Search results for: genetic markers

Authors: Evgeny A. Ermakov, Lyudmila P. Smirnova, Valentina N. Buneva

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Schizophrenia associated with dysregulation of neurotransmitter processes in the central nervous system and disturbances in the humoral immune system resulting in the formation of antibodies (Abs) to the various components of the nervous tissue. Abs to different neuronal receptors and DNA were detected in the blood of patients with schizophrenia. Abs hydrolyzing DNA were detected in pool of polyclonal autoantibodies in autoimmune and infectious diseases, such catalytic Abs were named abzymes. It is believed that DNA-hydrolyzing abzymes are cytotoxic, cause nuclear DNA fragmentation and induce cell death by apoptosis. Abzymes with DNAase activity are interesting because of the mechanism of formation and the possibility of use as diagnostic markers. Therefore, in our work we have set following goals: to determine the level anti-DNA Abs in the serum of patients with schizophrenia and to study DNA-hydrolyzing activity of IgG of patients with schizophrenia. Materials and methods: In our study there were included 41 patients with a verified diagnosis of paranoid or simple schizophrenia and 24 healthy donors. Electrophoretically and immunologically homogeneous IgGs were obtained by sequential affinity chromatography of the serum proteins on protein G-Sepharose and gel filtration. The levels of anti-DNA Abs were determined using ELISA. DNA-hydrolyzing activity was detected as the level of supercoiled pBluescript DNA transition in circular and linear forms, the hydrolysis products were analyzed by agarose electrophoresis followed by ethidium bromide stain. To correspond the registered catalytic activity directly to the antibodies we carried out a number of strict criteria: electrophoretic homogeneity of the antibodies, gel filtration (acid shock analysis) and in situ activity. Statistical analysis was performed in ‘Statistica 9.0’ using the non-parametric Mann-Whitney test. Results: The sera of approximately 30% of schizophrenia patients displayed a higher level of Abs interacting with single-stranded (ssDNA) and double-stranded DNA (dsDNA) compared with healthy donors. The average level of Abs interacting with ssDNA was only 1.1-fold lower than that for interacting with dsDNA. IgG of patient with schizophrenia were shown to possess DNA hydrolyzing activity. Using affinity chromatography, electrophoretic analysis of isolated IgG homogeneity, gel filtration in acid shock conditions and in situ DNAse activity analysis we proved that the observed activity is intrinsic property of studied antibodies. We have shown that the relative DNAase activity of IgG in patients with schizophrenia averaged 55.4±32.5%, IgG of healthy donors showed much lower activity (average of 9.1±6.5%). It should be noted that DNAase activity of IgG in patients with schizophrenia with a negative symptoms was significantly higher (73.3±23.8%), than in patients with positive symptoms (43.3±33.1%). Conclusion: Anti-DNA Abs of patients with schizophrenia not only bind DNA, but quite efficiently hydrolyze the substrate. The data show a correlation with the level of DNase activity and leading symptoms of patients with schizophrenia.

Keywords: anti-DNA antibodies, abzymes, DNA hydrolysis, schizophrenia

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Authors: Weaam Aldeeban, Majd Aljamali, Lama A. Youssef

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Background & Objective: Warfarin is considered a problematic drug due to its narrow therapeutic window and wide inter-individual response variations, which are attributed to demographic, environmental, and genetic factors, particularly single nucleotide polymorphism (SNPs) in the genes encoding VKORC1 and CYP2C9 involved in warfarin's mechanism of action and metabolism, respectively. CYP2C9*2rs1799853 and CYP2C9*3rs1057910 alleles are linked to reduced enzyme activity, as carriers of either or both alleles are classified as moderate or slow metabolizers, and therefore exhibit higher sensitivity of warfarin compared with wild type (CYP2C9*1*1). Our study aimed to assess the frequency of *1, *2, and *3 alleles in the CYP2C9 gene in a cohort of Syrian patients receiving a maintenance dose of warfarin for different indications, the impact of genotypes on warfarin dosing, and the frequency of adverse effects (i.e., bleedings). Subjects & Methods: This retrospective cohort study encompassed 94 patients treated with warfarin. Patients’ genotypes were identified by sequencing the polymerase chain reaction (PCR) specific products of the gene encoding CYP2C9, and the effects on warfarin therapeutic outcomes were investigated. Results: Sequencing revealed that 43.6% of the study population has the *2 and/or *3 SNPs. The mean weekly maintenance dose of warfarin was 37.42 ± 15.5 mg for patients with the wild-type allele (CYP2C9*1*1), whereas patients with one or both variants (*2 and/or *3) demanded a significantly lower dose (28.59 ±11.58 mg) of warfarin, (P= 0.015). A higher percentage (40.7%) of patients with allele *2 and/or *3 experienced hemorrhagic accidents compared with only 17.9% of patients with the wild type *1*1, (P = 0.04). Conclusions: Our study proves an association between *2 and *3 genotypes and higher sensitivity to warfarin and a tendency to bleed, which necessitates lowering the dose. These findings emphasize the significance of CYP2C9 genotyping prior to commencing warfarin therapy in order to achieve optimal and faster dose control and to ensure effectiveness and safety.

Keywords: warfarin, CYP2C9, polymorphisms, Syrian, hemorrhage

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Authors: Rui Wu

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In the volatile modern manufacturing environment, new orders randomly occur at any time, while the pre-emptive methods are infeasible. This leads to a real-time scheduling method that can produce a reasonably good schedule quickly. The dynamic Flexible Job Shop problem is an NP-hard scheduling problem that hybrid the dynamic Job Shop problem with the Parallel Machine problem. A Flexible Job Shop contains different work centres. Each work centre contains parallel machines that can process certain operations. Many algorithms, such as genetic algorithms or simulated annealing, have been proposed to solve the static Flexible Job Shop problems. However, the time efficiency of these methods is low, and these methods are not feasible in a dynamic scheduling problem. Therefore, a dynamic rule selection scheduling system based on the reinforcement learning method is proposed in this research, in which the dynamic Flexible Job Shop problem is divided into several parallel machine problems to decrease the complexity of the dynamic Flexible Job Shop problem. Firstly, the features of jobs, machines, work centres, and flexible job shops are selected to describe the status of the dynamic Flexible Job Shop problem at each decision point in each work centre. Secondly, a framework of reinforcement learning algorithm using a double-layer deep Q-learning network is applied to select proper composite dispatching rules based on the status of each work centre. Then, based on the selected composite dispatching rule, an available operation is selected from the waiting buffer and assigned to an available machine in each work centre. Finally, the proposed algorithm will be compared with well-known dispatching rules on objectives of mean tardiness, mean flow time, mean waiting time, or mean percentage of waiting time in the real-time Flexible Job Shop problem. The result of the simulations proved that the proposed framework has reasonable performance and time efficiency.

Keywords: dynamic scheduling problem, flexible job shop, dispatching rules, deep reinforcement learning

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Authors: Yahia Massinissa, Yahia Mouloud

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Cystic fibrosis (CF), the most common lethal genetic disease in the Europe population, is caused by mutations in the transmembrane conductance regulator gene (CFTR). It affects most organs including an epithelial tissue, base of hydroelectrolytic transepithelial transport, notably that aerial ways, the pancreas, the biliary ways, the intestine, sweat glands and the genital tractus. The gene whose anomalies are responsible of the cystic fibrosis codes for a protein Cl channel named CFTR (cystic fibrosis transmembrane conductance regulator) that exercises multiple functions in the cell, one of the most important in control of sodium and chlorine through epithelia. The deficient function translates itself notably by an abnormal production of viscous secretion that obstructs the execrator channels of this target organ: one observes then a dilatation, an inflammation and an atrophy of these organs. It also translates itself by an increase of the concentration in sodium and in chloride in sweat, to the basis of the sweat test. In order to do a phenotypical and genotypical diagnosis at a part of the Algerian population, our survey has been carried on 16 patients with evocative symptoms of the cystic fibrosis at that the clinical context has been confirmed by a sweat test. However, anomalies of the CFTR gene have been determined by electrophoresis in gel of polyacrylamide of the PCR products (polymerase chain reaction), after enzymatic digestion, then visualized to the ultraviolet (UV) after action of the ethidium bromide. All mutations detected at the time of our survey have already been identified at patients attained by this pathology in other populations of the world. However, the important number of found mutation with regard to the one of the studied patients testifies that the origin of this big clinical variability that characterizes the illness in the consequences of an enormous diversity of molecular defects of the CFTR gene.

