Search results for: pharmaceutical markers

Authors: Pankaj Prashar, Bimlesh Kumar, Ankita Sood, Anamika Gautam

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Pharmacovigilance (PV) is a rapidly growing discipline in pharmaceutical industries as an integral part of clinical research and drug development over the past few decades. PV carries a breadth of scope from drug manufacturing to its regulation with safer utilization. The fundamental steps of PV not only includes data collection and verification, coding of drugs with adverse drug reactions, causality assessment and timely reporting to the authorities but also monitoring drug manufacturing, safety issues, product quality and conduction of due diligence. Standardization of adverse event information, collaboration of multiple departments in different companies, preparation of documents in accordance to both governmental as well as non-governmental organizations (FDA, EMA, GVP, ICH) are the advancements in discipline of PV. De-harmonization, lack of predictive drug safety models, improper funding by government, non-reporting, and non-acceptability of ADRs by developing countries and reports directly from patients to the monitoring centres respectively are the major road backs of PV. Mandatory pharmacovigilance reporting, frequent inspections, funding by government, educating and training medical students, pharmacists and nurses in this segment can bring about empowerment in PV. This area needs to be addressed with a sense of urgency for the safe utilization of pharmaceuticals.

Keywords: pharmacovigilance, regulatory, adverse event, drug safety

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Authors: Farzad Khajavi

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Studies about the interaction between active pharmaceutical ingredients (API) and excipients are so important in the pre-formulation stage of development of all dosage forms. Analytical techniques such as differential scanning calorimetry (DSC), Thermal gravimetry (TG), and Furrier transform infrared spectroscopy (FTIR) are commonly used tools for investigating regarding compatibility and incompatibility of APIs with excipients. Sometimes the interpretation of data obtained from these techniques is difficult because of severe overlapping of API spectrum with excipients in their mixtures. Principal component analysis (PCA) as a powerful factor analytical method is used in these situations to resolve data matrices acquired from these analytical techniques. Binary mixtures of API and interested excipients are considered and produced. Peaks of FTIR, DSC, or TG of pure API and excipient and their mixtures at different mole ratios will construct the rows of the data matrix. By applying PCA on the data matrix, the number of principal components (PCs) is determined so that it contains the total variance of the data matrix. By plotting PCs or factors obtained from the score of the matrix in two-dimensional spaces if the pure API and its mixture with the excipient at the high amount of API and the 1:1mixture form a separate cluster and the other cluster comprise of the pure excipient and its blend with the API at the high amount of excipient. This confirms the existence of compatibility between API and the interested excipient. Otherwise, the incompatibility will overcome a mixture of API and excipient.

Keywords: API, compatibility, DSC, TG, interactions

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Authors: Rafat M. Mohareb

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Several D-ring alkylated estrone analogues display exceptionally high affinity for estrogen receptors. In particular, compounds in which an E-ring is formed are known to be involved in the inhibition of steroidogenic enzymes. Such compounds also have an effect on steroid dehydrogenase activity and the ability to inhibit the detrimental action of the steroid sulfatase enzyme. Generally, E-ring extended steroids have been accessed by modification of the C17-ketone in the D-ring by either arylimine or oximino formation, addition of a carbon nucleophile or hydrazone formation. Other approaches have included ketone reduction, silyl enol ether formation or ring-closing metathesis (giving five- or six-membered E-rings). Chemical modification of the steroid D-ring provides a way to alter the functional groups, sizes and stereochemistry of the D-ring, and numerous structure-activity relationships have been established by such synthetic alterations. Steroids bearing heterocycles fused to the D-ring of the steroid nucleus have been of pharmaceutical interest. In the present paper, we report on the efficient synthesis of estrone possessing pyran, pyrimidine and thiazole ring systems. This study focused on the synthesis and biochemical evaluation of newly synthesized heterocyclic compounds which were then subjected through inhibitory evaluations towards human cancer and normal cell lines.

Keywords: estrone, heterocyclization, cytotoxicity, biomedicine

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Authors: Balazs Patczai, Katalin Gocze, Gabriella Kiss, Dorottya Szabo, Tibor Mintal

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Introduction: Running has become increasingly popular on a global scale in the past decades. Amateur athletes are taking their sport to a new level in an attempt to surpass their performance goals. The aim of our study was to assess the musculoskeletal condition of amateur runners and the prevalence of injuries with a special focus on stress fracture risk. Methods: The cross sectional analysis included ankle mobility, hamstring and lower back flexibility, the use of Renne’s test for iliotibial band syndrome, functional tests for trunk and rotary stability, and measurements of bone density. Data was collected at 2 major half-marathon events in Hungary. Results: Participants (n=134) mean age was 41.76±8.57 years (males: 40.67±8.83, females: 42.08±8.56). Measures of hamstring and lower back flexibility fell into the category of good for both genders (males: 7.13±6.83cm, females: 10.17±6.67cm). No side asymmetry nor gender differences were characteristic in the case of ankle mobility. Trunk stability was significantly better for males than in females (p=0.004). Markers of bone health were in the low normal range for females and were significantly better for males (T-score: p=0.003, T-ratio: p=0.014, Z-score: p=0.034, Z-ratio: p=0.011). 5.2% of females had a previous stress fracture and 24.1% experienced irregular menstrual cycles during the past year. As for the knowledge on the possible association of energy deficiency, menstrual disturbances and their effect on bone health, Only 8.6% of females have heard of the female athlete triad either during their studies or from a health professional. Discussion: The overall musculoskeletal state was satisfactory for both genders both physically and functionally. More attention and effort should be placed on primary and secondary prevention of amateur runners. Very few active women are well informed about the effects of low energy availability and menstrual dysfunction and the negative impact these have on bone health.

