Search results for: biological traits

Authors: M. E. Abalaka, S. Y. Daniyan, S. O. Adeyemo, D. Damisa

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Gold nanoparticles (AuNPs) have gained increasing interest in recent times. This is greatly due to their special features, which include unusual optical and electronic properties, high stability and biological compatibility, controllable morphology and size dispersion, and easy surface functionalization. In typical synthesis, AuNPs were produced by reduction of gold salt AuCl4 in an appropriate solvent. A stabilizing agent was added to prevent the particles from aggregating. The antibacterial activity of different sizes of gold nanoparticles was investigated against Staphylococcus aureus, Salmonella typhi and Pseudomonas pneumonia using the disk diffusion method in a Müeller–Hinton Agar. The Au-NPs were effective against all bacteria tested. That the Au-NPs were successfully synthesized in suspension and were used to study the antibacterial activity of the two medicinal plants against some bacterial pathogens suggests that Au-NPs can be employed as an effective bacteria inhibitor and may be an effective tool in medical field. The study clearly showed that the Au-NPs exhibiting inhibition towards the tested pathogenic bacteria in vitro could have the same effects in vivo and thus may be useful in the medical field if well researched into.

Keywords: gold nanoparticles, Gomphrena celesioides, Prunus amygdalus, pathogens

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Authors: Bjorn-Ole Meyer, Dominik Marti, Peter E. Andersen

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A multimodal imaging system, combining spectrally resolved multiphoton microscopy (MPM) and optical coherence microscopy (OCM) is demonstrated. MPM and OCM are commonly integrated into multimodal imaging platforms to combine functional and morphological information. The MPM signals, such as two-photon fluorescence emission (TPFE) and signals created by second harmonic generation (SHG) are biomarkers which exhibit information on functional biological features such as the ratio of pyridine nucleotide (NAD(P)H) and flavin adenine dinucleotide (FAD) in the classification of cancerous tissue. While the spectrally resolved imaging allows for the study of biomarkers, using a spectrometer as a detector limits the imaging speed of the system significantly. To overcome those limitations, an OCM setup was added to the system, which allows for fast acquisition of structural information. Thus, after rapid imaging of larger specimens, navigation within the sample is possible. Subsequently, distinct features can be selected for further investigation using MPM. Additionally, by probing a different contrast, complementary information is obtained, and different biomarkers can be investigated. OCM images of tissue and cell samples are obtained, and distinctive features are evaluated using MPM to illustrate the benefits of the system.

Keywords: optical coherence microscopy, multiphoton microscopy, multimodal imaging, two-photon fluorescence emission

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Authors: Katsuyuki Umezawa, Makoto Nakazawa, Manabu Kobayashi, Yutaka Ishii, Michiko Nakano, Shigeichi Hirasawa

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This research aims to develop a self-study system equipped with an artificial teacher who gives advice to students by detecting the learners and to evaluate language learning in a unified framework. 'Detecting the learners' means that the system understands the learners' learning conditions, such as each learner’s degree of understanding, the difference in each learner’s thinking process, the degree of concentration or boredom in learning, and problem solving for each learner, which can be interpreted from learning behavior. In this paper, we propose a system to efficiently collect brain waves from learners by focusing on only the brain waves among the biological information for 'detecting the learners'. The conventional Electroencephalograph (EEG) measurement method during learning using a simple EEG has the following disadvantages. (1) The start and end of EEG measurement must be done manually by the experiment participant or staff. (2) Even when the EEG signal is weak, it may not be noticed, and the data may not be obtained. (3) Since the acquired EEG data is stored in each PC, there is a possibility that the time of data acquisition will be different in each PC. This time, we developed a system to collect brain wave data on the server side. This system overcame the above disadvantages.

Keywords: artificial teacher, e-learning, self-study system, simple EEG

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Authors: Sanja O. Podunavac-Kuzmanović, Strahinja Z. Kovačević, Lidija R. Jevrić, Stela Jokić

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The complexes of transition metals with various organic ligands have been extensively studied as models of some important pharmaceutical molecules. It was found that biological properties of different substituted organic molecules are improved when they are complexed by different metals. Therefore, it is of great importance for the development of coordination chemistry to explore the assembly of functional organic ligands with metal ion and to investigate the relationship between the structure and property. In the present work, we have bioassayed the antibacterial potency of benzimidazoles and their metal salts (Cu or Zn) against yeast Sarcina lutea. In order to validate our in vitro study, we performed in silico studies using molecular docking software. The investigated compounds and their metal complexes (Cu, Zn) showed good to moderate inhibitory activity against Sarcina lutea. In silico docking studies of the synthesized compounds suggested that complexed benzimidazoles have a greater binding affinity and improved antibacterial activity in comparison with non-complexed ligands. These results are part of the CMST COST Action No. 1105 "Functional metal complexes that bind to biomolecules".

Keywords: organometallic complexes, benzimidazoles, chemometric design, Sarcina lutea

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Authors: Rochelyn Dona, Serdar Satar

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The aim of this study was to evaluate the biological parameters of A. fabae Scopoli (Hemiptera: Aphididae). Developmental, survival, and reproductive data were collected for Aphis fabae reared on detached bean leaves (Phaseolus vulgaris L.) ‘pinto beans’ at five temperature regimes (12, 16, 20, 24, and 28 °C), 65% relative humidity (RH), relative and a photoperiod of 16:8 (LD) h. The developmental times of immature stages ranged from 16, 65 days at 12°C to 5.70 days at 24°C, but a slight increase again at 28°C (6.62 days). At 24°C from this study presented the developmental threshold for A. fabae slightly to 24°C. The average longevity of mature females significantly decreased from 42.32 days at 12°C to 16.12 days at 28°C. The reproduction rate per female was 62.27 at 16°C and 12.72 at 28°C. The mean generation period of the population ranged from 29.24 at 12°C to 11.50 at 28°C. The highest intrinsic rate of increase (rm = 0.41) were recorded at 24°C, the lowest at 12°C (rm = 0.15). It was evident that temperatures over 28°C augmented the development time, accelerated the death ratio of the nymphal stages, Shrunk Adult longevity, and reduced fecundity. The optimal range of temperature for the population growth of A. fabae on the bean was 16°C-24°C, according to this study.

