Search results for: vacuum induced freezing

Authors: Julie V. Logan, Preston T. Webster, Kevin B. Woller, Christian P. Morath, Michael P. Short

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Semiconductors, as the base of critical electronic systems, are exposed to damaging radiation while operating in space, nuclear reactors, and particle accelerator environments. What innate property allows some semiconductors to sustain little damage while others accumulate defects rapidly with dose is, at present, poorly understood. This limits the extent to which radiation tolerance can be implemented as a design criterion. To address this problem of determining the driver of semiconductor radiation tolerance, the first step is to generate a dataset of the relative radiation tolerance of a large range of semiconductors (exposed to the same radiation damage and characterized in the same way). To accomplish this, Rutherford backscatter channeling experiments are used to compare the displaced lattice atom buildup in InAs, InP, GaP, GaN, ZnO, MgO, and Si as a function of step-wise alpha particle dose. With this experimental information on radiation-induced incorporation of interstitial defects in hand, hybrid density functional theory electron densities (and their derived quantities) are calculated, and their gradient and Laplacian are evaluated to obtain key fundamental information about the interactions in each material. It is shown that simple, undifferentiated values (which are typically used to describe bond strength) are insufficient to predict radiation tolerance. Instead, the curvature of the electron density at bond critical points provides a measure of radiation tolerance consistent with the experimental results obtained. This curvature and associated forces surrounding bond critical points disfavors localization of displaced lattice atoms at these points, favoring their diffusion toward perfect lattice positions. With this criterion to predict radiation tolerance, simple density functional theory simulations can be conducted on potential new materials to gain insight into how they may operate in demanding high radiation environments.

Keywords: density functional theory, GaN, GaP, InAs, InP, MgO, radiation tolerance, rutherford backscatter channeling

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Authors: Malgorzata Rudnicka, Waldemar Karcz

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Introduction: Naphthoquinones are widely occurring organic compounds produced by bacteria, fungi, and plants. They can act as the functional components of biochemical systems (plastoquinone) as well as biologically active substances, which have a negative impact on environmental processes. Naphthoquinones seem to act through two mechanisms: a covalent modification of biological molecules at their nucleophilic sites or by generation of reactive oxygen species (ROS) connected with redox cycling. Investigating the effect of naphthoquinones (1,4-naphthoquinone, lawsone and naphthazarin) on the elongation growth, membrane potential and the level of oxidative stress of maize cells seems to be important due to the possibility of using these substances as herbicides. Methods: All experiments were performed on etiolated maize coleoptile segments. Simultaneous measurements of elongation growth and pH of the incubation medium were carried out using an angular position transducer, allowing a precise record of the growth kinetics. To compare the oxidative stress level induced by all tested naphthoquinones, the changes in malondialdehyde content, as well as superoxide dismutase and catalase activities were measured. In order to measure the membrane potential of parenchymal cells the standard electrophysiology technique was used. Results: Naphthoquinones such as: 1,4-naphthoquinone, lawsone and naphthazarin were studied. It was found that all of the naphthoquinones diminished the growth of the maize coleoptile cells depending on the type of naphthoquinones and their concentration. Interestingly, naphthazarin at the intermediate concentration was less toxic compared to the others. In addition, the effect of naphthoquinones on the oxidative stress was dependent on their concentration as well. Superoxide dismutase and catalase activities were changed in the presence of higher concentrations of naphthoquinones. Similar interrelations were observed for membrane potential changes. Conclusion: It can be concluded that naphthoquinones tested differ in their toxic effect on the growth of maize coleoptile cells. Furthermore, naphthoquinones can be distinguish considering the oxidative stress level and membrane potential changes. The results presented here give new insight into the possible opportunities of practical usage of naphthoquinones for herbicides improvement.

Keywords: growth rate, membrane potential, naphthoquinones, oxidative stress

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Authors: Marina G. Galitskaya, Svetlana G. Makarova, Andrey P. Fisenko.

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Children on medication-induced immunosuppression are at high risk of developing severe course infectious diseases. Therefore, preventive vaccination is especially important for these children. However, due to the immunosuppressive effects of treatment for the underlying disease, the effectiveness of vaccination may decrease below the protective level. In a multidisciplinary children's medical center, post-vaccination immunity was studied in 79 children aged 4-17 years. The children were divided into 2 groups: Group 1 (38 children) with kidney pathology (Nephrotic Syndrome) and Group 2 (41 children) with inflammatory bowel diseases (Ulcerative Colitis, Crohn's Disease). Both groups of children were vaccinated according to the national vaccination calendar and received immunosuppressive therapy (prednisolone, methotrexate, cyclosporine, and other drugs) for at least 1 year. Using the enzyme-linked immunosorbent assay method, specific IgG antibodies to vaccine-preventable infections were determined: measles, rubella, mumps, diphtheria, pertussis, tetanus, and hepatitis B. The study showed the percentage of children with positive IgG values for vaccine-preventable infections. The highest percentage of children had protective antibody levels to measles (84.2% in children with nephrotic syndrome and 92.6% in those with inflammatory bowel disease) and rubella (71% and 80.4%, respectively). The lowest percentage of children with protective antibodies was for hepatitis B (5.2% and 29.2% respectively). Antibodies to mumps, diphtheria, pertussis, and tetanus were found not in all children (from 39,4% to 82,9%). The remaining percentage of children did not have detectable IgG antibodies to vaccine-preventable infections. Not all children, despite the previous vaccination, preserved antibodies to vaccine-controlled infections and remained unprotected by specific IgG antibodies. The issue of a booster vaccine dose should be considered in children without contraindications to vaccination. Children receiving long-term immunosuppressive therapy require an individual vaccination approach, including a specific definition of the performed vaccination.

Keywords: immunosuppressive therapy, inflammatory bowel diseases, nephrotic syndrome, post-vaccination immunity, specific antibodies, vaccine-preventable infections.

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Authors: Adel Salem Bahakeem, Ahmad Jamal, Mir Md. Maruf Morshed, Elwaleed Awad Khidir

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Direct Current (DC) servo motors, or simply DC motors, play an important role in many industrial applications such as manufacturing of plastics, precise positioning of the equipment, and operating computer-controlled systems where speed of feed control, maintaining the position, and ensuring to have a constantly desired output is very critical. These parameters can be controlled with the help of control systems such as the Proportional Integral Derivative (PID) controller. The aim of the current work is to investigate the effects of Proportional (P) and Integral (I) controllers on the steady state and transient response of the DC motor. The controller gains are varied to observe their effects on the error, damping, and stability of the steady and transient motor response. The current investigation is conducted experimentally on a servo trainer CE 110 using analog PI controller CE 120 and theoretically using Simulink in MATLAB. Both experimental and theoretical work involves varying integral controller gain to obtain the response to a steady-state input, varying, individually, the proportional and integral controller gains to obtain the response to a step input function at a certain frequency, and theoretically obtaining the proportional and integral controller gains for desired values of damping ratio and response frequency. Results reveal that a proportional controller helps reduce the steady-state and transient error between the input signal and output response and makes the system more stable. In addition, it also speeds up the response of the system. On the other hand, the integral controller eliminates the error but tends to make the system unstable with induced oscillations and slow response to eliminate the error. From the current work, it is desired to achieve a stable response of the servo motor in terms of its angular velocity subjected to steady-state and transient input signals by utilizing the strengths of both P and I controllers.

