Search results for: fluorescence quenching effect

Authors: Prabhavathi Sundaram, Heidi Abrahamse

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In recent years, nanotechnology coupled with photodynamic therapy (PDT) has received considerable attention in terms of improving the effectiveness of drug delivery in cancer therapeutics. The development of functionalized single-walled carbon nanotubes (SWCNTs) has become revolutionary in targeted photosensitizers delivery since it improves the therapeutic index of drugs. The objective of this study was to prepare, characterize and evaluate the potential of functionalized SWCNTs using hyaluronic acid and loading it with photosensitizer and to effectively target colon cancer cells. The single-walled carbon nanotubes were covalently functionalized with hyaluronic acid and the loaded photosensitizer by non-covalent interaction. The photodynamic effect of SWCNTs is detected under laser irradiation in vitro. The hyaluronic acid-functionalized nanocomposites had a good affinity with CD44 receptors, and it avidly binds on to the surface of CACO-2 cells. The cellular uptake of nanocomposites was studied using fluorescence microscopy using lyso tracker. The anticancer activity of nanocomposites was analyzed in CACO-2 cells using different studies such as cell morphology, cell apoptosis, and nuclear morphology. The combined effect of nanocomposites and PDT improved the therapeutic effect of cancer treatment. The study suggested that the nanocomposites and PDT have great potential in the treatment of colon cancer.

Keywords: colon cancer, hyaluronic acid, single walled carbon nanotubes, photosensitizers, photodynamic therapy

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Authors: Indu Verma, Santanu Kumar Pal

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Interactions between DNA and adsorbed Poly (L-lysine) (PLL) on liquid crystal (LC) droplets were investigated using polarizing optical microcopy (POM) and epi-fluorescence microscopy. Earlier, we demonstrated that adsorption of PLL to the LC/aqueous interface resulted in homeotropic orientation of the LC and thus exhibited a radial configuration of the LC confined within the droplets. Subsequent adsorption of DNA (single stranded DNA/double stranded DNA) at PLL coated LC droplets was found to trigger a LC reorientation within the droplets leading to pre-radial/bipolar configuration of those droplets. To our surprise, subsequent exposure of complementary ssDNA (c-ssDNA) to ssDNA/ adsorbed PLL modified LC droplets did not cause the LC reorientation. This is likely due to the formation of polyplexes (DNA-PLL complex) as confirmed by fluorescence microscopy and atomic force microscopy. In addition, dsDNA adsorbed PLL droplets have been found to be effectively used to displace (controlled release) propidium iodide (a model drug) encapsulated within dsDNA over time. These observations suggest the potential for a label free droplet based LC detection system that can respond to DNA and may provide a simple method to develop DNA-based drug nano-carriers.

Keywords: DNA biosensor, drug delivery, interfaces, liquid crystal droplets

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Authors: Isil Gazioglu, Abdulselam Ertas

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Aflatoxins (AFs) are secondary toxic metabolites of Aspergillus flavus and A. parasiticus. AFs can be absorbed through the skin. Potent carcinogens like AFs should be completely absent from cosmetics, this can be achieved by careful quality control of the raw plant materials. Regulatory limits for aflatoxins have been established in many countries, and reliable testing methodology is needed to implement and enforce the regulatory limits. In this study, ten medicinal plant samples (Bundelia tournefortti, Capsella bursa-pastoris, Carduus tenuiflorus, Cardaria draba, Malva neglecta, Malvella sharardiana, Melissa officinalis, Sideritis libanotica, Stakys thirkei, Thymus nummularius) were investigated for aflatoxin (AF) contaminations by employing an HPLC assay for the determination of AFB1, B2, G1 and G2. The samples were extracted with 70% (v/v) methanol in water before further cleaned up with an immunoaffinity column and followed by the detection of AFs by using an electrochemically post-column derivatization with Kobra-Cell and fluorescence detector. The extraction procedure was optimized in order to obtain the best recovery. The method was successfully carried out with all medicinal plant samples. The results revealed that five (50%) of samples were contaminated with AFs. The association between particular samples and the AF contaminated could not be determined due to the low frequency of positive samples.

Keywords: aflatoxin B1, HPLC-FLD, KOBRA-Cell, mycotoxin

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Authors: Rohit Agarwal, Mrityunjay K. Singh, Soma Ghosh, Ramesh Shankar, Biswajit Ghosh, Vinay V. Mahashabde

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Thermo-mechanical treatment (TMT) of rebars is a critical process to impart sufficient strength and ductility to rebar. TMT rebars are produced by the Tempcore process, involves an 'in-line' heat treatment in which hot rolled bar (temperature is around 1080°C) is passed through water boxes where it is quenched under high pressure water jets (temperature is around 25°C). The quenching rate dictates composite structure consisting (four non-homogenously distributed phases of rebar microstructure) pearlite-ferrite, bainite, and tempered martensite (from core to rim). The ferrite and pearlite phases present at core induce ductility to rebar while martensitic rim induces appropriate strength. The TMT process is difficult to model as it brings multitude of complex physics such as heat transfer, highly turbulent fluid flow, multicomponent and multiphase flow present in the control volume. Additionally the presence of film boiling regime (above Leidenfrost point) due to steam formation adds complexity to domain. A coupled heat transfer and fluid flow model based on computational fluid dynamics (CFD) has been developed at product technology division of Tata Steel, India which efficiently predicts temperature profile and percentage martensite rim thickness of rebar during quenching process. The model has been validated with 16 mm rolling of New Bar mill (NBM) plant of Tata Steel Limited, India. Furthermore, based on the scenario analyses, optimal configuration of nozzles was found which helped in subsequent increase in rolling speed.

Keywords: boiling, critical heat flux, nozzles, thermo-mechanical treatment

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Authors: A. Jourani, C. Trevisiol, S. Bouvier

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The aim of this work is to study the coupled effect of microstructure and surface roughness on friction coefficient, wear resistance and wear mechanisms. Friction tests on 35NCD16 steel are performed under different normal loads (50-110 N) on a pin-on-plane configuration at cyclic sliding with abrasive silicon carbide grains ranging from 35 µm to 200 µm. To vary hardness and microstructure, the specimens are subjected to water quenching and tempering at various temperatures from 200°C to 600°C. The evolution of microstructures and wear mechanisms of worn surfaces are analyzed using scanning electron microscopy (SEM). For a given microstructure and hardness, the friction coefficient decreases with increasing of normal load and decreasing of the abrasive particle size. The wear rate increase with increasing of normal load and abrasive particle size. The results also reveal that there is a critical hardness Hcᵣᵢₜᵢcₐₗ around 430 Hv which maximizes the friction coefficient and wear rate. This corresponds to a microstructure transition from martensite laths to carbides and equiaxed grains, for a tempering around 400°C. Above Hcᵣᵢₜᵢcₐₗ the friction coefficient and the amount of material loss decrease with an increase of hardness and martensite volume fraction. This study also shows that the debris size and the space between the abrasive particles decrease with a reduction in the particle size. The coarsest abrasive grains lost their cutting edges, accompanied by particle damage and empty space due to the particle detachment from the resin matrix. The compact packing nature of finer abrasive papers implicates lower particle detachment and facilitates the clogging and the transition from abrasive to adhesive wear.

