Search results for: gene name recognition

Authors: G. Krishnamoorthy, S. Anandhakumar

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The effect of D-Lysine (D-Lys) on collagen with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide(EDC)/N-hydroxysuccinimide(NHS) initiated cross linking using experimental and modelling tools are evaluated. The results of the Coll-D-Lys-EDC/NHS scaffold also indicate an increase in the tensile strength (TS), percentage of elongation (% E), denaturation temperature (Td), and decrease the decomposition rate compared to L-Lys-EDC/NHS. Scanning electron microscopic (SEM) and atomic force microscopic (AFM) analyses revealed a well ordered with properly oriented and well-aligned structure of scaffold. The D-Lys stabilizes the scaffold against degradation by collagenase than L-Lys. The cell assay showed more than 98% fibroblast viability (NIH3T3) and improved cell adhesions, protein adsorption after 72h of culture when compared with native scaffold. Cell attachment after 74h was robust, with cytoskeletal analysis showing that the attached cells were aligned along the fibers assuming a spindle-shape appearance, despite, gene expression analyses revealed no apparent alterations in mRNA levels, although cell proliferation was not adversely affected. D-Lysine (D-Lys) plays a pivotal role in the self-assembly and conformation of collagen fibrils. The D-Lys assisted EDC/NHS initiated cross-linking induces the formation of an carboxamide by the activation of the side chain -COOH group, followed by aminolysis of the O-iso acylurea intermediates by the -NH2 groups are directly joined via an isopeptides bond. This leads to the formation of intra- and inter-helical cross links. Modeling studies indicated that D-Lys bind with collagen-like peptide (CLP) through multiple H-bonding and hydrophobic interactions. Orientational changes in collagenase on CLP-D-Lys are observed which may decrease its accessibility to degradation and stabilize CLP against the action of the former. D-Lys has lowest binding energy and improved fibrillar-assembly and staggered alignment without the undesired structural stiffness and aggregations. The proteolytic machinery is not well equipped to deal with Coll-D-Lys than Coll-L-Lys scaffold. The information derived from the present study could help in designing collagenolytically stable heterochiral collagen based scaffold for biomedical applications.

Keywords: collagen, collagenase, collagen like peptide, D-lysine, heterochiral collagen scaffold

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Authors: Charlotte Williams

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Yale University Professor Hiram Bingham’s discovery of Machu Picchu in 1911 spurred an international interest in the Inca Empire, and with it, a dispute with the Peruvian government over who had rightful jurisdiction and curatorship over Inca history. By 2011, the Peruvian government initiated a legal battle for the return of artifacts that Bingham had removed from Machu Picchu, successfully returning them not to the site of Machu Picchu, but to Cusco, employing the rationale that the ancient Inca capital housed descendants of the Inca empire. This conflation of the past and present can be traced to a largely unanalyzed study that accompanied Bingham’s expedition: an ethnographic analysis of Inca descendants, which at the time portrayed indigenous Peruvian Andean peoples as remnants of a lost civilization, using Cusco as an assumed repository for people with 'Inca' characteristics. This study draws from the original Yale Peruvian Expedition archives, the Cusco Library archives, and in-depth interviews with curators of the Inca Museum and Machu Picchu Museum to analyze both the political conflict that emerged as a reaction to the ethnographic study, and how the study articulated with an inflating tourism market attempting to define what it meant to be Inca to an international public. The construction of the modern Inca as both directors of tourism management and purveyors of their archaeological material culture points to a unique case in which modern Peruvian citizens could claim heritage to an Inca past despite a lack of recognition as a legally defined group. The result has far-reaching implications, since Bingham’s artifacts returned not necessarily to a traditional nation-state, but to an imagined one, broadening the conditions under which informal repatriations can occur.

Keywords: archaeology of memory, imagined communities, Incanismo, repatriation

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Authors: Eryati Darwin, Eka Fithra Elfi, Indria Hafizah

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Background: Physical exercise induces a pattern of hormonal and immunological responses that prevent endothelial dysfunction by maintaining the availability of nitric oxide (NO). Regular and moderate exercise stimulates NO release, that can be considered as protective factor of cardiovascular diseases, while strenuous exercise induces increased levels in a number of pro-inflammatory and anti-inflammatory cytokines. Pro-inflammatory cytokines tumor necrosis factor-α (TNF-α) triggers endothelial activation which results in an increased vascular permeability. Nuclear gene factor kappa B (NF-κB) activates biological effect of TNF-α. Aim of Study: To determine the effect of physical exercise on the endothelial and skeletal muscle, we measured the level of NF-κB on rats’ serum, macrophages, and myocytes after strenuous physical exercise. Methods: 30 male Rattus norvegicus in the age of eight weeks were randomly divided into five groups (each containing six), and there were treated groups (T) and control group (C). The treated groups obtain strenuous physical exercise by ran on treadmill at 32 m/minutes for 1 hour or until exhaustion. Blood samples, myocytes of gastrocnemius muscle, and intraperitoneal macrophages were collected sequentially. There were investigated immediately, 2 hours, 6 hours, and 24 hours (T1, T2, T3, and T4) after sacrifice. The levels of NF-κB were measured by ELISA methods. Results: From our study, we found that the levels of NF-κB on myocytes in treated group from which its specimen was taken immediately (T1), 2 hours after treadmill (T2), and 6 hours after treadmill (T3) were significantly higher than control group (p<0.05), while the group from which its specimen was taken 24 hours after treadmill, was no significantly different (p>0.05). Also on macrophages, NF-κB in treated groups T1, T2, and T3 was significantly higher than control group (p<0.05), but there was no difference between T4 and control group (p>0.05). The level of serum NF-κB was not significantly different between treatment group as well as compared to control group (p>0.05). Serum NF-κB was significantly higher than the level on macrophages and myocytes (p<0.05). Conclusion: This study demonstrated that strenuous physical exercise stimulates the activation of NF-κB that plays a role in vascular inflammation and muscular damage, and may be recovered after resting period.