Keywords: DC servo motor, proportional controller, integral controller, controller gain optimization, Simulink

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Authors: Abbas Ali Husseini, Ali Mohammad Yazdani, Fatemeh Ghadiri, Alper Şişman

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We developed a microchip device that uses surface acoustic waves for rapid lysis of low level of cell samples. The device incorporates sharp-edge glass microparticles for improved performance. We optimized the lysis conditions for high efficiency and evaluated the device's feasibility for point-of-care applications. The microchip contains a 13-finger pair interdigital transducer with a 30-degree focused angle. It generates high-intensity acoustic beams that converge 6 mm away. The microchip operates at a frequency of 16 MHz, exciting Rayleigh waves with a 250 µm wavelength on the LiNbO3 substrate. Cell lysis occurs when Candida albicans cells and glass particles are placed within the focal area. The high-intensity surface acoustic waves induce centrifugal forces on the cells and glass particles, resulting in cell lysis through lateral forces from the sharp-edge glass particles. We conducted 42 pilot cell lysis experiments to optimize the surface acoustic wave-induced streaming. We varied electrical power, droplet volume, glass particle size, concentration, and lysis time. A regression machine-learning model determined the impact of each parameter on lysis efficiency. Based on these findings, we predicted optimal conditions: electrical signal of 2.5 W, sample volume of 20 µl, glass particle size below 10 µm, concentration of 0.2 µg, and a 5-minute lysis period. Downstream analysis successfully amplified a DNA target fragment directly from the lysate. The study presents an efficient microchip-based cell lysis method employing acoustic streaming and microparticle collisions within microdroplets. Integration of a surface acoustic wave-based lysis chip with an isothermal amplification method enables swift point-of-care applications.

Keywords: cell lysis, surface acoustic wave, micro-glass particle, droplet

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Authors: Chih-Hsin Lin, Shyh-Yuan Lee, Yuan-Min Lin

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For many tissue engineering applications, an important goal is to create functional tissues in-vitro, and such tissues to be viable, they have to be vascularized. Endothelial cells (EC) and endothelial progenitor cells (EPC) are promising candidates for vascularization. However, both of them have limited expansion capacity and autologous cells currently do not exist for either ECs or EPCs. Therefore, we use bone marrow mesenchymal stem cells (MSC) as a source material for ECs. Growth supplements are commonly used to induce MSC differentiation, and further improvements in differentiation conditions can be made by modifying the cell's growth environment. An example is pre-treatment of the growth dish with gas plasma, in order to modify the surface functional groups of the material that the cells are seeded on. In this work, we compare the effects of different gas plasmas on the growth and differentiation of MSCs. We treat the dish with different plasmas (CO2, N2, and O2) and then induce MSC differentiation with endothelial growth medium-2 (EGM-2). We find that EGM-2 by itself upregulates EC marker CD31 mRNA expression, but not VEGFR2, CD34, or vWF. However, these additional EC marker expressions were increased for cells seeded on plasma treated substrates. Specifically, for EC markers, we found that N2 plasma treatment upregulated CD31 and VEGFR-2 mRNA expressions; CO2 plasma treatment upregulated CD34 and vWF mRNA expressions. The osteogenic markers ALP and osteopontin mRNA expressions were markedly enhanced on all plasma-treated dishes. We also found that plasma treatment in conjunction with EGM-2 growth medium can enhance MSCs differentiation into endothelial-like cells and osteogenic-like cells. Our work shows that the effect of the growth medium (EGM-2) on MSCs differentiation is influenced by the plasma modified surface chemistry of the substrate. In conclusion, plasma surface modification can enhance EGM-2 effectiveness and induced both endothelial and osteogenic differentiation. Our findings provide a method to enhance EGM-2 based cell differentiation, with consequences for tissue engineering and stem cell biology applications.

Keywords: endothelial differentiation, EGM-2, osteogenesis, plasma treatment, surface modification

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Authors: Mansouri Ouarda, Abdennour Cherif, Saidi Malika

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Since the industrial revolution, many anthropogenic activities have caused environmental, considerable and overall changes. The lead represents a very dangerous disruptive for the functioning of the body. In this context the current study aims at evaluating a natural therapy by the use of the plant grass in wheat (Triticum durum) against the toxicity of lead in rat wistar male. The rats were divided into three groups: the control group, the group treated with 600 mg /kg food of lead only (Pb) is the group treated with the combination of 600 mg/kg of food and 9g/rat /day of the plant grass in wheat (Pb-bl). The duration of the treatment is 6 weeks. The results of the biometrics of the organs (thyroid, kidney, testis and epididymis) show no significant difference between the three groups. The dosage of a few parameters and hormonal biochemical shows a decrease in the concentration of the hormone T3 and TSH levels among the group pb alone compared to the control and Pb-Bl. These results have been confirmed by the study of histological slices. A morphological changes represented by a shrinking volume of vesicles with the group treated with Pb alone. A return to the normal state of the structure of the follicles was observed. The concentration in serum testosterone, urea and creatinine was significantly increased among the group treated by Pb only in relation to the control and Pb-Bl. whereas the rate of glucose did not show any significant difference. The histology study of the kidney, testis and epididymal weights show no modification at the group Pb-bl comparing to the control. The parenchyma of the kidney shows a dilation of tubes distal and proximal causing a tubular nephropathy for the batch processed by Pb only. The testicles have marked a destruction or absence of germ cells and the light of some seminiferous are almost empty. Conclusion: The supplementation of the plant Triticum durum has caused a considerable improvement which ensures the return of parameters investigated in the normal state.

Keywords: creatinine, glucose, histological sections, T3, TSH, testosterone

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Authors: R. Fardid, R. Coppes

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Introduction: In mediastinal radiotherapy and to a lesser extend also in total-body irradiation (TBI) radiation exposure may lead to development of cardiac diseases. Radiation-induced heart disease is dose-dependent and it is characterized by a loss of cardiac function, associated with progressive heart cells degeneration. We aimed to determine the in-vivo radiation effects on fibronectin, ColaA1, ColaA2, galectin and TGFb1 gene expression levels in left ventricle heart tissues of rats after irradiation. Material and method: Four non-treatment adult Wistar rats as control group (group A) were selected. In group B, 4 adult Wistar rats irradiated to 20 Gy single dose of 150 Mev proton beam locally in heart only. In heart plus lung irradiate group (group C) 4 adult rats was irradiated by 50% of lung laterally plus heart radiation that mentioned in before group. At 8 weeks after radiation animals sacrificed and left ventricle heart dropped in liquid nitrogen for RNA extraction by Absolutely RNA® Miniprep Kit (Stratagen, Cat no. 400800). cDNA was synthesized using M-MLV reverse transcriptase (Life Technologies, Cat no. 28025-013). We used Bio-Rad machine (Bio Rad iQ5 Real Time PCR) for QPCR testing by relative standard curve method. Results: We found that gene expression of fibronectin in group C significantly increased compared to control group, but it was not showed significant change in group B compared to group A. The levels of gene expressions of Cola1 and Cola2 in mRNA did not show any significant changes between normal and radiation groups. Changes of expression of galectin target significantly increased only in group C compared to group A. TGFb1 expressions in group C more than group B showed significant enhancement compared to group A. Conclusion: In summary we can say that 20 Gy of proton exposure of heart tissue may lead to detectable damages in heart cells and may distribute function of them as a component of heart tissue structure in molecular level.