Keywords: bone health, flexibility, running, stress fracture

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Authors: M. Merabet-Khelassi, Z. Houiene, L. Aribi-Zouioueche, O. Riant

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Lipases (EC.3.1.1.3) are efficient biotools widely used for their remarkable chemo-, regio- and enantio-selectivity, especially, in kinetic resolution of racemates. They offer access to a large panel of enantiopure building blocks, such as secondary benzylic alcohols, commonly used as synthetic intermediates in pharmaceutical and agrochemical industries. Due to the stability of lipases in both water and organic solvents poor in water, they are able to catalyze both transesterifications of arylalkylcarbinols and hydrolysis of their corresponding acetates. The use of enzymatic hydrolysis in aqueous media still limited. In this presentation, we expose a practical methodology for the preparation of optically enriched acetates using a Candida antarctica lipase B-catalyzed hydrolysis in non-aqueous media in the presence of alkaline carbonate salts. The influence of several parameters which can intervene on the enzymatic efficiency such as the impact of the introduction of the carbonates salts, its amount and the nature of the alkaline earth metal are discussed. The obtained results show that the use of sodium carbonate with CAL-B enhances drastically both reactivity and selectivity of this immobilized lipase. In all cases, the resulting alcohols and remaining acetates are obtained in high ee values (up to > 99 %), and the selectivities reach (E > 500).

Keywords: alkaline-hydrolysis, enzymatic kinetic resolution, lipases, arylalkylcarbinol, non-aqueous media

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Authors: Peramachi Sathiyamoorthy, Jacop Gopas, Avi Golan Goldhirsh

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Corchorus olitorius is a leafy vegetable and an industrial crop. The herb has antioxidant, anti inflammatory, and anti-cancer properties. To assay the pharmaceutical properties, aqueous extracts of leaves and seeds from C. olitorius were tested against drug resistant melanoma cell line. The test showed LC50 of the extract was 0.08µg/ml. Aqueous seed extract exhibited higher melanoma inhibiting activity than leaf extract. Dialysis of seed extract showed that the active compound is less than 12 KDa. The compound with <3 KDa MW separated by microconcentration of seed extract showed 70.5 % inhibition of melanoma cell growth. Among the two fractions obtained by Gel filtration with G10 column, the first fraction at 1:2000 dilutions exhibited 100% inhibition of melanoma growth. The compound with Rf value 0.86 (MA4) isolated by TLC separation showed about 98% cytotoxicity against melanoma at 1: 1000 dilutions. Furthermore, HPLC separation of MA4 compound with Superdex 75 column resulted in 4 compounds. Out of 4, one compound showed melanoma inhibition. The active compound is identified by reagent methods as Strophanthidin. Further toxicological and clinical studies will lead to the development of a potential drug to treat drug resistant melanoma.

Keywords: corchorus olitorius, melanoma, drug development, strophanthidin

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Authors: Chia-Yu Lin, Chun-Ying Lee, Chih-Jer Lin

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Lemongrass (Cymbopogon citratus), grown in tropical and subtropical regions over the world, has many potential uses in pharmaceutical, cosmetics, food and flavor, and agriculture industries. In this study, because of its affinity to human body and friendliness to the environment, lemongrass extract was prepared from different processes to evaluate its effectiveness as mosquito repellent. Moreover, the supercritical fluid extraction method has been widely used as an effective and environmental friendly process in the preparation of a variety of compounds. Thus, both the extracts from lemongrass by the conventional hydrodistillation method and the supercritical CO₂ assisted method were compared. The effects of pressure, temperature and time duration on the supercritical CO₂ extraction were also investigated. The compositions of different extracts were examined using mass spectrometer. As for the experiment of mosquito repellence, the extract was placed inside a mosquito trap along with syrup. The mosquito counts in each trap with extracts prepared from different processes were employed in the quantitative evaluation. It was found that the extract from the supercritical CO₂ assisted process contained higher citronellol content than the conventional hydrodistillation method. The extract with higher citronellol content also demonstrated more effective as a mosquito repellent.

Keywords: lemongrass (Cymbopogon citratus), hydrodistillation, supercritical fluid extraction, mosquito repellent

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Authors: Manjusha C. Padole, Parag A. Deshpande

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Density functional theory calculations were carried out for identification of an active heterogeneous catalyst to carry out Heck coupling reaction which is of pharmaceutical importance. One of the carbonaceous nanomaterials, heterofullerene, was designed for the reaction. Stability and reactivity of the proposed heterofullerenes (C59M, M = Pd/Ni) were established with insights into the metal-carbon bond, electron affinity and chemical potential. Adsorbent potentials of both the heterofullerenes were examined from the adsorption study of four halobenzenes (C6H5F, C6H5Cl, C6H5Br and C6H5I). Oxidative addition activities of all four halobenzenes were investigated by developing free energy landscapes over both the heterofullerenes for rate determining step (oxidative addition). C6H5I showed a good catalytic activity for the rate determining step. Thus, C6H5I was proposed as a suitable halobenzene and complete free energy landscapes for Heck coupling reaction were developed over C59Pd and C59Ni. Smaller activation barriers observed over C59Pd in comparison with C59Ni put us in a position to propose C59Pd to be an efficient heterofullerene for carrying Heck coupling reaction.