Keywords: endothelial function, inflammation, NFkB, physical exercise

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Authors: Iffat Ara Ebu, Fahmida Islam, Mohammad Abdus Shahid Rafi, Mahfuzur Rahman, Umar Iqbal, John Ball

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The application of multi-sensor fusion for enhanced distance estimation accuracy in dynamic environments is crucial for advanced driver assistance systems (ADAS) and autonomous vehicles. Limitations of single sensors such as cameras or radar in adverse conditions motivate the use of combined camera and radar data to improve reliability, adaptability, and object recognition. A multi-sensor fusion approach using an extended Kalman filter (EKF) is proposed to combine sensor measurements with a dynamic system model, achieving robust and accurate distance estimation. The research utilizes the Mississippi State University Autonomous Vehicular Simulator (MAVS) to create a controlled environment for data collection. Data analysis is performed using MATLAB. Qualitative (visualization of fused data vs ground truth) and quantitative metrics (RMSE, MAE) are employed for performance assessment. Initial results with simulated data demonstrate accurate distance estimation compared to individual sensors. The optimal sensor measurement noise variance and plant noise variance parameters within the EKF are identified, and the algorithm is validated with real-world data from a Chevrolet Blazer. In summary, this research demonstrates that multi-sensor fusion with an EKF significantly improves distance estimation accuracy in dynamic environments. This is supported by comprehensive evaluation metrics, with validation transitioning from simulated to real-world data, paving the way for safer and more reliable autonomous vehicle control.

Keywords: sensor fusion, EKF, MATLAB, MAVS, autonomous vehicle, ADAS

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Authors: Sarah Barnard, John Arnold, Fehmidah Munir, Sara Bosley

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Gender issues in higher education continue to represent a complex issue as institutions grapple with the role that organisations can play in combatting inequality. Schemes like Athena SWAN and the Aurora leadership programme in the UK context are attempting to tackle some of the issues around representation and the recognition of women in the sector. This paper is the first of its kind in reporting findings from a mixed-methods longitudinal study on both professional services and academic women in higher education in the UK. Online surveys have been completed by over 2,000 women in the sector. The qualitative elements include interviews with women and their mentors, and diaries with a select group of women. So far results have shown that contrary to the stereotype of women lacking leadership skills or having no desire to go into higher roles, women in the sector consistently assessed their leadership abilities positively, especially but not only regarding interpersonal interaction and facilitation. Over 80% of women agreed that they felt confident about putting themselves forward for positions of responsibility at work. However, qualitative data shows that confidence remains a salient term for how women talk about the challenges they have faced at work. This suggests that the work needed to challenge systemic gender issues requires action to be driven above the individual level. Overall, academics reported more negative experiences than professional services staff. Similarly BAME women’s responses are more negative. Therefore, the study offers some information on the differential experiences of women. In conclusion, women in higher education are undertaking considerable ‘below the radar’ leadership activities in what they perceive to be a somewhat inhospitable hostile workplace culture. The significant amount of effort expended in the sector is affecting slow, partial impacts on gender inequalities.

Keywords: gender, higher education, leadership, longitudinal research

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Authors: Alexis Akinyemi, Laurene Houtin

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Measures of employee motivation at work are often based on the theory of self-determined motivation, which implies that human resources departments and managers seek to motivate employees in the most self-determined way possible and use strategies to achieve this goal. In practice, they often tend to assess employee motivation and then adapt management to the most important source of motivation for their employees, for example by financially rewarding an employee who is extrinsically motivated, and by rewarding an intrinsically motivated employee with congratulations and recognition. Thus, the use of motivation measures contradicts theoretical positioning: theory does not provide for the promotion of extrinsically motivated behaviour. In addition, a corpus of social psychology linked to fundamental needs makes it possible to personally address a person’s different sources of motivation (need for cognition, need for uniqueness, need for effects and need for closure). By developing a composite measure of motivation based on these needs, we provide human resources professionals, and in particular occupational psychologists, with a tool that complements the assessment of self-determined motivation, making it possible to precisely address the objective of adapting work not to the self-determination of behaviours, but to the motivational traits of employees. To develop such a model, we gathered the French versions of the cognitive needs scales (need for cognition, need for uniqueness, need for effects, need for closure) and conducted a study with 645 employees of several French companies. On the basis of the data collected, we conducted a confirmatory factor analysis to validate the model, studied the correlations between the various needs, and highlighted the different reference groups that could be used to use these needs as a basis for interviews with employees (career, recruitment, etc.). The results showed a coherent model and the expected links between the different needs. Taken together, these results make it possible to propose a valid and theoretically adjusted tool to managers who wish to adapt their management to their employees’ current motivations, whether or not these motivations are self-determined.

Keywords: motivation, personality, work commitment, cognitive needs

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Authors: Dongguk Im, Gunfeel Moon, Younwoo Nam, Kangwoo Chun

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Recognition of the needs about protecting environment throughout worldwide is widespread. In the shipping industry, International Maritime Organization (IMO) has been regulating pollutants emitted from ships by MARPOL 73/78. Recently, the Marine Environment Protection Committee (MEPC) of IMO, at its 68th session, approved the definition of Black Carbon (BC) specified by the following physical properties (light absorption, refractory, insolubility and morphology). The committee also agreed to the need for a protocol for any voluntary measurement studies to identify the most appropriate measurement methods. Filter Smoke Number (FSN) based on light absorption is categorized as one of the IMO relevant BC measurement methods. EUROMOT provided a FSN measurement data (measured by smoke meter) of 31 different engines (low, medium and high speed marine engines) of member companies at the 3rd International Council on Clean Transportation (ICCT) workshop on marine BC. From the comparison of FSN, the results indicated that BC emission from low speed marine diesel engines was ranged from 0.009 to 0.179 FSN and it from medium and high speed marine diesel engine was ranged 0.012 to 3.2 FSN. In consideration of measured the low FSN from low speed engine, an experimental study was conducted using both a low speed marine diesel engine (2 stroke, power of 7,400 kW at 129 rpm) and a high speed marine diesel engine (4 stroke, power of 403 kW at 1,800 rpm) under E3 test cycle. The results revealed that FSN was ranged from 0.01 to 0.16 and 1.09 to 1.35 for low and high speed engines, respectively. The measurement equipment (smoke meter) ranges from 0 to 10 FSN. Considering measurement range of it, FSN values from low speed engines are near the detection limit (0.002 FSN or ~0.02 mg/m3). From these results, it seems to be modulated the measurement range of the measurement equipment (smoke meter) for enhancing measurement accuracy of marine BC and evaluation on performance of BC abatement technologies.