Keywords: gene expression, heart damage, proton irradiation, radiotherapy

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Authors: Howaida I. Abd-Alla, Omnia Kutkat, Yassmin Moatasim, Magda T. Ibrahim, Marwa A. Mostafa, Mohamed GabAllah, Mounir M. El-Safty

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Artemisia judaica (known shih-balady), Azadirachta indica and Sophora tomentosa (known yellow necklace pod) are members of available medicinal plants well-known for their traditional medical use in Egypt which suggests that they probably harbor broad-spectrum antiviral, immunostimulatory and anti-inflammatory functions. Their ethyl acetate-dichloromethane (1:1, v/v) extracts were evaluated for the potential anti-Middle East respiratory syndrome-related coronavirus (anti-MERS-CoV) activity. Their cytotoxic activity was tested in Vero-E6 cells using 3-(4,-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method with minor modification. The plot of percentage cytotoxicity for each extract concentration has calculated the concentration which exhibited 50% cytotoxic concentration (TC50). A plaque reduction assay was employed using safe dose of extract to evaluate its effect on virus propagation. The highest inhibition percentage was recorded for the yellow necklace pod, followed by Shih-balady. The possible mode of action of virus inhibition was studied at three different levels viral replication, viral adsorption and virucidal activity. The necklace pod leaves have induced virucidal effects and direct effects on the replication of virus. Phytochemical investigation of the promising necklace pod led to the isolation and structure determination of nine compounds. The structure of each compound was determined by a variety of spectroscopic methods. Compounds 4-O-methyl sorbitol 1, 8-methoxy daidzin 6 and 6-methoxy apigenin-7-O-β-D-glucopyranoside 8 were isolated for the first time from the Sophora genus and the other six compounds were the first time that they were isolated from this species according to available works of literature. Generally, the highest anti-CoV 2 activity of S. tomentosa was associated with the crude ethanolic extract, indicating the possibility of synergy among the antiviral phytochemical constituents (1-9).

Keywords: coronavirus, MERS-CoV, mode of action, necklace pod, shih-balady

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Authors: Fei Sun, Meilan Xu, Jianghui Zhu, Maria Stefanie Dwiyanti, Cheolwoo Park, Fanjiang Kong, Baohui Liu, Tetsuya Yamada, Jun Abe

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Reduced or lack of sensitivity to long daylengths is a key character for soybean, a short-day crop, to adapt to higher latitudinal environments. However, the photoperiod-insensitivity often results in a reduction of the duration of vegetative growth and final yield. To overcome this limitation, a photoperiod insensitive line (RIL16) was developed in this study that delayed flowering from the recombinant inbred population derived from a cross between a photoperiod-insensitive cultivar AGS292 and a late-flowering Thai cultivar K3. Expression analyses under SD and LD conditions revealed that the expression levels of FLOWERING LOCUS T (FT) orthologues, FT2a and FT5a, were lowered in RIL16 relative to AGS292, although the expression of E1, a soybean-specific suppressor for FTs, was inhibited in both conditions. A soybean orthologue of TARGET OF EAT1 (TOE1), another suppressor of FT, showed an upregulated expression in RIL16, which appeared to reflect a lower expression of miR172a. Our data suggest that the delayed flowering of RIL16 most likely is controlled by genes involved in an age-dependent pathway in flowering. The QTL analysis based on 1,125 SNPs obtained from Restriction Site Associated DNA Sequencing revealed two major QTLs for flowering dates in Chromosome 16 and two minor QTLs in Chromosome 4, all of which accounted for 55% and 48% of the whole variations observed in natural day length and artificially-induced long day length conditions, respectively. The intervals of the major QTLs harbored FT2a and FT5a, respectively, on the basis of annotated genes in the Williams 82 reference genome. Sequencing analysis further revealed a nonsynonymous mutation in FT2a and an SNP in the 3′ UTR region of FT5a. A further study may elucidate a detailed mechanism underlying the QTL for late flowering. The alleles from K3 at the two QTLs can be used singly or in combination to retain an appropriate duration of vegetative growth to maximize the final yield of photoperiod-insensitive soybeans.

Keywords: FT genes, miR72a, photoperiod-insensitive, soybean flowering

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Authors: Rawia M. Khalil, Ahmed A. Abd El Rahman, Mahfouz A. Kassem, Mohamed S. El Ridi, Mona M. Abou Samra, Ghada E. A. Awad, Soheir S. Mansy

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Solid lipid nanoparticles (SLNs) have gained great attention for the topical treatment of skin associated fungal infection as they facilitate the skin penetration of loaded drugs. Our work deals with the preparation of nystatin loaded solid lipid nanoparticles (NystSLNs) using the hot homogenization and ultrasonication method. The prepared NystSLNs were characterized in terms of entrapment efficiency, particle size, zeta potential, transmission electron microscopy, differential scanning calorimetry, rheological behavior and in vitro drug release. A stability study for 6 months was performed. A microbiological study was conducted in male rats infected with Candida albicans, by counting the colonies and examining the histopathological changes induced on the skin of infected rats. The results showed that SLNs dispersions are spherical in shape with particle size ranging from 83.26±11.33 to 955.04±1.09 nm. The entrapment efficiencies are ranging from 19.73±1.21 to 72.46±0.66% with zeta potential ranging from -18.9 to -38.8 mV and shear-thinning rheological Behavior. The stability studies done for 6 months showed that nystatin (Nyst) is a good candidate for topical SLN formulations. A least number of colony forming unit/ ml (cfu/ml) was recorded for the selected NystSLN compared to the drug solution and the commercial Nystatin® cream present in the market. It can be fulfilled from this work that SLNs provide a good skin targeting effect and may represent promising carrier for topical delivery of Nyst offering the sustained release and maintaining the localized effect, resulting in an effective treatment of cutaneous fungal infection.

Keywords: candida infections, hot homogenization, nystatin, solid lipid nanoparticles, stability, topical delivery

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Authors: Yasmin Begum Mohammed

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Ketorolac tromethamine (KTM) is a non-steroidal anti-inflammatory drug with a potent analgesic and anti-inflammatory activity due to prostaglandin related inhibitory effect of drug. It is a non-selective cyclo-oxygenase inhibitor. The drug is currently used orally and intramuscularly in multiple divided doses, clinically for the management arthritis, cancer pain, post-surgical pain, and in the treatment of migraine pain. KTM has short biological half-life of 4 to 6 hours, which necessitates frequent dosing to retain the action. The frequent occurrence of gastrointestinal bleeding, perforation, peptic ulceration, and renal failure lead to the development of other drug delivery strategies for the appropriate delivery of KTM. The ideal solution would be to target the drug only to the cells or tissues affected by the disease. Drug targeting could be achieved effectively by liposomes that are biocompatible and biodegradable. The aim of the study was to develop a parenteral liposome formulation of KTM with improved efficacy while reducing side effects by targeting the inflammation due to arthritis. PEG-anchored (stealth) and non-PEG-anchored liposomes were prepared by thin film hydration technique followed by extrusion cycle and characterized for in vitro and in vivo. Stealth liposomes (SLs) exhibited increase in percent encapsulation efficiency (94%) and 52% percent of drug retention during release studies in 24 h with good stability for a period of 1 month at -20°C and 4°C. SLs showed about maximum 55% of edema inhibition with significant analgesic effect. SLs produced marked differences over those of non-SL formulations with an increase in area under plasma concentration time curve, t₁/₂, mean residence time, and reduced clearance. 0.3% of the drug was detected in arthritic induced paw with significantly reduced drug localization in liver, spleen, and kidney for SLs when compared to other conventional liposomes. Thus SLs help to increase the therapeutic efficacy of KTM by increasing the targeting potential at the inflammatory region.