Keywords: martensite, microstructure, friction, wear, surface roughness

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Authors: Azuraida Amat, Halimah Mohamed Kamari, Che Azurahanim Che Abdullah, Ishak Mansor

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The changes of the optical and structural properties of Bismuth-Boro-Tellurite glasses pre and post gamma irradiation were studied. Six glass samples, with different compositions [(TeO2)0.7 (B2O3)0.3]1-x (Bi2O3)x prepared by melt quenching method were irradiated with 25kGy gamma radiation at room temperature. The Fourier Transform Infrared Spectroscopy (FTIR) was used to explore the structural bonding in the prepared glass samples due to exposure, while UV-VIS Spectrophotometer was used to evaluate the changes in the optical properties before and after irradiation. Gamma irradiation causes a profound changes in the peak intensity as shown by FTIR spectra which is due to the breaking of the network bonding. Before gamma irradiation, the optical band gap, Eg value decreased from 2.44 eV to 2.15 eV with the addition of Bismuth content. The value kept decreasing (from 2.18 eV to 2.00 eV) following exposure to gamma radiation due to the increase of non-bridging oxygen (NBO) and the increase of defects in the glass. In conclusion, the glass with high content of Bi2O3 (0.30Bi) give the smallest Eg and show less changes in FTIR spectra after gamma irradiation, which indicate that this glass is more resistant to gamma radiation compared to other glasses.

Keywords: boro-tellurite, bismuth, gamma radiation, optical properties

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Authors: Jacques M. Berner, Lehlogonolo Maloma

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Context: Glyphosate, a widely used herbicide, has raised concerns about its impact on various crops. Amaranthus cruentus, an important grain crop species, is particularly susceptible to glyphosate. Understanding the specific disruptions caused by glyphosate on the photosynthetic process in Amaranthus cruentus is crucial for assessing its effects on crop productivity and ecological sustainability. Research Aim: This study aimed to investigate the dose-dependent impact of glyphosate on the photochemical efficiency of Amaranthus cruentus using the OJIP transient analysis. The goal was to assess the specific disruptions caused by glyphosate on key parameters of photosystem II. Methodology: The experiment was conducted in a controlled greenhouse environment. Amaranthus cruentus plants were exposed to different concentrations of glyphosate, including half, recommended, and double the recommended application rates. The photochemical efficiency of the plants was evaluated using non-invasive chlorophyll a fluorescence measurements and subsequent analysis of OJIP transients. Measurements were taken on 1-hour dark-adapted leaves using a Hansatech Handy PEA+ chlorophyll fluorimeter. Findings: The study's results demonstrated a significant reduction in the photochemical efficiency of Amaranthus cruentus following glyphosate treatment. The OJIP transients showed distinct alterations in the glyphosate-treated plants compared to the control group. These changes included a decrease in maximal fluorescence (FP) and a delay in the rise of the fluorescence signal, indicating impairment in the energy conversion process within the photosystem II. Glyphosate exposure also led to a substantial decrease in the maximum quantum yield efficiency of photosystem II (FV/FM) and the total performance index (PItotal), which reflects the overall photochemical efficiency of photosystem II. These reductions in photochemical efficiency were observed even at half the recommended dose of glyphosate. Theoretical Importance: The study provides valuable insights into the specific disruptions caused by glyphosate on the photochemical efficiency of Amaranthus cruentus. Data Collection and Analysis Procedures: Data collection involved non-invasive chlorophyll a fluorescence measurements using a chlorophyll fluorimeter on dark-adapted leaves. The OJIP transients were then analyzed to assess specific disruptions in key parameters of photosystem II. Statistical analysis was conducted to determine the significance of the differences observed between glyphosate-treated plants and the control group. Question Addressed: The study aimed to address the question of how glyphosate exposure affects the photochemical efficiency of Amaranthus cruentus, specifically examining disruptions in the photosynthetic electron transport chain and overall photochemical efficiency. Conclusion: The study demonstrates that glyphosate severely impairs the photochemical efficiency of Amaranthus cruentus, as indicated by the alterations in OJIP transients. Even at half the recommended dose, glyphosate caused significant reductions in photochemical efficiency. These findings highlight the detrimental effects of glyphosate on crop productivity and emphasize the need for further research to evaluate its long-term consequences and ecological implications in agriculture. The authors gratefully acknowledge the support from North-West University for making this research possible.

Keywords: glyphosate, amaranthus cruentus, ojip transient analysis, pitotal, photochemical efficiency, chlorophyll fluorescence, weeds

Procedia PDF Downloads 75

Authors: Azadeh Farzaneh, Mohammad Reza Abdi, A. Quaranta, Matteo Dalla Palma, Seyedshahram Mortazavi

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We report on the synthesis of cesium-iodide nanoparticles using sol-gel technique. The structural properties of CsI nanoparticles were characterized by X-ray diffraction and Scanning Electron Microscope (SEM) Also, optical properties were followed by optical absorption and UV–vis fluorescence. Intense photoluminescence is also observed, with some spectral tuning possible with ripening time getting a range of emission photon wavelength approximately from 366 to 350 nm. The size effect on CsI luminescence leads to an increase in scintillation light yield, a redshift of the emission bands of the on_center and off_center self_trapped excitons (STEs) and an increase in the contribution of the off_center STEs to the net intrinsic emission yield. The energy transfer from the matrix to CsI nanoparticles is a key characteristic for scintillation detectors. So the scintillation spectra to alpha particles of sample were monitored.