Keywords: black carbon, filter smoke number, international maritime organization, marine diesel engine (two and four stroke), particulate matter

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Authors: Mohammed Abdullah Alshahrani

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The research focuses on investigating the use of partial least squares (PLS) methodology for addressing challenges associated with high-dimensional correlated data. Recent technological advancements have led to experiments producing data characterized by a large number of variables compared to observations, with substantial inter-variable correlations. Such data patterns are common in chemometrics, where near-infrared (NIR) spectrometer calibrations record chemical absorbance levels across hundreds of wavelengths, and in genomics, where thousands of genomic regions' copy number alterations (CNA) are recorded from cancer patients. PLS serves as a widely used method for analyzing high-dimensional data, functioning as a regression tool in chemometrics and a classification method in genomics. It handles data complexity by creating latent variables (components) from original variables. However, applying PLS can present challenges. The study investigates key areas to address these challenges, including unifying interpretations across three main PLS algorithms and exploring unusual negative shrinkage factors encountered during model fitting. The research presents an alternative approach to addressing the interpretation challenge of predictor weights associated with PLS. Sparse estimation of predictor weights is employed using a penalty function combining a lasso penalty for sparsity and a Cauchy distribution-based penalty to account for variable dependencies. The results demonstrate sparse and grouped weight estimates, aiding interpretation and prediction tasks in genomic data analysis. High-dimensional data scenarios, where predictors outnumber observations, are common in regression analysis applications. Ordinary least squares regression (OLS), the standard method, performs inadequately with high-dimensional and highly correlated data. Copy number alterations (CNA) in key genes have been linked to disease phenotypes, highlighting the importance of accurate classification of gene expression data in bioinformatics and biology using regularized methods like PLS for regression and classification.

Keywords: partial least square regression, genetics data, negative filter factors, high dimensional data, high correlated data

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Authors: Paula Buldres, Jorge Toledo

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Objectives: This study aimed to characterize potential probiotic strains isolated from canine breast milk for use in dogs with enteropathies. Methodology: Six canine breast milk strains, one canine colostrum strain, and one control porcine breast milk strain were characterized. According to its functional properties of resistance to acids, different concentrations of bile salts, and pancreatin, its presumptive properties of safety and inhibitory effect on pathogens, non-cytotoxic characteristics, and adhesion to the intestine. The immunomodulatory effect of formulations with better probiotic characterization in vitro and in vivo was also analyzed. Results: Two strains characterized as potential probiotics were obtained, which corresponded to the canine strains (TUCO-16 and TUCO-17), presenting resistance to acidic pH, bile salts, and pancreatin, as well as an inhibitory effect on pathogenic Escherichia coli, Salmonella sp., and Clostridium perfringens. Strains TUCO-16 and TUCO-17 induced a significant increase in the expression of TNF-α and IL-8 in canine macrophages, respectively. Expression analyses of pattern recognition receptors in DH82 cells suggest that TUCO-16 and TUCO-17 might increase the TLR2 expression marker, and porcine strain (TUCO-4) increases the NOD2 expression marker. Based on the count obtained and the encapsulation yield, the best formulations correspond to FOS-Inulin for the TUCO-17 and TUCO-4 strains; Maltodextrin-Inulin for TUCO-16. All the strains are non-cytotoxic. The strain that showed the highest adhesion to intestinal epithelial cells was TUCO-17 with the FOS-Inulin formulation. On the other hand, the probiotics decreased the expression of pro-inflammatory markers in vivo, both in the intestine and in the spleen of mice. Conclusion: The combination of these three strains under study (TUCO-16, TUCO-17, and TUCO-4) would cover the probiotic properties in formulation and immunomodulation of all the markers under study.

Keywords: probiotics, gastrointestinal infec, dog, probiotic formulation, immunomodulatory probiotics

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Authors: Binaya Tamang, Buddhhi Raj Pokhrel, Narayan Gautam

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Background: There may be a connection between thyroid function and vitamin D status since both bind to similar nuclear hormone receptors and have similar response regions on gene promoters. The purpose of the current study was to investigate the relationship between thyroid hormones and vitamin D levels in females who were attending a tertiary care center in western Nepal and were either hypothyroid or euthyroid. Methods: This hospital-based cross-sectional study was carried out between March 2020 and March 2021 by the Biochemistry department of the Universal College of Medical Sciences (UCMS), Bhairahawa, Province No. 5, Nepal, in cooperation with Internal medicine. Prior to the study, institutional review committee approval (UCMS/IRC/008/20) was acquired from UCMS. Women who visited the Internal Medicine OPD of UCMS and were advised to get a thyroid function test (TFT) were included in the study population. Only those who were willing to participate in the study were enrolled after the goals and advantages of the study had been explained to them. Participants who had recently used vitamin D supplements and medications that affected thyroid hormones were excluded. The participants gave their consent verbally and in writing. After getting the consent, a convenient sample technique was applied. Serum was isolated after drawing 3 ml of blood in a plain vial. Chemiluminescence assay was used to analyze vitamin D and thyroid hormones (MAGLUMI 2000). SPSS version 16.0 for Windows was used to conduct the statistical analysis. Statistical significance was defined as a P-value < 0.05. Results: Majority of the study population (n=214, 71%) had insufficient serum vitamin D levels. Among the thyroid groups, the median Vitamin D levels were significantly lower in hypothyroid (16.88 ng/ml) as compared to the euthyroid groups (25.01 ng/ml) (P<0.001). Similarly, serum Vitamin D levels were considerably lower in the obese population (16.86 ng/ml) as compared to the normal BMI group (24.90 ng/ml) (P<0.001) as well as in the vegetarian (15.43 ng.ml) than mixed diet consumer (24.89 ng/ml) (P<0.01). Even after the adjustment for these variables, the Vitamin D levels were significantly lower in the hypothyroid population than in the euthyroid group (P<0.001). Conclusion: Comparing the hypothyroid population to the euthyroid, the median serum vitamin D levels were considerably lower. We were alarmed to see that the majority of euthyroid participants also had low levels of vitamin D. Therefore if left untreated, low vitamin D levels in hypothyroid patients could worsen their health further.

Keywords: vitamin D, thyroid hormones, euthyroid, hypothyroid, Nepal

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Authors: Euripides Antoniades

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In 1821, the Greek Revolution movement, under the political influence that arose from the French revolution, and the corresponding movements in Italy, Germany and America, demanded the liberation of the nation and the establishment of an independent national state. Published topics in the British press regarding the Greek Revolution, focused on: a)the right of the Greeks to claim their freedom from Turkish domination in order to establish an independent state based on the principle of national autonomy, b)criticism regarding Turkish rule as illegal and the power of the Ottoman Sultan as arbitrary, c)the recognition of the Greek identity and its distinction from the Turkish one and d)the endorsement Greeks as the descendants of ancient Greeks. The London Times is a print publication that presents, in chronological or thematic order, the news, opinions or announcements about the most important events that have occurred in a place during a specified period of time. A combination of qualitative and quantitative content analysis was applied. An attempt was made to record Greek Revolution references along with the usage of specific words and expressions that contribute to the representation of the historical events and their exposure to the reading public. Key finds of this research reveal that a)there was a frequency of passionate daily articles concerning the events in Greece, their length, and context in The Times of London, b)he British public opinion was influenced by this particular newspaper and c) he newspaper published various news about the revolution by adopting the role of animator of the Greek struggle. In fact, this type of news was the main substance of the The London Times’ structure, establishing a positive image about the Greek Revolution contributing to the European diplomatic development. These factors offered a change in the attitude of the British and Russians respectively assuming a positive approach towards Greece.