Keywords: cystic fibrosis, CFTR gene, polymorphism, algerian population, sweat test, genotypical diagnosis

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Authors: Irina Veselova, Maria Makedonskaya, Olga Eremina, Alexandr Sidorov, Eugene Goodilin, Tatyana Shekhovtsova

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Such catecholamines as dopamine, norepinephrine, and epinephrine are the principal neurotransmitters in the sympathetic nervous system. Catecholamines and their metabolites are considered to be important markers of socially significant diseases such as atherosclerosis, diabetes, coronary heart disease, carcinogenesis, Alzheimer's and Parkinson's diseases. Currently, neurotransmitters can be studied via electrochemical and chromatographic techniques that allow their characterizing and quantification, although these techniques can only provide crude spatial information. Besides, the difficulty of catecholamine determination in biological materials is associated with their low normal concentrations (~ 1 nM) in biomaterials, which may become even one more order lower because of some disorders. In addition, in blood they are rapidly oxidized by monoaminooxidases from thrombocytes and, for this reason, the determination of neurotransmitter metabolism indicators in an organism should be very rapid (15—30 min), especially in critical states. Unfortunately, modern instrumental analysis does not offer a complex solution of this problem: despite its high sensitivity and selectivity, HPLC-MS cannot provide sufficiently rapid analysis, while enzymatic biosensors and immunoassays for the determination of the considered analytes lack sufficient sensitivity and reproducibility. Fluorescent and SERS-sensors remain a compelling technology for approaching the general problem of selective neurotransmitter detection. In recent years, a number of catecholamine sensors have been reported including RNA aptamers, fluorescent ribonucleopeptide (RNP) complexes, and boronic acid based synthetic receptors and the sensor operated in a turn-off mode. In this work we present the fluorescent and SERS turn-on sensor systems based on the bio- or chemorecognizing nanostructured films {chitosan/collagen-Tb/Eu/Cu-nanoparticles-indicator reagents} that provide the selective recognition, visualization, and sensing of the above mentioned catecholamines on the level of nanomolar concentrations in biomaterials (cell cultures, tissue etc.). We have (1) developed optically transparent porous films and gels of chitosan/collagen; (2) ensured functionalization of the surface by molecules-'recognizers' (by impregnation and immobilization of components of the indicator systems: biorecognizing and auxiliary reagents); (3) performed computer simulation for theoretical prediction and interpretation of some properties of the developed materials and obtained analytical signals in biomaterials. We are grateful for the financial support of this research from Russian Foundation for Basic Research (grants no. 15-03-05064 a, and 15-29-01330 ofi_m).

Keywords: biomaterials, fluorescent and SERS-recognition, neurotransmitters, solid-phase turn-on sensor system

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Authors: Sharique Ahmed, Khushtar Anwar Salman

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Alpha-1-antitrypsin (AAT) is a major plasma serine protease inhibitor (SERPIN). Hereditary AAT deficiency is one of the common diseases in some part of the world. AAT is mainly produced in the liver and functions to protect the lung against proteolytic damage (e.g., from neutrophil elastase) acting as the major inhibitor for neutrophil elastase. α (1)-Antitrypsin (AAT) deficiency is an under recognized genetic condition that affects approximately 1 in 2,000 to 1 in 5,000 individuals and predisposes to liver disease and early-onset emphysema. Not only does α-1-antitrypsin deficiency lead to disabling syndrome of pulmonary emphysema, there are other disorders too which include ANCA (antineutrophilic cytoplasmic antibody) positive Wegener's granulomatosis, diffuse bronchiectasis, necrotizing panniculitis in α-1-antitrypsin phenotype (S), idiopathic pulmonary fibrosis and steroid dependent asthma. Augmentation therapy with alpha-1 antitrypsin (AAT) from human plasma has been available for specific treatment of emphysema due to AAT deficiency. Apart from this several observations have also suggested a role for endogenous suppressors of HIV-1, alpha-1 antitrypsin (AAT) has been identified to be one of those. In view of its varied important role in humans, serum from a mammalian source was chosen for the isolation and purification. Studies were performed on the homogeneous fraction. This study suggests that the buffalo serum α-1-antritrypsin has characteristics close to ovine, dog, horse and more importantly to human α-1-antritrypsin in terms of its hydrodynamic properties such as molecular weight, carbohydrate content, etc. The similarities in the hydrodynamic properties of buffalo serum α-1-antitrypsin with other sources of mammalian α-1-antitrypsin mean that it can be further studied and be a potential source for "augmentation therapy", as well as a source of AAT replacement therapy to raise serum levels above the protective threshold. Other parameters like the amino acid sequence, the effect of denaturants, and the thermolability or thermostability of the inhibitor will be the interesting basis of future studies on buffalo serum alpha-1 antitrypsin (AAT).

Keywords: α-1-antitrypsin, augmentation therapy , hydrodynamic properties, serine protease inhibitor

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Authors: Aswandi Anas Husin, Cut Rizlani Kholibrina

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Harvesting camphor resin has been carried out since the beginning of civilization on the west coast of Sumatra. Oil or crystals that containing borneol are harvested from the camphor tree (Dryobalanops aromatica). Non-timber forest products are utilized for the manufacture of fragrances, antiseptics, anti-inflammatory, analgesic as well as effective for the treatment of blocked arteries. Based on exploration on the west coast of Sumatra, these endemic tree species were found remaining growing in groups on small spots in the lowlands to the hills. Some populations are found at an altitude of 700 meters above sea level in Kadabuhan, Jongkong and Sultan Daulat in Subulussalam district, Singkohor and Lake Paris in Aceh Singkil district, and Sirandorung and Manduamas in the north of Barus, Central Tapanuli district. These multi-purpose tree species was also identified as being able to adapt to the Singkil Peat Swamp. The decline in tree population has a direct impact on reducing their productivity. Conventionally, the crystals are harvested by cutting and splitting the stem into wooden blocks. In this way about 1.5-2.5 kg of crystals are obtained with various qualities. Camphor retrieval can also be done by making a notch on a standing tree trunk and collecting liquid resin (ombil) that is removed from the injured resin channel. Twigs and leaves also contain borneol. The aromatic content in this section opens opportunities for the supply of borneol through the distillation process. Vegetative propagation technology is needed to overcome the limitations of available seeds. This breeding strategy for vulnerable species starts with gathering genetic material from various provenances which are then used to support the provision of basic populations, breeding populations, multiplication populations and production populations for extensive development of camphor tree plantations

Keywords: camphor, conservation, natural borneol, productivity, vulnerable species

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Authors: Zewdneh Zana Zate

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The study focuses on the evolutionary and ecological genomics of both wild and cultivated Coffea arabica L. at its center of origin, Ethiopia, aiming to uncover how this vital species may withstand future climate changes. Utilizing bioclimatic models, we project the future distribution of Arabica under varied climate scenarios for 2050 and 2080, identifying potential conservation zones and immediate risk areas. Through whole-genome resequencing of accessions from Ethiopian gene banks, this research assesses genetic diversity and divergence between wild and cultivated populations. It explores relationships, demographic histories, and potential hybridization events among Coffea arabica accessions to better understand the species' origins and its connection to parental species. This genomic analysis also seeks to detect signs of natural or artificial selection across populations. Integrating these genomic discoveries with ecological data, the study evaluates the current and future ecological and genomic vulnerabilities of wild Coffea arabica, emphasizing necessary adaptations for survival. We have identified key genomic regions linked to environmental stress tolerance, which could be crucial for breeding more resilient Arabica varieties. Additionally, our ecological modeling predicted a contraction of suitable habitats, urging immediate conservation actions in identified key areas. This research not only elucidates the evolutionary history and adaptive strategies of Arabica but also informs conservation priorities and breeding strategies to enhance resilience to climate change. By synthesizing genomic and ecological insights, we provide a robust framework for developing effective management strategies aimed at sustaining Coffea arabica, a species of profound global importance, in its native habitat under evolving climatic conditions.