Keywords: metal-substituted fullerene, density functional theory, electron affinity, oxidative addition, Heck coupling reaction

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Authors: Akbar Nawaz Khan, Abdul Ghaffar, Muhammad Naeem Riaz

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The study was conducted to investigate the phenotypic features, morphometry and production potentialities of indigenous naked neck chicken (NN) of Pakistan under intensive management condition. A total of 35 NN chicks were randomly selected, and the experiment was performed at Poultry and wildlife research section NARC Islamabad for a period of 22 weeks. The predominant plumage color was black and golden while skin color was observed white. The average shank length, leg length, thigh length, keel length, chest breadth, head width, wing space, wing length, body length, body girth, body height and pubic bone width in adult males and females were 69.19 ± 3.34mm, 117.93 ± 4.42mm, 117.93 ± 4.42mm, 90.87 ± 6.53mm, 95.03 ± 4.56mm, 49.77 ± 2.53mm, 30.63 ± 1.50cm, 27.24 ± 2.71cm, 18.88 ± 0.65cm, 17.77 ± 1.01cm, 25.96 ± 0.56cm, 47.81 ± 1.41cm and 35.69 ± 4.09mm respectively. The average age and live body weight of NN chicken at sexual maturity were recorded as 165.85 days and 1269.38 g. While hen-day egg production of NN was recorded as 45%. The present study was aimed to investigate the existence of polymorphism at IGF-I gene in indigenous naked neck chicken through PCR based Restriction Fragment Length Polymorphism. Based on restriction analysis using Hinf I restriction enzyme, three genotypes were detected designated as AA, AC, and CC. Restriction analysis of PCR amplified product showed the presence of DNA fragments of 622, 378, 244 and 191, (genotypes). The PCR-RFLP analysis is easy, cost effective method which permits the easy characterization of IGF-I gene. This showed the investigated IGF-I genes can serve as good molecular markers for marker assisted selection (MAS) concerning growth related traits in chicken.

Keywords: Desi chicken, naked neck, morphology, morphometry, production potential, egg traits, egg geometry, IGF-I, growth, PCR- RFLP, chicken

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Authors: Halima Albalushi, Mohadese Boroojerdi, Murtadha Alkhabori

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Introduction: Maintenance of stem cell properties during culture necessitates the recreation of the natural cell niche. Studies reported the promising outcome of mesenchymal stem cells (MSC) properties maintenance after using extracellular matrix such as CELLstart™, which is the recommended coating material for stem cells cultured in serum-free and xeno-free conditions. Laminin-521 is known as a crucial adhesion protein, which is found in natural stem cell niche, and plays an important role in facilitating the maintenance of self-renewal, pluripotency, standard morphology, and karyotype of human pluripotent stem cells (PSCs). The aim of this study is to investigate the effects of Laminin-521 on human umbilical cord-derived mesenchymal stem cells (UC-MSC) characteristics as a step toward clinical application. Methods: Human MSC were isolated from the umbilical cord via the explant method. Umbilical cord-derived-MSC were cultured in serum-free and xeno-free conditions in the presence of Laminin-521 for six passages. Cultured cells were evaluated by morphology and expansion index for each passage. Phenotypic characterization of UC-MSCs cultured on Laminin-521 was evaluated by assessment of cell surface markers. Results: Umbilical cord derived-MSCs formed small colonies and expanded as a homogeneous monolayer when cultured on Laminin-521. Umbilical cord derived-MSCs reached confluence after 4 days in culture. No statistically significant difference was detected in all passages when comparing the expansion index of UC-MSCs cultured on LN-521 and CELLstart™. Phenotypic characterization of UC-MSCs cultured on LN-521 using flow cytometry revealed positive expression of CD73, CD90, CD105 and negative expression of CD34, CD45, CD19, CD14 and HLA-DR.Conclusion: Laminin-521 is comparable to CELLstart™ in supporting UC-MSCs expansion and maintaining their characteristics during culture in xeno-free and serum-free culture conditions.

Keywords: mesenchymal stem cells, culture, laminin-521, xeno-free serum-free

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Authors: Leixuri Aguirre, Iñaki Milton-Laskibar, Elizabeth Hijona, Luis Bujanda, Agnes M. Rimando, Maria P. Portillo

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Introduction: In recent years great attention has been paid by scientific community to phenolic compounds as active biomolecules naturally present in foodstuffs due to their beneficial effects on health. Pterostilbene is a resveratrol dimethylether derivative which shows higher biodisponibility. Objective. To analyze the effects of two doses of pterostilbene on several markers of thermogenic capacity in a model of genetic obesity, which shows reduced thermogenesis. Methods: The experiment was conducted with thirty Zucker (fa/fa) rats that were distributed in 3 experimental groups, the control group and two groups orally administered with pterostilbene at 15 and 30 mg/kg body weight/day for 6 weeks. Gene expression of Ucp1, Pgc-1α, Cpt1b, Pparα, Nfr1, Tfam and Cox-2 were assessed by RT-PCR, protein expression of UCP1 and GLUT4 by western blot and enzyme activity of carnitine palmitoyl transferase 1b and citrate synthase by spectrophotometry in interscapular brown adipose tissue (iBAT). Statistical analysis was performed by using one way ANOVA and Newman-Keuls as post-hoc test. Results: Pterostilbene did not change gene expression of Pgc-1α. However, significant increases were found in the expression of Ucp1, Pparα, Nfr-1 and Cox-2. Protein expression of UCP1 and GLUT4 was increased in animals treated with pterostilbene, as well as the activities of CPT-1b and CS. These effects were observed with both doses of pterostilbene, without differences between them. Conclusions: These results show that pterostilbene increases thermogenic and oxidative capacity of brown adipose tissue in obese rats. Whether these effects effectively contribute to the anti-obesity properties of these compound needs further research. Acknowledgments: MINECO-FEDER (AGL2015-65719-R), Basque Government (IT-572-13), University of the Basque Country (ELDUNANOTEK UFI11/32), Institut of Health Carlos III (CIBERobn). Iñaki Milton is a fellowship from the Basque Government.