Keywords: nanoparticles, luminescence, sol gel, scintillator

Procedia PDF Downloads 581

Authors: David Oluigbo, Erik Hemberg, Nathan Shwatal, Wenqi Ding, Yin Yuan, Susanna Mierau

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Introduction: Calcium imaging is an established technique in neuroscience research for detecting activity in neural networks. Bursts of action potentials in neurons lead to transient increases in intracellular calcium visualized with fluorescent indicators. Manual identification of cell bodies and their contours by experts typically takes 10-20 minutes per calcium imaging recording. Our aim, therefore, was to design an automated pipeline to facilitate and optimize calcium imaging data analysis. Our pipeline aims to accelerate cell body and contour identification and production of graphical representations reflecting changes in neuronal calcium-based fluorescence. Methods: We created a Python-based pipeline that uses OpenCV (a computer vision Python package) to accurately (1) detect neuron contours, (2) extract the mean fluorescence within the contour, and (3) identify transient changes in the fluorescence due to neuronal activity. The pipeline consisted of 3 Python scripts that could both be easily accessed through a Python Jupyter notebook. In total, we tested this pipeline on ten separate calcium imaging datasets from murine dissociate cortical cultures. We next compared our automated pipeline outputs with the outputs of manually labeled data for neuronal cell location and corresponding fluorescent times series generated by an expert neuroscientist. Results: Our results show that our automated pipeline efficiently pinpoints neuronal cell body location and neuronal contours and provides a graphical representation of neural network metrics accurately reflecting changes in neuronal calcium-based fluorescence. The pipeline detected the shape, area, and location of most neuronal cell body contours by using binary thresholding and grayscale image conversion to allow computer vision to better distinguish between cells and non-cells. Its results were also comparable to manually analyzed results but with significantly reduced result acquisition times of 2-5 minutes per recording versus 10-20 minutes per recording. Based on these findings, our next step is to precisely measure the specificity and sensitivity of the automated pipeline’s cell body and contour detection to extract more robust neural network metrics and dynamics. Conclusion: Our Python-based pipeline performed automated computer vision-based analysis of calcium image recordings from neuronal cell bodies in neuronal cell cultures. Our new goal is to improve cell body and contour detection to produce more robust, accurate neural network metrics and dynamic graphs.

Keywords: calcium imaging, computer vision, neural activity, neural networks

Procedia PDF Downloads 71

Authors: Minghui Zhang, Sirine Fkaier, Sabri Fernana, Svetlana Kiseleva, Denis Kiselev

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The real-time identification of bacteria and fungi is difficult because they emit much weaker signals than pollen. In 2020, Plair developed Rapid-E+, which extends abilities of Rapid-E to detect smaller bioaerosols such as bacteria and fungal spores with diameters down to 0.3 µm, while keeping the similar or even better capability for measurements of large bioaerosols like pollen. Rapid-E+ enables simultaneous measurements of (1) time-resolved, polarization and angle dependent Mie scattering patterns, (2) fluorescence spectra resolved in 16 channels, and (3) fluorescence lifetime of individual particles. Moreover, (4) it provides 2D Mie scattering images which give the full information on particle morphology. The parameters of every single bioaerosol aspired into the instrument are subsequently analysed by machine learning. Firstly, pure species of microbes, e.g., Bacillus subtilis (a species of bacteria), and Penicillium chrysogenum (a species of fungal spores), were aerosolized in a bioaerosol chamber for Rapid-E+ training. Afterwards, we tested microbes under different concentrations. We used several steps of data analysis to classify and identify microbes. All single particles were analysed by the parameters of light scattering and fluorescence in the following steps. (1) They were treated with a smart filter block to get rid of non-microbes. (2) By classification algorithm, we verified the filtered particles were microbes based on the calibration data. (3) The probability threshold (defined by the user) step provides the probability of being microbes ranging from 0 to 100%. We demonstrate how Rapid-E+ identified simultaneously microbes based on the results of Bacillus subtilis (bacteria) and Penicillium chrysogenum (fungal spores). By using machine learning, Rapid-E+ achieved identification precision of 99% against the background. The further classification suggests the precision of 87% and 89% for Bacillus subtilis and Penicillium chrysogenum, respectively. The developed algorithm was subsequently used to evaluate the performance of microbe classification and quantification in real-time. The bacteria and fungi were aerosolized again in the chamber with different concentrations. Rapid-E+ can classify different types of microbes and then quantify them in real-time. Rapid-E+ enables classifying different types of microbes and quantifying them in real-time. Rapid-E+ can identify pollen down to species with similar or even better performance than the previous version (Rapid-E). Therefore, Rapid-E+ is an all-in-one instrument which classifies and quantifies not only pollen, but also bacteria and fungi. Based on the machine learning platform, the user can further develop proprietary algorithms for specific microbes (e.g., virus aerosols) and other aerosols (e.g., combustion-related particles that contain polycyclic aromatic hydrocarbons).

Keywords: bioaerosols, laser-induced fluorescence, Mie-scattering, microorganisms

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Authors: K. Akutsu, S. Mori, T. Hanashima

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Fluorescent probes are useful for the selective detection of trace amount of ions and biomolecular imaging in living cells. Various kinds of metal ion-selective fluorescent compounds have been developed, and some compounds have been applied as effective metal ion-selective fluorescent probes. However, because competition between the ligand and water molecules for the metal ion constitutes a major contribution to the stability of a complex in aqueous solution, it is difficult to develop a highly sensitive, selective, and stable fluorescent probe in aqueous solution. The micelles, these are formed in the surfactant aqueous solution, provides a unique hydrophobic nano-environment for stabilizing metal-organic complexes in aqueous solution. Therefore, we focused on the unique properties of micelles to develop a new fluorescence analysis system. We have been developed a fluorescence analysis system for Sr(II) by using a Sr(II) fluorescent sensor, N-(2-hydroxy-3-(1H-benzimidazol-2-yl)-phenyl)-1-aza-18-crown-6-ether (BIC), and studied its complexation behavior with Sr(II) in micellar solution. We revealed that the stability constant of Sr(II)-BIC complex was 10 times higher than that in aqueous solution. In addition, its detection limit value was also improved up to 300 times by this system. However, the mechanisms of these phenomena have remained obscure. In this study, we investigated the structure of Sr(II)-BIC complex in aqueous micellar solution by combining use the extended X-ray absorption fine structure (EXAFS) and neutron reflectivity (NR) method to understand the unique properties of the fluorescence analysis system from the view point of structural chemistry. EXAFS and NR experiments were performed on BL-27B at KEK-PF and on BL17 SHARAKU at J-PARC MLF, respectively. The obtained EXAFS spectra and their fitting results indicated that Sr(II) and BIC formed a Sr(18-crown-6-ether)-like complex in aqueous micellar solution. The EXAFS results also indicated that the hydrophilic head group of surfactant molecule was directly coordinated with Sr(II). In addition, the NR results also indicated that Sr(II)-BIC complex would interact with the surface of micelle molecules. Therefore, we concluded that Sr(II), BIC, and surfactant molecule formed a ternary complexes in aqueous micellar solution, and at least, it is clear that the improvement of the stability constant in micellar solution is attributed to the result of the formation of Sr(BIC)(surfactant) complex.