Keywords: developmental time, intrinsic rate, reproduction period, temperature dependence

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Authors: Ali Seirafi

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The purpose of this study was to investigate the effect and application of starch nanocrystals on physicochemical and microbial properties in the industrial production of probiotic yogurt. In this study, probiotic yoghurt was manufactured by industrial method with the optimization and control of the technological factors affecting the probabilistic biomass, using probiotic bacteria Lactobacillus acidophilus and Bifidobacterium bifidum with commonly used yogurt primers. Afterwards, the effects of different levels of fat (1.3%, 2.5 and 4%), as well as the effects of various perbiotic compounds include starch nanocrystals (0.5%, 1 and 1.5%), galactolegalosaccharide (0.5% 1 and 1.5%) and fructooligosaccharide (0.5%, 1 and 1.5%) were evaluated. In addition, the effect of packaging (polyethylene and glass) was studied, while the effect of pH changes and final acidity were studied at each stage. In this research, all experiments were performed in 3 replications and the results were analyzed in a completely randomized design with SAS version 9.1 software. The results of this study showed that the addition of starch nanocrystal compounds as well as the use of glass packaging had the most positive effects on the survival of Lactobacillus acidophilus bacteria and the addition of nano-crystals and the increase in the cooling rate of the product, had the most positive effects on the survival of bacteria Bifidobacterium bifidum.

Keywords: Bifidobacterium bifidum, Lactobacillus acidophilus, prebiotics, probiotic yogurt

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Authors: Aycan Gurluyer

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21st-century theatre has been shaped by lots of world-changing forces devastating human psychology and existence. Accepted as the greatest living English playwright, it is post-war British dramatist Edward Bond who uses a late-21st-century apocalyptic landscape as a weapon to question the human existence in his play 'Have I None?'. In this play, he tries to underline the degenerating and destructive effects of the society and environment on a couple whose lives are changed by an unexpected and annoying stranger. As victim of the society and the cultural corruption, the three vulnerable Bondian characters struggle for their expectations to find a place in this fictional world by sacrificing their own lives. Set in the 2077’s world, the play depicts that rigidly formed rules of the system/authority eliminates the existence of humans. According to Bond, the fascist practices of the governments/systems make people paralyzed in any way, so they choose to deny all realities by becoming biological beings or they gather to commit to suicide as troops. Our main aim is to underscore the questioning of the human existence by drawing the socio-political framework of the era, the capitalist system’s dehumanized individuals and their defence to survive, and what reality is in the 21st century, by focusing on Bond’s hallucinatory and tragic vision of the future in 'Have I None?'.

Keywords: Edward Bond, apocalyptic, existence, Have I None?

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Authors: L. Major, J. M. Lackner, M. Dyner, B. Major

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Development of new generation bio- tribological, multilayer coatings, opens an avenue for fabrication of future high- tech functional surfaces. In the presented work, nano- composite, Cr/CrN+[Cr/ a-C:H implanted by metallic nanocrystals] multilayer coatings have been developed for surface protection of medical tools. Thin films were fabricated by a hybrid Pulsed Laser Deposition technique. Complex microstructure analysis of nano- multilayer coatings, subjected to mechanical and biological tests, were performed by means of transmission electron microscopy (TEM). Microstructure characterization revealed the layered arrangement of Cr23C6 nanoparticles in multilayer structure. Influence of deposition conditions on bio- tribological properties of the coatings were studied. The bio-tests were used as a screening tool for the analyzed nano- multilayer coatings before they could be deposited on medical tools. Bio- medical tests were done using fibroblasts. The mechanical properties of the coatings were investigated by means of a ball-on-disc mechanical test. The microhardness was done using Berkovich indenter. The scratch adhesion test was done using Rockwell indenter. From the bio- tribological point of view, the optimal properties had the C106_1 material.

Keywords: bio- tribological coatings, cell- material interaction, hybrid PLD, tribology

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Authors: Hatem A. El-Mezayen, Ahmed Abdelmajeed, Fatehya Metwally, Usama Elsaly, Salwa Atef

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Tumor metastasis involves the dissemination of malignant cells into the basement membrane and vascular system contributes to the circulating pool of these markers. In this context our aim has been focused on development of a non-invasive. Circulating tumor cells (CTCs) represent a unique liquid biopsy carrying comprehensive biological information of the primary tumor. Herein, we sought to develop a novel score based on the combination of the most significant CTCs biomarkers with and routine laboratory tests for accurate detection of metastatic breast cancer. Methods: Cytokeratin 18 (CK18), Cytokeratin 19 (CK19), and CA15.3 were assayed in metastatic breast cancer (MBC) patients (75), non-MBC patients (50) and healthy control (20). Results: Areas under receiving operating curve (AUCs) were calculated and used for construction on novel score. A novel score named MBC-CTCs = CA15.3 (U/L) × 0.08 + CK 18 % × 2.9 + CK19 × 3.1– 510. That function correctly classified 87% of metastatic breast cancer at cut-off value = 0.55. (i.e great than 0.55 indicates patients with metastatic breast cancer and less than 0.55 indicates patients with non-metastatic breast cancer). Conclusion: MBC-CTCs is a novel, non-invasive and simple can applied to discriminate patients with metastatic breast cancer.