Keywords: coffea arabica, climate change adaptation, conservation strategies, genomic resilience

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Authors: Preethy Mary Donald, Renjith George

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Oral tori are localized non-neoplastic protuberances of maxilla and mandible. Torus palatinus (TP) is found on the midline of the roof of mouth existing as single growth or in clusters. Torus mandibularis(TM) is located on the lingual aspect of the mandible commonly between canine and premolar region. Etiology of their presence was not clear and was found to be multifactorial. Their variations in relation to age, gender, ethnicity and also the characteristics of TP and TM have become the interest of multiple studies. The objectives of this study were to determine the prevalence of torus palatinus (TP) and torus mandibularis (TM) among patients who have visited outpatient department, Faculty of Dentistry, Melaka Manipal Medical College. 108 patients were examined for the presence of oral tori at the outpatient department, Faculty of Dentistry, Melaka-Manipal Medical College. Factors such as age, gender, ethnicity of the patients and size, shape, location of the oral tori were studied. For TP, Malays (62.96%) have been found to have the highest prevalence than Chinese (43.3%) and Indians (35.71%). For TM, Chinese (7.46%) had predominated compared to Malays (7.41%) and Indians (0%). There is no significant association between occurrence of TP and TM with age, gender and ethnicity. For Torus palatinus, the most common size was Grade 1(1-3mm), most common location was molar region, and the most common shape was spindle. For Torus mandibularis, the most frequent location was canine premolar region and exists in unilateral single or bilateral single fashion. The overall prevalence rates were 47.2% for TP and 6.48% for TM. However, there is no significant association between occurrence of TP and TM with age, gender and ethnicity. The results showed variations in clinical characteristics and support the findings that occurrence of tori is a dynamic phenomenon which is multifactorial owing to the environmental factors such as stress from occlusion and dietary habits. It could be due to the genetic make-up of the individual.

Keywords: torus palatinus, torus mandibularis, age, gender

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Authors: Zintle Kolo, Ndiko Ludidi

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The enzymatic activity of p-coumarate 3-hydroxylase (C3H) synthesize caffeic acid from p-coumaric acid. We recently showed that exogenously applied caffeic acid confers salinity tolerance in soybean (Glycine max) by inducing antioxidant enzymatic activity to promote enhanced scavenging or reactive oxygen species, thus limiting salinity-induced oxidative stress. Recent evidence also establishes that pre-treatment of plants with exogenously supplied caffeic acid improves plant tolerance to osmotic stress by improving plant antioxidant capacity and enhancing biosynthesis of compatible solutes. We aimed to identify a C3H in maize (Zea mays) and evaluate the effect of drought on the spatial and temporal expression of the gene encoding the candidate maize C3H (ZmC3H). Primary sequence analysis shows that ZmC3H shares 71% identity with an Arabidopsis thaliana C3H that is implicated in the control of Arabidopsis cell expansion, growth, and responses to stress. In silico ZmC3H promoter analysis reveals the presence of cis-acting elements that interact with transcription factors implicated in plant responses to drought. Spatial expression analysis by semi-quantitative RT-PCR shows that ZmC3H is expressed in both leaves and roots under normal conditions. However, drought represses the expression of ZmC3H in leaves whereas it up-regulates its expression in roots. These changes in ZmC3H expression correlate with the changes in the content of caffeic acid in maize in response to drought. We illustrate the implications of these changes in the expression of the gene in relation to maize responses to drought and discuss the potential of regulating caffeic acid biosynthesis towards genetic improvement of maize tolerance to drought stress. These findings have implications for food security because of the potential of the implications of the study for drought tolerance in maize.

Keywords: caffeic acid, drought-responsive expression, maize drought tolerance, p-coumarate 3-hydroxylase

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Authors: Ji-Won Park, Jong-Wha Kim

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Triploid L. lancifolium have a wide geographic distribution. By contrast, diploid L. lancifolium have limited distributions in the islands and coastal regions of the South and West Korean Peninsula and northern Tsushima Island, Japan. L. lancifolium diploids and triploids are not sympatrically distributed with other lily species or ploidy lines in West Sea and South Sea Islands of the Korean Peninsula. This observation raises the following questions: 'Why have autotriploid L. lancifolium never been observed in those isolated islands?', 'What mechanism excludes the occurrence of autotriploids, if they arise?'. To determine the occurrence and survival of triploid plants in natural diploid populations of tiger lily (Lilium lancifolium), ploidy analysis was conducted on natural open-pollinated seeds produced from plants grown on isolated islands, and on hybrid seeds produced by artificial crossing between plant populations originating on different Korean islands. Normal seeds were classified into five grades depending on the ratio of embryo/endosperm lengths, including 5/5, 4/5, 3/5, 2/5, and 1/5. Triploids were not observed among seedlings produced from natural open pollinations on isolated islands. Triploids were detected only in seedlings of underdeveloped seed grades(3/5 and 2/5) from artificial crosses between populations from different isolated islands. The triploid occurrence frequency was calculated as 0.0 for natural open-pollinated seedlings and 0.000582 for artificial crosses(6 triploids from 10,303 seedlings). Triploids were produced from crosses between isolated populations located at least 70 km apart; no triploids were detected in inter-population crosses of plants originating on the same islands. Triploid seedlings have very low viability in soil. We analyzed factors affecting triploid occurrence and survival in natural diploid populations of L. lancifolium. The results suggest that triploids originate from fertilization between plants that are genetically isolated due to geographical isolation and/or genotypic differences.

Keywords: Lilium lancifolium, autotriploid, natural population, genetic distance, 2n female gamete

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Authors: Shilpa Mukundaraj, Nagaraju Shivaiah

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Amyloidogenic conditions, driven by protein aggregation into insoluble fibrils, which pose significant challenges in the clinical condition of diabetes management, particularly through the amyloidogenic LVEALYL sequence in insulin B-chain. This study explores a dual therapeutic strategy involving cysteine protease enzymes such as papain and ficin and inhibitory peptides to target insulin amyloidosis. Combining in silico, in vitro, and in vivo methodologies, the research aims to inhibit amyloid formation and degrade preformed fibrils. Inhibitory peptides were designed using structure-guided approaches in Rosetta to specifically target the LVEALYL sequence. Concurrently, cysteine protease enzymes, including papain and ficin, were evaluated for their fibril disassembly potential. In vitro experiments, utilizing SDS- PAGE and spectroscopic techniques, confirmed dose-dependent degradation 50 to 300ug in vitro and 60mg/kg in vivo of amyloid aggregates by these enzymes, with significant disaggregation observed at higher concentrations 20mg. Peptide inhibitors effectively reduced fibril formation, as evidenced by reduced Thioflavin T fluorescence and circular dichroism spectroscopy. Complementary in silico analyses, including molecular docking and dynamic simulations, provided structural insights into enzyme binding interactions with amyloidogenic regions. Key residues involved in substrate recognition and cleavage were identified, with computational findings aligning strongly with experimental data. These insights confirmed the specificity of papain and ficin in targeting insulin fibrils. For translational potential, an in vivo rat model was developed, involving subcutaneous insulin amyloid injections to induce localized amyloid deposits. Over six days of enzyme treatment, a marked reduction in amyloid burden was observed through histological findings and biochemical assay superoxide dismutase can provide insights into oxidative damage due to amyloid deposition. Furthermore, inflammatory markers IL-6, TNFα were significantly attenuated in treated groups, emphasizing the dual role of enzymes in amyloid clearance and inflammation modulation. This integrative study highlights the promise of cysteine protease enzymes and inhibitory peptides as complementary therapeutic strategies for managing insulin amyloidosis. By targeting both the formation and persistence of amyloid fibrils, this dual approach offers a novel and effective avenue for amyloidosis treatment.