Keywords: biodistribution, ketorolac tromethamine, stealth liposomes, thin film hydration technique

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Authors: Afshin Zahedi, Keivan Jamshidi

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This study was conducted to determine the neurotoxic effects of different doses of ACR on the thoracic axons of the spinal cord of rat. To evaluate this hypothesis in the thoracic axons, amino-cupric silver staining technique of the de Olmos was conducted to define the histopathologic characteristic (argyrophilia) of axonal damage following ACR exposure. For this purpose 60 adult male rats (Wistar, approximately 250 g) were selected. Rats were hosed in polycarbonate boxes as two per each. Randomly assigned groups of rats (10 rats per exposure group, total 5 exposure groups as A, B, C, D and E) were exposed to 0.5, 5, 50, 100 and 500 mg/kg per day×11days intraperitoneal injection (IP injection) respectively. The remaining 10 rats were housed in group (F) as control group. Control rats received daily injections of 0.9% saline (3ml/kg). As indices of developing neurotoxicity, weight gain, gait scores and landing hindlimb foot splay (LHF) were determined. Weight gains were measured daily prior to injection. Gait scoring involved observation of spontaneous open field locomotion, included evaluations of ataxia, hopping, rearing and hind foot placement, and hindlimb foot splay were determined 3-4 times per week. Gait score was assigned from 1-4. After 11 days, two rats for silver stain, were randomly selected, dissected and proper samples were collected from thoracic portion of the spinal cord of rat. Results did show no neurological behavior in groups A, B and F, whereas severe neurotoxicity was observed in groups C and D. Rats in groups E died within 1-2 hours due to severe toxemia. In histopathological studies based on the de Olmos technique no argyrophilic neurons or processes were observed in stained sections obtained from the thoracic portion of the spinal cord of rats belong to groups A, B and F, while moderate to severe argyrophilic changes were observed in different stained sections obtained from the thoracic portion of the spinal cord of rats belong to groups C and D.

Keywords: acrylamide, rat, axonopathy, argyrophily, de Olmos

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Authors: Q. B. Maria de la Luz Ortega Juárez, Saúl Rojas Hernández, Itzel Berenice Rodríguez Mera, María Maricela Carrasco Yépez, Mara Gutierrez Sánchez

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Naegleria fowleri is a free-living amoeba found mainly in temperate freshwater and is the etiologic agent of primary amebic meningoencephalitis (PAM), a fatal acute disease with a mortality rate greater than 95%. At present, there are no treatments available for MAP, and the development of effective vaccines that generate long-term immunological memory allowing protection against MAP would be of great importance. The objective of this work was to analyze the effector and memory immune response in BALB/c mice immunized with total extract of N. fowleri co-administered with cholera toxin. In this study, BALB/c mice were immunized four times intranasally with ET of N. fowleri adjuvanted with CT with or without booster at three months and were challenged or not with the lethal dose of N. fowleri, determining survival, the humoral, effector and memory response, by ELISA and flow cytometry techniques. The results obtained showed that the survival of mice immunized with booster had 60% protection compared to the group without booster, which obtained 20% protection. Evaluating the humoral response, it was found that both IgG and IgA levels were higher in sera than in nasal washes in both treatments. In the cellular response, the increase in the percentage of positive cells was found for effector T and B lymphocytes in the nasal passages (NP) in the group with boost and nasopharynx-associated lymphoid tissue (NALT) in the group without boost and lymphocytes only. B in both treatments, as well as in memory cells treatment with boost T lymphocytes in PN and NALT and without boost in cervical lymph nodes (CG) with respect to B lymphocytes, in PN, GC and NALT in treatment with boost and NALT in treatment without booster. Therefore, the involvement of the effector immune response and memory play a fundamental role for protection against N. fowleri and for the development of vaccine candidates.

Keywords: immune response, immunological memory, naegleria fowleri, primary amebic meningoencephalitis

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Authors: Shivdayal Patel, Suhail Ahmad

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Safety assurance and failure prediction of composite material component of an offshore structure due to low velocity impact is essential for associated risk assessment. It is important to incorporate uncertainties associated with material properties and load due to an impact. Likelihood of this hazard causing a chain of failure events plays an important role in risk assessment. The material properties of composites mostly exhibit a scatter due to their in-homogeneity and anisotropic characteristics, brittleness of the matrix and fiber and manufacturing defects. In fact, the probability of occurrence of such a scenario is due to large uncertainties arising in the system. Probabilistic finite element analysis of composite plates due to low-velocity impact is carried out considering uncertainties of material properties and initial impact velocity. Impact-induced damage of composite plate is a probabilistic phenomenon due to a wide range of uncertainties arising in material and loading behavior. A typical failure crack initiates and propagates further into the interface causing de-lamination between dissimilar plies. Since individual crack in the ply is difficult to track. The progressive damage model is implemented in the FE code by a user-defined material subroutine (VUMAT) to overcome these problems. The limit state function is accordingly established while the stresses in the lamina are such that the limit state function (g(x)>0). The Gaussian process response surface method is presently adopted to determine the probability of failure. A comparative study is also carried out for different combination of impactor masses and velocities. The sensitivity based probabilistic design optimization procedure is investigated to achieve better strength and lighter weight of composite structures. Chain of failure events due to different modes of failure is considered to estimate the consequences of failure scenario. Frequencies of occurrence of specific impact hazards yield the expected risk due to economic loss.

