to its inherently slower reaction kinetics with Sn–based solders. Investigation on the growth kinetics of phases in Ni–Sn system is reported in this study. Just for simplicity, Sn being major solder constituent is chosen. Ni–Sn electroplated diffusion couples are prepared by electroplating pure Sn on Ni substrate. Bulk diffusion couples prepared by the conventional method are also studied along with Ni–Sn electroplated diffusion couples. Diffusion couples are annealed for 25–1000 h at 50–215°C to study the phase evolutions and growth kinetics of various phases. The interdiffusion zone was analysed using field emission gun equipped scanning electron microscope (FE–SEM) for imaging. Indexing of selected area diffraction (SAD) patterns obtained from transmission electron microscope (TEM) and composition measurements done in electron probe micro−analyser (FE–EPMA) confirms the presence of various product phases grown across the interdiffusion zone. Time-dependent experiments indicate diffusion controlled growth of the product phase. The estimated activation energy in the temperature range 125–215°C for parabolic growth constants (and hence integrated interdiffusion coefficients) of the Ni₃Sn₄ phase shed light on the growth mechanism of the phase; whether its grain boundary controlled or lattice controlled diffusion. The location of the Kirkendall marker plane indicates that the Ni₃Sn₄ phase grows mainly by diffusion of Sn in the binary Ni–Sn system.

Keywords: diffusion, equilibrium phase, metastable phase, the Ni-Sn system

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Authors: Amir Seyfoori, Ehsan Samiei, Mohsen Akbari

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The use of microtechnology for detection and high yield isolation of circulating tumor cells (CTCs) has shown enormous promise as an indication of clinical metastasis prognosis and cancer treatment monitoring. The Immunomagnetic assay has been also coupled to microtechnology to improve the selectivity and efficiency of the current methods of cancer biomarker isolation. In this way, generation and configuration of the local high gradient magnetic field play essential roles in such assay. Additionally, considering the intrinsic heterogeneity of cancer cells, real-time analysis of isolated cells is necessary to characterize their responses to therapy. Totally, on-chip isolation and monitoring of the specific tumor cells is considered as a pressing need in the way of modified cancer therapy. To address these challenges, we have developed a bi-layer magnetic-based microfluidic chip for enhanced CTC detection and capturing. Micromagnet arrays at the bottom layer of the chip were fabricated using a new method of magnetic nanoparticle paste deposition so that they were arranged at the center of the chain microchannel with the lowest fluid velocity zone. Breast cancer cells labelled with EPCAM-conjugated smart microgels were immobilized on the tip of the micromagnets with greater localized magnetic field and stronger cell-micromagnet interaction. Considering different magnetic nano-powder usage (MnFe2O4 & gamma-Fe2O3) and micromagnet shapes (ellipsoidal & arrow), the capture efficiency of the systems was adjusted while the higher CTC capture efficiency was acquired for MnFe2O4 arrow micromagnet as around 95.5%. As a proof of concept of on-chip tumor cell monitoring, magnetic smart microgels made of thermo-responsive poly N-isopropylacrylamide-co-acrylic acid (PNIPAM-AA) composition were used for both purposes of targeted cell capturing as well as cell monitoring using antibody conjugation and fluorescent dye loading at the same time. In this regard, magnetic microgels were successfully used as cell tracker after isolation process so that by raising the temperature up to 37⁰ C, they released the contained dye and stained the targeted cell just after capturing. This microfluidic device was able to provide a platform for detection, isolation and efficient real-time analysis of specific CTCs in the liquid biopsy of breast cancer patients.

Keywords: circulating tumor cells, microfluidic, immunomagnetic, cell isolation

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Authors: Ratnakar Rao, Subair Khan, Hari Haran

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Posterior cruciate ligament (PCL) avulsion fractures is a rare condition and commonly mismanaged.Surgical reattachment has been shown to produce better result compared with conservative management.Only few techniques are reported in arthroscopic fixation of PCL Avulsion Fracture and they are complex.We describe a new technique in fixation of the PCL Avulsion fracture through a posterior trans septal portal using button fixation device (Mini Tight Rope). Eighteen patients with an isolated posterior cruciate ligament avulsion fracture were operated under arthroscopy. Standard Antero Medial Portal and Antero Lateral portals made and additional Postero Medial and Postero Lateral portals made and trans Septal portal established. Avulsion fracture identified, elevated, prepared. Reduction achieved using PCL Tibial guide (Arthrex) and fixation was achieved using Mini Tight Rope,Arthrex (2 buttons with a suture). Reduction confirmed using probe and Image intensifier. Postoperative assessment made clinically and radiologically. 15 patients had good to excellent results with no posterior sag or instability. The range of motion was normal. No complications were recorded per operatively. 2 patients had communition of the fragment while drilling, for one patient it was managed by suturing technique and the second patient PCL Reconstruction was done. One patient had persistent instability with poor outcome. Establishing trans septal portal helps in better visualization of the posterior compartment of the knee. Assessment of the bony fragment, preparation 0f the bone bed andit protects from injury to posterior neurovascular structures. Fixation using the button with suture (Mini Tight Rope) is stable and easily reproducible for PCL Avulsion fracture with single large fragment.

