Search results for: genetic diversity and viral proteins
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 4214

Search results for: genetic diversity and viral proteins

794 Understanding the Role of Nitric Oxide Synthase 1 in Low-Density Lipoprotein Uptake by Macrophages and Implication in Atherosclerosis Progression

Authors: Anjali Roy, Mirza S. Baig

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Atherosclerosis is a chronic inflammatory disease characterized by the formation of lipid rich plaque enriched with necrotic core, modified lipid accumulation, smooth muscle cells, endothelial cells, leucocytes and macrophages. Macrophage foam cells play a critical role in the occurrence and development of inflammatory atherosclerotic plaque. Foam cells are the fat-laden macrophages in the initial stage atherosclerotic lesion formation. Foam cells are an indication of plaque build-up, or atherosclerosis, which is commonly associated with increased risk of heart attack and stroke as a result of arterial narrowing and hardening. The mechanisms that drive atherosclerotic plaque progression remain largely unknown. Dissecting the molecular mechanism involved in process of macrophage foam cell formation will help to develop therapeutic interventions for atherosclerosis. To investigate the mechanism, we studied the role of nitric oxide synthase 1(NOS1)-mediated nitric oxide (NO) on low-density lipoprotein (LDL) uptake by bone marrow derived macrophages (BMDM). Using confocal microscopy, we found that incubation of macrophages with NOS1 inhibitor, TRIM (1-(2-Trifluoromethylphenyl) imidazole) or L-NAME (N omega-nitro-L-arginine methyl ester) prior to LDL treatment significantly reduces the LDL uptake by BMDM. Further, addition of NO donor (DEA NONOate) in NOS1 inhibitor treated macrophages recovers the LDL uptake. Our data strongly suggest that NOS1 derived NO regulates LDL uptake by macrophages and foam cell formation. Moreover, we also checked proinflammatory cytokine mRNA expression through real time PCR in BMDM treated with LDL and copper oxidized LDL (OxLDL) in presences and absences of inhibitor. Normal LDL does not evoke cytokine expression whereas OxLDL induced proinflammatory cytokine expression which significantly reduced in presences of NOS1 inhibitor. Rapid NOS-1-derived NO and its stable derivative formation act as signaling agents for inducible NOS-2 expression in endothelial cells, leading to endothelial vascular wall lining disruption and dysfunctioning. This study highlights the role of NOS1 as critical players of foam cell formation and would reveal much about the key molecular proteins involved in atherosclerosis. Thus, targeting NOS1 would be a useful strategy in reducing LDL uptake by macrophages at early stage of disease and hence dampening the atherosclerosis progression.

Keywords: atherosclerosis, NOS1, inflammation, oxidized LDL

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793 Development of Power System Stability by Reactive Power Planning in Wind Power Plant With Doubley Fed Induction Generators Generator

Authors: Mohammad Hossein Mohammadi Sanjani, Ashknaz Oraee, Oriol Gomis Bellmunt, Vinicius Albernaz Lacerda Freitas

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The use of distributed and renewable sources in power systems has grown significantly, recently. One the most popular sources are wind farms which have grown massively. However, ¬wind farms are connected to the grid, this can cause problems such as reduced voltage stability, frequency fluctuations and reduced dynamic stability. Variable speed generators (asynchronous) are used due to the uncontrollability of wind speed specially Doubley Fed Induction Generators (DFIG). The most important disadvantage of DFIGs is its sensitivity to voltage drop. In the case of faults, a large volume of reactive power is induced therefore, use of FACTS devices such as SVC and STATCOM are suitable for improving system output performance. They increase the capacity of lines and also passes network fault conditions. In this paper, in addition to modeling the reactive power control system in a DFIG with converter, FACTS devices have been used in a DFIG wind turbine to improve the stability of the power system containing two synchronous sources. In the following paper, recent optimal control systems have been designed to minimize fluctuations caused by system disturbances, for FACTS devices employed. For this purpose, a suitable method for the selection of nine parameters for MPSH-phase-post-phase compensators of reactive power compensators is proposed. The design algorithm is formulated ¬¬as an optimization problem searching for optimal parameters in the controller. Simulation results show that the proposed controller Improves the stability of the network and the fluctuations are at desired speed.

Keywords: renewable energy sources, optimization wind power plant, stability, reactive power compensator, double-feed induction generator, optimal control, genetic algorithm

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792 Indoor and Outdoor Forest Farming for Year-Round Food and Medicine Production, Carbon Sequestration, Soil-Building, and Climate Change Mitigation

Authors: Jerome Osentowski

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The objective at Central Rocky Mountain Permaculture Institute has been to put in practice a sustainable way of life while growing food, medicine, and providing education. This has been done by applying methods of farming such as agroforestry, forest farming, and perennial polycultures. These methods have been found to be regenerative to the environment through carbon sequestration, soil-building, climate change mitigation, and the provision of food security. After 30 years of implementing carbon farming methods, the results are agro-diversity, self-sustaining systems, and a consistent provision of food and medicine. These results are exhibited through polyculture plantings in an outdoor forest garden spanning roughly an acre containing about 200 varieties of fruits, nuts, nitrogen-fixing trees, and medicinal herbs, and two indoor forest garden greenhouses (one Mediterranean and one Tropical) containing about 50 varieties of tropical fruits, beans, herbaceous plants and more. While the climate zone outside the greenhouse is 6, the tropical forest garden greenhouse retains an indoor climate zone of 11 with near-net-zero energy consumption through the use of a climate battery, allowing the greenhouse to serve as a year-round food producer. The effort to source food from the forest gardens is minimal compared to annual crop production. The findings at Central Rocky Mountain Permaculture Institute conclude that agroecological methods are not only beneficial but necessary in order to revive and regenerate the environment and food security.

Keywords: agroecology, agroforestry, carbon farming, carbon sequestration, climate battery, food security, forest farming, forest garden, greenhouse, near-net-zero, perennial polycultures

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791 Congolese Wood in the Antwerp Interwar Interior

Authors: M. Jaenen, M. de Bouw, A. Verdonck, M. Leus

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During the interwar period artificial materials were often preferred, but many Antwerp architects relied on the application of wood for most of the interior finishing works and furnishings. Archival, literature and on site research of interwar suburban townhouses and the Belgian wood and furniture industry gave a new insight to the application of wood in the interwar interior. Many interwar designers favored the decorative values in all treatments of wood because of its warmth, comfort, good-wearing, and therefore, economic qualities. For the creation of a successful modern interior the texture and surface of the wood becomes as important as the color itself. This aesthetics valuation was the result of the modernization of the wood industry. The development of veneer and plywood gave the possibility to create strong, flat, long and plain wooden surfaces which are capable of retaining their shape. Also the modernization of cutting machines resulted in high quality and diversity in texture of veneer. The flat and plain plywood surfaces were modern decorated with all kinds of veneer-sliced options. In addition, wood species from the former Belgian Colony Congo were imported. Limba (Terminalia superba), kambala (Chlorophora excelsa), mubala (Pentaclethra macrophylla) and sapelli (Entandrophragma cylindricum) were used in the interior of many Antwerp interwar suburban town houses. From the thirties onwards Belgian wood firms established modern manufactures in Congo. There the local wood was dried, cut and prepared for exportation to the harbor of Antwerp. The presence of all kinds of strong and decorative Congolese wood products supported its application in the interwar interior design. The Antwerp architects combined them in their designs for doors, floors, stairs, built-in-furniture, wall paneling and movable furniture.