Keywords: composites, damage propagation, low velocity impact, probability of failure, uncertainty modeling

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Authors: Kirsten Wilson, Patrick M. Brauer, Sandra Babic, Diana Golubeva, Jessica Van Eyk, Tinya Wang, Avanti Karkhanis, Tim A. Le Fevre, Andy I. Kokaji, Allen C. Eaves, Sharon A. Louis, , Nooshin Tabatabaei-Zavareh

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Long-lived plasma cells are rare, non-proliferative B cells generated from antibody-secreting cells (ASCs) following an immune response to protect the host against pathogen re-exposure. Despite their therapeutic potential, the lack of in vitro protocols in the field makes it challenging to use B cells as a cellular therapeutic tool. As a result, there is a need to establish robust and reproducible methods for the generation of B cells. To address this, we have developed a culture system for generating B cells from hematopoietic stem and/or progenitor cells (HSPCs) derived from human umbilical cord blood (CB) or pluripotent stem cells (PSCs). HSPCs isolated from CB were cultured using the StemSpan™ B Cell Generation Kit and produced CD19+ B cells at a frequency of 23.2 ± 1.5% and 59.6 ± 2.3%, with a yield of 91 ± 11 and 196 ± 37 CD19+ cells per input CD34+ cell on culture days 28 and 35, respectively (n = 50 - 59). CD19+IgM+ cells were detected at a frequency of 31.2 ± 2.6% and were produced at a yield of 113 ± 26 cells per input CD34+ cell on culture day 35 (n = 50 - 59). The B cell receptor loci of CB-derived B cells were sequenced to confirm V(D)J gene rearrangement. ELISpot analysis revealed that ASCs were generated at a frequency of 570 ± 57 per 10,000 day 35 cells, with an average IgM+ ASC yield of 16 ± 2 cells per input CD34+ cell (n = 33 - 42). PSC-derived HSPCs were generated using the STEMdiff™ Hematopoietic - EB reagents and differentiated to CD10+CD19+ B cells with a frequency of 4 ± 0.8% after 28 days of culture (n = 37, 1 embryonic and 3 induced pluripotent stem cell lines tested). Subsequent culture of PSC-derived HSPCs increased CD19+ frequency and generated ASCs from 1 - 2 iPSC lines. This method is the first report of a serum- and feeder-free system for the generation of B cells from CB and PSCs, enabling further B lineage-specific research for potential future clinical applications.

Keywords: stem cells, B cells, immunology, hematopoiesis, PSC, differentiation

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Authors: Mona Alharbi

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Melioidosis has emerged as a lethal disease. Unfortunately, the molecular mechanisms of virulence and pathogenicity of Burkholderia pseudomallei remain unknown. However, proteomics research has selected putative targets in B. pseudomallei that might play roles in the B. pseudomallei virulence. BPSL 2418 putative protein has been predicted as a free methionine sulfoxide reductase and interestingly there is a link between the level of the methionine sulfoxide in pathogen tissues and its virulence. Therefore in this work, we describe the cloning expression, purification, and crystallization of BPSL 2418 and the solution of its 3D structure using X-ray crystallography. Also, we aimed to identify the substrate binding and reduced forms of the enzyme to understand the role of BPSL 2418. The gene encoding BPSL2418 from B. pseudomallei was amplified by PCR and reclone in pETBlue-1 vector and transformed into E. coli Tuner DE3 pLacI. BPSL2418 was overexpressed using E. coli Tuner DE3 pLacI and induced by 300μM IPTG for 4h at 37°C. Then BPS2418 purified to better than 95% purity. The pure BPSL2418 was crystallized with PEG 4000 and PEG 6000 as precipitants in several conditions. Diffraction data were collected to 1.2Å resolution. The crystals belonged to space group P2 21 21 with unit-cell parameters a = 42.24Å, b = 53.48Å, c = 60.54Å, α=γ=β= 90Å. The BPSL2418 binding MES was solved by molecular replacement with the known structure 3ksf using PHASER program. The structure is composed of six antiparallel β-strands and four α-helices and two loops. BPSL2418 shows high homology with the GAF domain fRMsrs enzymes which suggest that BPSL2418 might act as methionine sulfoxide reductase. The amino acids alignment between the fRmsrs including BPSL 2418 shows that the three cysteines that thought to catalyze the reduction are fully conserved. BPSL 2418 contains the three conserved cysteines (Cys⁷⁵, Cys⁸⁵ and Cys¹⁰⁹). The active site contains the six antiparallel β-strands and two loops where the disulfide bond formed between Cys⁷⁵ and Cys¹⁰⁹. X-ray structure of free methionine sulfoxide binding and native forms of BPSL2418 were solved to increase the understanding of the BPSL2418 catalytic mechanism.

Keywords: X-Ray Crystallography, BPSL2418, Burkholderia pseudomallei, Melioidosis

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Authors: Lintle Mohase, Ninikoe Lebusa, Mpho Stephen Mafa

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Wheat is an economically important cereal crop. However, the changing climatic conditions that intensify drought in production areas, and additional pest infestation, such as the Russian wheat aphid (RWA, Diuraphis noxia), severely hamper its production. Drought and pest management require an additional water supply through irrigation and applying inorganic nutrients (including silicon) as alternative strategies to mitigate the stress effects. Therefore, other approaches are needed to enhance wheat productivity during drought stress and aphid abundance. Two wheat cultivars were raised under greenhouse conditions, exposed to drought stress, and treated with silicon before infestation with the South African RWA biotype 2 (RWASA2). The morphological evaluations showed that severe drought or a combination of drought and infestation significantly reduced the plant height of wheat cultivars. Silicon treatment did not alleviate the growth reduction. The biochemical responses were measured using spectrophotometric assays with specific substrates. An evaluation of the enzyme activities associated with oxidative stress and defence responses indicated that drought stress increased NADPH oxidase activity, while silicon treatment significantly reduced it in drought-stressed and infested plants. At 48 and 72 hours sampling periods, a combination of silicon, drought and infestation treatment significantly increased peroxidase activity compared to drought and infestation treatment. The treatment also increased β-1,3-glucanase activity 72 hours after infestation. In addition, silicon and drought treatment increased glucose but reduced sucrose accumulation. Furthermore, silicon, drought, and infestation treatment combinations reduced the sucrose content. Finally, silicon significantly increased the trehalose content under severe drought and infestation, evident at 48 and 72-hour sampling periods. Our findings shed light on silicon’s ability to induce protective biochemical responses during drought and aphid infestation.

Keywords: drought, enzyme activity, silicon, soluble sugars, Russian wheat aphid, wheat

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Authors: Maxime Moreau, Silvère Baron, Jean-Marc Lobaccaro, Karine Charlet, Sébastien Menecier, Frédéric Perisse

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Cold Atmospheric Plasma (CAP) is an ionized gas generated at atmospheric pressure with the temperature of heavy particles (molecules, ions, atoms) close to the room temperature. Recent studies have shown that both in-vitro and in-vivo plasma exposition to many cancer cell lines are efficient to induce the apoptotic way of cell death. In some other works, normal cell lines seem to be less impacted by plasma than cancer cell lines. This is called selectivity of plasma. It is highly likely that the generated RNOS (Reactive Nitrogen Oxygen Species) in the plasma jet, but also in the medium, play a key-role in this selectivity. In this study, two CAP devices will be compared to electrical power, chemical species composition and their efficiency to kill cancer cells. A particular focus on the action of hydrogen peroxide will be made. The experiments will take place as described next for both devices: electrical and spectroscopic characterization for different voltages, plasma treatment of normal and cancer cells to compare the CAP efficiency between cell lines and to show that death is induced by an oxidative stress. To enlighten the importance of hydrogen peroxide, an inhibitor of H2O2 will be added in cell culture medium before treatment and a comparison will be made between the results of cell viability in this case and those from a simple plasma exposition. Besides, H2O2 production will be measured by only treating medium with plasma. Cell lines will also be exposed to different concentrations of hydrogen peroxide in order to characterize the cytotoxic threshold for cells and to make a comparison with the quantity of H2O2 produced by CAP devices. Finally, the activity of catalase for different cell lines will be quantified. This enzyme is an important antioxidant agent against hydrogen peroxide. A correlation between cells response to plasma exposition and this activity could be a strong argument in favor of the predominant role of H2O2 to explain the selectivity of plasma cancer treatment by cold atmospheric plasma.