Keywords: PCL avulsion, arthroscopy, transeptal, minitight rope technique

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Authors: Syed Beenish Rufai, Sarman Singh

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Background: Performance of Genotype MTBDRsl (Hain Life science GmbH Germany) for detection of mutations associated with second-line drug resistance is well known. However, less evidence regarding the association of mutations and genetic background of strains is known which, in the future, is essential for clinical management of anti-tuberculosis drugs in those settings where the probability of particular genotype is predominant. Material and Methods: During this retrospective study, a total of 259 MDR-TB isolates obtained from pulmonary TB patients were tested for second-line drug susceptibility testing (DST) using Genotype MTBDRsl VER 1.0 and compared with BACTEC MGIT-960 as a reference standard. All isolates were further characterized using spoligotyping. The spoligo patterns obtained were compared and analyzed using SITVIT_WEB. Results: Of total 259 MDR-TB isolates which were screened for second-line DST by Genotype MTBDRsl, mutations were found to be associated with gyrA, rrs and emb genes in 82 (31.6%), 2 (0.8%) and 90 (34.7%) isolates respectively. 16 (6.1%) isolates detected mutations associated with both FQ as well as to AG/CP drugs (XDR-TB). No mutations were detected in 159 (61.4%) isolates for corresponding gyrA and rrs genes. Genotype MTBDRsl showed a concordance of 96.4% for detection of sensitive isolates in comparison with second-line DST by BACTEC MGIT-960 and 94.1%, 93.5%, 60.5% and 50% for detection of XDR-TB, FQ, EMB, and AMK/CAP respectively. D94G was the most prevalent mutation found among (38 (46.4%)) OFXR isolates (37 FQ mono-resistant and 1 XDR-TB) followed by A90V (23 (28.1%)) (17 FQ mono-resistant and 6 XDR-TB). Among AG/CP resistant isolates A1401G was the most frequent mutation observed among (11 (61.1%)) isolates (2 AG/CP mono-resistant isolates and 9 XDR-TB isolates) followed by WT+A1401G (6 (33.3%)) and G1484T (1 (5.5%)) respectively. On spoligotyping analysis, Beijing strain (46%) was found to be the most predominant strain among pre-XDR and XDR TB isolates followed by CAS (30%), X (6%), Unique (5%), EAI and T each of 4%, Manu (3%) and Ural (2%) respectively. Beijing strain was found to be strongly associated with D94G (47.3%) and A90V mutations by (47.3%) and 34.8% followed by CAS strain by (31.6%) and 30.4% respectively. However, among AG/CP resistant isolates, only Beijing strain was found to be strongly associated with A1401G and WT+A1401G mutations by 54.5% and 50% respectively. Conclusion: Beijing strain was found to be strongly associated with the most prevalent mutations among pre-XDR and XDR TB isolates. Acknowledgments: Study was supported with Grant by All India Institute of Medical Sciences, New Delhi reference No. P-2012/12452.

Keywords: tuberculosis, line probe assay, XDR TB, drug susceptibility

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Authors: Yi Huang

Abstract:

Background: For the development of children, it is important for parents to encourage children not only on academic competence but also on children’s social competence. In the western cultural context, parents emphasize more heavily on female children’s social-behavioral development. However, whether the conclusion is correct in eastern culture and whether the parent’s gender affects such an emphasis remains unclear. And, more valuably, from the perspective of intervention, except for the nature factors - child’s gender and parent’s gender, it is also worth to probe whether the improvable factors, such as parent’s perspective taking, influence parent’s emphasis on child’s social competence. Aim: This study was aimed to investigate the impact of parent’s gender, child’s gender, and parent’s perspective-taking on parent’s attitudes of encouragement of the child’s social competence under the Chinese cultural context. Method: 461 Chinese parents whose children were in the first year of middle school during the research time participated in this study. Among all participants, there were 155 fathers and 306 mothers. The research adopted the self-report of perspective-taking, which is the sub-scale of the Interpersonal Reactivity Index and the self-report of the encouragement on a child’s social communication, which is the sub-scale of the Chinese version of The Children Rearing Practice Report. In this study, 291 parents reported regarding male children, and 170 parents reported regarding female children. Results: Contrary to the traditional western theory, which usually suggests parent puts more attention on social development and competence to girl the instead of the boy, in the Chinese context, parent emphasizes social competence more on the male child. Analogically, in China, compared to mother, father underscores the child’s social competence more heavily. By constructing the hierarchical regression model, the result indicated that after controlling the variables of the gender of child and the gender of parent, parent’s perspective-taking still explains for the variance of parent’s encouragement on child’s social competence, which means, parent’s perspective-taking predicts parent’s encouragement on child’s social competence after excluding the impact of the gender of parent and child. Conclusion: For Chinese parents, the ability of perspective-taking is beneficial to enhance their awareness of encouraging children’s social competence.