Keywords: insulin amyloidosis, peptide inhibitors, cysteine protease enzymes, amyloid degradation

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Authors: Norsuhana Omar, Amilia Aminuddina Zaiton Zakaria, Raifana Rosa Mohamad Sattar, Kalaivani Chellappan, Mohd Alauddin Mohd Ali, Norizam Salamt, Zanariyah Asmawi, Norliza Saari, Aini Farzana Zulkefli, Nor Anita Megat Mohd. Nordin

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Inflammation plays an important role in the pathogenesis of vascular dysfunction leading to arterial stiffness. Pulse wave velocity (PWV) and augmentation index (AS), as tools for the assessment of vascular damages are widely used and have been shown to predict cardiovascular disease (CVD). C-reactive protein (CRP) is a marker of inflammation. Several studies noted that regular exercise is associated with reduced arterial stiffness. The lack of exercise among Malaysians and the increasing CVD morbidity and mortality among young men are of concern. In Malaysia data on the workplace exercise intervention is scarce. A programme was designed to enable subjects to increase their level of walking as part of their daily work routine and self-monitored by using pedometers. The aim of this study to evaluate the reducing of inflammation by measuring CRP and improvement arterial stiffness measured by carotid femoral PWV (PWVCF) and AI. A total of 70 young men (20 - 40 years) who were sedentary, achieving less than 5,000 steps/day in casual walking with 2 or more cardiovascular risk factors were recruited in Institute of Vocational Skills for Youth (IKBN Hulu Langat). Subjects were randomly assigned to a control (CG) (n=34; no change in walking) and pedometer group (PG) (n=36; minimum target: 8,000 steps/day). The CRP was measured by using immunological method while PWVCF and AI were measured using Vicorder. All parameters were measured at baseline and after 12 weeks. Data for analysis was conducted using Statistical Package of Social Sciences Version 22 (SPSS Inc., Chicago, IL, USA). At post intervention, the CG step counts were similar (4983 ± 366vs 5697 ± 407steps/day). The PG increased step count from 4996 ± 805 to 10,128 ±511 steps/day (P<0.001). The PG showed significant improvement in anthropometric variables and lipid (time and group effect p<0.001). For vascular assessment, the PG showed significantly decreased for time and effect (p<0.001) for PWV (7.21± 0.83 to 6.42 ± 0.89) m/s; AI (11.88± 6.25 to 8.83 ± 3.7) % and CRP (pre= 2.28 ± 3.09, post=1.08± 1.37mg/L). However, no changes were seen in CG. As a conclusion, a pedometer-based walking programme may be an effective strategy for promoting increased daily physical activity which reduces cardiovascular risk markers and thus improve cardiovascular health in terms of inflammation and arterial stiffness. The community intervention for health maintenance has potential to adopt walking as an exercise and adopting vascular fitness index as the performance measuring tools.

Keywords: arterial stiffness, exercise, inflammation, pedometer

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Authors: Nguyen J. M., Lucas G., Ruan S., Digonnet H., Antonioli D.

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Introduction: Deep Learning (DL) is a very powerful tool for analyzing image data. But for tabular data, it cannot compete with machine learning methods like XGBoost. The research question becomes: can tabular data be transformed into images that can be analyzed by simple CNNs (Convolutional Neuron Networks)? Will DL be the absolute tool for data classification? All current solutions consist in repositioning the variables in a 2x2 matrix using their correlation proximity. In doing so, it obtains an image whose pixels are the variables. We implement a technology, DeepNIC, that offers the possibility of obtaining an image for each variable, which can be analyzed by simple CNNs. Material and method: The 'ROP' (Regression OPtimized) model is a binary and atypical decision tree whose nodes are managed by a new artificial neuron, the Neurop. By positioning an artificial neuron in each node of the decision trees, it is possible to make an adjustment on a theoretically infinite number of variables at each node. From this new decision tree whose nodes are artificial neurons, we created the concept of a 'Random Forest of Perfect Trees' (RFPT), which disobeys Breiman's concepts by assembling very large numbers of small trees with no classification errors. From the results of the RFPT, we developed a family of 10 statistical information criteria, Nguyen Information Criterion (NICs), which evaluates in 3 dimensions the predictive quality of a variable: Performance, Complexity and Multiplicity of solution. A NIC is a probability that can be transformed into a grey level. The value of a NIC depends essentially on 2 super parameters used in Neurops. By varying these 2 super parameters, we obtain a 2x2 matrix of probabilities for each NIC. We can combine these 10 NICs with the functions AND, OR, and XOR. The total number of combinations is greater than 100,000. In total, we obtain for each variable an image of at least 1166x1167 pixels. The intensity of the pixels is proportional to the probability of the associated NIC. The color depends on the associated NIC. This image actually contains considerable information about the ability of the variable to make the prediction of Y, depending on the presence or absence of other variables. A basic CNNs model was trained for supervised classification. Results: The first results are impressive. Using the GSE22513 public data (Omic data set of markers of Taxane Sensitivity in Breast Cancer), DEEPNic outperformed other statistical methods, including XGBoost. We still need to generalize the comparison on several databases. Conclusion: The ability to transform any tabular variable into an image offers the possibility of merging image and tabular information in the same format. This opens up great perspectives in the analysis of metadata.

Keywords: tabular data, CNNs, NICs, DeepNICs, random forest of perfect trees, classification

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Authors: Yong Min You

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The study on the torque ripple of Permanent Magnet Synchronous Motors (PMSMs) has been rapidly progressed, which effects on the noise and vibration of the electric vehicle. There are several ways to reduce torque ripple, which are the increase in the number of slots and poles, the notch of the rotor and stator teeth, and the skew of the rotor and stator. However, the conventional methods have the disadvantage in terms of material cost and productivity. The demagnetization characteristic of PMSMs must be attained for electric vehicle application. Due to rare earth supply issue, the demand for Dy-free permanent magnet has been increasing, which can be applied to PMSMs for the electric vehicle. Dy-free permanent magnet has lower the coercivity; the demagnetization characteristic has become more significant. To improve the torque ripple as well as the demagnetization characteristics, which are significant parameters for electric vehicle application, an unequal air-gap model is proposed for a PMSM. A shape optimization is performed to optimize the design variables of an unequal air-gap model. Optimal design variables are the shape of an unequal air-gap and the angle between V-shape magnets. An optimization process is performed by Latin Hypercube Sampling (LHS), Kriging Method, and Genetic Algorithm (GA). Finite element analysis (FEA) is also utilized to analyze the torque and demagnetization characteristics. The torque ripple and the demagnetization temperature of the initial model of 45kW PMSM with unequal air-gap are 10 % and 146.8 degrees, respectively, which are reaching a critical level for electric vehicle application. Therefore, the unequal air-gap model is proposed, and then an optimization process is conducted. Compared to the initial model, the torque ripple of the optimized unequal air-gap model was reduced by 7.7 %. In addition, the demagnetization temperature of the optimized model was also increased by 1.8 % while maintaining the efficiency. From these results, a shape optimized unequal air-gap PMSM has shown the usefulness of an improvement in the torque ripple and demagnetization temperature for the electric vehicle.