Keywords: brown adipose tissue, pterostilbene, thermogenesis, uncoupling protein 1

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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: Lucja Rodzik, Joanna Lewandowska-Lancucka, Michal Szuwarzynski, Krzysztof Szczubialka, Maria Nowakowska

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The analysis of nucleobases is of great importance for bioscience since their abnormal concentration in body fluids suggests the deficiency and mutation of the immune system, and it is considered to be an important parameter for diagnosis of various diseases. The presented conjugate meets the need for development of the effective, selective and highly sensitive sensor for nucleobase/nucleoside detection. The novel, highly fluorescent cadmium telluride quantum dots (CdTe QDs) functionalized with thymine and stabilized with thioglycolic acid (TGA) conjugates has been developed and thoroughly characterized. Successful formation of the material was confirmed by elemental analysis, and UV–Vis fluorescence and FTIR spectroscopies. The crystalline structure of the obtained product was characterized with X-ray diffraction (XRD) method. The composition of CdTe QDs and their thymine conjugate was also examined using X-ray photoelectron spectroscopy (XPS). The size of the CdTe-thymine was 3-6 nm as demonstrated using atomic force microscopy (AFM) and high resolution transmission electron microscopy (HRTEM) imaging. The plasmon resonance fluorescence band at 540 nm on excitation at 351 nm was observed for these nanoparticles. The intensity of this band increased with the increase in the amount of conjugated thymine with no shift in its position. Based on the fluorescence measurements, it was found that the CdTe-thymine conjugate interacted efficiently and selectively not only with adenine, a nucleobase complementary to thymine, but also with nucleosides and adenine-containing modified nucleosides, i.e., 5′-deoxy-5′-(methylthio)adenosine (MTA) and 2’-O-methyladenosine, the urinary tumor markers which allow monitoring of the disease progression. The applicability of the CdTe-thymine sensor for the real sample analysis was also investigated in simulated urine conditions. High sensitivity and selectivity of CdTe-thymine fluorescence towards adenine, adenosine and modified adenosine suggest that obtained conjugate can be potentially useful for development of the biosensor for complementary nucleobase/nucleoside detection.

Keywords: CdTe quantum dots, conjugate, sensor, thymine

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Authors: Flávio A. F. da Ponte, Jackson Q. Malveira, José A. S. Ramos Filho, Monica C. G. Albuquerque

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The vegetable oils have many applications in industrial processes and due to this potential have constantly increased the demand for the use of low-quality oils, mainly in the production of biofuel. This work aims to the physicochemical evaluation of babassu oil (Orbinya speciosa) and coconut (Cocos nucifera) of low quality, as well the obtaining the free fatty acids 6 to 16 carbon atoms, with intention to be used as raw material for the biofuels production. The babassu oil and coconut low quality, as well the fatty acids obtained from these oils were characterized as their physicochemical properties and fatty acid composition (using gas chromatography coupled to mass). The NMR technique was used to assess the efficiency of fractional distillation under reduced pressure to obtain the intermediate carbonic chain fatty acids. The results showed that the bad quality in terms of physicochemical evaluation of babassu oils and coconut oils interfere directly in industrial application. However the fatty acids of intermediate carbonic chain (C6 to C16) may be used in cosmetic, pharmaceutical and particularly as the biokerosene fuel. The chromatographic analysis showed that the babassu oil and coconut oil have as major fatty acids are lauric acid (57.5 and 38.6%, respectively), whereas the top phase from distillation of coconut oil showed caprylic acid (39.1%) and major fatty acid.

Keywords: babassu oil (Orbinya speciosa), coconut oil (Cocos nucifera), fatty acids, biomass

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Authors: Yaghoubi N., Moore Z., Van Der Veen S. M., Pidcoe P. E., Thomas J. S., Dexheimer B.

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Optoelectronic 3D motion capture systems, such as the Vicon kinematic system, are widely utilized in biomedical research to track joint motion. These systems are considered powerful and accurate measurement tools with <2 mm average error. However, these systems are costly and may be difficult to implement and utilize in a clinical setting. 3D virtual reality (VR) is gaining popularity as an affordable and accessible tool to investigate motor control and perception in a controlled, immersive environment. The HTC Vive VR system includes puck-style trackers that seamlessly integrate into its VR environments. These affordable, wireless, lightweight trackers may be more feasible for clinical kinematic data collection. However, the accuracy of HTC Vive Trackers (3.0), when compared to optoelectronic 3D motion capture systems, remains unclear. In this preliminary study, we compared the HTC Vive Tracker system to a Vicon kinematic system in a simulated lumbar flexion task. A 6-DOF robot arm (SCORBOT ER VII, Eshed Robotec/RoboGroup, Rosh Ha’Ayin, Israel) completed various reaching movements to mimic increasing levels of hip flexion (15°, 30°, 45°). Light reflective markers, along with one HTC Vive Tracker (3.0), were placed on the rigid segment separating the elbow and shoulder of the robot. We compared position measures simultaneously collected from both systems. Our preliminary analysis shows no significant differences between the Vicon motion capture system and the HTC Vive tracker in the Z axis, regardless of hip flexion. In the X axis, we found no significant differences between the two systems at 15 degrees of hip flexion but minimal differences at 30 and 45 degrees, ranging from .047 cm ± .02 SE (p = .03) at 30 degrees hip flexion to .194 cm ± .024 SE (p < .0001) at 45 degrees of hip flexion. In the Y axis, we found a minimal difference for 15 degrees of hip flexion only (.743 cm ± .275 SE; p = .007). This preliminary analysis shows that the HTC Vive Tracker may be an appropriate, affordable option for gross motor motion capture when the Vicon system is not available, such as in clinical settings. Further research is needed to compare these two motion capture systems in different body poses and for different body segments.