Keywords: metastatic breast cancer, circulating tumor cells, cytokeratin, EpiCam

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Authors: Anupuma Raina, Ajay P. Balayan, Prateek Pandya, Pankaj Shrivastava, Uma Kanga, Tulika Seth

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DNA fingerprinting analysis aids in personal identification for forensic purposes and has always been a driving motivation for law enforcement agencies in almost all countries since its inception. The introduction of DNA markers (Y-STR) has allowed for greater precision and higher discriminatory power in forensic testing. A criminal/ person committing crime after bone marrow transplantation is a rare situation but not an impossible one. Keeping such a situation in mind, a study was carried out to find out the best biological sample to be used for personal identification, especially in forensic situation. We choose a female patient (recipient) and a male donor. The pre transplant sample (blood) and post transplant samples (blood, buccal swab, hair roots) were collected from the recipient (patient). The same were compared with the blood sample of the donor using DNA FP technique. Post transplant samples were collected at different interval of time (15, 30, 60, and 90 days). The study was carried out using Y-STR kit at 23 loci. The results determined discusses the phenomenon of chimerism and its impact on Y-STR. Hair sample was found the most suitable sample which had no donor DNA profiling up to 90 days.

Keywords: bone marrow transplantation, chimerism, DNA profiling, Y-STR

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Authors: Sanjay Bari, Vinod Ugale, Kamalkishor Patil

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Over the past several years the emergence of micro-organisms resistant to nearly all the class of antimicrobial agents has become a serious public health concern. In the present research, we report the synthesis and in-vitro antimicrobial activity of a new series of novel quinazolino-thiadiazoles 3 (a-j). The synthesized compounds were confirmed by melting point, IR, 1H-NMR, 13C NMR and Mass spectroscopy. In general, the results of the in-vitro antibacterial activity are encouraging, as out of 10 compounds tested, Compound 3f and 3i with a 4-chloro phenyl and 4-nitro phenyl at C-2 of thiadiazolyl of quinazolino-thiadiazoles, displayed the excellent antibacterial and antifungal activities against all the tested microorganisms (Bacterial and Fungal strain) with MIC values of 62.5 μg/mL. It is worth to mention that the combination of two biologically active moieties quinazoline and thiadiazole profoundly influences the biological activity. While evaluating the antimicrobial activity, it was observed that compounds having electron withdrawing groups on thiazole has shown profound activity in comparison to compounds having electron releasing groups. As a result of this study, it can be concluded that halogen substituent on thiazole ring increases antimicrobial activity. Possible improvements in the antimicrobial activity can be further achieved by slight modifications in the substituent’s and/or additional structural activity investigations to have good antimicrobial activity.

Keywords: antifungal, antimicrobial, quinazolino-thiazoles, synthesis

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Authors: Dilek Ozturk, Narin Ozturk, Zeliha Pala Kara, Engin Kaptan, Serap Sancar Bas, Nurten Ozsoy, Alper Okyar

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Most physiological processes oscillate in a rhythmic manner in mammals including metabolism and energy homeostasis, locomotor activity, hormone secretion, immune and endocrine system functions. Endocrine body rhythms are tightly regulated by the circadian timing system. The hypothalamic-pituitary-thyroid (HPT) axis is under circadian control at multiple levels from hypothalamus to thyroid gland. Since circadian timing system controls a variety of biological functions in mammals, circadian rhythms of biological functions may modify the drug tolerability/toxicity depending on the dosing time. Selective mTOR (mammalian target of rapamycin) inhibitor everolimus is an immunosuppressant and anticancer agent that is active against many cancers. It was also found to be active in medullary thyroid cancer. The aim of this study was to investigate the dosing time-dependent toxicity of everolimus on the thyroid gland and hormones in mice. Healthy C57BL/6J mice were synchronized with 12h:12h Light-Dark cycle (LD12:12, with Zeitgeber Time 0 – ZT0 – corresponding to Light onset). Everolimus was administered to male (5 mg/kg/day) and female mice (15 mg/kg/day) orally at ZT1-rest period- and ZT13-activity period- for 4 weeks; body weight loss, clinical signs and possible changes in serum thyroid hormone levels (TSH and free T4) were examined. Histological alterations in the thyroid gland were evaluated according to the following criteria: follicular size, colloid density and viscidity, height of the follicular epithelium and the presence of necrotic cells. The statistical significance between differences was analyzed with ANOVA. Study findings included everolimus-related diarrhea, decreased activity, decreased body weight gains, alterations in serum TSH levels, and histopathological changes in thyroid gland. Decreases in mean body weight gains were more evident in mice treated at ZT1 as compared to ZT13 (p < 0.001, for both sexes). Control tissue sections of thyroid glands exhibited well-organized histoarchitecture when compared to everolimus-treated groups. Everolimus caused histopathological alterations in thyroid glands in male (5 mg/kg, slightly) and female mice (15 mg/kg; p < 0.01 for both ZT as compared to their controls) irrespective of dosing-time. TSH levels were slightly decreased upon everolimus treatment at ZT13 in both males and females. Conversely, increases in TSH levels were observed when everolimus treated at ZT1 in both males (5 mg/kg; p < 0.05) and females (15 mg/kg; slightly). No statistically significant alterations in serum free T4 levels were observed. TSH and free T4 is clinically important thyroid hormones since a number of disease states have been linked to alterations in these hormones. Serum free T4 levels within the normal ranges in the presence of abnormal serum TSH levels in everolimus treated mice may suggest subclinical thyroid disease which may have repercussions on the cardiovascular system, as well as on other organs and systems. Our study has revealed the histological damage on thyroid gland induced by subacute everolimus administration, this effect was irrespective of dosing time. However, based on the body weight changes and clinical signs upon everolimus treatment, tolerability for the drug was best following dosing at ZT13 in both male and females. Yet, effects of everolimus on thyroid functions may deserve further studies regarding their clinical importance and chronotoxicity.