Keywords: Antwerp, congo, furniture, interwar

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790 The Gray Dance - An Analysis of Ageism in Dance

Authors: Paula Higa

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This paper briefly examines age and its impact on a dancer’s performance career. An investigation into the possible reasons why audiences don’t regularly see veteran dancers on stage (termed as the Gray Dancer) supports the research. This analysis reflects some of the social dynamics that shape perceptions of the aging body in the U.S., as well as the correlation between the meaning of old and decay in Western culture. The primary question addressed in this research asks, who has the prerogative to determine when a dancer should stop dancing - society or the dancer themselves The aging process can significantly shorten a performer's professional career. The body has less endurance and is more susceptible to injuries, fatigue, etc. It also becomes less flexible and loses muscular strength and tone. A reduction in physical skills may usher gray dancers to embrace an ideology of shorter careers. However, in today’s age of diversity, equity, and inclusion, the realm of dance performance should reflect the times in which it is rooted; a multi-generational environment where people interact and participate in all of life's events. Overall, this study champions the inclusion of gray dancers as representations of mastery and wisdom akin to those traits associated with age and experience across various professions. Dance is an art form that transcends the assumptions of youthful beauty and physical ability. It serves as a conduit for conveying a lifetime of experiences, emotions, and ideas through the expressive vehicle of the body. Furthermore, it presents audiences with a medium to perceive and comprehend both themselves and life itself, echoing Noverre's insightful contemplation. The essay underscores the importance of valuing, sensing, and appreciating the richness that gray dancers bring to the stage by delving into segments of dance history and analyzing the possible influence of curators, directors, audiences, and society in general on ageism in dance.

Keywords: dance, ageism, politics in dance, curatorial process

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789 Immuno-Protective Role of Mucosal Delivery of Lactococcus lactis Expressing Functionally Active JlpA Protein on Campylobacter jejuni Colonization in Chickens

Authors: Ankita Singh, Chandan Gorain, Amirul I. Mallick

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Successful adherence of the mucosal epithelial cells is the key early step for Campylobacter jejuni pathogenesis (C. jejuni). A set of Surface Exposed Colonization Proteins (SECPs) are among the major factors involved in host cell adherence and invasion of C. jejuni. Among them, constitutively expressed surface-exposed lipoprotein adhesin of C. jejuni, JlpA, interacts with intestinal heat shock protein 90 (hsp90α) and contributes in disease progression by triggering pro-inflammatory response via activation of NF-κB and p38 MAP kinase pathway. Together with its ability to express in the bacterial surface, higher sequence conservation and predicted predominance of several B cells epitopes, JlpA protein reserves its potential to become an effective vaccine candidate against wide range of Campylobacter sps including C. jejuni. Given that chickens are the primary sources for C. jejuni and persistent gut colonization remain as major cause for foodborne pathogenesis to humans, present study explicitly used chickens as model to test the immune-protective efficacy of JlpA protein. Taking into account that gastrointestinal tract is the focal site for C. jejuni colonization, to extrapolate the benefit of mucosal (intragastric) delivery of JlpA protein, a food grade Nisin inducible Lactic acid producing bacteria, Lactococcus lactis (L. lactis) was engineered to express recombinant JlpA protein (rJlpA) in the surface of the bacteria. Following evaluation of optimal surface expression and functionality of recombinant JlpA protein expressed by recombinant L. lactis (rL. lactis), the immune-protective role of intragastric administration of live rL. lactis was assessed in commercial broiler chickens. In addition to the significant elevation of antigen specific mucosal immune responses in the intestine of chickens that received three doses of rL. lactis, marked upregulation of Toll-like receptor 2 (TLR2) gene expression in association with mixed pro-inflammatory responses (both Th1 and Th17 type) was observed. Furthermore, intragastric delivery of rJlpA expressed by rL. lactis, but not the injectable form, resulted in a significant reduction in C. jejuni colonization in chickens suggesting that mucosal delivery of live rL. lactis expressing JlpA serves as a promising vaccine platform to induce strong immune-protective responses against C. jejuni in chickens.

Keywords: chickens, lipoprotein adhesion of Campylobacter jejuni, immuno-protection, Lactococcus lactis, mucosal delivery

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788 Phytochemistry and Alpha-Amylase Inhibitory Activities of Rauvolfia vomitoria (Afzel) Leaves and Picralima nitida (Stapf) Seeds

Authors: Oseyemi Omowunmi Olubomehin, Olufemi Michael Denton

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Diabetes mellitus is a disease that is related to the digestion of carbohydrates, proteins and fats and how this affects the blood glucose levels. Various synthetic drugs employed in the management of the disease work through different mechanisms. Keeping postprandial blood glucose levels within acceptable range is a major factor in the management of type 2 diabetes and its complications. Thus, the inhibition of carbohydrate-hydrolyzing enzymes such as α-amylase is an important strategy in lowering postprandial blood glucose levels, but synthetic inhibitors have undesirable side effects like flatulence, diarrhea, gastrointestinal disorders to mention a few. Therefore, it is necessary to identify and explore the α-amylase inhibitors from plants due to their availability, safety, and low costs. In the present study, extracts from the leaves of Rauvolfia vomitoria and seeds of Picralima nitida which are used in the Nigeria traditional system of medicine to treat diabetes were tested for their α-amylase inhibitory effect. The powdered plant samples were subjected to phytochemical screening using standard procedures. The leaves and seeds macerated successively using n-hexane, ethyl acetate and methanol resulted in the crude extracts which at different concentrations (0.1, 0.5 and 1 mg/mL) alongside the standard drug acarbose, were subjected to α-amylase inhibitory assay using the Benfield and Miller methods, with slight modification. Statistical analysis was done using ANOVA, SPSS version 2.0. The phytochemical screening results of the leaves of Rauvolfia vomitoria and the seeds of Picralima nitida showed the presence of alkaloids, tannins, saponins and cardiac glycosides while in addition Rauvolfia vomitoria had phenols and Picralima nitida had terpenoids. The α-amylase assay results revealed that at 1 mg/mL the methanol, hexane, and ethyl acetate extracts of the leaves of Rauvolfia vomitoria gave (15.74, 23.13 and 26.36 %) α-amylase inhibitions respectively, the seeds of Picralima nitida gave (15.50, 30.68, 36.72 %) inhibitions which were not significantly different from the control at p < 0.05, while acarbose gave a significant 56 % inhibition at p < 0.05. The presence of alkaloids, phenols, tannins, steroids, saponins, cardiac glycosides and terpenoids in these plants are responsible for the observed anti-diabetic activity. However, the low percentages of α-amylase inhibition by these plant samples shows that α-amylase inhibition is not the major way by which both plants exhibit their anti-diabetic effect.

Keywords: alpha-amylase, Picralima nitida, postprandial hyperglycemia, Rauvolfia vomitoria

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787 Establishing a Genetic Link between Fat Mass and Obesity Associated and Vitamin D Receptor Gene Polymorphisms and Obesity in the Emirati Population

Authors: Saad Mahmud Khan, Sarah El Hajj Chehadeh, Mehera Abdulrahman, Wael Osman, Habiba Al Safar

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Obesity is a non-communicable disease that is widely prevalent with approximately 600 million people classified as obese worldwide. Its etiology is multifactorial and involves a complex interplay between genes and the environment. Over the past few decades, obesity rates among the Emirati population have been increasing. The aim of this study was to investigate the association of candidate gene single nucleotide polymorphisms (SNPs), namely the fat mass and obesity associated (FTO) gene SNP rs9939609 and Vitamin D Receptor (VDR) gene SNP rs1544410, with obesity in the UAE population. Methods: This is a case-control study in which 414 individuals were enrolled during their routine visit to endocrinology clinics in Abu Dhabi, United Arab Emirates between the period of June 2012 and December 2013. Several biochemical tests and clinical assessments along with a lifestyle questionnaire for each participant were completed at the clinic. Genomic DNA was extracted from saliva samples of 201 obese, 114 overweight and 99 normal subjects. Genotyping for the variants was performed using TaqMan assay. Results: The mean Body Mass Index (BMI) ± SD for the obese, overweight, and normal subjects was 35.76 ± 4.54, 27.53 ± 1.45 and 22.69 ± 1.84 kg/m2, respectively. Increasing BMI values were associated with an increase in values for systolic blood pressure, diastolic blood pressure, HbA1c, and triglycerides. The SNP rs9939609 in the FTO gene was found to be significantly associated with the BMI (p=0.028), with the minor allele A having a clear additive effect on BMI values. No significant association was detected between BMI and rs1544410 of the VDR gene. Conclusions: Our study findings indicate that the minor allele A of the rs9939609 has a significant association with increasing BMI values. In addition, our findings support the fact that increasing BMI is associated with increasing risks of other comorbidities such as higher blood pressure, poorer glycemic control and higher triglycerides.