Keywords: parent; child, gender differences, perspective-taking, social development

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Authors: Eriselda Simoni, Leonard Simoni, Endri Paparisto, Laureta Flaga, Silvana Bara, Edit Xhajanka, Arjan Harxhi

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Background: Some pathologies are encountered more often in HIV/ AIDS, such as those with bacterial, fungal, viral, and neoplastic causes, but what has been more noticeable in recent years is the increased and more aggressive manifestation of periodontal disease and oral caries. Our purpose is to investigate the correlation between the HIV/AIDS stage and CD4 level with oral health, dentition, and periodontal status. Materials and Methods: We conducted a prospective observational study that included 35 patients newly diagnosed with HIV/AIDS and underwent an oral examination at the University Dental Clinic in Tirana, Albania, in the period April - July 2024. This study evaluated the basic demographic, laboratory characteristics, oral hygiene, and the presence of oral lesions. The dentition status was assessed with the values DT (decay teeth), FT (filled teeth), and MT (missing teeth) presented as DMFT. The periodontal status was evaluated through a periodontal probe measuring CPI (community periodontal index) and LOA (loss of attachment) as recommended by the WHO Oral Health Assessment Form 2013. The Pearson Correlation Coefficient (r) was used to evaluate the relationship between levels of CD4+ and DMF, CD4+ and CPI, and CD4+ and LOA. The P value ≤ 0.05 was considered statistically significant. Results: 80% of patients included were males with a mean age of 35.8 years. 8.6% of patients were categorized as HIV stage I, 28.6% as stage II, and 62.8% as HIV stage III/AIDS. The mean level value of CD4+ was 266.2 cells/mm3 and the rapport CD4+/ WBC (White Blood Cells) was 15.7%. Most patients (57.2%) used toothbrushes less than 1 time a day. An important negative correlation was found between CD4+ and dentition and periodontal status. A lower level of CD4+ was correlated with a higher DMFT, CPI, and LOA, respectively coefficient (r) for CD4/DMFT = -0.52, p =0.01, (r) for CD4/CPI= - 0.38, p=0.024 and (r) for CD4/LOA= - 0.37, p=0.029. Conclusions: In our study, it was documented that patients with HIV/AIDS had worse oral health, an important negative correlation between CD4+ and dentition and periodontal status. A lower level of CD4+ was correlated with a worse dentition status (higher DMFT), and poor periodontal health (higher CPI and LOA). The monitoring and treatment of oral pathologies can be important in early HIV/AIDS diagnoses and treatment.

Keywords: HIV/AIDS, oral health, dentition, periodontal

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Authors: Amjad I. Aqib, Shanza R. Khan, Tanveer Ahmad, Syed A. R. Shah, Muhammad A. Naseer, Muhammad Shoaib, Iqra Sarwar, Muhammad F. A. Kulyar, Zeeshan A. Bhutta, Mumtaz A. Khan, Mahboob Ali, Khadija Yasmeen

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The current study focused on resistance modulation of dairy linked epidemic mec A positive S. aureus for resistance modulation by plant extract (Eucalyptus globolus, Calotropis procera), NSAIDs, and star like microparticles. Zinc oxide {ZnO}c and {Zn (OH)₂} microparticles were synthesized by solvothermal method and characterized by calcination, X-ray diffraction (XRD), and scanning electron microscope (SEM). Plant extracts were prepared by the Soxhlet extraction method. The study found 34% of subclinical samples (n=200) positive for S. aureus from dairy milk having significant (p < 0.05) association of assumed risk factors with pathogen. The antimicrobial assay showed 55, 42, 41, and 41% of S. aureus resistant to oxacillin, ciprofloxacin, streptomycin, and enoxacin. Amoxicillin showed the highest percentage of increase in zone of inhibitions (ZOI) at 100mg of Calotropis procera extract (31.29%) followed by 1mg/mL (28.91%) and 10mg/mL (21.68%) of Eucalyptus globolus. Amoxicillin increased ZOI by 42.85, 37.32, 29.05, and 22.78% in combination with 500 ug/ml with each of diclofenac, aspirin, ibuprofen, and meloxicam, respectively. Fractional inhibitory concentration indices (FICIs) showed synergism of amoxicillin with diclofenac and aspirin and indifferent synergy with ibuprofen and meloxicam. The preliminary in vitro finding of combination of microparticles with amoxicillin proved to be synergistic, giving rise to 26.74% and 14.85% increase in ZOI of amoxicillin in combination with zinc oxide and zinc hydroxide, respectively. The modulated antimicrobial resistance incurred by NSAIDs, plant extracts, and microparticles against pathogenic S. aureus invite immediate attention to probe alternative antimicrobial sources.