Keywords: lumbar, vivetracker, viconsystem, 3dmotion, ROM

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Authors: Yasser Gaber, Mohamed Ismail, Serena Bisagni, Mohamad Takwa, Rajni Hatti-Kaul

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Enzymes are safe and selective catalysts. They skillfully catalyze chemical reactions; however, the native form is not usually suitable for industrial applications. Enzymes are therefore engineered by several techniques to meet the required catalytic task. Clopidogrel is recorded among the five best selling pharmaceutical in 2010 under the brand name Plavix. The commonly used route for production of the drug on an industrial scale is the synthesis of the racemic mixture followed by diastereomeric resolution to obtain the pure S isomer. The process consumes a lot of solvents and chemicals. We have evaluated a biocatalytic cleaner approach for asymmetric hydrolysis of racemic clopidogrel. Initial screening of a selected number of hydrolases showed only one enzyme EST to exhibit activity and selectivity towards the desired stereoisomer. As the crude EST is a mixture of several isoenzymes, a homology model of EST-1 was used in molecular dynamic simulations to study the interaction of the enzyme with R and S isomers of clopidogrel. Analysis of the geometric hindrances of the tetrahedral intermediates revealed a potential site for mutagenesis in order to improve the activity and the selectivity. Single point mutation showed dramatic increase in activity and inversion of the enantioselectivity (400 fold change in E value).

Keywords: biocatalysis, biotechnology, enzyme, protein engineering, molecular modeling

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Authors: Sakshi Tomar, Renu Chadha

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Cocrystallization comprises a selective route to the intensive design of pharmaceutical products with desired physiochemical and pharmacokinetic properties. The present study is focused on the preparation, characterization, and evaluation of etodolac (ET) co-crystals with coformers nicotinamide (ETNI) and Glutaric acid (ETGA), using cocrystallization approach. Preliminarily examination of the prepared co-crystal was done by differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FT-IR), powder X-ray diffraction (PXRD). DSC thermographs of ETNI and ETGA cocrystals showed single sharp melting endotherms at 144°C and 135°C, respectively, which were different from the melting of drugs and coformers. FT-IR study points towards carbonyl-acid interaction sandwiched between the involving molecules. The emergence of new peaks in the PXRD pattern confirms the formation of new crystalline solid forms. Both the cocrystals exhibited better apparent solubility, and 3.8-5.0 folds increase in IDR were established, as compared to pure etodolac. Evaluations of these solid forms were done using anti-osteoarthritic activities. All the results indicate that etodolac cocrystals possess better anti-osteoarthritic efficacy than free drug. Thus loom of cocrystallization has been found to be a viable approach to resolve the solubility and bioavailability issues that circumvent the use of potential antiosteoarthritic molecules.

Keywords: bioavailability, etodolac, nicotinamide, osteoarthritis

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Authors: Gina Cecilia Pistol, Loredana Calin, Mariana Stancu, Veronica Chedea, Ionelia Taranu

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Inflammatory-associated diseases have an increased trend in the past decades. The pharmacological strategies aimed to treat these inflammatory diseases are very expensive and with non-beneficial results. The current trend is to find alternative strategies to counteract or to control inflammatory component of diseases. The grape by-products either seeds or pomace are rich in bioactive compounds (e.g. polyphenols) which may be beneficial in prevention of inflammation associated with cancer progression and other pathologies with inflammatory component. The in vivo models are very useful for studying the immune and inflammatory status. The domestic pig (Sus scrofa domesticus) is related to human from anatomic and physiologic point of view, representing a feasible model for studying the human inflammatory pathologies. Starting from these data, we evaluated the effect of a diet containing 5% grape seed cakes (GS) on piglets blood biochemical parameters and immune pro- and anti-inflammatory biomarkers (IL-1 beta, IL-8, TNF-alpha, IL-6, IFN-gamma, IL-10, IL-4) in spleen and lymph nodes. 12 weaned piglets were fed for 30 days with a control diet or an experimental diet containing 5% GS. At the end of trial, plasma and tissue samples (spleen and lymph nodes) were collected and the biochemical and inflammatory markers were analysed by using biochemistry analyser and ELISA techniques. Our results showed that diet included 5% GS did not influence the health status determined by plasma biochemical parameters. Only a tendency for a slight increase of the biochemical parameters associated with energetic profile (glucose, cholesterol, triglycerides) was observed. Also, GS diet had no effect on pro- and anti-inflammatory cytokines content in spleen and lymph nodes tissue. Further experiments are needed in order to investigate other rate of dietary inclusion which could provide more evidence about the effect of grape bioactive compounds on pigs used as animal model.

Keywords: animal model, inflammation, grape seed by-product, immune organs

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Authors: Jiraporn Rojtinnakorn, Mallika Supa-Aksorn, Sudaporn Tongsiri, Prachaub Chaibu

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Marble goby is one of high demand consuming in Southeast Asia. However, the product was from riparian fisheries because of low yield in aquaculture, especially in nursery stage. Therefore, we studied for herb supplement in pellet feed of marble goby fingering. Garlic, a common herb and illustrated novel pharmaceutical and medical effectiveness, was considered. Garlic extracts with water (DW), 50% EtOH (50E), 95% EtOH (95E) and diethyl ether (DE) were subjected for feed additive to induce immune response in marble goby fingering for 0 (control), 0.3, 0.5, 1.0, 3.0 and 5.0 % (w/w). After seven days of feeding, blood was collected for analysis of blood composition; i.e. haematocrit (HCT), red blood cells (RBC), white blood cells (WBC) and humoral immune responses; i.e. lysozyme activity (Lys). It was resulted that values of HCT, WBC and Lys in all garlic fed group were significantly different from control (p < 0.05). For HCT, the highest values belonged to 5% DW and 0.5% 95E. For WBC and Lys, the highest values were 5% DW. For RBC, there was not obviously significant (p < 0.05). There were only 3 groups; 0.5% 95E, 1% and 5% DW, showed distinct statistical significance from the other groups. It was concluded that garlic extracts showed satisfy bioactivity to enhancing innate immune response in marble goby fingering. This result will be valuable for specific feed formula of marble goby nursery.