Keywords: body mass index, FTO gene, obesity, rs9939609, United Arab Emirates

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786 Enhancement Effect of Compound 4-Hydroxybenzoic Acid from Petung Bamboo (Dendrocalamus Asper) Shoots on α1β2γ2S of GABA (A) Receptor Expressed in Xenopus laevis Oocytes- Preliminary Study on Its Anti-Epileptic Potential

Authors: Muhammad Bilal, Amelia Jane Llyod, Habsah Mohamad, Jia Hui Wong, Abdul Aziz Mohamed Yusoff, Jafri Malin Abdullah, Jingli Zhang

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Epilepsy is one of the major brain afflictions occurs with uncontrolled excitation of cortex; disturbed 50 million of world’s population. About 25 percent of patients subjected to adverse effects from antiepileptic drugs (AEDs) such as depression, nausea, tremors, gastrointestinal symptoms, osteoporosis, dizziness, weight change, drowsiness, fatigue are commonly observed indications; therefore, new drugs are required to cure epilepsy. GABA is principle inhibitory neurotransmitter, control excitation of the brain. Mutation or dysfunction of GABA receptor is one of the primary causes of epilepsy, which is confirmed from many acquired models of epilepsy like traumatic brain injury, kindling, and status epilepticus models of epilepsy. GABA receptor has 3 distinct types such as GABA (A), GABA (B), GABA(C).GABA (A) receptor has 20 different subunits, α1β2γ2 subunits composition of GABA (A) receptor is the most used combination of subunits for screening of compounds against epilepsy. We expressed α1β2γ2s subunits of GABA (A) Receptor in Xenopus leavis oocytes and examined the enhancement potential of 4-Hydroxybenzoic acid compound on GABA (A) receptor via two-electrode voltage clamp current recording technique. Bamboo shoots are the young, tender offspring of bamboo, which are usually harvested after a cultivating period of 2 weeks. Proteins, acids, fat, starch, carbohydrate, fatty acid, vitamin, dietary fiber, and minerals are the major constituent found systematically in bamboo shoots. These shoots reported to have anticancer, antiviral, antibacterial activity, also possess antioxidant properties due to the presence of phenolic compounds. Student t-test analysis suggested that 4- hydroxybenzoic acid positively allosteric GABA (A) receptor, increased normalized current amplitude to 1.0304±0.0464(p value 0.032) compared with vehicle. 4-Hydrobenzoic acid, a compound from Dendrocalamus Asper bamboo shoot gives new insights for future studies on bamboo shoots with motivation for extraction of more compounds to investigate their effects on human and rodents against epilepsy, insomnia, and anxiety.

Keywords: α1β2γ2S, antiepileptic, bamboo shoots, epilepsy GABA (A) receptor, two-microelectrode voltage clamp, xenopus laevis oocytes

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785 Effects of Plasma Technology in Biodegradable Films for Food Packaging

Authors: Viviane P. Romani, Bradley D. Olsen, Vilásia G. Martins

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Biodegradable films for food packaging have gained growing attention due to environmental pollution caused by synthetic films and the interest in the better use of resources from nature. Important research advances were made in the development of materials from proteins, polysaccharides, and lipids. However, the commercial use of these new generation of sustainable materials for food packaging is still limited due to their low mechanical and barrier properties that could compromise the food quality and safety. Thus, strategies to improve the performance of these materials have been tested, such as chemical modifications, incorporation of reinforcing structures and others. Cold plasma is a versatile, fast and environmentally friendly technology. It consists of a partially ionized gas containing free electrons, ions, and radicals and neutral particles able to react with polymers and start different reactions, leading to the polymer degradation, functionalization, etching and/or cross-linking. In the present study, biodegradable films from fish protein prepared through the casting technique were plasma treated using an AC glow discharge equipment. The reactor was preliminary evacuated to ~7 Pa and the films were exposed to air plasma for 2, 5 and 8 min. The films were evaluated by their mechanical and water vapor permeability (WVP) properties and changes in the protein structure were observed using Scanning Electron Microscopy (SEM) and X-ray diffraction (XRD). Potential cross-links and elimination of surface defects by etching might be the reason for the increase in tensile strength and decrease in the elongation at break observed. Among the times of plasma application tested, no differences were observed when higher times of exposure were used. The X-ray pattern showed a broad peak at 2θ = 19.51º that corresponds to the distance of 4.6Å by applying the Bragg’s law. This distance corresponds to the average backbone distance within the α-helix. Thus, the changes observed in the films might indicate that the helical configuration of fish protein was disturbed by plasma treatment. SEM images showed surface damage in the films with 5 and 8 min of plasma treatment, indicating that 2 min was the most adequate time of treatment. It was verified that plasma removes water from the films once weight loss of 4.45% was registered for films treated during 2 min. However, after 24 h in 50% of relative humidity, the water lost was recovered. WVP increased from 0.53 to 0.65 g.mm/h.m².kPa after plasma treatment during 2 min, that is desired for some foods applications which require water passage through the packaging. In general, the plasma technology affects the properties and structure of fish protein films. Since this technology changes the surface of polymers, these films might be used to develop multilayer materials, as well as to incorporate active substances in the surface to obtain active packaging.

Keywords: fish protein films, food packaging, improvement of properties, plasma treatment

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784 The Immunology Evolutionary Relationship between Signal Transducer and Activator of Transcription Genes from Three Different Shrimp Species in Response to White Spot Syndrome Virus Infection

Authors: T. C. C. Soo, S. Bhassu

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Unlike the common presence of both innate and adaptive immunity in vertebrates, crustaceans, in particular, shrimps, have been discovered to possess only innate immunity. This further emphasizes the importance of innate immunity within shrimps in pathogenic resistance. Under the study of pathogenic immune challenge, different shrimp species actually exhibit varying degrees of immune resistance towards the same pathogen. Furthermore, even within the same shrimp species, different batches of challenged shrimps can have different strengths of immune defence. Several important pathways are activated within shrimps during pathogenic infection. One of them is JAK-STAT pathway that is activated during bacterial, viral and fungal infections by which STAT(Signal Transducer and Activator of Transcription) gene is the core element of the pathway. Based on theory of Central Dogma, the genomic information is transmitted in the order of DNA, RNA and protein. This study is focused in uncovering the important evolutionary patterns present within the DNA (non-coding region) and RNA (coding region). The three shrimp species involved are Macrobrachium rosenbergii, Penaeus monodon and Litopenaeus vannamei which all possess commercial significance. The shrimp species were challenged with a famous penaeid shrimp virus called white spot syndrome virus (WSSV) which can cause serious lethality. Tissue samples were collected during time intervals of 0h, 3h, 6h, 12h, 24h, 36h and 48h. The DNA and RNA samples were then extracted using conventional kits from the hepatopancreas tissue samples. PCR technique together with designed STAT gene conserved primers were utilized for identification of the STAT coding sequences using RNA-converted cDNA samples and subsequent characterization using various bioinformatics approaches including Ramachandran plot, ProtParam and SWISS-MODEL. The varying levels of immune STAT gene activation for the three shrimp species during WSSV infection were confirmed using qRT-PCR technique. For one sample, three biological replicates with three technical replicates each were used for qRT-PCR. On the other hand, DNA samples were important for uncovering the structural variations within the genomic region of STAT gene which would greatly assist in understanding the STAT protein functional variations. The partially-overlapping primers technique was used for the genomic region sequencing. The evolutionary inferences and event predictions were then conducted through the Bayesian Inference method using all the acquired coding and non-coding sequences. This was supplemented by the construction of conventional phylogenetic trees using Maximum likelihood method. The results showed that adaptive evolution caused STAT gene sequence mutations between different shrimp species which led to evolutionary divergence event. Subsequently, the divergent sites were correlated to the differing expressions of STAT gene. Ultimately, this study assists in knowing the shrimp species innate immune variability and selection of disease resistant shrimps for breeding purpose. The deeper understanding of STAT gene evolution from the perspective of both purifying and adaptive approaches not only can provide better immunological insight among shrimp species, but also can be used as a good reference for immunological studies in humans or other model organisms.