Keywords: Greece, revolution, press, the london times, great britain, mass media

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Authors: Salina Yahya Saddick

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Ovarian cancer remains the most lethal gynecological malignancy due to the lack of highly sensitive and specific screening tools for detection of early-stage disease. The OSE provides the progenitor cells for 90% of human ovarian cancers. Recent morphologic, immunohistochemical and molecular genetic studies have led to the development of a new paradigm for the pathogenesis and origin of epithelial ovarian cancer (EOC) based on a ualistic model of carcinogenesis that divides EOC into two broad categories designated Types I and II which are characterized by specific mutations, including KRAS, BRAF, ERBB2, CTNNB1, PTEN PIK3CA, ARID1A, and PPPR1A, which target specific cell signaling pathways. Type 1 tumors rarely harbor TP53. type I tumors are relatively genetically stable and typically display a variety of somatic sequence mutations that include KRAS, BRAF, PTEN, PIK3CA CTNNB1 (the gene encoding beta catenin), ARID1A and PPP2R1A but very rarely TP53 . The cancer stem cell (CSC) hypothesis postulates that the tumorigenic potential of CSCs is confined to a very small subset of tumor cells and is defined by their ability to self-renew and differentiate leading to the formation of a tumor mass. Potential protein biomarker miRNA, are promising biomarkers as they are remarkably stable to allow isolation and analysis from tissues and from blood in which they can be found as free circulating nucleic acids and in mononuclear cells. Recently, genomic anaylsis have identified biomarkers and potential therapeutic targets for ovarian cancer namely, FGF18 which plays an active role in controlling migration, invasion, and tumorigenicity of ovarian cancer cells through NF-κB activation, which increased the production of oncogenic cytokines and chemokines. This review summarizes update information on epithelial ovarian cancers and point out to the most recent ongoing research.

Keywords: epithelial ovarian cancers, somatic sequence mutations, cancer stem cell (CSC), potential protein, biomarker, genomic analysis, FGF18 biomarker

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Authors: Emna Ketata, Wafa Farhat

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Introduction: Hyponatremia is defined by a blood sodium level of ≤ 136 mmol/L; it is associated with a high risk of morbidity and mortality in the emergency room. This was explained by transit disorders, including diarrhea and inappropriate antidiuretic hormone secretion (Syndrome of inappropriate antidiuretic hormone secretion). Pneumonia can cause dyspnea, stress-causing SIADH and digestive symptoms (diarrhea and vomiting). Aim: The purpose of this study was to determine the link between pneumonia and hyponatremia as a predictor of patient’s prognosis and intra-hospital mortality. Methodology: This is a prospective observational study over a period of 3 years in the emergency department. Inclusion :patients (age > 14 years), with clinical signs in favor of pneumonia. Natremia was measured. Natremia was classified as mild to moderate with a blood sodium level between 121 and 135 mmol/L and as severe with a blood sodium level ≤ 120 mmol/L. Results: This study showed an average serum sodium value of 135 mmol/L (range 114–159 mmol/L) in these patients. Hyponatremia was observed in 123 patients (43.6%), 115 patients (97,8%) had mild to moderate hyponatremia and 2,8% had severe hyponatremia. The mean age was 65±17 years with a sex ratio of 1.05. The main reason for consultation in patients with hyponatremia was cough in 58 patients (47.2%), and digestive symptoms were present in 25 patients (20.3. An altered state of consciousness was observed in 11 patients (3%). Patients with hyponatremia had greater heart rate (p=0.02),white blood cell count (p=0.009) , plasmatic lactate (p=0.002) and higher rate of pneumonia recurrence (p=0.001) .In addition, 80% of them have a positive CURB65 score (>=2). hyponatremia had higher rates of use of oxygen therapy compared to patients with normo-natremia (54% vs. 45%). The analytical study showed that hyponatremia is significantly associated with intra-hospital mortality with( p=0.01), severe hyponatremia p=0.04. Conclusion: Hyponatremia is a predictor of mortality and worse prognosis. Recognition of the pathophysiological mechanisms of hyponatremia in pneumonia will probably allow better management of it.

Keywords: oxygenotherapy, mortality, recurrence, positif curb65

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Authors: Mwafaq Ibdah, Mossab, Yahyaa, Dor Rachmany, Yoram Gerchman, Doron Holland, Liora Shaltiel-Harpaz

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Pear Psylla is the most important pest of pear in all pear-growing regions, in Asian, European, and the USA. Pear psylla damages pears in several ways: high-density populations of these insects can cause premature leaf and fruit drop, diminish plant growth, and reduce fruit size. In addition, their honeydew promotes sooty mold on leaves and russeting on fruit. Pear psyllas are also considered vectors of pear pathogens such as Candidatus Phytoplasma pyri causing pear decline that can lead to loss of crop and tree vigor, and sometimes loss of trees. Psylla control is a major obstacle to efficient integrated pest management. Recently we have identified two naturally resistance pear accessions (Py.760-261 and Py.701-202) in the Newe Ya’ar live collection. GC-MS volatile metabolic profiling identified several volatile compounds common in these accessions but lacking, or much less common, in a sensitive accession, the commercial Spadona variety. Among these volatiles were styrene and its derivatives. When the resistant accessions were used as inter-stock, the volatile compounds appear in commercial Spadona scion leaves, and it showed reduced susceptibility to pear psylla. Laboratory experiments and applications of some of these volatile compounds were very effective against psylla eggs, nymphs, and adults. The genes and enzymes involved in the specific reactions that lead to the biosynthesis of styrene in plant are unknown. We have identified a phenolic acid decarboxylase that catalyzes the formation of p-hydroxystyrene, which occurs as a styrene analog in resistant pear genotypes. The His-tagged and affinity chromatography purified E. coli-expressed pear PyPAD1 protein could decarboxylate p-coumaric acid and ferulic acid to p-hydroxystyrene and 3-methoxy-4-hydroxystyrene. In addition, PyPAD1 had the highest activity toward p-coumaric acid. Expression analysis of the PyPAD gene revealed that its expressed as expected, i.e., high when styrene levels and psylla resistance were high.