Keywords: garlic extract, innate immune, marble goby, Oxyeleotris marmoratus

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Authors: Marwa Elkalla, E. Ali Abdelfadil, Ali. Mohamed. M. Sami, Ali M. Abdel-Monem

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To assess the impact of the blood collection technique on clinical pathology markers and to establish reference intervals, a study was performed using normal, healthy C57BL/6 mice. Both sexes were employed, and they were randomly assigned to different groups depending on the phlebotomy technique used. The blood was drawn in one of four ways: intracardiac (IC), caudal vena cava (VC), caudal vena cava (VC) plus a peritoneal collection of any extravasated blood, or retroorbital phlebotomy (RO). Several serum biochemistries, such as a liver function test, a complete blood count with differentials, and a platelet count, were analysed from the blood and serum samples analysed. Red blood cell count, haemoglobin (p >0.002), hematocrit, alkaline phosphatase, albumin, total protein, and creatinine were all significantly greater in female mice. Platelet counts, specific white blood cell numbers (total, neutrophil, lymphocyte, and eosinophil counts), globulin, amylase, and the BUN/creatinine ratio were all greater in males. The VC approach seemed marginally superior to the IC approach for the characteristics under consideration and was linked to the least variation among both sexes. Transaminase levels showed the greatest variation between study groups. The aspartate aminotransferase (AST) values were linked with decreased fluctuation for the VC approach, but the alanine aminotransferase (ALT) values were similar between the IC and VC groups. There was a lot of diversity and range in transaminase levels between the MC and RO groups. We found that the RO approach, the only one tested that allowed for repeated sample collection, yielded acceptable ALT readings. The findings show that the test results are significantly affected by the phlebotomy technique and that the VC or IC techniques provide the most reliable data. When organising a study and comparing data to reference ranges, the ranges supplied here by collection method and sex can be utilised to determine the best approach to data collection. The authors suggest establishing norms based on the procedures used by each individual researcher in his or her own lab.

Keywords: clinical, pathology, blood, effect

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Authors: Daniel Faraj, Arabella Raupach, Charlotte Hespe, Helen Wilcox, Kristie-Lee Anning

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Aim: International child sponsorship programs comprise a considerable proportion of global aid accessible to the general population. Team Philippines (TP), a healthcare and welfare initiative run in association with the University of Notre Dame Sydney since 2013, leads a holistic sponsorship program for thirty children from Calauan, Philippines. To date, empirical research has not been performed on the overall success and impact of the TP child sponsorship program. As such, this study aims to evaluate its effectiveness in improving pediatric outcomes. Methods: Study cohorts comprised thirty sponsored and twenty-nine age- and gender-matched non-sponsored children. Data were extracted from the TP Medical Director database and lifestyle questionnaires for July-November 2019. Outcome measures included anthropometry, markers of medical health, dental health, exercise, and diet. Statistical analyses were performed in SPSS. Results: Sponsorship resulted in fewer medical diagnoses and prescription medications, superior dental health, and improved diet. Further, sponsored children may show a clinically significant trend toward improved physical health. Sponsorship did not affect growth and development metrics or levels of physical activity. Conclusions: The TP child sponsorship program significantly impacts positive pediatric health outcomes in the Calauan community. The strength of the program lies in its holistic, sustainable, and community-based model, which is enabled by effective international child sponsorship. This study further supports the relationship between supporting early livelihood and improved health in the pediatric population.

Keywords: child health, public health, health status disparities, healthcare disparities, social determinants of health, morbidity, community health services, culturally competent care, medically underserved areas, population health management, Philippines

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Authors: Sasan Khademnematolahi, Kevine Kamga Silihe, Katarína Pružinská, Martina Chrastina, Elisabeth Louise Ndjengue Mindang, František Dráfi, Katarína Bauerová

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Background: In ethnomedicine, plant parts and compounds are traditionally utilized to treat many disorders. Azadirachta indica, known as Neem, has been traditionally used in medicinal practices. Neem has various pharmaceutical activities, such as antioxidant and anti-inflammatory, due to the content of bioactive compounds like nimbolide, azadirachtin, and gedunin.Through its effect on pathological inflammatory processes, supplementation with it could alleviate the symptoms of rheumatoid arthritis (RA). Methods: This research aimed to assess Neem seed oil's impact on rats with adjuvant arthritis. Three doses in monotherapy and two in combination with methotrexate (MTX) have been studied and their effect was compared. Neem p.o. doses of 100, 200, and 300 mg/kg and MTX p.o. doses of 0.3 mg/kg were examined. After clinical parameters assessment, biochemical analysis was performed in plasma. Results: During the acute phase of the experimental arthritis (Day21), levels of MMP-9, MCP-1 and cytokines IL-1beta and IL-17A were measured. The positive results of inflammatory mediators evaluation in plasma encourage additional analysis also in related tissues to prove if Neem seed oil can be used as an adjuvant therapy for RA. Conclusion: In this study, the combination therapy of Neem with MTX was most effective from all therapies investigated.

Keywords: adjuvant, neem, methotrexate, arthritis

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Authors: Nawel Ouennoughi, Kamel Daoud

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During recent years, new pharmaceutical dosage forms were developed in the research laboratories and which consists of encapsulating one or more active molecules in a polymeric envelope. Several techniques of encapsulation allow obtaining the microparticles or the nanoparticles containing one or several polymers. In the industry, microencapsulation is implemented to fill the following objectives: to ensure protection, the compatibility and the stabilization of an active matter in a formulation, to carry out an adapted working, to improve the presentation of a product, to mask a taste or an odor, to modify and control the profile of release of an active matter to obtain, for example, prolonged or started effect. To this end, we focus ourselves on the encapsulation of the antidiabetic. It is an oral hypoglycemic agent belonging to the second generation of sulfonylurea’s commonly employed in the treatment of type II non-insulin-dependent diabetes in order to improve profile them dissolution. Our choice was made on the technique of encapsulation by complex coacervation with two types of polymers (gelatin and the gum Arabic) which is a physicochemical process. Several parameters were studied at the time of the formulation of the microparticles and the nanoparticles: temperature, pH, ratio of polymers etc. The microparticles and the nanoparticles obtained were characterized by microscopy, laser granulometry, FTIR and UV-visible spectrophotometry. The profile of dissolution obtained for the microparticles showed an improvement of the kinetics of dissolution compared to that obtained for the active ingredient.