Keywords: gene evolution, JAK-STAT pathway, immunology, STAT gene

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783 The Effects of Myelin Basic Protein Charge Isomers on the Methyl Cycle Metabolites in Glial Cells

Authors: Elene Zhuravliova, Tamar Barbakadze, Irina Kalandadze, Elnari Zaalishvili, Lali Shanshiashvili, David Mikeladze

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Background: Multiple sclerosis (MS) is an inflammatory, neurodegenerative disease, which is accompanied by demyelination and autoimmune response to myelin proteins. Among post-translational modifications, which mediate the modulation of inflammatory pathways during MS, methylation is the main one. The methylation of DNA, also amino acids lysine and arginine, occurs in the cell. It was found that decreased trans-methylation is associated with neuroinflammatory diseases. Therefore, abnormal regulation of the methyl cycle could induce demyelination through the action on PAD (peptidyl-arginine-deiminase) gene promoter. PAD takes part in protein citrullination and targets myelin basic protein (MBP), which is affected during demyelination. To determine whether MBP charge isomers are changing the methyl cycle, we have estimated the concentrations of methyl cycle metabolites in MBP-activated primary astrocytes and oligodendrocytes. For this purpose, the action of the citrullinated MBP- C8 and the most cationic MBP-C1 isomers on the primary cells were investigated. Methods: Primary oligodendrocyte and astrocyte cell cultures were prepared from whole brains of 2-day-old Wistar rats. The methyl cycle metabolites, including homocysteine, S-adenosylmethionine (SAM), and S-adenosylhomocysteine (SAH), were estimated by HPLC analysis using fluorescence detection and prior derivatization. Results: We found that the action of MBP-C8 and MBP-C1 induces a decrease in the concentration of both methyl cycle metabolites, S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH), in astrocytes compared to the control cells. As for oligodendrocytes, the concentration of SAM was increased by the addition of MBP-C1, while MBP-C8 has no significant effect. As for SAH, its concentration was increased compared to the control cells by the action of both MBP-C1 and MBP-C8. A significant increase in homocysteine concentration was observed by the action of the MBP-C8 isomer in both oligodendrocytes and astrocytes. Conclusion: These data suggest that MBP charge isomers change the concentration of methyl cycle metabolites. MBP-C8 citrullinated isomer causes elevation of homocysteine in astrocytes and oligodendrocytes, which may be the reason for decreased astrocyte proliferation and increased oligodendrocyte cell death which takes place in neurodegenerative processes. Elevated homocysteine levels and subsequent abnormal regulation of methyl cycles in oligodendrocytes possibly change the methylation of DNA that activates PAD gene promoter and induces the synthesis of PAD, which in turn provokes the process of citrullination, which is the accompanying process of demyelination. Acknowledgment: This research was supported by the SRNSF Georgia RF17_534 grant.

Keywords: myelin basic protein, astrocytes, methyl cycle metabolites, homocysteine, oligodendrocytes

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782 Engineering of Reagentless Fluorescence Biosensors Based on Single-Chain Antibody Fragments

Authors: Christian Fercher, Jiaul Islam, Simon R. Corrie

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Fluorescence-based immunodiagnostics are an emerging field in biosensor development and exhibit several advantages over traditional detection methods. While various affinity biosensors have been developed to generate a fluorescence signal upon sensing varying concentrations of analytes, reagentless, reversible, and continuous monitoring of complex biological samples remains challenging. Here, we aimed to genetically engineer biosensors based on single-chain antibody fragments (scFv) that are site-specifically labeled with environmentally sensitive fluorescent unnatural amino acids (UAA). A rational design approach resulted in quantifiable analyte-dependent changes in peak fluorescence emission wavelength and enabled antigen detection in vitro. Incorporation of a polarity indicator within the topological neighborhood of the antigen-binding interface generated a titratable wavelength blueshift with nanomolar detection limits. In order to ensure continuous analyte monitoring, scFv candidates with fast binding and dissociation kinetics were selected from a genetic library employing a high-throughput phage display and affinity screening approach. Initial rankings were further refined towards rapid dissociation kinetics using bio-layer interferometry (BLI) and surface plasmon resonance (SPR). The most promising candidates were expressed, purified to homogeneity, and tested for their potential to detect biomarkers in a continuous microfluidic-based assay. Variations of dissociation kinetics within an order of magnitude were achieved without compromising the specificity of the antibody fragments. This approach is generally applicable to numerous antibody/antigen combinations and currently awaits integration in a wide range of assay platforms for one-step protein quantification.

Keywords: antibody engineering, biosensor, phage display, unnatural amino acids

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781 Features of the Functional and Spatial Organization of Railway Hubs as a Part of the Urban Nodal Area

Authors: Khayrullina Yulia Sergeevna, Tokareva Goulsine Shavkatovna

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The article analyzes the modern major railway hubs as a main part of the Urban Nodal Area (UNA). The term was introduced into the theory of urban planning at the end of the XX century. Tokareva G.S. jointly with Gutnov A.E. investigated the structure-forming elements of the city. UNA is the basic unit, the "cell" of the city structure. Specialization is depending on the position in the frame or the fabric of the city. This is related to feature of its organization. Spatial and functional features of UNA proposed to investigate in this paper. The base object for researching are railway hubs as connective nodes of inner and extern-city communications. Research used a stratified sampling type with the selection of typical objects. Research is being conducted on the 14 railway hubs of the native and foreign experience of the largest cities with a population over 1 million people located in one and close to the Russian climate zones. Features of the organization identified in the complex research of functional and spatial characteristics based on the hypothesis of the existence of dual characteristics of the organization of urban nodes. According to the analysis, there is using the approximation method that enable general conclusions of a representative selection of the entire population of railway hubs and it development’s area. Results of the research show specific ratio of functional and spatial organization of UNA based on railway hubs. Based on it there proposed typology of spaces and urban nodal areas. Identification of spatial diversity and functional organization’s features of the greatest railway hubs and it development’s area gives an indication of the different evolutionary stages of formation approaches. It help to identify new patterns for the complex and effective design as a prediction of the native hub’s development direction.