Keywords: micell, fluorescent probe, neutron reflectivity, EXAFS

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Authors: Sylwia Orzechowska, Andrzej Wróbel, Eugeniusz Rokita

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The mineralization is a curious problem of connective tissues. Factors which may play a decisive role in the regulation of the yellow ligaments (YL) mineralization are still open questions. The aim of the studies was a detailed description of the chemical composition and morphology of mineral deposits in the human yellow ligaments. Investigations of the structural features of deposits were used to explain the impact of various factors on mineralization process. The studies were carried out on 24 YL samples, surgically removed from patients suffer from spinal canal stenosis and the patients who sustained a trauma. The micro-computed tomography was used to describe the morphology of mineral deposits. The X-ray fluorescence method and Fourier transform infrared spectroscopy were applied to determine the chemical composition of the samples. In order to eliminate the effect of blur in microtomographic images, the correction method of partial volume effect was used. The mineral deposits appear in 60% of YL samples, both in patients with a stenosis and following injury. The mineral deposits have a heterogeneous structure and they are a mixture of the tissue and mineral grains. The volume of mineral grains amounts to (1.9 ± 3.4)*10-3 mm3 while the density distribution of grains occurs in two distinct ranges (1.75 - 2.15 and 2.15-2.5) g/cm3. Application of the partial volume effect correction allows accurate calculations by eliminating the averaging effect of gray levels in tomographic images. The B-type carbonate-containing hydroxyapatite constitutes the mineral phase of majority YLs. The main phase of two samples was calcium pyrophosphate dihydrate (CPPD). The elemental composition of minerals in all samples is almost identical. This pathology may be independent on the spine diseases and it does not evoke canal stenosis. The two ranges of grains density indicate two stages of grains growth and the degree of maturity. The presence of CPPD crystals may coexist with other pathologies.

Keywords: FTIR, micro-tomography, mineralization, spinal ligaments

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Authors: Joanna Cabaj, Sylwia Baluta, Karol Malecha

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Catecholamines are vital neurotransmitters that mediate a variety of central nervous system functions, such as motor control, cognition, emotion, memory processing, and endocrine modulation. Dysfunctions in catecholamine neurotransmission are induced in some neurologic and neuropsychiatric diseases. Changeable neurotransmitters level in biological fluids can be a marker of several neurological disorders. Because of its significance in analytical techniques and diagnostics, sensitive and selective detection of neurotransmitters is increasingly attracting a lot of attention in different areas of bio-analysis or biomedical research. Recently, optical techniques for the detection of catecholamines have attracted interests due to their reasonable cost, convenient control, as well as maneuverability in biological environments. Nevertheless, with the observed need for a sensitive and selective catecholamines sensor, the development of a convenient method for this neurotransmitter is still at its basic level. The manipulation of nanostructured materials in conjunction with biological molecules has led to the development of a new class of hybrid-modified enzymatic sensors in which both enhancement of charge transport and biological activity preservation may be obtained. Immobilization of biomaterials on electrode surfaces is the crucial step in fabricating electrochemical as well as optical biosensors and bioelectronic devices. Continuing systematic investigation in manufacturing of enzyme–conducting sensitive systems, here is presented a convenient fluorescence as well as electrochemical sensing strategy for catecholamines detection.

Keywords: biosensors, catecholamines, fluorescence, enzymes

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Authors: Kai-Cheng Yang, Yen-Ling Chen, Er-Chieh Cho, Kuen-Chan Lee

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Titanium dioxide nanomaterials offer superior protection for human skin against the full spectrum of ultraviolet light. However, some literature reviews indicated that it might be associated with adverse effects such as cytotoxicity or reactive oxygen species (ROS) due to their nanoscale. The surface of fullerene is covered with π electrons constituting aromatic structures, which can effectively scavenge large amount of radicals. Unfortunately, fullerenes are poor solubility in water, severe aggregation, and toxicity in biological applications when dispersed in solvent have imposed the limitations to the use of fullerenes. Carboxyfullerene acts as the scavenger of radicals for several years. Some reports indicate that carboxyfullerene not only decrease the concentration of free radicals in ambience but also prevent cells from reducing the number or apoptosis under UV irradiation. The aim of this study is to decorate fullerene –C70-carboxylic acid (C70-COOH) on the surface of titanium dioxide nanoparticles (P25) for the purpose of scavenging ROS during the irradiation. The modified material is prepared through the esterification of C70-COOH with P25 (P25/C70-COOH). The binding edge and structure are studied by using Transmission electron microscope (TEM) and Fourier transform infrared (FTIR). The diameter of P25 is about 30 nm and C70-COOH is found to be conjugated on the edge of P25 in aggregation morphology with the size of ca. 100 nm. In the next step, the FTIR was used to confirm the binding structure between P25 and C70-COOH. There are two new peaks are shown at 1427 and 1720 cm-1 for P25/C70-COOH, resulting from the C–C stretch and C=O stretch formed during esterification with dilute sulfuric acid. The IR results further confirm the chemically bonded interaction between C70-COOH and P25. In order to provide the evidence of scavenging radical ability of P25/C70-COOH, we chose pyridoxine (Vit.B6) and terephthalic acid (TA) to react with singlet oxygen and hydroxyl radicals. We utilized these chemicals to observe the radicals scavenging statement via detecting the intensity of ultraviolet adsorption or fluorescence emission. The UV spectra are measured by using different concentration of C70-COOH modified P25 with 1mM pyridoxine under UV irradiation for various duration times. The results revealed that the concentration of pyridoxine was increased when cooperating with P25/C70-COOH after three hours as compared with control (only P25). It indicates fewer radicals could be reacted with pyridoxine because of the absorption via P25/C70-COOH. The fluorescence spectra are observed by measuring P25/C70-COOH with 1mM terephthalic acid under UV irradiation for various duration times. The fluorescence intensity of TAOH was decreased in ten minutes when cooperating with P25/C70-COOH. Here, it was found that the fluorescence intensity was increased after thirty minutes, which could be attributed to the saturation of C70-COOH in the absorption of radicals. However, the results showed that the modified P25/C70-COOH could reduce the radicals in the environment. Therefore, we expect that P25/C70-COOH is a potential materials in using for antioxidant.

Keywords: titanium dioxide, fullerene, radical scavenging activity, antioxidant

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Authors: Mark Osborn

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Clustered, regularly interspaced short palindromic repeat (CRISPR/Cas) proteins can be designed to bind specified DNA and RNA sequences and hold great promise for the accurate detection of nucleic acids for diagnostics. Commercially available reagents were integrated into a CRISPR/Cas9-based lateral flow assay that can detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) sequences with single-base specificity. This approach requires minimal equipment and represents a simplified platform for field-based deployment. A rapid, multiplex fluorescence CRISPR/Cas9 nuclease cleavage assay capable of detecting and differentiating SARS-CoV-2, influenza A and B, and respiratory syncytial virus in a single reaction was also developed. These findings provide proof of principle for CRISPR/Cas9 point-of-care diagnosis that can detect specific SARS-CoV-2 strain(s). Further, Cas9 cleavage allows for a scalable fluorescent platform for identifying respiratory viral pathogens with overlapping symptomology. Collectively, this approach is a facile platform for diagnostics with broad application to user-defined sequence interrogation and detection.