Keywords: permanent magnet synchronous motor, optimal design, finite element method, torque ripple

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Authors: H. K. Ratre, M. Roy, S. Roy, M. S. Parmar, V. Bhagat

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Mastitis is a common problem of dairy industries. Reduction in milk production and an irreparable damage to the udder associated with the disease are common causes of culling of dairy cows. Milk from infected animals is not suitable for drinking and for making different milk products. So, it has a major economic importance in dairy cattle. The aims of this study were to investigate the bacteriological panorama in milk from udder quarters with subclinical mastitis and to carried out for the molecular characterization of the major isolated organisms, from subclinical mastitis-affected cows in and around Durg and Rajnandgaon district of Chhattisgarh. Isolation and identification of bacteria from the milk samples of subclinical mastitis-affected cows were done by standard and routine culture procedures. A total of 78 isolates were obtained from cows and among the various bacteria isolated, Staphylococcus spp. occupied prime position with occurrence rate of 51.282%. However, other bacteria isolated includeStreptococcus spp. (20.512%), Micrococcus spp. (14.102%), E. coli (8.974%), Klebsiela spp. (2.564%), Salmonella spp. (1.282%) and Proteus spp. (1.282%). Staphylococcus spp. was isolated as the major causative agent of subclinical mastitis in the studied area. Molecular characterization of Staphylococus aureusisolates was done for genetic expression of the virulence genes like ‘nuc’ encoding thermonucleaseexoenzyme, coa and spa by PCR amplification of the respective genes in 25 Staphylococcus isolates. In the present study, 15 isolates (77.27%) out of 20 coagulase positive isolates were found to be genotypically positive for ‘nuc’ where as 20 isolates (52.63%) out of 38 CNS expressed the presence of the same virulence gene. In the present study, three Staphylococcus isolates were found to be genotypically positive for coa gene. The Amplification of the coa gene yielded two different products of 627, 710 bp. The amplification of the gene segment encoding the IgG binding region of protein A (spa) revealed a size of 220 and 253bp in twostaphylococcus isolates. The X-region binding of the spa gene produced an amplicon of 315 bp in one Staphylococcal isolates. Staphylococcus aureus was found to be major isolate (51.28%) responsible for causing subclinical mastitis in cows which also showed expression of virulence genesnuc, coa and spa.

Keywords: mastitis, bacteria, characterization, expression, gene

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Authors: AbdelRahman A. Faragalla, Mohamed H. Alqhtani, Mohamed M. M.Ahmed

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Saudi Arabia has currently been beset by a barrage of bizarre assemblages of subterranean termite fauna, inflicting heavy catastrophic havocs on human valued properties in various homes, storage facilities, warehouses, agricultural and horticultural crops including okra, sweet pepper, tomatoes, sorghum, date palm trees, citruses and many forest domains and green lush desert oases. The most pressing urgent priority is to use modern technologies to alleviate the painstaking obstacle of taxonomic identification of these injurious noxious pests that might lead to effective pest control in both infested agricultural commodities and field crops. Our study has indicated the use of DNA fingerprinting technologies, in order to generate basic information of the genetic similarity between 3 predominant families containing the most destructive termite species. The methodologies included extraction and DNA isolation from members of the major families and the use of randomly selected primers and PCR amplifications with the nucleotide sequences. GC content and annealing temperatures for all primers, PCR amplifications and agarose gel electrophoresis were also conducted in addition to the scoring and analysis of Random Amplification Polymorphic DNA-PCR (RAPDs). A phylogenetic analysis for different species using statistical computer program on the basis of RAPD-DNA results, represented as a dendrogram based on the average of band sharing ratio between different species. Our study aims to shed more light on this intriguing subject, which may lead to an expedited display of the kinship and relatedness of species in an ambitious undertaking to arrive at correct taxonomic classification of termite species, discover sibling species, so that a logistic rational pest management strategy could be delineated.

Keywords: DNA fingerprinting, Western Saudi Arabia, DNA primers, RAPD

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Authors: Reham Algheshairy, Khaled Tayeb, Christopher Smith, Rebecca Gregg, Haruna Musa

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Background: Diabetes is a medical condition that refers to the pancreas’ inability to secrete sufficient insulin levels, a hormone responsible for controlling glucose levels in the body. Any surplus glucose in the blood stream is excreted through the urinary system. Insulin resistance in blood cells can also cause this condition despite the fact that the pancreas is producing the required amount of insulin A number of researchers claim that the prevalence of diabetes in Saudi Arabia has reached epidemic proportions, although one study did observe one positive in the rise in the awareness of diabetes, possibly indicative of Saudi Arabia’s improving healthcare system. While a number of factors can cause diabetes, the ever-increasing incidence of the disease in Saudi Arabia has been blamed primarily on low levels of physical activity and high levels of obesity. Objectives: The project has two aims. The first aim of the project is to investigate the regulatory effects of consumption of Nigella seeds and Ajwah dates on blood glucose levels in diabetic patients with type 2 diabetes. The second aim of the project is to investigate whether these dietary factors may have potentially beneficial effects in controlling the complications that associated with type 2 diabetes. Methods: This use a random-cross intervention trail of 75 Saudi male and female with type 2 diabetes in Al-Noor hospital in Makkah ( KSA) aged between 18 and 70 years were divided into 3 groups. Group 1 will consume 2g of Nigella Sativa seeds daily along with a modified diet for 12 weeks, group 2 will be given Ajwah dates daily with a modified diet for 12 weeks and group 3 will follow a modified diet for 12 weeks. Anthropometric measurements were taken at baseline, along with bloods for HbA1c, fasting blood sugar and at the end of 12 weeks. Results: This study found significant decrease in blood level (FBG & 2PPBG) and HbA1c in the groups with diet and Nigella seeds) compared to Ajwa date. However, there is no significant change were found in HbA1c, FBG and 2hrpp regarding Ajwa group. Conclusion: This study illustrated a significant improvement in some markers of glycaemia following 2 g of Ns and diet for 12 weeks. The dose of 2g/day of consumed Nigella seeds was found to be more effective in controlling BGL and HbA1c than control and Ajwa groups. This suggests that Nigella seeds and following a diet may have a potential effect (a role in controlling outcomes for type 2 diabetes and controlling the disease). Further research is needed on a large scale to determine the optimum dose and duration of Nigella and Ajwa in order to achieve the desired results.

Keywords: type 2 diabetes, Nigella seeds, Ajwa dates, fasting blood glucose, control

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Authors: Thao Nguyen, Maximiliano Hyon, Sany Rajagukguk, Anna Melkonyan

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Introduction: Type 2 Diabetes is a well-established risk factor for severe SARS-CoV-2 infection. Uncontrolled hyperglycemia in patients with established or newly diagnosed diabetes is associated with poor outcomes, including increased mortality and hospital length of stay. Objectives: Our study aims to compare three different glycemic management strategies and their association with clinical outcomes in patients hospitalized for moderate to severe SARS-CoV-2 infection. Identifying optimal glycemic management strategies will improve the quality of patient care and improve their outcomes. Method: This is a retrospective observational study on patients hospitalized at Adventist Health White Memorial with severe SARS-CoV-2 infection from 11/1/2020 to 02/28/2021. The following inclusion criteria were used: positive SARS-CoV-2 PCR test, age >18 yrs old, diabetes or random glucose >200 mg/dL on admission, oxygen requirement >4L/min, and treatment with glucocorticoids. Our exclusion criteria included: ICU admission within 24 hours, discharge within five days, death within five days, and pregnancy. The patients were divided into three glycemic management groups: Group 1, managed solely by the Primary Team, Group 2, by Pharmacy; and Group 3, by Endocrinologist. Primary outcomes were average glucose on Day 5, change in glucose between Days 3 and 5, and average insulin dose on Day 5 among groups. Secondary outcomes would be upgraded to ICU, inpatient mortality, and hospital length of stay. For statistics, we used IBM® SPSS, version 28, 2022. Results: Most studied patients were Hispanic, older than 60, and obese (BMI >30). It was the first CV-19 surge with the Delta variant in an unvaccinated population. Mortality was markedly high (> 40%) with longer LOS (> 13 days) and a high ICU transfer rate (18%). Most patients had markedly elevated inflammatory markers (CRP, Ferritin, and D-Dimer). These, in combination with glucocorticoids, resulted in severe hyperglycemia that was difficult to control. Average glucose on Day 5 was not significantly different between groups primary vs. pharmacy vs. endocrine (220.5 ± 63.4 vs. 240.9 ± 71.1 vs. 208.6 ± 61.7 ; P = 0.105). Change in glucose from days 3 to 5 was not significantly different between groups but trended towards favoring the endocrinologist group (-26.6±73.6 vs. 3.8±69.5 vs. -32.2±84.1; P= 0.052). TDD insulin was not significantly different between groups but trended towards higher TDD for the endocrinologist group (34.6 ± 26.1 vs. 35.2 ± 26.4 vs. 50.5 ± 50.9; P=0.054). The endocrinologist group used significantly more preprandial insulin compared to other groups (91.7% vs. 39.1% vs. 65.9% ; P < 0.001). The pharmacy used more basal insulin than other groups (95.1% vs. 79.5% vs. 79.2; P = 0.047). There were no differences among groups in the clinical outcomes: LOS, ICU upgrade, or mortality. Multivariate regression analysis controlled for age, sex, BMI, HbA1c level, renal function, liver function, CRP, d-dimer, and ferritin showed no difference in outcomes among groups. Conclusion: Given high-risk factors in our population, despite efforts from the glycemic management teams, it’s unsurprising no differences in clinical outcomes in mortality and length of stay.