Keywords: urban nodal area, railway hubs, features of structural, functional organization

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780 The Effect of Principled Human Resource Management and Training Based on Existing Standards in Order to Improve the Quality of Construction Projects

Authors: Arsalan Salahi

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Today, the number of changes in the construction industry and urban mass house building is increasing, which makes you need to pay more attention to targeted planning for human resource management and training. The human resources working in the construction industry have various problems and deficiencies, and in order to solve these problems, there is a need for basic management and training of these people in order to lower the construction costs and increase the quality of the projects, especially in mass house building projects. The success of any project in reaching short and long-term professional goals depends on the efficient combination of work tools, financial resources, raw materials, and most importantly, human resources. Today, due to the complexity and diversity of each project, specialized management fields have emerged to maximize the potential benefits of each component of that project. Human power is known as the most important resource in construction projects for its successful implementation, but unfortunately, due to the low cost of human power compared to other resources, such as materials and machinery, little attention is paid to it. With the correct management and training of human resources, which depends on its correct planning and development, it is possible to improve the performance of construction projects. In this article, the training and motivation of construction industry workers and their effects on the effectiveness of projects in this industry have been researched. In this regard, some barriers to the training and motivation of construction workers and personnel have been identified and solutions have been provided for construction companies. Also, the impact of workers and unskilled people on the efficiency of construction projects is investigated. The results of the above research show that by increasing the use of correct and basic training for human resources, we will see positive results and effects on the performance of construction projects.

Keywords: human resources, construction industry, principled training, skilled and unskilled workers

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779 Stereological Evaluation of Liver of Rabbit Fetuses After Transplantation of Human Wharton’s Jelly-Derived Mesenchymal Stromal/Stem Cells

Authors: Zahra Khodabandeh, Leila Rezaeian, Mohammad Amin Edalatmanesh, Asghar Mogheiseh, Nader Tanideh, Mehdi Dianatpour, Shahrokh Zare, Hossein Bordbar, Neda Baghban, Amin Tamadon

Abstract:

Background: In-utero xenotransplantation of stem cells in abnormal fetuses effectively treats several genetic illnesses. Objective: The current research aimed to evaluate structural and morphological alterations in the liver of rabbit fetuses following xenotransplantation of human Wharton’s jelly-derived mesenchymal stromal cells (hWJ-MSCs) using a stereological technique. Methods: hWJ-MSCs were isolated from the human umbilical cord, and their authenticity was established by flow cytometry and differentiation. At gestational day 14, the rabbits were anesthetized, and hWJ-MSCs were injected into the uteri of 24 fetuses. Twenty-two fetuses were born successfully. Ten rabbit liver specimens were prepared from injected fetuses, including eight rabbits on day three following birth and two rabbits on the 21st post-natal day. The non-injected fetuses were considered positive controls. The livers of the control and hWJ-MSCs-treated rabbits were fixed, processed, stained, and examined through stereological approaches. Results: In the hWJ-MSCs-treated group, the mean liver weight and volume increased by 42% and 78% compared to the control group. The total volume of the hepatocytes increased by 63% and that of sinusoids by threefold in the treated rabbits. The total volume of the central veins increased by 70%. The total number corresponding to hepatocytes in the experimental group increased by 112% compared to the rabbits in the control. The total volume of the hepatocyte nuclei in the experimental group increased by 117% compared to the rabbits in the control. Conclusion: After xenotransplantation of human MSCs, host tissue microenvironments (here, the rabbit liver) were altered, and these included quantitative factors corresponding to the liver tissue and hepatocyte morphometric indices.

Keywords: xenotransplantation, mesenchymal stromal, stem cell, Wharton ‘s jelly, liver

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778 Human Leukocyte Antigen Class 1 Phenotype Distribution and Analysis in Persons from Central Uganda with Active Tuberculosis and Latent Mycobacterium tuberculosis Infection

Authors: Helen K. Buteme, Rebecca Axelsson-Robertson, Moses L. Joloba, Henry W. Boom, Gunilla Kallenius, Markus Maeurer

Abstract:

Background: The Ugandan population is heavily affected by infectious diseases and Human leukocyte antigen (HLA) diversity plays a crucial role in the host-pathogen interaction and affects the rates of disease acquisition and outcome. The identification of HLA class 1 alleles and determining which alleles are associated with tuberculosis (TB) outcomes would help in screening individuals in TB endemic areas for susceptibility to TB and to predict resistance or progression to TB which would inevitably lead to better clinical management of TB. Aims: To be able to determine the HLA class 1 phenotype distribution in a Ugandan TB cohort and to establish the relationship between these phenotypes and active and latent TB. Methods: Blood samples were drawn from 32 HIV negative individuals with active TB and 45 HIV negative individuals with latent MTB infection. DNA was extracted from the blood samples and the DNA samples HLA typed by the polymerase chain reaction-sequence specific primer method. The allelic frequencies were determined by direct count. Results: HLA-A*02, A*01, A*74, A*30, B*15, B*58, C*07, C*03 and C*04 were the dominant phenotypes in this Ugandan cohort. There were differences in the distribution of HLA types between the individuals with active TB and the individuals with LTBI with only HLA-A*03 allele showing a statistically significant difference (p=0.0136). However, after FDR computation the corresponding q-value is above the expected proportion of false discoveries (q-value 0.2176). Key findings: We identified a number of HLA class I alleles in a population from Central Uganda which will enable us to carry out a functional characterization of CD8+ T-cell mediated immune responses to MTB. Our results also suggest that there may be a positive association between the HLA-A*03 allele and TB implying that individuals with the HLA-A*03 allele are at a higher risk of developing active TB.

Keywords: HLA, phenotype, tuberculosis, Uganda

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777 DNA Methylation Changes in Response to Ocean Acidification at the Time of Larval Metamorphosis in the Edible Oyster, Crassostrea hongkongensis

Authors: Yong-Kian Lim, Khan Cheung, Xin Dang, Steven Roberts, Xiaotong Wang, Vengatesen Thiyagarajan

Abstract:

Unprecedented rate of increased CO₂ level in the ocean and the subsequent changes in carbonate system including decreased pH, known as ocean acidification (OA), is predicted to disrupt not only the calcification process but also several other physiological and developmental processes in a variety of marine organisms, including edible oysters. Nonetheless, not all species are vulnerable to those OA threats, e.g., some species may be able to cope with OA stress using environmentally induced modifications on gene and protein expressions. For example, external environmental stressors, including OA, can influence the addition and removal of methyl groups through epigenetic modification (e.g., DNA methylation) process to turn gene expression “on or off” as part of a rapid adaptive mechanism to cope with OA. In this study, the above hypothesis was tested through testing the effect of OA, using decreased pH 7.4 as a proxy, on the DNA methylation pattern of an endemic and a commercially important estuary oyster species, Crassostrea hongkongensis, at the time of larval habitat selection and metamorphosis. Larval growth rate did not differ between control pH 8.1 and treatment pH 7.4. The metamorphosis rate of the pediveliger larvae was higher at pH 7.4 than those in control pH 8.1; however, over one-third of the larvae raised at pH 7.4 failed to attach to an optimal substrate as defined by biofilm presence. During larval development, a total of 130 genes were differentially methylated across the two treatments. The differential methylation in the larval genes may have partially accounted for the higher metamorphosis success rate under decreased pH 7.4 but with poor substratum selection ability. Differentially methylated loci were concentrated in the exon regions and appear to be associated with cytoskeletal and signal transduction, oxidative stress, metabolic processes, and larval metamorphosis, which implies the high potential of C. hongkongensis larvae to acclimate and adapt through non-genetic ways to OA threats within a single generation.