Keywords: pear Psylla, volatile, GC-MS, resistance

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Authors: N. Umasuthan, S. D. N. K. Bathige, W. S. Thulasitha, I. Whang, J. Lee

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The MyD88 is an evolutionarily conserved host-expressed adaptor protein that is essential for proper TLR/ IL1R immune-response signaling. A previously identified complete cDNA (1626 bp) of OfMyD88 comprised an ORF of 867 bp encoding a protein of 288 amino acids (32.9 kDa). The gDNA (3761 bp) of OfMyD88 revealed a quinquepartite genome organization composed of 5 exons (with the sizes of 310, 132, 178, 92 and 155 bp) separated by 4 introns. All the introns displayed splice signals consistent with the consensus GT/AG rule. A bipartite domain structure with two domains namely death domain (24-103) coded by 1st exon, and TIR domain (151-288) coded by last 3 exons were identified through in silico analysis. Moreover, homology modeling of these two domains revealed a similar quaternary folding nature between human and rock bream homologs. A comprehensive comparison of vertebrate MyD88 genes showed that they possess a 5-exonic structure. In this structure, the last three exons were strongly conserved, and this suggests that a rigid structure has been maintained during vertebrate evolution. A cluster of TATA box-like sequences were found 0.25 kb upstream of cDNA starting position. In addition, putative 5'-flanking region of OfMyD88 was predicted to have TFBS implicated with TLR signaling, including copies of NFB1, APRF/ STAT3, Sp1, IRF1 and 2 and Stat1/2. Using qPCR technique, a ubiquitous mRNA expression was detected in liver and blood. Furthermore, a significantly up-regulated transcriptional expression of OfMyD88 was detected in head kidney (12-24 h; >2-fold), spleen (6 h; 1.5-fold), liver (3 h; 1.9-fold) and intestine (24 h; ~2-fold) post-Fla challenge. These data suggest a crucial role for MyD88 in antibacterial immunity of teleosts.

Keywords: MyD88, innate immunity, flagellin, genomic analysis

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Authors: Vichayada Laohapiboolkul

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Introduction: As the legal use of cannabis is being widely accepted throughout the world, medical cannabis has been explored in order to become an alternative cure for patients. Tetrahydrocannabinol (THC) and Cannabidiol (CBD) are natural cannabinoids found in the Cannabis plant which is proved to have positive treatment for various diseases and symptoms such as chronic pain, neuropathic pain, spasticity resulting from multiple sclerosis, reduce cancer-associated pain, autism spectrum disorders (ASD), dementia, cannabis and opioid dependence, psychoses/schizophrenia, general social anxiety, posttraumatic stress disorder, anorexia nervosa, attention-deficit hyperactivity disorder, and Tourette's disorder. Regardless of all the medical benefits, THC, if not metabolized, can lead to mild up to severe adverse drug reactions (ADR). The enzyme CYP2C19 was found to be one of the metabolizers of THC. However, the suballele CYP2C19*2 manifests as a poor metabolizer which could lead to higher levels of THC than usual, possibly leading to various ADRs. Objective: The aim of this study was to investigate the distribution of CYP2C19, specifically CYP2C19*2, genes in Thai patients treated with medical cannabis along with adverse drug reactions. Materials and Methods: Clinical data and EDTA whole blood for DNA extraction and genotyping were collected from patients for this study. CYP2C19*2 (681G>A, rs4244285) genotyping was conducted using the Real-time PCR (ABI, Foster City, CA, USA). Results: There were 42 medical cannabis-induced ADRs cases and 18 medical cannabis tolerance controls who were included in this study. A total of 60 patients were observed where 38 (63.3%) patients were female and 22 (36.7%) were male, with a range of age approximately 19 - 87 years. The most apparent ADRs for medical cannabis treatment were dry mouth/dry throat (76.7%), followed by tachycardia (70%), nausea (30%) and a few arrhythmias (10%). In the total of 27 cases, we found a frequency of 18 CYP2C19*1/*1 alleles (normal metabolizers, 66.7%), 8 CYP2C19*1/*2 alleles (intermediate metabolizers, 29.6%) and 1 CYP2C19*2/*2 alleles (poor metabolizers, 3.7%). Meanwhile, 63.6% of CYP2C19*1/*1, 36.3% and 0% of CYP2C19*1/*2 and *2/*2 in the tolerance controls group, respectively. Conclusions: This is the first study to confirm the distribution of CYP2C19*2 allele and the prevalence of poor metabolizer genes in Thai patients who received medical cannabis for treatment. Thus, CYP2C19 allele might serve as a pharmacogenetics marker for screening before initiating treatment.

Keywords: medical cannabis, adverse drug reactions, CYP2C19, tetrahydrocannabinol, poor metabolizer

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Authors: Gulnaz Khan, Meha F. Aftab, Munazza Murtaza, Rizwana S. Waraich

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Advanced glycation end products (AGEs) precursor and its abnormal accumulation cause damage to various tissues and organs. AGEs have pathogenic implication in several diseases including diabetes. Existing AGEs inhibitors are not in clinical use, and there is a need for development of novel inhibitors. The present investigation aimed at identifying the novel AGEs inhibitors and assessing their mechanism of action for treating insulin resistance in mice model of diabetes. Novel derivatives of benzylidene of indan-1,3-dione were synthesized. The compounds were selected to study their action mechanism in improving insulin resistance, in vitro, in human hepatocytes and murine adipocytes and then, in vivo, in mice genetic model of diabetes (db/db). Mice were treated with novel derivatives of benzylidene of indane 1,3-dione. AGEs mediated ROS production was measured by dihydroethidium fluorescence assay. AGEs level in the serum of treated mice was observed by ELISA. Gene expression of receptor for AGEs (RAGE), PPAR-gamma, TNF-alpha and GLUT-4 was evaluated by RT-PCR. Glucose uptake was measured by fluorescent method. Microscopy was used to analyze glycogen synthesis in muscle. Among several derivatives of benzylidene of indan-1,3-dione, IDD-24, demonstrated highest inhibition of AGESs. IDD-24 significantly reduced AGEs formation and expression of receptor for advanced glycation end products (RAGE) in fat, liver of db/db mice. Suppression of AGEs mediated ROS production was also observed in hepatocytes and fat cell, after treatment with IDD-24. Glycogen synthesis was increased in muscle tissue of mice treated with IDD-24. In adipocytes, IDD-24 prevented AGEs induced reduced glucose uptake. Mice treated with IDD-24 exhibited increased glucose tolerance, serum adiponectin levels and decreased insulin resistance. The result of present study suggested that IDD-24 can be a possible treatment target to address glycotoxins induced insulin resistance.