Keywords: cold atmospheric plasma, hydrogen peroxide, prostate cancer, selectivity

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Authors: Nurul Aliaa Idris, David A. Collings

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Phi thickenings are specialized secondary cell wall thickenings that are found in the cortex of the roots in a wide range of plant species, including orchids. The role of phi thickenings in the root is still under debate through research have linked environmental conditions, particularly abiotic stresses such as water stress, heavy metal stress and salinity to their induction in the roots. It has also been suggested that phi thickenings may act as a barrier to regulate solute uptake, act as a physical barrier against fungal hyphal penetration due to its resemblance to the Casparian strip and play a mechanical role to support cortical cells. We have investigated phi thickening function in epiphytic orchids of the genus Miltoniopsis through induction experiment against factors such as soil compaction and water stress. The permeability of the phi thickenings in Miltoniopsis was tested through uptake experiments using the fluorescent tracer dyes Calcofluor white, Lucifer yellow and Propidium iodide then viewed with wide-field or confocal microscopy. To test whether phi thickening may prevent fungal colonization in the root cell, fungal re-infection experiment was conducted by inoculating isolated symbiotic fungus to sterile in vitro Miltoniopsis explants. As the movement of fluorescent tracers through the apoplast was not blocked by phi thickenings, and as phi thickenings developed in the roots of sterile cultures in the absence of fungus and did not prevent fungal colonization of cortical cells, the phi thickenings in Miltoniopsis do not function as a barrier. Phi thickenings were found to be absent in roots grown on agar and remained absent when plants were transplanted to moist soil. However, phi thickenings were induced when plants were transplanted to well-drained media, and by the application of water stress in all soils tested. It is likely that phi thickenings stabilize the root cortex during dehydration. Nevertheless, the varied induction responses present in different plant species suggest that the phi thickenings may play several adaptive roles, instead of just one, depending on species.

Keywords: abiotic stress, Miltoniopsis, orchid, phi thickening

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Authors: Seham Samir Soliman, Ahmed A.Suliman, Khaled Fathy, Ahmed A. Sedik

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Cyclophosphamide (CP), an antineoplastic drug, has been found to induce reproductive damage. It is essential to develop approaches aimed at safeguarding ovarian tissue integrity in women experiencing reproductive toxicity as a result of chemotherapy. The current study was conducted to assess the impact of an extract derived from Moringa oleifera (M. oleifera) leaves on ovarian damage produced by CP. A total of 32 female Wistar Albino rats, which were in a healthy cycling state, were randomly separated into 4 groups, with every group contains 8 rats. The first group was administered intraperitoneal (i.p.) saline. The second group was administered a solitary intraperitoneal dosage of cyclophosphamide (200 mg/kg). The third one received M. oleifera extract (150 mg/kg orally) for 20 days, followed by i.p. of CP on the last day of the experiment. The fourth group received M. oleifera extract (250 mg/kg orally) for 20 days, followed by i.p. of CP on the last day of the experiment. Hormonal assessments, including luteinizing hormone (LH), estrogen (ES), and follicle-stimulating hormone (FSH), were performed 24 hours after CP administration. In addition, evaluating the antioxidant status and inflammatory response against CP. Moreover, conducting detailed histopathological and ultra-structural pictures of the ovary. Our findings reported that rats intoxicated with CP exhibited elevated levels of FSH, LH, malondialdehyde (MDA), tumor necrosis factor-alpha (TNF-α), and a decrease in E₂, and glutathione (GSH) levels. Pre-treatment with M. oleifera extract (250 mg/kg orally) ameliorated the disturbance in hormonal changes, oxidative stress indices, and the levels of pro-inflammatory mediators. Also, the histopathological and ultra-structural pictures of the ovaries were improved significantly in rats. In conclusion, M. oleifera extract possesses a significant protective role against CP-induced acute reproductive toxicity via modulating the values of FSH, LH, E₂ and quenching the release of reactive oxygen species and inflammatory mediators in female rats.

Keywords: cyclophosphamide, Moringa oleifera, ovarian function, oxidative stress, pro-inflammatory mediators

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Authors: Rohollah Ghasemi, Arvin Ghorbani

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This study investigates the effect of austempering temperature on microstructure and scratch behavior of the austempered heat-treated compacted graphite irons. The as-cast was used as base material for heat treatment practices. The samples were extracted from as-cast ferritic CGI pieces and were heat treated under austenitising temperature of 900°C for 60 minutes which followed by quenching in salt-bath at different austempering temperatures of 275°C, 325°C and 375°C. For all heat treatments, an austempering holding time of 30 minutes was selected for this study. Light optical microscope (LOM) and scanning electron microscope (SEM) and electron back scattered diffraction (EBSD) analysis confirmed the ausferritic matrix formed in all heat-treated samples. Microscratches were performed under the load of 200, 600 and 1000 mN using a sphero-conical diamond indenter with a tip radius of 50 μm and induced cone angle 90° at a speed of 10 μm/s at room temperature ~25°C. An instrumented nanoindentation machine was used for performing nanoindentation hardness measurement and microscratch testing. Hardness measurements and scratch resistance showed a significant increase in Brinell, Vickers, and nanoindentation hardness values as well as microscratch resistance of the heat-treated samples compared to the as-cast ferritic sample. The increase in hardness and improvement in microscratch resistance are associated with the formation of the ausferrite matrix consisted of carbon-saturated retained austenite and acicular ferrite in austempered matrix. The maximum hardness was observed for samples austempered at 275°C which resulted in the formation of very fine acicular ferrite. In addition, nanohardness values showed a quite significant variation in the matrix due to the presence of acicular ferrite and carbon-saturated retained austenite. It was also observed that the increase of austempering temperature resulted in increase of volume of the carbon-saturated retained austenite and decrease of hardness values.

Keywords: austempered CGI, austempering, scratch testing, scratch plastic deformation, scratch hardness

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Authors: Jan Bohata, Stanislav Zvanovec, Matej Komanec, Jakub Jaros, David Hruby

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Even though there is a rapid development in new optical networks, still optical communication infrastructures remain composed of thousands of kilometers of aging optical cables. Many of them are located in a harsh environment which contributes to an increased attenuation or induced birefringence of the fibers leading to the increase of polarization mode dispersion (PMD). In this paper, we report experimental results from environmental optical cable tests and characterization in the climate chamber. We focused on the evaluation of optical network reliability in a harsh environment. For this purpose, a special thermal chamber was adopted, targeting to the large temperature changes between -60 °C and 160 C° with defined humidity. Single mode optical cable 230 meters long, having six tubes and a total number of 72 single mode optical fibers was spliced together forming one fiber link, which was afterward tested in the climate chamber. The main emphasis was put to the polarization mode dispersion (PMD) changes, which were evaluated by three different PMD measuring methods (general interferometry technique, scrambled state-of-polarization analysis and polarization optical time domain reflectometer) in order to fully validate obtained results. Moreover, attenuation and chromatic dispersion (CD), as well as the PMD, were monitored using 17 km long single mode optical cable. Results imply a strong PMD dependence on thermal changes, imposing the exceeding 200 % of its value during the exposure to extreme temperatures and experienced more than 20 dB insertion losses in the optical system. The derived statistic is provided in the paper together with an evaluation of such as optical system reliability, which could be a crucial tool for the optical network designers. The environmental tests are further taken in context to our previously published results from long-term monitoring of fundamental parameters within an optical cable placed in a harsh environment in a special outdoor testbed. Finally, we provide a correlation between short-term and long-term monitoring campaigns and statistics, which are necessary for optical network safety and reliability.