Keywords: antimicrobial resistance, dairy milk, nanoparticles, NSIDs, plant extracts, resistance modulation, S. aureus

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Authors: Astrid Tannert, Anuradha Ramoji, Christina Ebert, Frederike Gladigau, Lorena Tuchscherr, Jürgen Popp, Ute Neugebauer

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Staphylococcus aureus is a facultative intracellular pathogen, which by entering host cells may evade immunologic host response as well as antimicrobial treatment. In that way, S. aureus can cause persistent intracellular infections which are difficult to treat. Depending on the strain, S. aureus may persist at different intracellular locations like the phagolysosome. The first barrier invading pathogens from the blood stream that they have to cross are the endothelial cells lining the inner surface of blood and lymphatic vessels. Upon proceeding from an acute to a chronic infection, intracellular pathogens undergo certain biochemical and structural changes including a deceleration of metabolic processes to adopt for long-term intracellular survival and the development of a special phenotype designated as small colony variant. In this study, the endothelial cell line Ea.hy 926 was used as a model for acute and chronic S. aureus infection. To this end, Ea.hy 926 cells were cultured on QIAscout™ Microraft Arrays, a special graded cell culture substrate that contains around 12,000 microrafts of 200 µm edge length. After attachment to the substrate, the endothelial cells were infected with GFP-expressing S. aureus for 3 weeks. The acute infection and the development of persistent bacteria was followed by confocal laser scanning microscopy, scanning the whole Microraft Array for the presence and for detailed determination of the intracellular location of fluorescent intracellular bacteria every second day. After three weeks of infection representative microrafts containing infected cells, cells with protruded infections and cells that did never show any infection were isolated and fixed for Raman micro-spectroscopic investigation. For comparison, also microrafts with acute infection were isolated. The acquired Raman spectra are correlated with the fluorescence microscopic images to give hints about a) the molecular alterations in endothelial cells during acute and chronic infection compared to non-infected cells, and b) metabolic and structural changes within the pathogen when entering a mode of persistence within host cells. We thank Dr. Ruth Kläver from QIAGEN GmbH for her support regarding QIAscout technology. Financial support by the BMBF via the CSCC (FKZ 01EO1502) and from the DFG via the Jena Biophotonic and Imaging Laboratory (JBIL, FKZ PO 633/29-1, BA 1601/10-1) is highly acknowledged.

Keywords: correlative image analysis, intracellular infection, pathogen-host adaption, Raman micro-spectroscopy

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Authors: Tania Vasquez Mata, Luis A. Herrera, Cristian Arriaga Canon

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Background: Locally advanced breast cancer of the luminal B phenotype, poses challenges due to its variable response to neoadjuvant chemotherapy. A predictive biomarker is needed to identify patients who will not respond to treatment, allowing for alternative therapies. This study aims to validate the use of the lncRNA GATA3-AS1, as a predictive biomarker using RNA in situ hybridization. Research aim: The aim of this study is to determine if GATA3-AS1 can serve as a biomarker for resistance to neoadjuvant chemotherapy in patients with locally advanced luminal B breast cancer. Methodology: The study utilizes RNA in situ hybridization with predesigned probes for GATA3-AS1 on Formalin-Fixed Paraffin-Embedded tissue sections. The samples underwent pretreatment and protease treatment to enable probe penetration. Chromogenic detection and signal evaluation were performed using specific criteria. Findings: Patients who did not respond to neoadjuvant chemotherapy showed a 3+ score for GATA3-AS1, while those who had a complete response had a 1+ score. Theoretical importance: This study demonstrates the potential clinical utility of GATA3-AS1 as a biomarker for resistance to neoadjuvant chemotherapy. Identifying non-responders early on can help avoid unnecessary treatment and explore alternative therapy options. Data collection and analysis procedures: Tissue samples from patients with locally advanced luminal B breast cancer were collected and processed using RNA in situ hybridization. Signal evaluation was conducted under a microscope, and scoring was based on specific criteria. Questions addressed: Can GATA3-AS1 serve as a predictive biomarker for neoadjuvant chemotherapy response in locally advanced luminal B breast cancer? Conclusion: The lncRNA GATA3-AS1 can be used as a biomarker for resistance to neoadjuvant chemotherapy in patients with locally advanced luminal B breast cancer. Its identification through RNA in situ hybridization of tissue obtained from the initial biopsy can aid in treatment decision-making.

Keywords: biomarkers, breast neoplasms, genetics, neoadjuvant therapy, tumor

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