Keywords: adaptive plasticity, DNA methylation, larval metamorphosis, ocean acidification

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776 How to Incorporate Vernacular Architecture into Practice for Sustainable Development: Case Studies from Kashmir and Kerala, India

Authors: Debanjana Chatterjee

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Vernacular settlements in India often take the form that is dictated by the climate they are in. India, with its vast cultural diversity and various climatic regions, offers a wide range of vernacular architecture. This paper focuses on two main geographical regions: Kashmir and Kerala. They bring together myriad challenges of climatic and social characteristics to incorporate into their vernacular architectures, which are still relevant despite the advent of globalization and modernization. Scholars like William Wurster and Catherine Bauer even claimed that all the traditional buildings in these places have the kind of urbanity, which is dignified and elegant but also lively and human that every architect would like to achieve if they knew how. With modernization, and with a greater ease of construction, a reduction in labor, and the apparent robustness of contemporary construction techniques, people have, however, become increasingly tentative in respect of vernacular architecture. And yet modern architecture has typically led to energize intensive structures without much consideration to the location and surroundings of the structure itself. In contrary, Laurie Baker, the British-born Indian architect, had shown us the way to integrate the knowledge of vernacular when he developed his designs based on the traditional architecture of Kerala, respecting the local climate and environment. This paper also explores his technical creativity in his design of Center for Development Studies (CDS) in Trivandrum. Hence, in order to protect and conserve our rich cultural and architectural heritage, the elements of vernacular should be incorporated into the contemporary planning and architecture for sustainable building design. The provision should be made to incorporate vernacular architecture and traditional knowledge in the policies. Ultimately, the policymakers, planners, and architects should consider this incorporation of traditional vernacular and contemporary sustainability in their work for the betterment of society now.

Keywords: vernacular, architecture, sustainable development, Kashmir and Kerala, climate, Laurie Baker

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775 Electrospun Conducting Polymer/Graphene Composite Nanofibers for Gas Sensing Applications

Authors: Aliaa M. S. Salem, Soliman I. El-Hout, Amira Gaber, Hassan Nageh

Abstract:

Nowadays, the development of poisonous gas detectors is considered to be an urgent matter to secure human health and the environment from poisonous gases, in view of the fact that even a minimal amount of poisonous gas can be fatal. Of these concerns, various inorganic or organic sensing materials have been used. Among these are conducting polymers, have been used as the active material in the gassensorsdue to their low-cost,easy-controllable molding, good electrochemical properties including facile fabrication process, inherent physical properties, biocompatibility, and optical properties. Moreover, conducting polymer-based chemical sensors have an amazing advantage compared to the conventional one as structural diversity, facile functionalization, room temperature operation, and easy fabrication. However, the low selectivity and conductivity of conducting polymers motivated the doping of it with varied materials, especially graphene, to enhance the gas-sensing performance under ambient conditions. There were a number of approaches proposed for producing polymer/ graphene nanocomposites, including template-free self-assembly, hard physical template-guided synthesis, chemical, electrochemical, and electrospinning...etc. In this work, we aim to prepare a novel gas sensordepending on Electrospun nanofibers of conducting polymer/RGO composite that is the effective and efficient expectation of poisonous gases like ammonia, in different application areas such as environmental gas analysis, chemical-,automotive- and medical industries. Moreover, our ultimate objective is to maximize the sensing performance of the prepared sensor and to check its recovery properties.

Keywords: electro spinning process, conducting polymer, polyaniline, polypyrrole, polythiophene, graphene oxide, reduced graphene oxide, functionalized reduced graphene oxide, spin coating technique, gas sensors

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774 Changing Emphases in Mental Health Research Methodology: Opportunities for Occupational Therapy

Authors: Jeffrey Chase

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Historically the profession of Occupational Therapy was closely tied to the treatment of those suffering from mental illness; more recently, and especially in the U.S., the percentage of OTs identifying as working in the mental health area has declined significantly despite the estimate that by 2020 behavioral health disorders will surpass physical illnesses as the major cause of disability worldwide. In the U.S. less than 10% of OTs identify themselves as working with the mentally ill and/or practicing in mental health settings. Such a decline has implications for both those suffering from mental illness and the profession of Occupational Therapy. One reason cited for the decline of OT in mental health has been the limited research in the discipline addressing mental health practice. Despite significant advances in technology and growth in the field of neuroscience, major institutions and funding sources such as the National Institute of Mental Health (NIMH) have noted that research into the etiology and treatment of mental illness have met with limited success over the past 25 years. One major reason posited by NIMH is that research has been limited by how we classify individuals, that being mostly on what is observable. A new classification system being developed by NIMH, the Research Domain Criteria (RDoc), has the goal to look beyond just descriptors of disorders for common neural, genetic, and physiological characteristics that cut across multiple supposedly separate disorders. The hope is that by classifying individuals along RDoC measures that both reliability and validity will improve resulting in greater advances in the field. As a result of this change NIH and NIMH will prioritize research funding to those projects using the RDoC model. Multiple disciplines across many different setting will be required for RDoC or similar classification systems to be developed. During this shift in research methodology OT has an opportunity to reassert itself into the research and treatment of mental illness, both in developing new ways to more validly classify individuals, and to document the legitimacy of previously ill-defined and validated disorders such as sensory integration.

Keywords: global mental health and neuroscience, research opportunities for ot, greater integration of ot in mental health research, research and funding opportunities, research domain criteria (rdoc)

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773 Effectiveness of Active Learning in Social Science Courses at Japanese Universities

Authors: Kumiko Inagaki

Abstract:

In recent, years, Japanese universities have begun to face a dilemma: more than half of all high school graduates go on to attend an institution of higher learning, overwhelming Japanese universities accustomed to small student bodies. These universities have been forced to embrace qualitative changes to accommodate the increased number and diversity of students who enter their establishments, students who differ in their motivations for learning, their levels of eagerness to learn, and their perspectives on the future. One of these changes is an increase in awareness among Japanese educators of the importance of active learning, which deepens students’ understanding of course material through a range of activities, including writing, speaking, thinking, and presenting, in addition to conventional “passive learning” methods such as listening to a one-way lecture.  The purpose of this study is to examine the effectiveness of the teaching method adapted to improve active learning. A teaching method designed to promote active learning was implemented in a social science course at one of the most popular universities in Japan. A questionnaire using a five-point response format was given to students in 2,305 courses throughout the university to evaluate the effectiveness of the method based on the following measures: ① the ratio of students who were motivated to attend the classes, ② the rate at which students learned new information, and ③ the teaching method adopted in the classes. The results of this study show that the percentage of students who attended the active learning course eagerly, and the rate of new knowledge acquired through the course, both exceeded the average for the university, the department, and the subject area of social science. In addition, there are strong correlations between teaching method and student motivation and between teaching method and knowledge acquisition rate. These results indicate that the active learning teaching method was effectively implemented and that it may improve student eagerness to attend class and motivation to learn.

Keywords: active learning, Japanese university, teaching method, university education

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772 Psychosocial Consequences of Discovering Misattributed Paternity in Adulthood: Insider Action Research

Authors: Alyona Cerfontyne, Levita D'Souza, Lefteris Patlamazoglou

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Unlike adoption and donor-assisted reproduction, misattributed paternity occurring within the context of spontaneous conception and outside of formally recognised practices of having a child remains largely an understudied phenomenon. In adulthood, to discover misattributed paternity, i.e., that the man you call your father is not related to you genetically, can have profound implications for everyone affected. Until the advent of direct-to-consumer DNA testing 20 years ago, such discoveries were relatively rare. Despite the growing number of individuals uncovering their biogenetic paternity through genetic testing, there is very limited research on misattributed paternity from the perspective of adult children affected by it. No research exists on how to support these individuals through counselling post-discovery. Framed as insider action research, this study aimed to explore the perceived psychosocial consequences of misattributed paternity discoveries and coping strategies used by individuals who discover their misattributed paternity status in adulthood. In total, 12 individuals with misattributed paternity participated in semi-structured interviews in July-August 2022. The collected data was analysed using reflexive thematic analysis. The study’s results indicate that discovering misattributed paternity in adulthood can be likened to a watershed moment forever changing the trajectory of one’s life. Psychological experiences consistent with trauma, as well as grief and loss, re-evaluation of close family relationships, reestablishment of one’s identity, as well as experiencing a profound need to belong are the key themes emerging from the analysis of psychosocial experiences. Post-discovery, individuals with misattributed paternity employ a wide range of emotional and problem-focused coping strategies, amongst which seeking connection with those who understand, searching for information on the new biogenetic family and finding new meanings to life are most prominent. The study contributes both to the academic and practical knowledge of experiences of misattributed paternity and highlights the importance of further research on the topic.