Keywords: circadian rhythm, chronotoxicity, everolimus, thyroid gland, thyroid hormones

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Authors: Xiao Zhou, Jianlin Cheng

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A single amino acid mutation can have a significant impact on the stability of protein structure. Thus, the prediction of protein stability change induced by single site mutations is critical and useful for studying protein function and structure. Here, we presented a deep learning network with the dropout technique for predicting protein stability changes upon single amino acid substitution. While using only protein sequence as input, the overall prediction accuracy of the method on a standard benchmark is >85%, which is higher than existing sequence-based methods and is comparable to the methods that use not only protein sequence but also tertiary structure, pH value and temperature. The results demonstrate that deep learning is a promising technique for protein stability prediction. The good performance of this sequence-based method makes it a valuable tool for predicting the impact of mutations on most proteins whose experimental structures are not available. Both the downloadable software package and the user-friendly web server (DNpro) that implement the method for predicting protein stability changes induced by amino acid mutations are freely available for the community to use.

Keywords: bioinformatics, deep learning, protein stability prediction, biological data mining

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Authors: Cristina S. Decano, Vitaliana U. Malamug, Melissa E. Agulto, Helen F. Gavino

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This study aimed to introduce new natural fiber to be used in the production of geotextile net for mitigation of soil erosion. Fiber extraction from the stalks was the main challenge faced during the processing of stalks to ropes. Thus, an investigation on the extraction procedures of corn (Zea mays L.) stalk under biological and chemical retting was undertaken. Results indicated significant differences among percent fiber yield as affected by the retting methods used with values of 15.07%, 12.97%, 11.60%, and 9.01%, for dew, water, chemical (1 day after harvest and15 days after harvest), respectively, with the corresponding average extracting duration of 70, 82, 89, and 94 minutes. Physical characterization of the developed corn stalk geotextile net resulted to average mass per unit area of 806.25 g/m² and 241% water absorbing capacity. The effect of corn stalk geotextile net in mitigating soil erosion was evaluated in a laboratory experiment for 30^o and 60^o inclinations with three treatments: bare soil (A₁), corn stalk geotextile net (A₂) and combined cornstalk geotextile net and vegetation cover (A₃). Results revealed that treatment A₂ and A₃ significantly decreased sediment yield and an increase in terms of soil loss reduction efficiency. The cost of corn stalk geotextile net is Php 62.41 per square meter.

Keywords: corn stalk, natural geotextile, retting, soil erosion

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Authors: Clarissa L. de Araujo, Kátia N. Suzuki, Wilson T. V. Machado, Luis F. Bellido, Alfredo V.B. Bellido

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This study aims at contributing to the characterization of the process of biological incorporation of chromium by two benthonic species, the macroalgae Ulva sp. and the aquatic macrophyte Ruppia maritima, to subsidize future activities of monitoring the contamination of aquatic biota. This study is based on laboratory experiments to characterize the incorporation kinetics of the radiotracer ⁵¹Cr in two oxidation states (III and VI), under different salinities (7, 15, and 21 ‰). Samples of two benthonic species were collected on the margins of Rodrigo de Freitas Lagoon (Rio de Janeiro, Brazil), acclimated in the laboratory and subsequently subjected to experiments. In tests with 51Cr (III and IV), it was observed that accumulation of the metal in Ulva sp. has inverse relationship with salinity, while for R. maritima, the maximum accumulation occurs in salinity 21‰. In experiments with Cr(III), increases in the uptake of ion by both species were verified. The activity of Cr(III) was up to 19 times greater than the Cr(VI). As regards the potential for accumulation of metals, a better sensitivity of Ulva sp. for any chromium tri or hexavalent forms was verified, while for the Cr(VI) it will require low salinities and longer exposure (>24h). For R. maritima, the results showed the uptake of Cr(VI) increase along with time (>20h), because this species is more resistant for the hexavalent form and useful for any salinity as well.

Keywords: chromium, Cr-51, macroalgae, macrophyte, uptake

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Authors: Narin Ozturk, Xiao-Mei Li, Sylvie Giachetti, Francis Levi, Alper Okyar

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P-glycoprotein (P-gp, MDR1, ABCB1) is a transmembrane protein acting as an ATP-dependent efflux pump and functions as a biological barrier by extruding drugs and xenobiotics out of cells in healthy tissues especially in intestines, liver and brain as well as in tumor cells. The circadian timing system controls a variety of biological functions in mammals including xenobiotic metabolism and detoxification, proliferation and cell cycle events, and may affect pharmacokinetics, toxicity and efficacy of drugs. Selective mTOR (mammalian target of rapamycin) inhibitor everolimus is an immunosuppressant and anticancer drug that is active against many cancers, and its pharmacokinetics depend on P-gp. The aim of this study was to investigate the dosing time-dependent toxicity of everolimus with respect to the intestinal P-gp expression rhythms in mdr1a::Luc mice using Real Time-Biolumicorder (RT-BIO) System. Mdr1a::Luc male mice were synchronized with 12 h of Light and 12 h of Dark (LD12:12, with Zeitgeber Time 0 – ZT0 – corresponding Light onset). After 1-week baseline recordings, everolimus (5 mg/kg/day x 14 days) was administered orally at ZT1-resting period- and ZT13-activity period- to mdr1a::Luc mice singly housed in an innovative monitoring device, Real Time-Biolumicorder units which let us monitor real-time and long-term gene expression in freely moving mice. D-luciferin (1.5 mg/mL) was dissolved in drinking water. Mouse intestinal mdr1a::Luc oscillation profile reflecting P-gp gene expression and locomotor activity pattern were recorded every minute with the photomultiplier tube and infrared sensor respectively. General behavior and clinical signs were monitored, and body weight was measured every day as an index of toxicity. Drug-induced body weight change was expressed relative to body weight on the initial treatment day. Statistical significance of differences between groups was validated with ANOVA. Circadian rhythms were validated with Cosinor Analysis. Everolimus toxicity changed as a function of drug timing, which was least following dosing at ZT13, near the onset of the activity span in male mice. Mean body weight loss was nearly twice as large in mice treated with 5 mg/kg everolimus at ZT1 as compared to ZT13 (8.9% vs. 5.4%; ANOVA, p < 0.001). Based on the body weight loss and clinical signs upon everolimus treatment, tolerability for the drug was best following dosing at ZT13. Both rest-activity and mdr1a::Luc expression displayed stable 24-h periodic rhythms before everolimus and in both vehicle-treated controls. Real-time bioluminescence pattern of mdr1a revealed a circadian rhythm with a 24-h period with an acrophase at ZT16 (Cosinor, p < 0.001). Mdr1a expression remained rhythmic in everolimus-treated mice, whereas down-regulation was observed in P-gp expression in 2 of 4 mice. The study identified the circadian pattern of intestinal P-gp expression with an unprecedented precision. The circadian timing depending on the P-gp expression rhythms may play a crucial role in the tolerability/toxicity of everolimus. The circadian changes in mdr1a genes deserve further studies regarding their relevance for in vitro and in vivo chronotolerance of mdr1a-transported anticancer drugs. Chronotherapy with P-gp-effluxed anticancer drugs could then be applied according to their rhythmic patterns in host and tumor to jointly maximize treatment efficacy and minimize toxicity.