Keywords: CRISPR/Cas9, lateral flow assay, SARS-Co-V2, single-nucleotide resolution

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Authors: Yacine Babou, Nitesh Ranjan, Branimir Radisavljevic , Martin Seeger, Daniel Over, Torsten Votteler, Bernardo Galletti, Paulo Cristini

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In the frame of replacement of Sulfur hexafluoride (SF6) gas as insulating and switching medium, diverse alternative gases, offering acceptable Global Warming Potential and fulfilling requirements in terms of heat dissipation, insulation and arc quenching performances are currently investigated for High Voltage Circuit Breaker applications. Among the potential gases, CO₂ seems a promising candidate for replacing SF6, because on one hand it is environmentally friendly, harmless, non-toxic, non-corrosive, non-flammable and on the other hand previous studies have demonstrated its fair interruption capabilities. The present study aims at investigating the performance of CO₂ for the thermal interruption in high voltage self-blast circuit breakers. In particular, the correlation between thermal interruption performance and arc voltage is considered and the effect of the arc-network interaction on the performance is rigorously analyzed. For the considered designs, the thermal interruption was evaluated by varying the slope at current zero (i.e., di/dt) for which the breaker could interrupt. Besides, the characteristics of the post-arc current are examined in detail for various rated voltages and currents. The outcome of these experimental investigations will be reported and analyzed.

Keywords: current zero measurement, high voltage circuit breaker, thermal arc discharge, thermal interruption

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Authors: Maria Lopez-Ibarra, Ana Velazquez-Wong, Lucelli Yañez-Gutierrez, Maria Araujo-Solis, Fabio Salamanca-Gomez, Alfonso Mendez-Tenorio, Haydeé Rosas-Vargas

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Congenital heart diseases (CHD) are the most common congenital abnormalities. These conditions can occur as both an element of distinct chromosomal malformation syndromes or as non-syndromic forms. Their etiology is not fully understood. Genetic variants such copy number variants have been associated with CHD. The aim of our study was to analyze these genomic variants in peripheral blood from Mexican children diagnosed with non-syndromic CHD. We included 16 children with atrial and ventricular septal defects and 5 healthy subjects without heart malformations as controls. To exclude the most common heart disease-associated syndrome alteration, we performed a fluorescence in situ hybridization test to identify the 22q11.2, responsible for congenital heart abnormalities associated with Di-George Syndrome. Then, a microarray based comparative genomic hybridization was used to identify global copy number variants. The identification of copy number variants resulted from the comparison and analysis between our results and data from main genetic variation databases. We identified copy number variants gain in three chromosomes regions from pediatric patients, 4q13.2 (31.25%), 9q34.3 (25%) and 20q13.33 (50%), where several genes associated with cellular, biosynthetic, and metabolic processes are located, UGT2B15, UGT2B17, SNAPC4, SDCCAG3, PMPCA, INPP6E, C9orf163, NOTCH1, C20orf166, and SLCO4A1. In addition, after a hierarchical cluster analysis based on the fluorescence intensity ratios from the comparative genomic hybridization, two congenital heart disease groups were generated corresponding to children with atrial or ventricular septal defects. Further analysis with a larger sample size is needed to corroborate these copy number variants as possible biomarkers to differentiate between heart abnormalities. Interestingly, the 20q13.33 gain was present in 50% of children with these CHD which could suggest that alterations in both coding and non-coding elements within this chromosomal region may play an important role in distinct heart conditions.

Keywords: aCGH, bioinformatics, congenital heart diseases, copy number variants, fluorescence in situ hybridization

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Authors: Busra Gundede, Ozal Mutlu, Nagihan Gulsoy

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Background: Over the years, material research has led to the development of numerous metals and alloys for using in biomedical applications. One of the major tasks of biomaterial research is the functionalization of the material surface to improve the biocompatibility according to a specific application. 316L and 316L alloys are excellent for various bio-applications. This research was investigated the effect of titanium (Ti), niobium (Nb), and zirconium (Zr) additives on injection molded austenitic grade 316L stainless steels in vitro biocompatibility. For this purpose, cytotoxic tests were performed to evaluate the potential biocompatibility of the specimens. Materials and Methods: 3T3 fibroblast were cultivated in DMEM supplemented with 10% fetal bovine serum and %1 penicillin-streptomycin at 37°C with 5% CO2 and 95%humidity. Trypsin/EDTA solution was used to remove cells from the culture flask. Cells were reseeded at a density of 1×105cell in 25T flasks. The medium change took place every 3 days. The trypan blue assay was used to determine cell viability. Cell viability is calculated as the number of viable cells divided by the total number of cells within the grids on the cell counter machine counted the number of blue staining cells and the number of total cells. Cell viability should be at least 95% for healthy log-phase cultures. MTT assay was assessed for 96-hours. Cells were cultivated in 6-well flask within 5 ml DMEM and incubated as same conditions. 0,5mg/ml MTT was added for 4-hours and then acid-isoprohanol was added for solubilize to formazan crystals. Cell morphology after 96h was investigated by SEM. The medium was removed, samples were washed with 0.15 M PBS buffer and fixed for 12h at 4- 8°C with %2,5 gluteraldehyte. Samples were treated with 1% osmium tetroxide. Samples were then dehydrated and dried, mounted on appropriate stubs with colloidal silver and sputter-coated with gold. Images were collected using a scanning electron microscope. ROS assay is a cell viability test for in vitro studies. Cells were grown for 96h, ROS solution added on cells in 6 well plate flask and incubated for 1h. Fluorescence signal indicates ROS generation by cells. Results: Trypan Blue exclusion assay results were 96%, 92%, 95%, 90%, 91% for negative control group, 316L, 316L-Ti, 316L-Nb and 316L-Zr, respectively. Results were found nearly similar to each other when compared with control group. Cell viability from MTT analysis was found to be 100%, 108%, 103%, 107%, and 105% for the control group, 316L, 316L-Ti, 316L-Nb and 316L-Zr, respectively. Fluorescence microscopy analysis indicated that all test groups were same as the control group in ROS assay. SEM images demonstrated that the attachment of 3T3 cells on biomaterials. Conclusion: We, therefore, concluded that Ti, Nb and Zr additives improved physical properties of 316L stainless. In our in vitro experiments showed that these new additives did not modify the cytocompatibility of stainless steel and these additives on 316L might be useful for biomedical applications.