Keywords: advance glycation end product, hyperglycemia, indan-1, 3-dione, insulin resistance

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Authors: Gaurav Kumar Sinha

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In an era marked by increasing concerns about energy sustainability, this research endeavors to address the pressing challenge of energy consumption in buildings and industries. This study delves into the transformative potential of AWS services in optimizing energy efficiency. The research is founded on the recognition that effective management of energy consumption is imperative for both environmental conservation and economic viability. Buildings and industries account for a substantial portion of global energy use, making it crucial to develop advanced techniques for analysis and reduction. This study sets out to explore the integration of AWS services with big data analytics to provide innovative solutions for energy consumption analysis. Leveraging AWS's cloud computing capabilities, scalable infrastructure, and data analytics tools, the research aims to develop efficient methods for collecting, processing, and analyzing energy data from diverse sources. The core focus is on creating predictive models and real-time monitoring systems that enable proactive energy management. By harnessing AWS's machine learning and data analytics capabilities, the research seeks to identify patterns, anomalies, and optimization opportunities within energy consumption data. Furthermore, this study aims to propose actionable recommendations for reducing energy consumption in buildings and industries. By combining AWS services with metrics-driven insights, the research strives to facilitate the implementation of energy-efficient practices, ultimately leading to reduced carbon emissions and cost savings. The integration of AWS services not only enhances the analytical capabilities but also offers scalable solutions that can be customized for different building and industrial contexts. The research also recognizes the potential for AWS-powered solutions to promote sustainable practices and support environmental stewardship.

Keywords: energy consumption analysis, big data analytics, AWS services, energy efficiency

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Authors: Nima Samie, Batoul Sadat Haerian, Sekaran Muniandy, M. S. Kanthimathi

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Colon and breast cancers are categorized as the most prevalent types of cancer worldwide. Recently, the broad clinical application of metal complex compounds has led to the discovery of potential therapeutic drugs. The aim of this study was to evaluate the cytotoxic action of a selected nickel complex compound (NCC) against human colon and breast cancer cells. In this context, we determined the potency of the compound in the induction of apoptosis, cell cycle arrest, and cytoskeleton rearrangement. HT-29, WiDr, CCD-18Co, MCF-7 and Hs 190.T cell lines were used to determine the IC50 of the compound using the MTT assay. Analysis of apoptosis was carried out using immunofluorescence, acridine orange/ propidium iodide double staining, Annexin-V-FITC assay, evaluation of the translocation of NF-kB, oxygen radical antioxidant capacity, quenching of reactive oxygen species content , measurement of LDH release, caspase-3/-7, -8 and -9 assays and western blotting. The cell cycle arrest was examined using flowcytometry and gene expression was assessed using qPCR array. Results showed that our nickel complex compound displayed a potent suppressive effect on HT-29, WiDr, MCF-7 and Hs 190.T after 24 h of treatment with IC50 value of 2.02±0.54, 2.13±0.65, 3.76±015 and 3.14±0.45 µM respectively. This cytotoxic effect on normal cells was insignificant. Dipping in the mitochondrial membrane potential and increased release of cytochrome c from the mitochondria indicated induction of the intrinsic apoptosis pathway by the nickel complex compound. Activation of this pathway was further evidenced by significant activation of caspase 9 and 3/7.The nickel complex compound (NCC) was also shown activate the extrinsic pathways of apoptosis by activation of caspase-8 which is linked to the suppression of NF-kB translocation to the nucleus. Cell cycle arrest in the G1 phase and up-regulation of glutathione reductase, based on excessive ROS production were also observed. The results of this study suggest that the nickel complex compound is a potent anti-cancer agent inducing both intrinsic and extrinsic pathways as well as cell cycle arrest in colon and breast cancer cells.

Keywords: nickel complex, apoptosis, cytoskeletal rearrangement, colon cancer, breast cancer

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Authors: Sweta Pandey, Samridhi Dhyani, Susmita Chaudhuri

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Klebsiella pneumoniae causes a wide range of infections, including urinary tract infections, sepsis, bacteremia, pneumonia, and liver abscesses. The emergence of multi-drug resistance in this bacterium led to a major setback for clinical management. WHO also endorsed a need for finding alternative therapy to antibiotics for the treatment of these infections. Development of vaccines and passive antibody therapy has been proven as a potent alternative to antibiotics in the case of MDR, XDR, and PDR Klebsiella infections. Siderophore receptors have been demonstrated to be overexpressed for the internalization of iron siderophore complexes during infections in most Gram-negative bacteria. For the present study, immune response to siderophore receptors to establish this protein as a potential immunogen for the development of therapeutic leads was explored. Clinical strains of Klebsiella pneumoniae were grown in iron-deficient conditions, and the iron-regulated outer membrane proteins were extracted and characterized through mass spectrometry for specific identification. The gene for identified protein was cloned in pET- 28a vector and expressed in E. coli. The native protein and the recombinant protein were isolated and purified and used as antigens for the generation of immune response in BALB/c mice. The native protein of Klebsiella pneumoniae grown in iron-deficient conditions was identified as FepA (Ferrienterobactin receptor) and other siderophore receptors. This 80 kDa protein generated an immune response in BALB/c mice. The antiserum from mice after subsequent booster doses was collected and showed binding with FepA protein in western blot and phagocytic uptake of the K. pneumoniae in the presence antiserum from immunized mice also observed from the animal studies after bacterial challenge post immunisation in mice have shown bacterial clearance. The antiserum from mice showed binding and clearance of the Klebsiella pneumoniae bacteria in vitro and in vivo. These antigens used for generating an active immune response in mice can further be used for therapeutic monoclonal antibody development against Klebsiella pneumoniae infections.

Keywords: antiserum, FepA, Klebsiella pneumoniae, multi drug resistance, siderophore receptor

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Authors: Ghebremedhin Tsegay, Weldu Tesfagaber, Yuanmao Zhu, Xijun He, Wan Wang, Zhenjiang Zhang, Encheng Sun, Jinya Zhang, Yuntao Guan, Fang Li, Renqiang Liu, Zhigao Bu, Dongming Zhao*

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African swine fever (ASF) is a highly infectious viral disease of pigs, resulting in significant economic loss worldwide. As there is no approved vaccines and treatments, the control of ASF entirely depends on early diagnosis and culling of infected pigs. Thus, highly specific and sensitive diagnostic assays are required for accurate and early diagnosis of ASF virus (ASFV). Currently, only a few recombinant proteins have been tested and validated for use as reagents in ASF diagnostic assays. The most promising ones for ASFV antibody detection were p72, p30, p54, and pp62. So far, three ELISA kits based on these recombinant proteins have been commercialized. Due to the complex nature of the virus and variety forms of the disease, robust serodiagnostic assays are still required. ASFV p22 protein, encoded by KP177R gene, is located in the inner membrane of viral particle and appeared transiently in the plasma membrane early after virus infection. The p22 protein interacts with numerous cellular proteins, involved in processes of phagocytosis and endocytosis through different cellular pathways. However, p22 does not seem to be involved in virus replication or swine pathogenicity. In this study, E.coli expressed recombinant p22 protein was used to generate a monoclonal antibody (mAb), and its potential use for the development of blocking ELISA (bELISA) was evaluated. A total of 806 pig serum samples were tested to evaluate the bELISA. Acording the ROC (Reciever operating chracteristic) analysis, 100% sensitivity and 98.10% of specificity was recorded when the PI cut-off value was set at 47%. The novel assay was able to detect the antibodies as early as 9 days post infection. Finaly, a highly sensitive, specific and rapid novel p22-mAb based bELISA assay was developed, and optimized for detection of antibodies against genotype I and II ASFVs. It is a promising candidate for an early and acurate detection of the antibodies and is highly expected to have a valuable role in the containment and prevention of ASF.