Keywords: discovery of misattributed paternity, misattributed paternity, paternal discrepancy, psychosocial consequences, coping

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771 Phytoplankton Assemblage and Physicochemical Parameters of a Perturbed Tropical Manmade Lake, Southwestern Nigeria

Authors: Adedolapo Ayoade, John the Beloved Dada

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This study identified the phytoplankton assemblage of the Dandaru Lake (that received effluents from a zoological garden and hospital) as bioindicators of water quality. Physicochemical parameters including Dissolved Oxygen (DO), biochemical oxygen demand, nitrate, phosphate and heavy metals were also determined. Samples of water and plankton were collected once monthly from April to September, 2015 at five stations (I – V). The mean physicochemical parameters were within the limits of National Environmental Standards and Regulations Enforcement Agency (NESREA) and USEPA except Lead, 0.02 ± 0.08 mg/ L; Manganese, 0.46 ± 1.00 mg/ L and Zinc, 0.05 ± 0.17 mg/ L. Means of DO, alkalinity, and phosphate were significantly different between the stations at p < 0.05. While highest mean DO (6.88 ± 1.34 mg/L) was recorded in station I with less anthropogenic activities, highest phosphate concentration (0.28 ± 0.28 mg/L) occurred in station II, the entry point of wastewater from hospital and zoological garden. The 147 phytoplankton species found in the lake belonged to six classes: Chlorophyceae (50), Euglenophyceae (40), Bacillariophyceae (37), Cyanophyceae (17), Xanthophyceae and Chrysophyceae (3). The order of abundance for phytoplankton was Euglenophyceae (49.77%) > Bacillariophyceae (18.00%) > Cyanophyceae (17.39%) > Chlorophyceae (13.7%) > Xanthophyceae (1.06%) > Chrysophyceae (0.02%). The stations impacted with effluents were dominated by members of Euglenophyceae (Station III, 77.09%; IV, 50.55%) and Cyanophyceae (Station II, 27.7%; V, 32.57%). While station I was dominated by diatoms (57.98%). The species richness recorded was 0.32 – 4.49. Evenness index was highest in station I and least in station III. Generally, pollution tolerant species (Microcystis, Oscillatoria, Scenedesmus, Anabaena, and Euglena) showed greater density in areas impacted by human activities. The phytoplankton assemblage and comparatively low biotic diversity in Dandaru Lake could be attributed to perturbations in the water column that exerted selective effects on the biological assemblage.

Keywords: manmade lake, Nigeria, phytoplankton, water quality

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770 Serum Neurotrophins in Different Metabolic Types of Obesity

Authors: Irina M. Kolesnikova, Andrey M. Gaponov, Sergey A. Roumiantsev, Tatiana V. Grigoryeva, Alexander V. Laikov, Alexander V. Shestopalov

Abstract:

Background. Neuropathy is a common complication of obesity. In this regard, the content of neurotrophins in such patients is of particular interest. Neurotrophins are the proteins that regulate neuron survival and neuroplasticity and include brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF). However, the risk of complications depends on the metabolic type of obesity. Metabolically unhealthy obesity (MUHO) is associated with a high risk of complications, while this is not the case with metabolically healthy obesity (MHO). Therefore, the aim of our work was to study the effect of the obesity metabolic type on serum neurotrophins levels. Patients, materials, methods. The study included 134 healthy donors and 104 obese patients. Depending on the metabolic type of obesity, the obese patients were divided into subgroups with MHO (n=40) and MUHO (n=55). In the blood serum, the concentration of BDNF and NGF was determined. In addition, the content of adipokines (leptin, asprosin, resistin, adiponectin), myokines (irisin, myostatin, osteocrin), indicators of carbohydrate, and lipid metabolism were measured. Correlation analysis revealed the relationship between the studied parameters. Results. We found that serum BDNF concentration was not different between obese patients and healthy donors, regardless of obesity metabolic type. At the same time, in obese patients, there was a decrease in serum NGF level versus control. A similar trend was characteristic of both MHO and MUHO. However, MUHO patients had a higher NGF level than MHO patients. The literature indicates that obesity is associated with an increase in the plasma concentration of NGF. It can be assumed that in obesity, there is a violation of NGF storage in platelets, which accelerates neurotrophin degradation. We found that BDNF concentration correlated with irisin levels in MUHO patients. Healthy donors had a weak association between NGF and VEGF levels. No such association was found in obese patients, but there was an association between NGF and leptin concentrations. In MHO, the concentration of NHF correlated with the content of leptin, irisin, osteocrin, insulin, and the HOMA-IR index. But in MUHO patients, we found only the relationship between NGF and adipokines (leptin, asprosin). It can be assumed that in patients with MHO, the replenishment of serum NGF occurs under the influence of muscle and adipose tissue. In the MUHO patients only the effect of adipose tissue on NGF was observed. Conclusion. Obesity, regardless of metabolic type, is associated with a decrease in serum NGF concentration. We showed that muscle and adipose tissues make a significant contribution to the serum NGF pool in the MHO patients. In MUHO there is no effect of muscle on the NGF level, but the effect of adipose tissue remains.

Keywords: neurotrophins, nerve growth factor, NGF, brain-derived neurotrophic factor, BDNF, obesity, metabolically healthy obesity, metabolically unhealthy obesity

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769 The Application of Whole-Cell Luminescent Biosensors for Assessing Bactericidal Properties of Medicinal Plants

Authors: Yuliya Y. Gavrichenko

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Background and Aims: The increasing bacterial resistance to almost all the available antibiotics has encouraged scientists to search for alternative sources of antibacterial agents. Nowadays, it is known that many plant secondary metabolites have diverse biological activity. These compounds can be potentially active against human bacterial and viral infections. Extended research has been carried out to explore the use of the luminescent bacterial test as a rapid, accurate and inexpensive method to assess the antibacterial properties and to predict the biological activity spectra for plant origin substances. Method: Botanical material of fifteen species was collected from their natural and cultural habitats on the Crimean peninsula. The aqueous extracts of following plants were tested: Robinia pseudoacacia L., Sideritis comosa, Cotinus coggygria Scop., Thymus serpyllum L., Juglans regia L., Securigera varia L., Achillea millefolium L., Phlomis taurica, Corylus avellana L., Sambucus nigra L., Helichrysum arenarium L., Glycyrrhiza glabra L., Elytrigia repens L., Echium vulgare L., Conium maculatum L. The test was carried out using luminous strains of marine bacteria Photobacterium leiognathi, which was isolated from the Sea of Azov as well as four Escherichia coli MG1655 recombinant strains harbouring Vibrio fischeri luxCDABE genes. Results: The bactericidal capacity of plant extracts showed significant differences in the study. Cotinus coggygria, Phlomis taurica, Juglans regia L. proved to be the most toxic to P. leiognathi. (EC50 = 0.33 g dried plant/l). Glycyrrhiza glabra L., Robinia pseudoacacia L., Sideritis comosa and Helichrysum arenarium L. had moderate inhibitory effects (EC50 = 3.3 g dried plant/l). The rest of the aqueous extracts have decreased the luminescence of no more than 50% at the lowest concentration (16.5 g dried plant/l). Antibacterial activity of herbal extracts against constitutively luminescent E. coli MG1655 (pXen7-lux) strain was observed at approximately the same level as for P. leiognathi. Cotinus coggygria and Conium maculatum L. extracts have increased light emission in the mutant E. coli MG1655 (pFabA-lux) strain which is associated with cell membranes damage. Sideritis comosa, Phlomis taurica, Juglans regia induced SOS response in E. coli (pColD-lux) strain. Glycyrrhiza glabra L. induced protein damage response in E. coli MG1655 (pIbpA-lux) strain. Conclusion: The received results have shown that the plants’ extracts had nonspecific antimicrobial effects against both E. coli (pXen7-lux) and P. leiognathi biosensors. Mutagenic, cytotoxic and protein damage effects have been observed. In general, the bioluminescent inhibition test result correlated with the traditional use of screened plants. It leads to the following conclusion that whole-cell luminescent biosensors could be the indicator of overall plants antibacterial capacity. The results of the investigation have shown a possibility of bioluminescent method in medicine and pharmacy as an approach to research the antibacterial properties of medicinal plants.