Keywords: coacervation, gum Arabic, microencapsulation, gelatin

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Authors: Li Yang, Yang Song, Martin Y. M. Chiang

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Objective: To experimentally substantiate the micromechanical compatibility between cell and scaffold, in the regenerative medicine approach for restoring bone volume, is essential for phenotypic transitions Methods: Through nanotechnology and electrospinning process, nanofibrous scaffolds were fabricated to host dental follicle stem cells (DFSCs). Blends (50:50) of polycaprolactone (PCL) and silk fibroin (SF), mixed with various content of cellulose nanocrystals (CNC, up to 5% in weight), were electrospun to prepare nanofibrous scaffolds with heterogeneous microstructure in terms of fiber size. Colloidal probe atomic force microscopy (AFM) and conventional uniaxial tensile tests measured the scaffold stiffness at the micro-and macro-scale, respectively. The cell elastic modulus and cell-scaffold adhesive interaction (i.e., a chemical function) were examined through single-cell force spectroscopy using AFM. The quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to determine if the mechanotransduction signal (i.e., Yap1, Wwr2, Rac1, MAPK8, Ptk2 and Wnt5a) is upregulated by the scaffold stiffness at the micro-scale (cellular scale). Results: The presence of CNC produces fibrous scaffolds with a bimodal distribution of fiber diameter. This structural heterogeneity, which is CNC-composition dependent, remarkably modulates the mechanical functionality of scaffolds at microscale and macroscale simultaneously, but not the chemical functionality (i.e., only a single material property is varied). In in vitro tests, the osteogenic differentiation and gene expression associated with mechano-sensitive cell markers correlate to the degree of micromechanical compatibility between DFSCs and the scaffold. Conclusion: Cells require compliant scaffolds to encourage energetically favorable interactions for mechanotransduction, which are converted into changes in cellular biochemistry to direct the phenotypic evolution. The micromechanical compatibility is indeed important to the efficacy of regenerative medicine.

Keywords: phenotype transition, scaffold stiffness, electrospinning, cellulose nanocrystals, single-cell force spectroscopy

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Authors: Disaya Jaroensattayatham

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Nowadays, infectious diseases are prevalent and severe health problems as they render the increment of casualty, illness, and global economic recession. Along with the emergence of antimicrobial resistance, the potency of typically used antibiotics can be affected to a considerable degree. As a result, unorthodox antibiotics have become an urgent issue in the pharmaceutical field. Piper betle L., known as betle leaf, has been used for many purposes, such as a traditional home remedy, and has shown its ability in inhibiting bacteria as well as fungus. Thus, in this study, the investigation of antimicrobial activities of the Piper betle L. extracts was carried out using the Agar disk-diffusion method and Broth microdilution, aiming to evaluate and determine its efficacy to inhibit bacterial and fungal growth of Staphylococcus aureus, Salmonella typhi, and Candida albicans. In the agar disk-diffusion test, the extracts of Piper betle L. gave the maximum zone of inhibition of 15.1 mm (S. aureus), 7.7 mm (S. typhi), and 11.7 mm (C. albicans), while its MIC values were 1000 µg/ml in S. aureus and greater than 2000 µg/ml in S. typhi and C. albicans. According to the results, the Piper betle L. obtains an antimicrobial activity and shows a higher effect towards gram-positive bacteria than gram-negative bacteria. To determine the mechanism behind its ability, more research is needed to be performed in the future.

Keywords: antimicrobial activity, Candida albicans, Piper betle L., Salmonella typhi, Staphylococcus aureus

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Authors: Mirza Mujtaba Baig

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Artificial Intelligence (AI) is evolving rapidly, and one of the areas in which this field has influenced is automation. The automobile, healthcare, education, and robotic industries deploy AI technologies constantly, and the automation of tasks is beneficial to allow time for knowledge-based tasks and also introduce convenience to everyday human endeavors. The paper reviews the challenges faced with the current implementations of autonomous self-driving cars by exploring the machine learning, robotics, and artificial intelligence techniques employed for the development of this innovation. The controversy surrounding the development and deployment of autonomous machines, e.g., vehicles, begs the need for the exploration of the configuration of the programming modules. This paper seeks to add to the body of knowledge of research assisting researchers in decreasing the inconsistencies in current programming modules. Blockchain is a technology of which applications are mostly found within the domains of financial, pharmaceutical, manufacturing, and artificial intelligence. The registering of events in a secured manner as well as applying external algorithms required for the data analytics are especially helpful for integrating, adapting, maintaining, and extending to new domains, especially predictive analytics applications.