Keywords: optical fiber, polarization mode dispersion, harsh environment, aging

Procedia PDF Downloads 363

Authors: Saroj Pandey

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COVID-19, declared by the World Health Organization a pandemic (WHO,2020), has created an unprecedented crisis world over endangering the human survival itself. Corona induced lockdowns forced approximately 140 million students of 190 countries at various levels of education from preprimary to higher education to remain confined to their homes. In India, approximately 360 million students were affected by the forced shut down of schools due to the countrywide lockdown in March 2020 and resultant disruption of education. After the initial shock and anxiety the Indian polity and education system bounced back with a number of initiatives, and online education came as a major rescuer for the education system of the country. The distance and online mode of learning that was treated as the poor cousin of conventional mode and often criticized for its quality became the major crusader overnight changing the entire ecosystem of traditional teaching -leaning towards the virtual mode. Teachers who were averse to technology were forced to remodel their educational pedagogies and reorient themselves overnight to use various online platforms such as Zoom, Google meet, and other such platforms to reach the learners. This metamorphosis through ensured students was meaningfully engaged in their studies during the lockdown period but it has its own set of challenges. This paper deals with the government initiatives, and teachers' self-efforts to keep the channel of teaching learning on providing academic and socio emotional support to students during the most difficult period of their life as well as the digital divide between the rich and poor, rural and urban, and boys and girls in India and resultant challenges. It also provides an overview of few significant self-initiatives of teachers to reach their students during the crisis period, who did not have internet and smartphone facilities as well as the initiatives being taken at the government level to address the learning needs and mitigate the learning gaps of learners, bridge the digital divide, strategic planning and upskilling of teachers to overcome the effect of COVID-19 crisis.

Keywords: COVID-19, online education, initiatives, challenges

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Authors: E. T. Carvalho Filho, J. T. N. Medeiros, L. G. Martinez

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Residual stresses are self equilibrating in a rigid body that acts on the microstructure of the material without application of an external load. They are elastic stresses and can be induced by mechanical, thermal and chemical processes causing a deformation gradient in the crystal lattice favoring premature failure in mechanicals components. The search for measurements with good reliability has been of great importance for the manufacturing industries. Several methods are able to quantify these stresses according to physical principles and the response of the mechanical behavior of the material. The diffraction X-ray technique is one of the most sensitive techniques for small variations of the crystalline lattice since the X-ray beam interacts with the interplanar distance. Being very sensitive technique is also susceptible to variations in measurements requiring a study of the factors that influence the final result of the measurement. Instrumental, operational factors, form deviations of the samples and geometry of analyzes are some variables that need to be considered and analyzed in order for the true measurement. The aim of this work is to analyze the sources of errors inherent to the residual stress measurement process by X-ray diffraction technique making an interlaboratory comparison to verify the reproducibility of the measurements. In this work, two specimens were machined, differing from each other by the surface finishing: grinding and polishing. Additionally, iron powder with particle size less than 45 µm was selected in order to be a reference (as recommended by ASTM E915 standard) for the tests. To verify the deviations caused by the equipment, those specimens were positioned and with the same analysis condition, seven measurements were carried out at 11Ψ tilts. To verify sample positioning errors, seven measurements were performed by positioning the sample at each measurement. To check geometry errors, measurements were repeated for the geometry and Bragg Brentano parallel beams. In order to verify the reproducibility of the method, the measurements were performed in two different laboratories and equipments. The results were statistically worked out and the quantification of the errors.

Keywords: residual stress, x-ray diffraction, repeatability, reproducibility, error analysis

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Authors: Zhala Ahmad, Zainab Lazim, Haider Hamzah

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Bacterial resistance is a pressing global health issue, with multidrug-resistant (MDR), extensively drug-resistant (XDR), and pandrug-resistant (PDR) strains to pose a serious threat. In this context, researchers are investigating effective, safe, and affordable metabolites to combat these pathogens. This study focuses on gum essential oil (GEO) extracted from Pistacia atlantica and its activity and the mechanism of action against XDR Gram-negative Acinetobacter baumannii. GEO was extracted by hydrodistillation and analyzed using GC-MS. Eleven A. baumannii isolates were collected from the ward environment of Burn and Plastic Surgery Hospital in Al Sulaymaniyah City, Iraq. They were identified using the VITEK 2 system, 16S rRNA gene, and confirmed with the blaₒₓₐ₋₅₁ gene; A. baumannii ATCC 19606 was used as a reference strain. The isolates were identified as resistant to twelve different antibiotics spanning six distinct antibiotic classes while showing susceptibility to tetracycline and trimethoprim. Over 40 chemical constituents were detected in the gum's essential oils, with α-pinene being the most abundant. GEO was found to inhibit the growth of A. baumannii isolates; the minimum inhibitory concentration (MIC) of GEO was 2.5 µl/ml. GEO induced protein leakage, phosphate, and potassium ion efflux, distorted cell morphology, and cell death in the tested bacteria. GEO exhibited bacterial clearance and anti-adhesion activity using Band-Aids. This study's findings suggest that GEO could be used as a potential alternative treatment for infectious diseases caused by XRD pathogens, shedding further light on the importance of GEO in biomedical applications. Future studies must focus on generating clinically feasible sources of GEO for testing in small animal models before proceeding to human trials, ensuring safe and effective translation from the laboratory to the clinic.

Keywords: antibiotic resistance, Acinetobacter baumannii, essential oils, Pistacia atlantica, alpha-pinene

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Authors: Theresa Steiner, Markus Pajones, Christian Haider

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Traffic is being induced by companies due to their economic behavior. Basically, two different types of traffic occur at company sites: freight traffic and commuting traffic. Due to the fact that these traffic types are connected to each other in different kinds, an integrated approach to manage them is useful. Mobility management is a proved method for companies, to handle the traffic processes caused by their business activities. According to recent trend analysis in Austria, the freight traffic as well as the individual traffic, as part of the commuting traffic, will continue to increase. More traffic jams, as well as negative environmental impacts, are expected impacts for the future. Mobility management is a tool to control the traffic behavior with the scope to reduce emissions and other negative effects which are caused by traffic. Until now, mobility management is mainly used for optimizing commuting traffic without taking the freight logistics processes into consideration. However, the method of mobility management can be used to improve the freight traffic area of a company as well. The focus of this paper will be particularly laid on analyzing to what extent companies are already using mobility management to influence not only the commuting traffic they produce but also their processes of freight logistics. A further objective is to acquire knowledge about the motivating factors which persuade companies to introduce and apply mobility management. Additionally, advantages and disadvantages of this tool will be defined as well as limitations and factors of success, with a special focus on freight logistics, will be depicted. The first step of this paper is to conduct a literature review on the issue of mobility management with a special focus on freight logistics processes. To compare the theoretical findings with the practice, interviews, following a structured interview guidline, with mobility managers of different companies in Upper Austria will be undertaken. A qualitative analysis of these surveys will in a first step show the motivation behind using mobility management to improve traffic processes and how far this approach is already being used to especially influence the freight traffic of the companies. An evaluation to what extent the method of mobility management is already being approached at Upper Austrian companies to regulate freight logistics processes will be one outcome of this publication. Furthermore, the results of the theoretical and practical analysis will reveal not only the possibilities but also the limitations of using mobility management to influence the processes of freight logistics.