Keywords: ASFV, blocking ELISA, diagnosis, monoclonal antibodies, sensitivity, specificity

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Authors: Sara Vora (Hoxha)

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Defamation, invasion of privacy, and cyberbullying have all increased in tandem with the growth of the internet. European international private law may struggle to deal with such transgressions if they occur in many jurisdictions. The current study examines how effectively the legal system of European international private law addresses abuses of personality rights in cyberspace. The study starts by discussing how established legal frameworks are being threatened by online personality rights abuses. The article then looks into the rules and regulations of European international private law that are in place to handle overseas lawsuits. This article examines the different elements that courts evaluate when deciding which law to use in a particular case, focusing on the concepts of jurisdiction, choice of law, and recognition and execution of foreign judgements. Next, the research analyses the function of the European Union in preventing and punishing online personality rights abuses. Key pieces of law that control the collecting and processing of personal data on the Internet, including the General Data Protection Regulation (GDPR) and the e-Commerce Directive, are discussed. In addition, this article investigates how the ECtHR handles cases involving the infringement of personal freedoms, including privacy and speech. The article finishes with an assessment of how well the legal framework of European international private law protects individuals' right to privacy online. It draws attention to problems with the present legal structure, such as the inability to enforce international judgements, the inconsistency between national laws, and the necessity for stronger measures to safeguard people' rights online. This paper concludes that while European international private law provides a useful framework for dealing with violations of personality rights online, further harmonisation and stronger enforcement mechanisms are necessary to effectively protect individuals' rights in the digital age.

Keywords: European international private law, personality rights, internet, jurisdiction, cross-border disputes, data protection

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Authors: Osvaldo N. Oliveira Jr.

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The control of molecular architecture inherent in some experimental methods to produce nanostructured films has had great impact on devices of various types, including sensors and biosensors. The self-assembly monolayers (SAMs) and the electrostatic layer-by-layer (LbL) techniques, for example, are now routinely used to produce tailored architectures for biosensing where biomolecules are immobilized with long-lasting preserved activity. Enzymes, antigens, antibodies, peptides and many other molecules serve as the molecular recognition elements for detecting an equally wide variety of analytes. The principles of detection are also varied, including electrochemical methods, fluorescence spectroscopy and impedance spectroscopy. In this presentation an overview will be provided of biosensors made with nanostructured films to detect antibodies associated with tropical diseases and HIV, in addition to detection of analytes of medical interest such as cholesterol and triglycerides. Because large amounts of data are generated in the biosensing experiments, use has been made of computational and statistical methods to optimize performance. Multidimensional projection techniques such as Sammon´s mapping have been shown more efficient than traditional multivariate statistical analysis in identifying small concentrations of anti-HIV antibodies and for distinguishing between blood serum samples of animals infected with two tropical diseases, namely Chagas´ disease and Leishmaniasis. Optimization of biosensing may include a combination of another information visualization method, the Parallel Coordinate technique, with artificial intelligence methods in order to identify the most suitable frequencies for reaching higher sensitivity using impedance spectroscopy. Also discussed will be the possible convergence of technologies, through which machine learning and other computational methods may be used to treat data from biosensors within an expert system for clinical diagnosis.

Keywords: clinical diagnosis, information visualization, nanostructured films, layer-by-layer technique

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Authors: Ghada Esheba, Ebtisam Aljerayan, Afnan Al-Ghamdi, Atheer Alsharif, Hanan alzahrani

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Background: Primary ovarian neoplasms comprise a heterogeneous group of tumors of three main subtypes: surface epithelial, germ cell, and sex cord-stromal. The wide morphological variation within and between these groups can result in diagnostic difficulties. Gonadal sex cord-stromal tumors (SCST) represent one of the most heterogeneous categories of human neoplasms, because they may contain various combinations of different gonadal sex cord and stromal element. Aim: The aim of this work is to highlight the clinicopathological characteristics of SCST and to assess the value of alpha-inhibin and calretinin in the distinction between SCST and their mimics. Material and methods: This study was carried out on 100 cases using full tissue sections; 70 cases were SCST and 30 cases were histological mimics of SCST. The cases were studied using immunohistochemically using alpha-inhibin. In addition, an ovarian tissue microarray containing 170 benign and malignant ovarian neoplasms was also studied immunohistochemically for calretinin expression. The ovarian microarray included 14 SCST, 59 ovarian serous borderline tumors, 17 mucinous borderline tumors, 10 mucinous adenocarcinomas, 32 endometrioid adenocarcinomas, 34 clear cell carcinomas, and 4 germ cell tumors. Results: 99% of SCST examined using full tissue sections exhibited positive cytoplasmic staining for inhibin. On the contrary, only 7% of the histological mimics (P value < 0.0001). 86% of SCST in the tissue microarray were positive for calretinin with nuclear and/or cytoplasmic staining compared to only 7% of the other tumor types (P value < 0.0001). Conclusions: SCST have characteristic clinicopathological and immunohistochemical features and their recognition is crucial for proper diagnosis and treatment. Alpha-inhibin and calretinin are of great help in the diagnosis of sex cord-stromal tumors.

Keywords: calretinin, granulosa cell tumor, inhibin, sex cord-stromal tumors

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Authors: Amanda J. Burridge, Keith J. Edwards, Paul A. Wilkinson, Tom Batstone, Erik H. Murchie, Lorna McAusland, Ana Elizabete Carmo-Silva, Ivan Jauregui, Tracy Lawson, Silvere R. M. Vialet-Chabrand