Keywords: antibacterial property, bioluminescence, medicinal plants, whole-cell biosensors

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768 Structure Clustering for Milestoning Applications of Complex Conformational Transitions

Authors: Amani Tahat, Serdal Kirmizialtin

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Trajectory fragment methods such as Markov State Models (MSM), Milestoning (MS) and Transition Path sampling are the prime choice of extending the timescale of all atom Molecular Dynamics simulations. In these approaches, a set of structures that covers the accessible phase space has to be chosen a priori using cluster analysis. Structural clustering serves to partition the conformational state into natural subgroups based on their similarity, an essential statistical methodology that is used for analyzing numerous sets of empirical data produced by Molecular Dynamics (MD) simulations. Local transition kernel among these clusters later used to connect the metastable states using a Markovian kinetic model in MSM and a non-Markovian model in MS. The choice of clustering approach in constructing such kernel is crucial since the high dimensionality of the biomolecular structures might easily confuse the identification of clusters when using the traditional hierarchical clustering methodology. Of particular interest, in the case of MS where the milestones are very close to each other, accurate determination of the milestone identity of the trajectory becomes a challenging issue. Throughout this work we present two cluster analysis methods applied to the cis–trans isomerism of dinucleotide AA. The choice of nucleic acids to commonly used proteins to study the cluster analysis is two fold: i) the energy landscape is rugged; hence transitions are more complex, enabling a more realistic model to study conformational transitions, ii) Nucleic acids conformational space is high dimensional. A diverse set of internal coordinates is necessary to describe the metastable states in nucleic acids, posing a challenge in studying the conformational transitions. Herein, we need improved clustering methods that accurately identify the AA structure in its metastable states in a robust way for a wide range of confused data conditions. The single linkage approach of the hierarchical clustering available in GROMACS MD-package is the first clustering methodology applied to our data. Self Organizing Map (SOM) neural network, that also known as a Kohonen network, is the second data clustering methodology. The performance comparison of the neural network as well as hierarchical clustering method is studied by means of computing the mean first passage times for the cis-trans conformational rates. Our hope is that this study provides insight into the complexities and need in determining the appropriate clustering algorithm for kinetic analysis. Our results can improve the effectiveness of decisions based on clustering confused empirical data in studying conformational transitions in biomolecules.

Keywords: milestoning, self organizing map, single linkage, structure clustering

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767 [Keynote Talk]: Study of Cooperative Career Education between Universities and Companies

Authors: Azusa Katsumata

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Where there is collaboration between universities and companies in the educational context, companies seek ‘knowledge’ from universities and provide a ‘place of practice’ to them. Several universities have introduced activities aimed at the mutual enlightenment of a diversity of people in career education. However, several programs emphasize on delivering results, and on practicing the prepared materials as planned. Few programs focus on unexpected failures and setbacks. This way of learning is important in career education so that classmates can help each other, overcome difficulties, draw out each other’s strengths, and learn from them. Seijo University in Tokyo offered Tokyo Tourism, a Project-Based Learning course, as a first-year career education course until 2016. In cooperation with a travel agency, students participate in planning actual tourism products for foreigners visiting Japan, undertake tours serving as guides. This paper aims to study the 'learning platform' created by a series of processes such as the fieldwork, planning tours, the presentation, selling the tourism products, and guiding the tourists. We conducted a questionnaire to measure the development of work-related skills in class. From the results of the questionnaire, we can see, in the example of this class, that students demonstrated an increased desire to be pro-active and an improved motivation to learn. Students have not, however, acquired policy or business skills. This is appropriate for first-year careers education, but we need to consider how this can be incorporated into future courses. In the questionnaire filled out by the students after the class, the following results were found. Planning and implementing travel products while learning from each other, and helping the teams has led to improvements in the student workforce. This course is a collaborative project between Japanese universities and the 2020 Tokyo Olympics and Paralympic Games committee.

Keywords: university career education, platform of learning, project-based learning, collaboration between university and company

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766 Re-Constructing the Research Design: Dealing with Problems and Re-Establishing the Method in User-Centered Research

Authors: Kerem Rızvanoğlu, Serhat Güney, Emre Kızılkaya, Betül Aydoğan, Ayşegül Boyalı, Onurcan Güden

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This study addresses the re-construction and implementation process of the methodological framework developed to evaluate how locative media applications accompany the urban experiences of international students coming to Istanbul with exchange programs in 2022. The research design was built on a three-stage model. The research team conducted a qualitative questionnaire in the first stage to gain exploratory data. These data were then used to form three persona groups representing the sample by applying cluster analysis. In the second phase, a semi-structured digital diary study was carried out on a gamified task list with a sample selected from the persona groups. This stage proved to be the most difficult to obtaining valid data from the participant group. The research team re-evaluated the design of this second phase to reach the participants who will perform the tasks given by the research team while sharing their momentary city experiences, to ensure the daily data flow for two weeks, and to increase the quality of the obtained data. The final stage, which follows to elaborate on the findings, is the “Walk & Talk,” which is completed with face-to-face and in-depth interviews. It has been seen that the multiple methods used in the research process contribute to the depth and data diversity of the research conducted in the context of urban experience and locative technologies. In addition, by adapting the research design to the experiences of the users included in the sample, the differences and similarities between the initial research design and the research applied are shown.

Keywords: digital diary study, gamification, multi-model research, persona analysis, research design for urban experience, user-centered research, “Walk & Talk”

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765 Bacterial Interactions of Upper Respiratory Tract Microbiota

Authors: Sarah Almuhayya, Andrew Mcbain, Gavin Humphreys

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Background. The microbiome of the upper respiratory tract (URT) has received less research attention than other body sites. This study aims to investigate the microbial ecology of the human URT with a focus on the antagonism between the corynebacteria and staphylococci. Methods. Mucosal swabs were collected from the anterior nares and nasal turbinates of 20 healthy adult subjects. Genomic DNA amplification targeting the (V4) of the 16Sr RNA gene was conducted and analyzed using QIIME. Nasal swab isolates were cultured and identified using near full-length sequencing of the 16S rRNA gene. Isolates identified as corynebacteria or staphylococci were typed using (rep-PCR). Antagonism was determined using an agar-based inhibition assay. Results. Four major bacterial phyla (Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria) were identified from all volunteers. The typing of cultured staphylococci and corynebacteria suggested that intra-individual strain diversity was limited. Analysis of generated nasal microbiota profiles suggested an inverse correlation in terms of relative abundance between staphylococci and corynebacteria. Despite the apparent antagonism between these genera, it was limited when investigated on agar. Of 1000 pairwise interactions, observable zones of inhibition were only reported between a single strain of C.pseudodiphtheriticum and S.aureus. Imaging under EM revealed this effect to be bactericidal with clear lytic effects on staphylococcal cell morphology. Conclusion. Nasal microbiota is complex, but culturable staphylococci and corynebacteria were limited in terms of clone type. Analysis of generated nasal microbiota profiles suggested an inverse correlation in terms of relative abundance between these genera suggesting an antagonism or competition between these taxonomic groups.

Keywords: nasal, microbiota, S.aureus, microbioal interaction

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