Keywords: circadian rhythm, chronotoxicity, everolimus, mdr1a::Luc mice, p-glycoprotein

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Authors: Adriana Muñoz-Talavera, Ana Rosa Rincón-Sánchez, Abraham Escobedo-Moratilla, María Cristina Islas-Carbajal, Miguel Ángel Gómez-Lim

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The highest rates of diabetes incidence and prevalence worldwide will increase the number of diabetic patients requiring insulin or insulin analogues. Then, current production systems would not be sufficient to meet the future market demands. Therefore, developing efficient expression systems for insulin and insulin analogues are needed. In addition, insulin analogues with better pharmacokinetics and pharmacodynamics properties and without mitogenic potential will be required. SCI-57 (single chain insulin-57) is an insulin analogue having 10 times greater affinity to the insulin receptor, higher resistance to thermal degradation than insulin, native mitogenicity and biological effect. Plants as expression platforms have been used to produce recombinant proteins because of their advantages such as cost-effectiveness, posttranslational modifications, absence of human pathogens and high quality. Immunoglobulin production with a yield of 50% has been achieved by transient expression in Nicotiana benthamiana (Nb). The aim of this study is to produce SCI-57 by transient expression in Nb. Methodology: DNA sequence encoding SCI-57 was cloned in pICH31070. This construction was introduced into Agrobacterium tumefaciens by electroporation. The resulting strain was used to infiltrate leaves of Nb. In order to isolate SCI-57, leaves from transformed plants were incubated 3 hours with the extraction buffer therefore filtrated to remove solid material. The resultant protein solution was subjected to anion exchange chromatography on an FPLC system and ultrafiltration to purify SCI-57. Detection of SCI-57 was made by electrophoresis pattern (SDS-PAGE). Protein band was digested with trypsin and the peptides were analyzed by Liquid chromatography tandem-mass spectrometry (LC-MS/MS). A purified protein sample (20µM) was analyzed by ESI-Q-TOF-MS to obtain the ionization pattern and the exact molecular weight determination. Chromatography pattern and impurities detection were performed using RP-HPLC using recombinant insulin as standard. The identity of the SCI-57 was confirmed by anti-insulin ELISA. The total soluble protein concentration was quantified by Bradford assay. Results: The expression cassette was verified by restriction mapping (5393 bp fragment). The SDS-PAGE of crude leaf extract (CLE) of transformed plants, revealed a protein of about 6.4 kDa, non-present in CLE of untransformed plants. The LC-MS/MS results displayed one peptide with a high score that matches SCI-57 amino acid sequence in the sample, confirming the identity of SCI-57. From the purified SCI-57 sample (PSCI-57) the most intense charge state was 1069 m/z (+6) on the displayed ionization pattern corresponding to the molecular weight of SCI-57 (6412.6554 Da). The RP-HPLC of the PSCI-57 shows the presence of a peak with similar retention time (rt) and UV spectroscopic profile to the insulin standard (SCI-57 rt=12.96 and insulin rt=12.70 min). The collected SCI-57 peak had ELISA signal. The total protein amount in CLE from transformed plants was higher compared to untransformed plants. Conclusions: Our results suggest the feasibility to produce insulin analogue SCI-57 by transient expression in Nicotiana benthamiana. Further work is being undertaken to evaluate the biological activity by glucose uptake by insulin-sensitive and insulin-resistant murine and human cultured adipocytes.

Keywords: insulin analogue, mass spectrometry, Nicotiana benthamiana, transient expression

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Authors: Aouina Nabila Yasmina, Chaoui Zine El Abiddine

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This study presents a comprehensive investigation into the elastic mean free path (EMFP) of electrons colliding with pyrimidine, a precursor to the pyrimidine bases in DNA, employing the Screen Corrected Additivity Rule (SCAR) method. The SCAR method is introduced as a novel approach that combines classical and quantum mechanical principles to elucidate the interaction of electrons with pyrimidine. One of the most fundamental properties characterizing the propagation of a particle in the nuclear medium is its mean free path. Knowledge of the elastic mean free path is essential to accurately predict the effects of radiation on biological matter, as it contributes to the distances between collisions. Additionally, the mean free path plays a role in the interpretation of almost all experiments in which an excited electron moves through a solid. Pyrimidine, the precursor of the pyrimidine bases of DNA, has interesting physicochemical properties, which make it an interesting molecule to study from a fundamental point of view. These include a relatively large dipole polarizability and dipole moment and an electronic charge cloud with a significant spatial extension, which justifies its choice in this present study.