Keywords: 316L stainles steel, biocompatibility, cell culture, Ti, Nb, Zr

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Authors: Hawon Lee, Young-Pil Kim

Abstract:

Visualizing matrix metalloproteinases (MMPs) activity is necessary for understanding cancer metastasis because they are implicated in cell migration and invasion by degrading the extracellular matrix (ECM). While much effort has been made to sense the MMP activity, but extracellularly long-term monitoring of MMP activity still remains challenging. Here, we report a collagen-bound fluorescent bioprobe for the detection of MMP-2 activity in the extracellular environment. This bioprobe consists of ECM-immobilized part (including collagen-bound protein) and MMP-sensing part (including peptide substrate linked with fluorescence resonance energy transfer (FRET) coupler between donor green fluorescent protein (GFP) and acceptor TAMRA dye), which was constructed through intein-mediated self-splicing conjugation. Upon being immobilized on the collagen-coated surface, this bioprobe enabled efficient long-lasting observation of MMP-2 activity in the cultured cells without affecting cell growth and viability. As a result, the FRET ratio (acceptor/donor) decreased as the MMP2 activity increased in cultured cancer cells. Furthermore, unlike wild-type MMP-2, mutated MMP-2 expression (Y580A in the hemopexin region) gave rise to lowering the secretion of MMP-2 in HeLa. Conclusively, our method is anticipated to find applications for tracing and visualizing enzyme activity.

Keywords: collagen, ECM, FRET, MMP

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Authors: Noha Ibrahim, Eman S. Elzanfaly, Said A. Hassan, Ahmed E. El Gendy

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Objectives: The purpose of the study is to develop a sensitive synchronous spectrofluorimetric method with multivariate calibration after studying and optimizing the different variables affecting the native fluorescence intensity of amlodipine and olmesartan implementing an experimental design approach. Method: In the first step, the fractional factorial design used to screen independent factors affecting the intensity of both drugs. The objective of the second step was to optimize the method performance using a Central Composite Face-centred (CCF) design. The optimal experimental conditions obtained from this study were; a temperature of (15°C ± 0.5), the solvent of 0.05N HCl and methanol with a ratio of (90:10, v/v respectively), Δλ of 42 and the addition of 1.48 % surfactant providing a sensitive measurement of amlodipine and olmesartan. The resolution of the binary mixture with a multivariate calibration method has been accomplished mainly by using partial least squares (PLS) model. Results: The recovery percentage for amlodipine besylate and atorvastatin calcium in tablets dosage form were found to be (102 ± 0.24, 99.56 ± 0.10, for amlodipine and Olmesartan, respectively). Conclusion: Method is valid according to some International Conference on Harmonization (ICH) guidelines, providing to be linear over a range of 200-300, 500-1500 ng mL⁻¹ for amlodipine and Olmesartan. The methods were successful to estimate amlodipine besylate and olmesartan in bulk powder and pharmaceutical preparation.

Keywords: amlodipine, central composite face-centred design, experimental design, fractional factorial design, multivariate calibration, olmesartan

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Authors: Mahamuda Sk, Srinivasa Rao Allam, Vijaya Prakash G.

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The different concentrations of Eu3+ ions doped in Oxy-fluoroborate glasses of composition 60 B2O3-10 BaF2-10 CaF2-15 CaF2- (5-x) Al2O3 -x Eu2O3 where x = 0.1, 0.5, 1.0 and 2.0 mol%, have been prepared by conventional melt quenching technique and are characterized through absorption and photoluminescence (PL), decay, color chromaticity and Confocal measurements. The absorption spectra of all the glasses consists of six peaks corresponding to the transitions 7F0→5D2, 7F0→5D1, 7F1→5D1, 7F1→5D0, 7F0→7F6 and 7F1→7F6 respectively. The experimental oscillator strengths with and without thermal corrections have been evaluated using absorption spectra. Judd-Ofelt (JO) intensity parameters (Ω2 and Ω4) have been evaluated from the photoluminescence spectra of all the glasses. PL spectra of all the glasses have been recorded at excitation wavelengths 395 nm (conventional excitation source) and 410 nm (diode laser) to observe the intensity variation in the PL spectra. All the spectra consists of five emission peaks corresponding to the transitions 5D0→7FJ (J = 0, 1, 2, 3 and 4). Surprisingly no concentration quenching is observed on PL spectra. Among all the glasses the glass with 2.0 mol% of Eu3+ ion concentration possesses maximum intensity for the transition 5D0→7F2 (612 nm) in bright red region. The JO parameters derived from the photoluminescence spectra have been used to evaluate the essential radiative properties such as transition probability (A), radiative lifetime (τR), branching ratio (βR) and peak stimulated emission cross-section (σse) for the 5D0→7FJ (J = 0, 1, 2, 3 and 4) transitions of the Eu3+ ions. The decay rates of the 5D0 fluorescent level of Eu3+ ions in the title glasses are found to be single exponential for all the studied Eu3+ ion concentrations. A marginal increase in lifetime of the 5D0 level has been noticed with increase in Eu3+ ion concentration from 0.1 mol% to 2.0 mol%. Among all the glasses, the glass with 2.0 mol% of Eu3+ ion concentration possesses maximum values of branching ratio, stimulated emission cross-section and quantum efficiency for the transition 5D0→7F2 (612 nm) in bright red region. The color chromaticity coordinates are also evaluated to confirm the reddish luminescence from these glasses. These color coordinates exactly fall in the bright red region. Confocal images also recorded to confirm reddish luminescence from these glasses. From all the obtained results in the present study, it is suggested that the glass with 2.0 mol% of Eu3+ ion concentration is suitable to emit bright red color laser.