Keywords: glycemic management, strategies, hospitalized, SARS-CoV-2, outcomes

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Authors: Oluwafunmibi Omotayo Fasanya, Augustine Kena Adjei

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Background and Aims: This meta-analysis aimed to evaluate the effect of vitamin D supplementation on key metabolic and cardiovascular parameters, such as glycated hemoglobin (HbA1C), fasting blood sugar (FBS), low-density lipoprotein (LDL), high-density lipoprotein (HDL), systolic blood pressure (SBP), and total vitamin D levels in patients with Type 2 diabetes mellitus (T2DM). Methods: A systematic search was performed across databases, including PubMed, Scopus, Embase, Web of Science, Cochrane Library, and ClinicalTrials.gov, from January 1990 to January 2024. A total of 4,177 relevant studies were initially identified. Using an unsupervised K-means clustering algorithm, publications were grouped based on common text features. Maximum entropy classification was then applied to filter studies that matched a pre-identified training set of 139 potentially relevant articles. These selected studies were manually screened for relevance. A parallel manual selection of all initially searched studies was conducted for validation. The final inclusion of studies was based on full-text evaluation, quality assessment, and meta-regression models using random effects. Sensitivity analysis and publication bias assessments were also performed to ensure robustness. Results: The unsupervised K-means clustering algorithm grouped the patients based on their responses to vitamin D supplementation, using key biomarkers such as HbA1C, FBS, LDL, HDL, SBP, and total vitamin D levels. Two primary clusters emerged: one representing patients who experienced significant improvements in these markers and another showing minimal or no change. Patients in the cluster associated with significant improvement exhibited lower HbA1C, FBS, and LDL levels after vitamin D supplementation, while HDL and total vitamin D levels increased. The analysis showed that vitamin D supplementation was particularly effective in reducing HbA1C, FBS, and LDL within this cluster. Furthermore, BMI, weight gain, and disease duration were identified as factors that influenced cluster assignment, with patients having lower BMI and shorter disease duration being more likely to belong to the improvement cluster. Conclusion: The findings of this machine learning-assisted meta-analysis confirm that vitamin D supplementation can significantly improve glycemic control and reduce the risk of cardiovascular complications in T2DM patients. The use of automated screening techniques streamlined the process, ensuring the comprehensive evaluation of a large body of evidence while maintaining the validity of traditional manual review processes.

Keywords: HbA1C, T2DM, SBP, FBS

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Authors: Suganiya Rama Rao, Yoon-Yen Yow, Thor-Seng Liew, Shyamala Ratnayeke

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Invasive snails in the genus Pomacea have spread across Southeast Asia including Peninsular Malaysia. Apart from significant economic costs to wetland crops, very little is known about the snails’ effects on native species, and wetland function through their alteration of macrophyte communities. This study was conducted to establish diagnostic characteristics of Pomacea species in the Malaysian environment using genetic and morphological criteria. Snails were collected from eight localities in northern and central regions of Peninsular Malaysia. The mitochondrial COI gene of 52 adult snails was amplified and sequenced. Maximum likelihood analysis was used to analyse species identity and assess phylogenetic relationships among snails from different geographic locations. Shells of the two species were compared using geometric morphometric analysis and covariance analyses. Shell height accounted for most of the observed variation between P. canaliculata and P. maculata, with the latter possessing a smaller mean ratio of shell height: aperture height (p < 0.0001) and shell height to shell width (give p < 0.0001). Genomic and phylogenetic analysis demonstrated the presence of two monophyletic taxa, P. canaliculata and P. maculata, in Peninsular Malaysia samples. P. maculata co-occurred with P. canaliculata in 5 localities, but samples from 3 localities contained only P. canaliculata. This study is the first to confirm the presence of two of the most invasive species of Pomacea in Peninsular Malaysia using a genomic approach. P. canaliculata appears to be the more widespread species. Despite statistical differences, both quantitative and qualitative morphological characteristics demonstrate much interspecific overlap and intraspecific variability; thus morphology alone cannot reliably verify species identity. Molecular techniques for distinguishing between these two highly invasive Pomacea species are needed to understand their specific ecological niches and develop effective protocols for their management.

Keywords: Pomacea canaliculata, Pomacea maculata, invasive species, phylog enetic analysis, geometric morphometric analysis

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Authors: Altilio Rosa, Chirico Francesco, Foglia Goffredo

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In recent years, the number of Unmanned Aerial Vehicles (UAVs) has significantly increased. The rapid development of commercial and recreational drones makes them an important part of our society. Despite the growing list of their applications, these vehicles pose a huge threat to civil and military installations: detection, classification and neutralization of such flying objects become an urgent need. Radar is an effective remote sensing tool for detecting and tracking flying objects, but scenarios characterized by the presence of a high number of tracks related to flying birds make especially challenging the drone detection task: operator PPI is cluttered with a huge number of potential threats and his reaction time can be severely affected. Flying birds compared to UAVs show similar velocity, RADAR cross-section and, in general, similar characteristics. Building from the absence of a single feature that is able to distinguish UAVs and birds, this paper uses a multiple features approach where an original feature selection technique is developed to feed binary classifiers trained to distinguish birds and UAVs. RADAR tracks acquired on the field and related to different UAVs and birds performing various trajectories were used to extract specifically designed target movement-related features based on velocity, trajectory and signal strength. An optimization strategy based on a genetic algorithm is also introduced to select the optimal subset of features and to estimate the performance of several classification algorithms (Neural network, SVM, Logistic regression…) both in terms of the number of selected features and misclassification error. Results show that the proposed methods are able to reduce the dimension of the data space and to remove almost all non-drone false targets with a suitable classification accuracy (higher than 95%).

Keywords: birds, classification, machine learning, UAVs

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Authors: C. Villamizar, M. Serrato

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In the pursuit of athletic performance, the role of physical training which is determined by a number of charges or taxes on physiological stress and musculoskeletal systems of the human body generated by the intensity and duration is fundamental. Given the physical demands of these activities both training and competitive must take into account the optimal relationship with a straining process recovery post favoring the process of overcompensation which aims to facilitate the return and rising energy potential and protein synthesis also of different tissues. Allowing muscle function returns to baseline or pre-exercise states. If this recovery process is not performed or is not allowed in a proper way, will result in an increased state of fatigue. Active recovery, is one of the strategies implemented in the sport for a return to pre-exercise physiological states. However, there are some adverse assumptions regarding the negative effects, as is the possibility of increasing the degradation of muscle glycogen and thus delaying the synthesis thereof. For them, it is necessary to investigate what would be the effects generated application made at different times after the effort. The aim of this study was to determine the effects of active recovery post effort made at three different times: immediately, at 12 and 24 hours on biochemical markers creatine kinase in youth soccer player’s categories. A randomized controlled trial with allocation to three groups was performed: A. active recovery immediately after the effort; B. active recovery performed at 12 hours after the effort; C. active recovery made at 24 hours after the effort. This study included 27 subjects belonging to a Colombian soccer team of the second division. Vital signs, weight, height, BMI, the percentage of muscle mass, fat mass percentage, personal medical history, and family were valued. The velocity, explosive force and Creatin Kinase (CK) in blood were tested before and after interventions. SAFT 90 protocol (Soccer Field specific Aerobic Test) was applied to participants for generating fatigue. CK samples were taken one hour before the application of the fatigue test, one hour after the fatigue protocol and 48 of the initial CK sample. Mean age was 18.5 ± 1.1 years old. Improvements in jumping and speed recovery the 3 groups (p < 0.05), but no statistically significant differences between groups was observed after recuperation. In all participants, there was a significant increment of CK when applied SAFT 90 in all the groups (median 103.1-111.1). The CK measurement after 48 hours reflects a recovery in all groups, however the group C, a decline below baseline levels of -55.5 (-96.3 /-20.4) which is a significant find. Other research has shown that CK does not return quickly to their baseline, but our study shows that active recovery favors the clearance of CK and also to perform recovery 24 hours after the effort generates higher clearance of this biomarker.