Keywords: elastic mean free path, elastic collision, pyrimidine, SCAR

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Authors: Mahnoor Khan, Bashir Ahmad, Khizar Hayat, Saad Ahmad Khan, Laiba Ahmad, Shumaila Bashir, Abid Ali Khan

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The objective of this study was to synthesize the smallest possible size of ZnO NPs using a modified wet chemical synthesis method and to prepare core shell using polyethylene glycol (PEG) as shell material. Advanced and sophisticated techniques were used to confirm the synthesis, size, and shape of these NPs. Rounded, clustered NPs of size 5.343 nm were formed. Both the plain and core shell NPs were tested against MDR bacteria (E. cloacae, E. amnigenus, Shigella, S. odorifacae, Citrobacter, and E. coli). Both of the NPs showed excellent antibacterial properties, whereas E. cloacae showed maximum zone of inhibition of 16 mm, 27 mm, and 32 mm for 500 μg/ml, 1000 μg/ml, and 1500 μg/ml, respectively for plain ZnO NPs and 18 mm, 28 mm and 35 mm for 500 μg/ml, 1000 μg/ml and 1500 μg/ml for core shell NPs. These NPs were also biocompatible on human red blood cells showing little hemolysis of only 4% for 70 μg/ml for plain NPs and 1.5% for 70 μg/ml for core shell NPs. Core shell NPs were highly biocompatible because of the PEG. Their therapeutic effect as photosensitizers in photodynamic therapy (PDT) for cancer treatment was also monitored. The cytotoxicity of ZnO and PEG-ZnO was evaluated using MTT assay. Our results demonstrated that these NPs could generate ROS inside tumor cells after irradiation which in turn initiates an apoptotic pathway leading to cell death hence proving to be an effective candidate for PDT.

Keywords: ZnO, hemolysis, cytotoxiciy assay, photodynamic therapy, antibacterial

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Authors: Wonil Nam, Xiang Ren, Inyoung Kim, Masoud Agah, Wei Zhou

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Despite its ultrasensitive detection capability, surface-enhanced Raman spectroscopy (SERS) faces challenges as a quantitative biochemical analysis tool due to the significant dependence of local field intensity in hotspots on nanoscale geometric variations of plasmonic nanostructures. Therefore, despite enormous progress in plasmonic nanoengineering of high-performance SERS devices, it is still challenging to quantitatively correlate the measured SERS signals with the actual molecule concentrations at hotspots. A significant effort has been devoted to developing SERS calibration methods by introducing internal standards. It has been achieved by placing Raman tags at plasmonic hotspots. Raman tags undergo similar SERS enhancement at the same hotspots, and ratiometric SERS signals for analytes of interest can be generated with reduced dependence on geometrical variations. However, using Raman tags still faces challenges for real-world applications, including spatial competition between the analyte and tags in hotspots, spectral interference, laser-induced degradation/desorption due to plasmon-enhanced photochemical/photothermal effects. We show that electronic Raman scattering (ERS) signals from metallic nanostructures at hotspots can serve as the internal calibration standard to enable quantitative SERS analysis and improve biostatistical analysis. We perform SERS with Au-SiO₂ multilayered metal-insulator-metal nano laminated plasmonic nanostructures. Since the ERS signal is proportional to the volume density of electron-hole occupation in hotspots, the ERS signals exponentially increase when the wavenumber is approaching the zero value. By a long-pass filter, generally used in backscattered SERS configurations, to chop the ERS background continuum, we can observe an ERS pseudo-peak, IERS. Both ERS and SERS processes experience the |E|⁴ local enhancements during the excitation and inelastic scattering transitions. We calibrated IMRS of 10 μM Rhodamine 6G in solution by IERS. The results show that ERS calibration generates a new analytical value, ISERS/IERS, insensitive to variations from different hotspots and thus can quantitatively reflect the molecular concentration information. Given the calibration capability of ERS signals, we performed label-free SERS analysis of living biological systems using four different breast normal and cancer cell lines cultured on nano-laminated SERS devices. 2D Raman mapping over 100 μm × 100 μm, containing several cells, was conducted. The SERS spectra were subsequently analyzed by multivariate analysis using partial least square discriminant analysis. Remarkably, after ERS calibration, MCF-10A and MCF-7 cells are further separated while the two triple-negative breast cancer cells (MDA-MB-231 and HCC-1806) are more overlapped, in good agreement with the well-known cancer categorization regarding the degree of malignancy. To assess the strength of ERS calibration, we further carried out a drug efficacy study using MDA-MB-231 and different concentrations of anti-cancer drug paclitaxel (PTX). After ERS calibration, we can more clearly segregate the control/low-dosage groups (0 and 1.5 nM), the middle-dosage group (5 nM), and the group treated with half-maximal inhibitory concentration (IC50, 15 nM). Therefore, we envision that ERS calibrated SERS can find crucial opportunities in label-free molecular profiling of complicated biological systems.

Keywords: cancer cell drug efficacy, plasmonics, surface-enhanced Raman spectroscopy (SERS), SERS calibration

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Authors: B. R. Madushan, S. B. Navaratne, I. Wickramasinge

Abstract:

Toddy sediment (TS) was cultured in a PDA medium to determine initial yeast load, and also it was undergone sun, shade, solar, dehumidified cold air (DCA) and hot air oven (at 400, 500 and 60oC) drying with a view to preserve viability of yeast. Thereafter, this study was conducted according to two factor factorial design in order to determine best preservation method. Therein the dried TS from the best drying method was taken and divided into two portions. One portion was mixed with 3: 7 ratio of TS: rice flour and the mixture was divided in to two again. While one portion was kept under in house condition the other was in a refrigerator. Same procedure was followed to the rest portion of TS too but it was at the same ratio of corn flour. All treatments were vacuum packed in triple laminate pouches and the best preservation method was determined in terms of leavening index (LI). The TS obtained from the best preservation method was used to make foods (bread and hopper) and organoleptic properties of it were evaluated against same of ordinary foods using sensory panel with a five point hedonic scale. Results revealed that yeast load or fresh TS was 58×106 CFU/g. The best drying method in preserving viability of yeast was DCA because LI of this treatment (96%) is higher than that of other three treatments. Organoleptic properties of foods prepared from best preservation method are as same as ordinary foods according to Duo trio test.