Keywords: Europium, Judd-Ofelt parameters, laser, luminescence

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Authors: Simran Baweja, Bhavika Kalal, Surajit Maity

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Photoinduced tautomerization reactions have been the centre of attention among scientific community over past several decades because of their significance in various biological systems. 7-azaindole (7AI) is considered as a model system for DNA base pairing and to understand the role of such tautomerization reactions in mutations. To the best of our knowledge, extensive studies have been carried on 7-azaindole and its solvent clusters exhibiting proton/ hydrogen transfer in both solution as well as gas phase. Derivatives of above molecule, like 2,7- and 2,6-diazaindoles are proposed to have even better photophysical properties due to the presence of -aza group on the 2nd position. However, there are a few studies in the solution phase which suggest the relevance of these molecules, but there are no experimental studies reported in the gas phase yet. In our current investigation, we present the first gas phase spectroscopic data of 2,7-diazaindole (2,7-DAI) and its solvent cluster (2,7-DAI-H2O). In this, we have employed state-of-the-art laser spectroscopic methods such as fluorescence excitation (LIF), dispersed fluorescence (DF), resonant two-photon ionization time of flight mass spectrometry (2C-R2PI), photoionization efficiency spectroscopy (PIE), IR-UV double resonance spectroscopy i.e. fluorescence-dip infrared spectroscopy (FDIR) and resonant ion-dip infrared spectroscopy (IDIR) to understand the electronic structure of the molecule. The origin band corresponding to S1 ← S0 transition of the bare 2,7-DAI is found to be positioned at 33910 cm-1 whereas the origin band corresponding to S1 ← S0 transition of the 2,7-DAI-H2O is positioned at 33074 cm-1. The red shifted transition in case of solvent cluster suggests the enhanced feasibility of excited state hydrogen/ proton transfer. The ionization potential for the 2,7-DAI molecule is found to be 8.92 eV, which is significantly higher that the previously reported 7AI (8.11 eV) molecule, making it a comparatively complex molecule to study. The ionization potential is reduced by 0.14 eV in case of 2,7-DAI-H2O (8.78 eV) cluster compared to that of 2,7-DAI. Moreover, on comparison with the available literature values of 7AI, we found the origin band of 2,7-DAI and 2,7-DAI-H2O to be red shifted by -729 and -280 cm-1 respectively. The ground and excited state N-H stretching frequencies of the 27DAI molecule were determined using fluorescence-dip infrared spectra (FDIR) and resonant ion dip infrared spectroscopy (IDIR), obtained at 3523 and 3467 cm-1, respectively. The lower value of vNH in the electronic excited state of 27DAI implies the higher acidity of the group compared to the ground state. Moreover, we have done extensive computational analysis, which suggests that the energy barrier in excited state reduces significantly as we increase the number of catalytic solvent molecules (S= H2O, NH3) as well as the polarity of solvent molecules. We found that the ammonia molecule is a better candidate for hydrogen transfer compared to water because of its higher gas-phase basicity. Further studies are underway to understand the excited state dynamics and photochemistry of such N-rich chromophores.

Keywords: photoinduced tautomerization reactions, gas phse spectroscopy, ), IR-UV double resonance spectroscopy, resonant two-photon ionization time of flight mass spectrometry (2C-R2PI)

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Authors: Yao Xing, Hong Ling Liu, Wei Dong Yu

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With the increase of global population, it is of importance for the safe water supply, while, the water-monitoring method with the capability of rapidness, low-cost, green and robustness remains unsolved. In this paper, gold nanoclusters-wool keratin is added into copper ions measured by fluorescent method in order to probe copper ions in aqueous solution. The fluorescent results show that gold nanoclusters-wool keratin exhibits high stability of pHs, while it is sensitive to temperature and time. Based on Gauss fitting method, the results exhibit that the slope of gold nanoclusters-wool keratin with pH resolution under acidic condition is higher compared to it under alkaline solutions. Besides, gold nanoclusters-wool keratin added into copper ions shows a fluorescence turn-off response transferring from red to blue under UV light, leading to the dramatically decreased fluorescent intensity of gold nanoclusters-wool keratin solution located at 690 nm. Moreover, the limited concentration of copper ions tested by gold nanoclusters-wool keratin system is around 1 µmol/L, which meets the need of detection standards. The fitting slope of Stern-Volmer plot at low concentration of copper ions is larger than it at high concentrations, which indicates that aggregated gold nanoclusters are from small amounts to large numbers with the increasing concentration of copper ions. It is expected to provide novel method and materials for copper ions testing with low cost, high efficiency, and easy operability.

Keywords: gold nanoclusters, copper ions, wool keratin, fluorescence

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Authors: Olushola Ayanda, Simphiwe Nelana, Eliazer Naidoo

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In the present study, the kinetics, equilibrium and thermodynamics of the adsorption of As(III) ions from aqueous solution onto fly ash (FA) was investigated in batch adsorption system. Prior to the adsorption studies, the FA was characterized by means of x-ray fluorescence (XRF), x-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM) and Brunauer-Emmett-Teller (BET) surface area determination. The effect of contact time, initial As(III) concentration, FA dosage, stirring speed, solution pH and temperature was examined on the adsorption rate. Experimental results showed a very good compliance with the pseudo-second-order equation, while the equilibrium study showed that the sorption of As(III) ions onto FA fitted the Langmuir and Freundlich isotherms. The adsorption process is endothermic and spontaneous, moreover, the maximum percentage removal of As(III) achieved with approx. 2.5 g FA mixed with 25 mL of 100 mg/L As(III) solution was 65.4 % at pH 10, 60 min contact time, temperature of 353 K and a stirring speed of 120 rpm.

Keywords: arsenic, fly ash, kinetics, isotherm, thermodynamics

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Authors: Sharon Yunger, Liat Altman, Yuval Garini, Yaron Shav-Tal

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Understanding the dynamics of transcription in living cells has improved since the development of quantitative fluorescence-based imaging techniques. We established a method for following transcription from a single copy gene in living cells. A gene tagged with MS2 repeats, used for mRNA tagging, in its 3' UTR was integrated into a single genomic locus. The actively transcribing gene was detected and analyzed by fluorescence in situ hybridization (FISH) and live-cell imaging. Several cell clones were created that differed in the promoter regulating the gene. Thus, comparative analysis could be obtained without the risk of different position effects at each integration site. Cells in S/G2 phases could be detected exhibiting two adjacent transcription sites on sister chromatids. A sharp reduction in the transcription levels was observed as cells progressed along the cell cycle. We hypothesized that a change in chromatin structure acts as a general mechanism during the cell cycle leading to down-regulation in the activity of some genes. We addressed this question by treating the cells with chromatin decondensing agents. Quantifying and imaging the treated cells suggests that chromatin structure plays a role both in regulating transcriptional levels along the cell cycle, as well as in limiting an active gene from reaching its maximum transcription potential at any given time. These results contribute to understanding the role of chromatin as a regulator of gene expression.