Keywords: artificial intelligence, automation, big data, self-driving cars, machine learning, neural networking algorithm, blockchain, business intelligence

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Authors: C. Flynn, Q. Yuan, C. Reinhardt

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Alzheimer’s Disease (AD) is a progressive neurodegenerative disorder affecting broad areas of the cerebral cortex and hippocampus. More than 95% of AD cases are representative of sporadic AD, where both genetic and environmental risk factors play a role. The main pathological features of AD include the widespread deposition of amyloid-beta and neurofibrillary tau tangles in the brain. The earliest brain pathology related to AD has been defined as hyperphosphorylated soluble tau in the noradrenergic locus coeruleus (LC) neurons, characterized by Braak. However, the cause of this pathology and the ultimate progression of AD is not understood. Increasing research points to a connection between the gut microbiota and the brain, and mounting evidence has shown that there is a bidirectional interaction between the two, known as the gut-brain axis. This axis can allow for bidirectional movement of neuroinflammatory cytokines and pathogenic misfolded proteins, as seen in AD. Prebiotics and probiotics have been shown to have a beneficial effect on gut health and can strengthen the gut-barrier as well as the blood-brain barrier, preventing the spread of these pathogens across the gut-brain axis. Our laboratory has recently established a pretangle tau rat model, in which we selectively express pseudo-phosphorylated human tau (htauE14) in the LC neurons of TH-Cre rats. LC htauE14 produced pathological changes in rats resembling those of the preclinical AD pathology (reduced olfactory discrimination and LC degeneration). In this work, we will investigate the effects of pre/probiotic ingestion on AD behavioral deficits, blood inflammation/cytokines, and various brain markers in our experimental rat model of AD. Rats will be infused with an adeno-associated viral vector containing a human tau gene pseudophosphorylated at 14 sites (common in LC pretangles) into 2-3 month TH-Cre rats. Fecal and blood samples will be taken at pre-surgery, and various post-surgery time points. A collection of behavioral tests will be performed, and immunohistochemistry/western blotting techniques will be used to observe various biomarkers. This work aims to elucidate the relationship between gut health and AD progression by strengthening gut-brain relationship and aims to observe the overall effect on tau formation and tau pathology in AD brains.

Keywords: alzheimer’s disease, aging, gut microbiome, neurodegeneration

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Authors: Shilu Deepa Thomas, P. Jayaprakash, Dorota Łazewska, Katarzyna Kieć-Kononowicz, B. Sadek

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A large body of evidence suggests the involvement of cognitive impairment, increased levels of inflammation and oxidative stress in the pathogenesis of autism spectrum disorder (ASD). ASD commonly coexists with psychiatric conditions like anxiety and cognitive challenges, and individuals with ASD exhibit significant levels of inflammation and immune system dysregulation. Previous Studies have identified elevated levels of pro-inflammatory markers such as IL-1β, IL-6, IL-2 and TNF-α, particularly in young children with ASD. The current therapeutic options for ASD show limited effectiveness, signifying the importance of exploring an efficient drugs to address the core symptoms. The role of histamine H3 receptors (H3Rs) in memory and the prospective role of H3R antagonists in pharmacological control of neurodegenerative disorders, e.g., ASD, is well-accepted. Hence, the effects of chronic systemic administration of H3R antagonist E159 on autistic-like repetitive behaviors, social deficits, memory and anxiety parameters, as well as neuroinflammation in Black and Tan BRachyury (BTBR) mice, were evaluated using Y maze, Barnes maze, self-grooming, open field and three chamber social test. E159 (2.5, 5 and 10 mg/kg, i.p.) dose-dependently ameliorated repetitive and compulsive behaviors by reducing the increased time spent in self-grooming and improved reduced spontaneous alternation in BTBR mice. Moreover, treatment with E159 attenuated disturbed anxiety levels and social deficits in tested male BTBR mice. Furthermore, E159 attenuated oxidative stress by significantly increasing GSH, CAT, and SOD and decreasing the increased levels of MDA in the cerebellum as well as the hippocampus. In addition, E159 decreased the elevated levels of proinflammatory cytokines (tumor necrosis factor (TNF-α), interleukin-1β (IL-1β), and IL-6). The observed results show that H3R antagonists like E159 may represent a promising novel pharmacological strategy for the future treatment of ASD.

Keywords: histamine H3 receptors, antagonist E159, autism, behaviors, mice

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Authors: Michael Lousis

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Mathematical competence is the great aim of every mathematical teaching and learning endeavour. This can be defined as an idealised conceptualisation of the quality of cognition and the ability of implementation in practice of the mathematical subject matter, which is included in the curriculum, and is displayed only through performance of doing mathematics. The present study gives a clear definition of mathematical competence in the domains of Arithmetic and Algebra that stems from the explanation of the learners’ errors in these domains. The learners, whose errors are explained, were Greek and English participants of a large, international, longitudinal, comparative research program entitled the Kassel Project. The participants’ errors emerged as results of their work in dealing with mathematical questions and problems of the tests, which were presented to them. The construction of the tests was such as only the outcomes of the participants’ work was to be encompassed and not their course of thinking, which resulted in these outcomes. The intention was that the tests had to provide undeviating comparable results and simultaneously avoid any probable bias. Any bias could stem from obtaining results by involving so many markers from different countries and cultures, with so many different belief systems concerning the assessment of learners’ course of thinking. In this way the validity of the research was protected. This fact forced the implementation of specific research methods and theoretical prospects to take place in order the participants’ erroneous way of thinking to be disclosed. These were Methodological Pragmatism, Symbolic Interactionism, Philosophy of Mind and the ideas of Computationalism, which were used for deciding and establishing the grounds of the adequacy and legitimacy of the obtained kinds of knowledge through the explanations given by the error analysis. The employment of this methodology and of these theoretical prospects resulted in the definition of the learners’ mathematical competence, which is the thesis of the present study. Thus, learners’ mathematical competence is depending upon three key elements that should be developed in their minds: appropriate representations, appropriate meaning, and appropriate developed schemata. This definition then determined the development of appropriate teaching practices and interventions conducive to the achievement and finally the entailment of mathematical competence.

Keywords: representations, meaning, appropriate developed schemata, computationalism, error analysis, explanations for the probable causes of the errors, Kassel Project, mathematical competence

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

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

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

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