Keywords: biological leavening agent, coconut toddy, fermentation, yeast

Procedia PDF Downloads 342

Authors: Mohammad Ahmad Ahmad Odah

Abstract:

Aging is a complex biological process characterized by the progressive decline of physiological functions and increased vulnerability to age-related diseases. Epigenetic changes, particularly DNA methylation alterations, play a critical role in the aging process by influencing gene expression and genomic stability. This study explores the potential of epigenetic reprogramming as a strategy to reverse aging phenotypes in human fibroblasts. Using CRISPR-Cas9 gene editing and small molecule inhibitors targeting DNA methylation and histone acetylation, we successfully induced significant changes in DNA methylation and gene expression profiles. Our results demonstrate a global reduction in DNA methylation levels and the identification of differentially methylated regions (DMRs) associated with cellular senescence and DNA repair. Additionally, treated fibroblasts exhibited enhanced proliferative capacity, reduced cellular senescence, and improved differentiation potential. These findings suggest that epigenetic reprogramming could be a promising approach for regenerative medicine, offering potential therapeutic strategies to counteract age-related decline and extend healthy lifespan.

Keywords: epigenetic reprogramming, aging, regenerative medicine, DNA methylation, cellular rejuvenation, CRISPR-Cas9, senescence

Procedia PDF Downloads 36

Authors: Amarjyoti Das Mahapatra, Umesh Kumar, Bhaskar Datta

Abstract:

A pseudo peptide can mimic the biological or structural properties of natural peptides. They have become an increasing attention in medicinal chemistry because of their interesting advantages like more bioavailability and less biodegradation than compare to the physiologically active native peptides which increase their therapeutic applications. Many biologically active compounds contain urea as functional groups, and they have improved pharmacokinetic properties because of their bioavailability and metabolic stability. Recently we have reported a single-step synthesis of sulfonyl urea and sulfonyltriuret from sulfonyl chloride and sodium cyanate. But the yield of sulfonyltriuret was less around 40-60% because of the formation of other products like sulfonamide and sulfonylureas. In the present work, we mainly focused on the selective synthesis of sulfonyltriuret using tetrabutylammonium cyanate and sulfonyl chloride. More precisely, we are interested in the controlled synthesis of oligomeric urea mainly sulfonyltriuret as a new class of pseudo peptide and their application as protease modulators. The distinctive architecture of these molecules in the form of their pseudo-peptide backbone offers promise as a potential pharmacophore. The synthesized molecules have been screened on trypsin enzyme, and we observed that these molecules are the efficient modulator of trypsin enzyme.

Keywords: pseudo peptide, pharmacophore, sulfonyltriuret, trypsin

Procedia PDF Downloads 167

Authors: Hammad Khan, Wookeun Bae

Abstract:

The objective of this study was to optimize and control the highly loaded and efficient nitrite production having suitability for autotrophic and heterotrophic denitritation. A lab scale CSTR for partial and full nitritation was operated to treat the livestock manure digester liquor having an ammonium concentration of ~600 mg-NH4+-N/L and biodegradable contents of ~0.35 g-COD/L. The experiments were performed at 30°C, pH: 8.0, DO: 1.5 mg/L and SRT ranging from 7-20 days. After 125 days operation, >95% nitrite buildup having the ammonium loading rate of ~3.2 kg-NH4+-N/m3-day was seen with almost complete ammonium conversion. On increasing the loading rate further (i-e, from 3.2-6.2 kg-NH4+-N/m3-day), stability of the system remained unaffected. On decreasing the pH from 8 to 7.5 and further 7.2, removal rate can be easily controlled as 95%, 75%, and even 50%. Results demonstrated that nitritation stability and desired removal rates are controlled by a balance of simultaneous inhibition by FA & FNA, pH effect and DO limitation. These parameters proved to be effective even to produce an appropriate influent for anammox. In addition, a mathematical model, identified through the occurring biological reactions, is proposed to optimize the full and partial nitritation process. The proposed model present relationship between pH, ammonium and produced nitrite for full and partial nitritation under the varying concentrations of DO, and simultaneous inhibition by FA and FNA.

Keywords: stable nitritation, high loading, autrophic denitritation, hetrotrophic denitritation

Procedia PDF Downloads 312

Authors: Elizabeth A. Haruna, Bulus Kpame, Kankanala Venkateswarlu

Abstract:

The interaction of health and athletic performance with biologic aging has been an interesting and intriguing area for research. There has been a general acknowledgement of its importance to major public health and elite performance outcomes. There are many questions unanswered about the mechanisms of effects and dose-response changes. An attempt has been made in this paper to highlight potentially positive effects of regular training on the aging process and its effects on health. Age associated decline in health and performance results from the combination of the aging process itself, inactive lifestyle and primary diseases. An attempt is made in this paper to critically review what is known and what is unknown about evidence based changes, common to disuse and aging. Mechanisms responsible for the slowing decline in muscle mass and muscle force (sarcopenia) down of age – associated, weakness and fatigability due to year round athletic training have been discussed. It is in this regard we have attempted to share our views on advances made so far in understanding the impact of aging on health. We also attempted to explain how the biological effects of aging are minimized during appropriate year round athletic training. On the basis of available research evidence it was concluded that exercise training significantly slow down the deleterious effects of aging on health.