Keywords: active recuperation, creatine phosphokinase, post training, young soccer players

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Authors: Saule Mussabekova

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Recent advances in genetics have allowed increasing acutely the capacities of the formation of reliable evidence in conducting forensic examinations. Thus, traces of biological origin are important sources of information about a crime. Currently, around the world, sexual offenses have increased, and among them are those in which the criminals use various detergents to remove traces of their crime. A feature of modern synthetic detergents is the presence of biological additives - enzymes. Enzymes purposefully destroy stains of biological origin. To study the nature and extent of the impact of modern washing powders on saliva stains on the physical evidence, specially prepared test specimens of different types of tissues to which saliva was applied have been examined. Materials and Methods: Washing machines of famous manufacturers of household appliances have been used with different production characteristics and advertised brands of washing powder for test washing. Over 3,500 experimental samples were tested. After washing, the traces of saliva were identified using modern research methods of forensic medicine. Results: The influence was tested and the dependence of the use of different washing programs, types of washing machines and washing powders in the process of establishing saliva trace and identify of the stains on the physical evidence while washing was revealed. The results of experimental and practical expert studies have shown that in most cases it is not possible to draw the conclusions in the identification of saliva traces on physical evidence after washing. This is a consequence of the effect of biological additives and other additional factors on traces of saliva during washing. Conclusions: On the basis of the results of the study, the feasibility of saliva traces of the stains on physical evidence after washing is established. The use of modern molecular genetic methods makes it possible to partially solve the problems arising in the study of unlaundered evidence. Additional study of physical evidence after washing facilitates detection and investigation of sexual offenses against women and children.

Keywords: saliva research, modern synthetic detergents, laundry detergents, forensic medicine

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Authors: Hannah S. Elizabeth, D. Gnanasekaran, M. R. Manju Gowda, Antony George

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Phytoremediation a new technology of remediating the contaminated soil, water and air using plants and serves as a green technology with environmental friendly approach. The main aim of this technique is cleansing and detoxifying of organic compounds, organo-phosphorous pesticides, heavy metals like arsenic, iron, cadmium, gold, radioactive elements which cause teratogenic and life threatening diseases to mankind and animal kingdom when consume the food crops, vegetables, fruits, cerals, and millets obtained from the contaminated soil. Also, directly they may damage the genetic materials thereby alters the biosynthetic pathways of secondary metabolites and other phytoconstituents which may have different pharmacological activities which lead to lost their efficacy and potency as well. It would reflect in mutagenicity, drug resistance and affect other antagonistic properties of normal metabolism. Is the technology for real clean-up of contaminated soils and the contaminants which are potentially toxic. It reduces the risks produced by a contaminated soil by decreasing contaminants using plants as a source. The advantages are cost-effectiveness and less ecosystem disruption. Plants may also help to stabilize contaminants by accumulating and precipitating toxic trace elements in the roots. Organic pollutants can potentially be chemically degraded and ultimately mineralized into harmless biological compounds. Hence, the use of plants to revitalize contaminated sites is gaining more attention and preferred for its cost-effective when compared to other chemical methods. The introduction of harmful substances into the environment has been shown to have many adverse effects on human health, agricultural productivity, and natural ecosystems. Because the costs of growing a crop are minimal compared to those of soil removal and replacement, the use of plants to remediate hazardous soils is seen as having great promise.

Keywords: cost effective, eco-friendly, phytoremediation, secondary metabolites

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Authors: Ajay Gogia, Svs Deo, Dn Sharma, Atul Batra, Ashutash Mishra

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Background and Aims: Breast cancer in women aged less than 25 years (defined as very young breast cancer, VYBC) is rare and accounts for 0.25% of all breast cancer in the West. There is no data available on VYBC from developing countries. The aim of this study was to analyze the clinical, pathological, and prognostic factors and outcomes in VYBC. Methods: This retrospective analysis was performed on 80 patients aged 25 years or less (screened 8000 files of female BC) who were registered at All India Institute of Medical Sciences (AIIMS), New Delhi, India, over a 15-year period between 2011 and 2023. Results: The median age was 21.5 years (range 16-25). A positive family history (siblings and parents) was elicited in 30% of cases, and breast cancer gene (BRCA1/2) mutation was found in 33% of cases patients. Ten patients (12.5%) patients have pregnancy-associated breast cancer (BC detected during pregnancy or 1 year after postpartum period). The TNM stage distribution was Stage I was 0, stage II -30%, stage III –60% and Stage IV -10 %patients. Seventy percent of tumors were high grade, and 90% had pathological node-positive disease. Estrogen, Progesterone, and human epidermal growth factor receptor 2 (HER2)/neu positivity were 25%,25% and 35%, respectively. Triple-negative breast cancer constituted 40% of patients. With a median follow-up of 42 months, 3 years, relapse-free survival (nonmetastatic disease), progression-free survival (metastatic disease) and overall survival were 30%, 15% and 50%, respectively. Conclusions: Very young women constituted 1% of all breast cancer cases. Advanced disease at presentation and high-risk pathological features result in poor outcomes. One-third of VYBCs are associated with BRCA mutation, which requires genetic counseling and risk reduction surgery if required. Due to the aggressive behavior of BC in this age group, need early diagnosis and prompt treatment

Keywords: very young, breast cancer, outcome, developing country, India

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Authors: Titus A. Beu

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Many modern gene-delivery protocols rely on condensed complexes of DNA with polycations to introduce the genetic payload into cells by endocytosis. In particular, polyethyleneimine (PEI) stands out by a high buffering capacity (enabling the efficient condensation of DNA) and relatively simple fabrication. Realistic computational studies can offer essential insights into the formation process of DNA-PEI polyplexes, providing hints on efficient designs and engineering routes. We present comprehensive computational investigations of solvated PEI and DNA-PEI polyplexes involving calculations at three levels: ab initio, all-atom (AA), and coarse-grained (CG) molecular mechanics. In the first stage, we developed a rigorous AA CHARMM (Chemistry at Harvard Macromolecular Mechanics) force field (FF) for PEI on the basis of accurate ab initio calculations on protonated model pentamers. We validated this atomistic FF by matching the results of extensive molecular dynamics (MD) simulations of structural and dynamical properties of PEI with experimental data. In a second stage, we developed a CG MARTINI FF for PEI by Boltzmann inversion techniques from bead-based probability distributions obtained from AA simulations and ensuring an optimal match between the AA and CG structural and dynamical properties. In a third stage, we combined the developed CG FF for PEI with the standard MARTINI FF for DNA and performed comprehensive CG simulations of DNA-PEI complex formation and condensation. Various technical aspects which are crucial for the realistic modeling of DNA-PEI polyplexes, such as options of treating electrostatics and the relevance of polarizable water models, are discussed in detail. Massive CG simulations (with up to 500 000 beads) shed light on the mechanism and provide time scales for DNA polyplex formation independence of PEI chain size and protonation pattern. The DNA-PEI condensation mechanism is shown to primarily rely on the formation of DNA bundles, rather than by changes of the DNA-strand curvature. The gained insights are expected to be of significant help for designing effective gene-delivery applications.

Keywords: DNA condensation, gene-delivery, polyethylene-imine, molecular dynamics.