Keywords: freight logistics processes, freight traffic, mobility management, passenger traffic

Procedia PDF Downloads 298

Authors: Wan-Ting Kuo, Yi-Wen Liu, Hsiao-Sheng Liu

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Annual incidence of bladder cancer increases in the world and occurs frequently in the male. Most common type is transitional cell carcinoma (TCC) which is treated by transurethral resection followed by intravesical administration of agents. In clinical treatment of bladder cancer, chemotherapeutic drugs-induced apoptosis is always used in patients. However, cancers usually develop resistance to chemotherapeutic drugs and often lead to aggressive tumors with worse clinical outcomes. Approximate 70% TCC recurs and 30% recurrent tumors progress to high-grade invasive tumors, indicating that new therapeutic agents are urgently needed to improve the successful rate of overall treatment. Nonapoptotic program cell death may assist to overcome worse clinical outcomes. Autophagy which is one of the nonapoptotic pathways provides another option for bladder cancer patients. Autophagy is reported as a potent anticancer therapy in some cancers. First of all, we established a mouse orthotopic bladder tumor formation model in order to create a similar tumor microenvironment. IVIS system and micro-ultrasound were utilized to noninvasively monitor tumor formation. In addition, we carried out intravesical treatment in our animal model to be consistent with human clinical treatment. In our study, we carried out intravesical instillation of the autophagy inducer in mouse orthotopic bladder tumor to observe tumor formation by noninvasive IVIS system and micro-ultrasound. Our results showed that bladder tumor formation is suppressed by the autophagy inducer, and there are no significant side effects in the physiology of mice. Furthermore, the autophagy inducer upregulated autophagy in bladder tissues of the treated mice was confirmed by Western blot, immunohistochemistry, and immunofluorescence. In conclusion, we reveal that a novel autophagy inducer with low side effects suppresses bladder tumor formation in our mouse orthotopic bladder tumor model, and it provides another therapeutic approach in bladder cancer patients.

Keywords: bladder cancer, transitional cell carcinoma, orthotopic bladder tumor formation model, autophagy

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Authors: Tarana Siddika, Ilka U. Heinemann, Patrick O’Donoghue

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Protein kinase B (AKT1) is a serine/threonine kinase and central transducer of cell survival pathways. Typical approaches to study AKT1 biology in cells rely on growth factor or insulin stimulation that activates AKT1 via phosphorylation at two key regulatory sites (Threonine308, Serine473), yet cell stimulation also activates many other kinases and fails to differentiate the effect of the two main activating sites of AKT1 on downstream substrate phosphorylation and cell growth. While both AKT1 activating sites are associated with disease and used as clinical markers, in some cancers, high levels of Threonine308 phosphorylation are associated with poor prognosis while in others poor survival correlates with high Serine473 levels. To produce cells with specific AKT1 activity, a system was developed to deliver active AKT1 to human cells. AKT1 phospho-variants were produced from Escherichia coli with programmed phosphorylation by genetic code expansion. Tagging of AKT1 with an N-terminal cell penetrating peptide tag derived from the human immunodeficiency virus trans-activator of transcription (TAT) helped to enter AKT1 proteins in mammalian cells. The TAT-tag did not alter AKT1 kinase activity and was necessary and sufficient to rapidly deliver AKT1 protein variants that persisted in human cells for 24 h without the need to use transfection reagents. TAT-pAKT1T308, TAT-pAKT1S473 and TAT-pAKT1T308S473 proteins induced selective phosphorylation of the known AKT1 substrate GSK-3αβ, and downstream stimulation of the AKT1 pathway as evidenced by phosphorylation of ribosomal protein S6 at Serine240/244 in transfected cells. Increase in cell growth and proliferation was observed due to the transfection of different phosphorylated AKT1 protein variants compared to cells with TAT-AKT1 protein. The data demonstrate efficient delivery of AKT1 with programmed phosphorylation to human cells, thus establishing a cell-based model system to investigate signaling that is dependent on specific AKT1 activity and phosphorylation.

Keywords: cell penetrating peptide, cell signaling, protein kinase b (AKT1), phosphorylation

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Authors: KuiDong Kang, Hye Bin Yim, Su Ah Kim

Abstract:

Purpose: Tectorigenin is an isoflavone derived from the rhizome of Belamacanda chinensis. In this study, oxygen-induced retinopathy was used to characterize the anti-angiogenic properties of tectorigenin in mice. Methods: ICR neonatal mice were exposed to 75% oxygen from postnatal day P7 until P12 and returned to room air (21% oxygen) for five days (P12 to P17). Mice were subjected to daily intraperitoneal injection of tectorigenin (1 mg/kg, 10 mg/kg) and vehicle from P12 to P17. Retro-orbital injection of FITC-dextran was performed and retinal flat mounts were viewed by fluorescence microscopy. The Central avascular area was quantified from the digital images in a masked fashion using image analysis software (NIH ImageJ). Neovascular tufts were quantified by using SWIFT_NV and neovascular lumens were quantified from a histologic section in a masked fashion. Immunohistochemistry and Western blot analysis were also performed to demonstrate the anti-angiogenic activity of this compound in vivo. Results: In the retina of tectorigenin injected mouse (10mg/kg), the central non-perfusion area was significantly decreased compared to the vehicle injected group (1.76±0.5 mm2 vs 2.85±0.6 mm2, P<0.05). In vehicle-injected group, 33.45 ± 5.51% of the total retinal area was avascular, whereas the retinas of pups treated with high-dose (10 mg/kg) tectorigenin showed avascular retinal areas of 21.25 ±4.34% (P<0.05). High dose of tectorigenin also significantly reduced the number of vascular lumens in the histologic section. Tectorigenin (10 mg/kg) significantly reduced the expression of vascular endothelial growth factor (VEGF), matrix metalloproteinase-2 (MMP-2), MMP-9, and angiotensin II compared to the vehicle injected group. Tectorigenin did not affect CD31 abundance at any tested dose. Conclusions: Our results show that tectorigenin possesses powerful anti-angiogenic properties and can attenuate new vessel formation in the retina after systemic administration. These results imply that this compound can be considered as a candidate substance for therapeutic inhibition of retinal angiogenesis.

Keywords: tectorigenin, anti-angiogenic, retinopathy, Belamacanda chinensis

Procedia PDF Downloads 249