Keywords: cell cycle, living cells, nucleus, transcription

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Authors: Sarim Ahmad

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The Single Cell Gel Electrophoresis Assay (SCGE, known as comet assay) is a potential, uncomplicated, sensitive and state-of-the-art technique for quantitating DNA damage at individual cell level and repair from in vivo and in vitro samples of eukaryotic cells and some prokaryotic cells, being popular in its widespread use in various areas including human biomonitoring, genotoxicology, ecological monitoring and as a tool for research into DNA damage or repair in different cell types in response to a range of DNA damaging agents, cancer risk and therapy. The method involves the encapsulation of cells in a low-melting-point agarose suspension, lysis of the cells in neutral or alkaline (pH > 13) conditions, and electrophoresis of the suspended lysed cells, resulting in structures resembling comets as observed by fluorescence microscopy; the intensity of the comet tail relative to the head reflects the number of DNA breaks. The likely basis for this is that loops containing a break lose their supercoiling and become free to extend towards the anode. This is followed by visual analysis with staining of DNA and calculating fluorescence to determine the extent of DNA damage. This can be performed by manual scoring or automatically by imaging software. The assay can, therefore, predict an individual’s tumor sensitivity to radiation and various chemotherapeutic drugs and further assess the oxidative stress within tumors and to detect the extent of DNA damage in various cancerous and precancerous lesions of oral cavity.

Keywords: comet assay, single cell gel electrophoresis, DNA damage, early detection test

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Authors: Han Xiao

Abstract:

The development of nanomedicines has recently achieved several breakthroughs in the field of cancer treatment; however, the biocompatibility and targeted burst release of these medications remain a limitation, which leads to serious side effects and significantly narrows the scope of their applications. The self-assembly of intermediate filament protein (IFP) peptides was triggered by a hydrophobic cation drug 7-amino actinomycin D (7-AAD) to synthesize pH-activatable nanoparticles (NPs) that could simultaneously locate tumors and produce antitumor effects. The designed IFP peptide included a target peptide (arginine–glycine–aspartate), a negatively charged region, and an α-helix sequence. It also possessed the ability to encapsulate 7-AAD molecules through the formation of hydrogen bonds and hydrophobic interactions by a one-step method. 7-AAD molecules with excellent near-infrared fluorescence properties could be target delivered into tumor cells by NPs and released immediately in the acidic environments of tumors and endosome/lysosomes, ultimately inducing cytotoxicity by arresting the tumor cell cycle with inserted DNA. It is noteworthy that the IFP/7-AAD NPs tail vein injection approach demonstrated not only high tumor-targeted imaging potential, but also strong antitumor therapeutic effects in vivo. The proposed strategy may be used in the delivery of cationic antitumor drugs for precise imaging and cancer therapy.

Keywords: 7-amino actinomycin D, intermediate filament protein, nanoparticle, tumor image

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Authors: Mufath Zorgani, Carlos Garcia-Mateo, Mohammad Jahazi

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In this study, the influence of pre-strained austenite on the extent of isothermal bainite transformation in medium-carbon, high-silicon steel was investigated. Different amounts of deformations were applied at 600°C on the austenite right before quenching to the region, where isothermal bainitic transformation is activated. Four different temperatures of 325, 350, 375, and 400°C considering similar holding time 1800s at each temperature, were selected to investigate the extent of isothermal bainitic transformation. The results showed that the deformation-free austenite transforms to the higher volume fraction of CFB bainite when the isothermal transformation temperature reduced from 400 to 325°C, the introduction of plastic deformation in austenite prior to the formation of bainite invariably involves a delay of the same or identical isothermal treatment. On the other side, when the isothermal transformation temperature and deformation increases, the volume fraction and the plate thickness of bainite decreases and the amount of retained austenite increases. The shape of retained austenite is mostly representing blocky-shape one due to the less amount of transformed bainite. Moreover, the plate-like shape bainite cannot be resolved when the deformation amount reached 30%, and the isothermal transformation temperatures are of 375 and 400°C. The amount of retained austenite and the percentage of its transformation to martensite during the final cooling stage play a significant role in the variation of hardness level for different thermomechanical regimes.

Keywords: ausforming, carbide free bainite, dilatometry, microstructure

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Authors: Shanawaz Patil, Mohamed Haneef, K. S. Narayanaswamy

Abstract:

In the recent years, aluminum metal matrix composites were most widely used, which are finding wide applications in various field such as automobile, aerospace defense etc., due to their outstanding mechanical properties like low density, light weight, exceptional high levels of strength, stiffness, wear resistance, high temperature resistance, low coefficient of thermal expansion and good formability. In the present work, an effort is made to study the effect of heat treatment on mechanical properties of aluminum 7075 alloy reinforced with constant weight percentage of naturally occurring mineral beryl and varying weight percentage of graphene. The hybrid composites are developed with 0.5 wt. %, 1wt.%, 1.5 wt.% and 2 wt.% of graphene and 6 wt.% of beryl  by stir casting liquid metallurgy route. The cast specimens of unreinforced aluminum alloy and hybrid composite samples were prepared for heat treatment process and subjected to solutionizing treatment (T6) at a temperature of 490±5 ^oC for 8 hours in a muffle furnace followed by quenching in boiling water. The microstructure analysis of as cast and heat treated hybrid composite specimens are examined by scanning electron microscope (SEM). The tensile test and hardness test of unreinforced aluminum alloy and hybrid composites are examined. The wear behavior is examined by pin-on disc apparatus. The results of as cast specimens and heat treated specimens were compared. The heat treated Al7075-Beryl-Graphene hybrid composite had better properties and significantly improved the ultimate tensile strength, hardness and reduced wear loss when compared to aluminum alloy and  as cast hybrid composites.

Keywords: beryl, graphene, heat treatment, mechanical properties

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Authors: Riksa Parikrama, Bremmy Laksono, Dadang S. H. Effendi

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Disorder of sexual development (DSD) covers various conditions with a specific term such as Klinefelter syndrome, Turner syndrome, androgen insensitivity syndrome, and many more. The techniques to accurately diagnose those conditions has developed extensively. However, conventional karyotype and fluorescence in situ hybridization (FISH) are still widely used in many genetic laboratories as the basic method to determine chromosomal condition of DSD patients. Cytogenetic study was conducted on 36 DSD patients in Cell Culture and Cytogenetics Laboratory, Faculty of Medicine Universitas Padjadjaran, Indonesia. Most of the patients referred to the laboratory diagnosed with primary amenorrhea, hypospadias, micropenis, genitalia ambiguity, or congenital adrenal hyperplasia. The study used G-banding technique to acquire complete karyotype and followed by FISH as either confirmation or comparison method. Among 36 patients, G-banding karyotype and FISH results showed that two were diagnosed with 45, X (Turner syndrome); three with 47, XXY (Klinefelter syndrome); five with 46, XX DSD; 22 with 46, XY DSD; and four with 46,XY complete androgen insensitivity syndrome. G-banding karyotype analysis were paired with FISH using X and Y chromosome probe produced similar results. The present analysis showed that FISH is a reliable method to attain a rapid and accurate chromosome analysis result of DSD patients. Nevertheless, conventional karyotype technique is still vital if other condition appeared in DSD patients in order to get more detailed karyotype result which FISH method cannot achieve.

Keywords: chromosome, DSD, FISH, karyotype

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