Keywords: aging, atrophy, sarcopenia, plyometric training

Procedia PDF Downloads 415

Authors: M. S. Mahmoud, Samah A. Mohamed, Neama A. Sobhy

Abstract:

For color removal from wastewater containing organic contaminants, biological treatment systems have been widely used such as physical and chemical methods of flocculation, coagulation. Fungal decolorization of dye containing wastewater is one of important goal in industrial wastewater treatment. This work was aimed to characterize Aspergillus niger strain for dye removal from aqueous solution and from raw textile wastewater. Batch experiments were studied for removal of color using fungal isolate biomass under different conditions. Environmental conditions like pH, contact time, adsorbent dose and initial dye concentration were studied. Influence of the pH on the removal of azo dye by Aspergillus niger was carried out between pH 1.0 and pH 11.0. The optimum pH for red dye decolonization was 9.0. Results showed the decolorization of dye was decreased with the increase of its initial dye concentration. The adsorption data was analyzed based on the models of equilibrium isotherm (Freundlich model and Langmuir model). During the adsorption isotherm studies; dye removal was better fitted to Freundlich model. The isolated fungal biomass was characterized according to its surface area both pre and post the decolorization process by Scanning Electron Microscope (SEM) analysis. Results indicate that the isolated fungal biomass showed higher affinity for dye in decolorization process.

Keywords: biomass, biosorption, dye, isotherms

Procedia PDF Downloads 305

Authors: D. Suman, B. Nikitha, J. Sarvani, V. Archana

Abstract:

Histological slides provide clinical diagnostic information about the subjects from the ancient times. Even with the advent of high resolution imaging cameras the image tend to have some background noise which makes the analysis complex. A study of the histological slides is done by using a nonlinear transfer function based image enhancement method. The method processes the raw, color images acquired from the biological microscope, which, in general, is associated with background noise. The images usually appearing blurred does not convey the intended information. In this regard, an enhancement method is proposed and implemented on 50 histological slides of human tissue by using nonlinear transfer function method. The histological image is converted into HSV color image. The luminance value of the image is enhanced (V component) because change in the H and S components could change the color balance between HSV components. The HSV image is divided into smaller blocks for carrying out the dynamic range compression by using a linear transformation function. Each pixel in the block is enhanced based on the contrast of the center pixel and its neighborhood. After the processing the V component, the HSV image is transformed into a colour image. The study has shown improvement of the characteristics of the image so that the significant details of the histological images were improved.

Keywords: HSV space, histology, enhancement, image

Procedia PDF Downloads 329

Authors: Hammad Khan, Wookeun Bae

Abstract:

The objective of this study was to optimize, achieve and control the highly loaded and efficient nitrite production having suitability for autotrophic and heterotrophic denitritation. A lab scale CSTR for partial and full nitritation was operated to treat the livestock manure digester liquor having an ammonium concentration of ~600 mg-NH4+-N/L and biodegradable contents of ~0.35 g-COD/L. The experiments were performed at 30°C, pH: 8.0, DO: 1.5 mg/L and SRT ranging from 7-20 days. After 125 days operation, >95% nitrite buildup having the ammonium loading rate of ~3.2 kg-NH4+-N/m3-day was seen with almost complete ammonium conversion. On increasing the loading rate further (i-e, from 3.2-6.2 kg-NH4+-N/m3-day), stability of the system remained unaffected. On decreasing the pH from 8 to7.5 and further 7.2, removal rate can be easily controlled as 95%, 75%, and even 50%. Results demonstrated that nitritation stability and desired removal rates are controlled by a balance of simultaneous inhibition by FA & FNA, pH affect and DO limitation. These parameters proved to be effective even to produce an appropriate influent for anammox. In addition, a mathematical model, identified through the occurring biological reactions, is proposed to optimize the full and partial nitritation process. The proposed model present relationship between pH, ammonium and produced nitrite for full and partial nitritation under the varying concentrations of DO, and simultaneous inhibition by FA and FNA.

Keywords: stable nitritation, high loading, autrophic denitritation, CSTR

Procedia PDF Downloads 240

Authors: Andreas Rüdiger

Abstract:

The selection of appropriate wastewater treatment technology for decentralized coastal tourist areas is an important engineering challenge. The local situation in coastal tourist cities and villages is characterized by important daily and seasonal fluctuations in hydraulic flow and pollution, high annual temperature variations, scarcity of building area and high housing density. At the same time, coastal zones have to meet stringent effluent limits all over the year and need simple and easy technologies to operate. This article presents the innovative technology of standardized modular aerated up-flow biofiltration SMBF as an adapted solution for decentralized wastewater treatment in sensitive touristic coastal areas. As modular technology with several biofiltration units, the system is able to treat low and high loads with low energy consumption and low demands for operators. The article focuses on the climatic and tourist situation in Croatia. Full-scale plants in Eastern Europe and Croatia have presented as well as dimensioning parameters and outlet concentrations. Energy consumption as a function of load is demonstrated.

Keywords: wastewater treatment, biofiltration, touristic areas, energy saving

Procedia PDF Downloads 91

Authors: Wenting Ye

Abstract:

The biological materials evolved in nature generally exhibit interpenetrating network structures, which may offer useful inspiration for the architectural design of wear-resistant composites. Here, a strategy for designing self-lubricating medium entropy alloy (MEA) composites with high strength and excellent anti-wear performance was proposed through quasi-continuously networked in-situ carbides and graphene nanosheets. The discontinuous coating of graphene on the MEA powder surface inhibits continuous metallurgy bonding of the MEA powders during sintering, generating the typical quasi-continuously networked architecture. A good combination of mechanical properties with high fracture strength over 2 GPa and large compressive plasticity over 30% benefits from metallurgy bonding that prevents crack initiation and extension. The wear rate of an order of 10-6 m3N-1m-1 ascribing to an amorphous-crystalline nanocomposite surface, tribo-film induced by graphene, as well as the gradient worn subsurface during friction was achieved by the MEA composite, which is an order of magnitude lower than the unreinforced MEA matrix.

Keywords: in-situ carbide, tribological behavior, medium entropy alloy matrix composite, graphene

Procedia PDF Downloads 32