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Authors: Ipsita Biswas, Arnab Sarkar, Ashikur Rahaman, Gopeswar Mukherjee, Subhrangsu Chatterjee, Shamee Bhattacharjee, Deba Prasad Mandal

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One of the major reasons for the high mortality rate of lung cancer is the substantial delays in disease detection at late metastatic stages. It is of utmost importance to understand the detailed molecular signaling and detect the molecular markers that can be used for the early diagnosis of cancer. Several studies explored the emerging roles of long noncoding RNAs (lncRNAs) in various cancers as well as lung cancer. A long non-coding RNA LINC00273 was recently discovered to promote cancer cell migration and invasion, and its positive correlation with the pathological stages of metastasis may prove it to be a potential target for inhibiting cancer cell metastasis. Comparing real-time expression of LINC00273 in various human clinical cancer tissue samples with normal tissue samples revealed significantly higher expression in cancer tissues. This long intergenic noncoding RNA was found to be highly expressed in human liver tumor-initiating cells, human gastric adenocarcinoma AGS cell line, as well as human non-small cell lung cancer A549 cell line. SiRNA and shRNA-induced knockdown of LINC00273 in both in vitro and in vivo nude mice significantly subsided AGS and A549 cancer cell migration and invasion. LINC00273 knockdown also reduced TGF-β induced SNAIL, SLUG, VIMENTIN, ZEB1 expression, and metastasis in A549 cells. Plenty of reports have suggested the role of microRNAs of the miR200 family in reversing epithelial to mesenchymal transition (EMT) by inhibiting ZEB transcription factors. In this study, hsa-miR-200a-3p was predicted via IntaRNA-Freiburg RNA tools to be a potential target of LINC00273 with a negative free binding energy of −8.793 kcal/mol, and this interaction was verified as a confirmed target of LINC00273 by RNA pulldown, real-time PCR and luciferase assay. Mechanistically, LINC00273 accelerated TGF-β induced EMT by sponging hsa-miR-200a-3p which in turn liberated ZEB1 and promoted prometastatic functions in A549 cells in vitro as verified by real-time PCR and western blotting. The similar expression patterns of these EMT regulatory pathway molecules, viz. LINC00273, hsa-miR-200a-3p, ZEB1 and TGF-β, were also detected in various clinical samples like breast cancer tissues, oral cancer tissues, lung cancer tissues, etc. Overall, this LINC00273 mediated EMT regulatory signaling can serve as a potential therapeutic target for the prevention of lung cancer metastasis.

Keywords: epithelial to mesenchymal transition, long noncoding RNA, microRNA, non-small-cell lung carcinoma

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Authors: Z. Karahaliloğlu, E. Yalçın, M. Demirbilek, E.B. Denkbaş

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Critical-sized bone defects caused by trauma, bone diseases, prosthetic implant revision or tumor excision cannot be repaired by physiological regenerative processes. Current orthopedic applications for critical-sized bone defects are to use autologous bone grafts, bone allografts, or synthetic graft materials. However, these strategies are unable to solve completely the problem, and motivate the development of novel effective biological scaffolds for tissue engineering applications and regenerative medicine applications. In particular, scaffolds combined with a variety of bio-agents as fundamental tools emerge to provide the regeneration of damaged bone tissues due to their ability to promote cell growth and function. In this study, a magnetic silk fibroin (SF) hydrogel scaffold was prepared by electrogelation process of the concentrated Bombxy mori silk fibroin (8 %wt) aqueous solution. For enhancement of osteoblast-like cells (SaOS-2) growth and adhesion, basal fibroblast growth factor (bFGF) were conjugated physically to the HSA-coated magnetic nanoparticles (Fe3O4) and magnetic SF e-gel scaffolds were prepared by incorporation of Fe3O4, HSA (human serum albumin)=Fe3O4 and HSA=Fe3O4-bFGF nanoparticles. HSA=Fe3O4, HSA=Fe3O4-bFGF loaded and bare SF e-gels scaffolds were characterized using scanning electron microscopy (SEM.) For cell studies, human osteoblast-like cell line (SaOS-2) was used and an MTT assay was used to assess the cytotoxicity of magnetic silk fibroin e-gel scaffolds and cell density on these surfaces. For the evaluation osteogenic activation, ALP (alkaline phosphatase), the amount of mineralized calcium, total protein and collagen were studied. Fe3O4 nanoparticles were successfully synthesized and bFGF was conjugated to HSA=Fe3O4 nanoparticles with %97.5 of binding yield which has a particle size of 71.52±2.3 nm. Electron microscopy images of the prepared HSA and bFGF incorporated SF e-gel scaffolds showed a 3D porous morphology. In terms of water uptake results, bFGF conjugated HSA=Fe3O4 nanoparticles has the best water absorbability behavior among all groups. In the in-vitro cell culture studies realized using SaOS-2 cell line, the coating of Fe3O4 nanoparticles surface with a protein enhance the cell viability and HSA coating and bFGF conjugation, the both have an inductive effect in the cell proliferation. One of the markers of bone formation and osteoblast differentiation, according to the ALP activity and total protein results, HSA=Fe3O4-bFGF loaded SF e-gels had significantly enhanced ALP activity. Osteoblast cultured HSA=Fe3O4-bFGF loaded SF e-gels deposited more calcium compared with SF e-gel. The proposed magnetic scaffolds seem to be promising for bone tissue regeneration and used in future work for various applications.

Keywords: basic fibroblast growth factor (bFGF), e-gel, iron oxide nanoparticles, silk fibroin

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Authors: Antonio Munar-Frau, Sascha Klismoch, Manfred Schmolz, Federico Hernandez-Alfaro, Jordi Caballe-Serrano

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Introduction: Biocompatibility of biomaterials has been proposed as one of the main criteria for treatment success. For guided bone regeneration (GBR), barrier membranes present a conflict given the number of origins and modifications of these materials. The biologic response to biomaterials is orchestrated by a series of events leading to the integration or rejection of the biomaterial, posing questions such as if a longer occlusive property may trigger an inflammatory reaction. Whole blood cultures are a solution to study the immune response to drugs or biomaterials during the first 24-48 hours. The aim of this study is to determine the early immune response of different origins and chemical modifications of barrier membranes. Materials & Methods: 5 different widely used barrier membranes were included in this study: Acellular dermal matrix (AlloDerm, LifeCell®), Porcine Peritoneum (BioGide, Geistlich Pharma®), Porcine Pericardium (Jason, Botiss Biomaterials GmbH®), Porcine Cross-linked collagen (Ossix Plus, Datum Dental®) and d-PTFE (Cytoplast TXT, Osteogenics Biomedical®). Blood samples were extracted from 3 different healthy donors and incubated with the different samples of barrier membranes for 24 hours. After the incubation time, serum samples were obtained and analyzed by means of biocompatibility assays taking into account 42 markers. Results: In an early stage of the inflammatory response, the Acellular dermal matrix, porcine peritoneum and porcine cross-linked collagen expressed similar patterns of cytokine expression with a great manifestation of ENA 78. Porcine pericardium and d-PTFE presented similar cytokine activation, especially for MMP-3 and MMP-9, although other cytokines were highlighted with lower expression. For the later immune response, Porcine peritoneum and acellular dermal matrix MCP-1 and IL-15 were evident. Porcine pericardium, porcine cross-linked collagen and d-PTFE presented a high expression of IL-16 and lower manifestation of other cytokines. Different behaviors depending on an earlier or later stage of the inflammation process were observed. Barrier membrane inflammatory expression does not only differ depending on the origin, variables such as treatment of the collagen and polymers may also have a great impact on the cytokine expression of the studied barrier membranes during inflammation. Conclusions: Surface treatment and modifications might affect the biocompatibility of the membranes, as different cytokine expressions were evidently depending on the origin of the biomaterial. This study is only a brushstroke regarding the biocompatibility of materials, as it is one of the pioneer studies for ex vivo barrier membranes assays. Studies regarding surface modification are needed in order to clarify mystifications of barrier membrane science.

Keywords: biomaterials, bone regeneration, biocompatibility, inflammation

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