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The rate of genetic progress in wheat production must be improved to meet global food security targets. However, past selection for domestication traits has reduced the genetic variation in modern wheat cultivars, a fact that could severely limit the future rate of genetic gain. The genetic variation in agronomically important traits for the wild relatives and progenitors of wheat is far greater than that of the current domesticated cultivars, but transferring these traits into modern cultivars is not straightforward. Between the elite cultivars of wheat, photosynthetic capacity is a key trait for which there is limited variation. Early screening of wheat wild relative and progenitors has shown differences in photosynthetic capacity and efficiency not only between wild relative species but marked differences between the accessions of each species. By identifying wild relative accessions with improved photosynthetic traits and characterising the genetic variation responsible, it is possible to incorporate these traits into advanced breeding programmes by wide crossing and introgression programmes. To identify the potential variety of photosynthetic capacity and efficiency available in the secondary and tertiary genepool, a wide scale survey was carried out for over 600 accessions from 80 species including those from the genus Aegilops, Triticum, Thinopyrum, Elymus, and Secale. Genotype data were generated for each accession using a ‘Wheat Wild Relative’ Single Nucleotide Polymorphism (SNP) genotyping array composed of 35,000 SNP markers polymorphic between wild relatives and elite hexaploid wheat. This genotype data was combined with phenotypic measurements such as gas exchange (CO₂, H₂O), chlorophyll fluorescence, growth, morphology, and RuBisCO activity to identify potential breeding material with enhanced photosynthetic capacity and efficiency. The data and associated analysis tools presented here will prove useful to anyone interested in increasing the genetic diversity in hexaploid wheat or the application of complex genotyping data to plant breeding.

Keywords: wheat, wild relatives, pre-breeding, genomics, photosynthesis

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Authors: Sarah Hebbouch

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Two decades ago, research on diasporic women’s participation within Sufi circles would have been inconceivable, not only because of a general lack of recognition of their contribution to Sufism but due to the intimacy of the rituals, often taking place in confined spaces, like zawiyas (Sufi lodges). Recent scholarly attention to female spiritual experience owes to a digital awareness and interest in exploring diasporic community reproduction of those experiences. Within a context where female disciples of a Sufi convent undergo a physical separation from the saint’s sanctuary -because of immigration from the homeland to the host country- technology becomes a social hub accounting for Sufis’ ritual commitment and preservation of cultural capital in the diaspora. This paper elucidates how female Sufi immigrants affiliating with the Boudchichi brotherhood (Morocco-based) maintain ‘a relational network’ and strong social online relationships with their female compatriots in Morocco through the use of online platforms. Sufi communities living in the diaspora find the internet an open interactive space that serves to kindle their distance of spiritual participation and corroborate their transnational belonging. The current paper explores the implications of the use of a digital baseline named “Tariqa Info,” the convent’s digital online platform, and how it mediates everyday ritual performance, the promotion of digital connection, and the communication of ideas and discourses. Such a platform serves the bolstering emotional bonds for transnational female disciples and inclusion within online communities in the homeland. Assisted by an ethnographic lens, this paper discusses the research findings of participatory field observation of Sufi women’s online communities, informed by the need to trace the many ostensible aspects of interconnectedness and divergences.

Keywords: digital connection, Sufi convent, social online relationship, transnational female disciples

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Authors: Beatriz Mella Lira, Karla Yohannessen, Robin Hickman

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The aim of the paper is recognising inequalities through the self-assessment of health-related factors, in the context of daily mobilities in Santiago de Chile. Human capabilities will be used as the theoretical basis for the recognition and assessment of these factors regarding the functioning (what people are currently able to do) and capabilities (what people want to achieve and what is valuable for them), reflecting differences across social groups and among types of transport users. The self-assessment of health-related factors considers perceptions of stress, physical effort, proximity to other transport users, pollution, safety, and comfort. The types of transport users are classified as: private (cars, taxis, colectivos, motos), public (buses and metro) and active (bicycles and walking). The methodology follows a capability-based questionnaire, which was applied in different areas of Santiago de Chile, considering concepts extracted from the human capabilities list. The self-assessment of these health-related factors examines the context of peoples’ mobilities for performing their daily activities, considering socioeconomic differences as income, age, gender, disabilities, residence location and primary mode choice. The paper uses Relative Importance Index (RII) for weighting the relative influence or valuation of the factors. The respondents were asked to rate the importance of each factor on a scale from 1 to 5, in an ascending order of importance. The results suggest that these health-related factors impact not just the perceptions of users, but their well-being and their propensity for achieving their capabilities and the things they value in life. The paper is focused on the development of an applicable approach, measuring factors that should be included in transport project appraisal, as a more comprehensive and complementary method.

Keywords: active transport, health, human capabilities, Santiago de Chile, transport inequalities, transportation planning, urban planning

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Authors: Ashokkumar Subbiah

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This study investigates the impact of factors that influenced the success of construction planning for a major construction project in Qatar. An approach of participant observation is adopted which is informed by the principles of ethnography: one that reports the participants’ view of their world rather than imposing an artificial theoretical framework upon it. As participant observant, key factors were observed and identified that had an impact on the management and execution of the construction planning. It is found that a ‘shadow culture’ exists between the project participants which, it is argued, is only observable from the perspective of an embedded participant observer. The shadow culture acts to enable the management of the planning process, and its efficacy relates to the ‘quality’ of human inter-relationships amongst immediate stakeholders. Whilst this study uses the concept of shadow culture, it is treated as both a methodological stance and one of the findings of this research in the context of the major construction project in Qatar. The concept of shadow culture is not imposed upon the findings, but instead is used as a research tool: respondents report their own worldview and this is reported from the view of a participant observant in a manner that is understandable and useful to those who are not part of the construction project. The findings of this study identify similar factors influencing the planning process of the Qatar project, but the shadow culture predominantly influences these factors towards the failure of planning process. The research concludes by questioning the assumption that construction planning is a mechanistic process that has to be conducted solely by the planning team. Instead, it is a highly social phenomenon in which the seemingly mechanistic process is made workable by the quality of relationships that exist in the project. Drawing on this the final section provides a series of recommendations that may be helpful in enhancing the efficacy of project planning; these include better training/education at the pre-construction phase; recognition of the importance of shadow processes at management levels, and better appreciation of the impact of contract type and chosen procurement route.

Keywords: construction planning, participant observation, project participants, shadow culture

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Authors: Lukas J. Evans, Danilo D. Fernando

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Shrub willows (Salix spp.) have many characteristics which make them suitable for a variety of applications such as riparian zone buffers, environmental contaminant sequestration, living snow fences, and biofuel production. In some cases, these functions are limited due to physical or financial obstacles associated with the number of individuals needed to reasonably satisfy that purpose. One way to increase the efficiency of willows is to bioengineer them with the genetic improvements suitable for the desired use. To accomplish this goal, an optimized in vitro transformation protocol via Agrobacterium tumefaciens is necessary to reliably express genes of interest. Therefore, the aim of this study is to observe the influence of tissue culture with different willow cultivars, hormones, and explants on the percentage of calli expressing reporter gene green florescent protein (GFP) to find ideal transformation conditions. Each callus was produced from 1 month old open-pollinated seedlings of three Salix miyabeana cultivars (‘SX61’, ‘WT1’, and ‘WT2’) from three different explants (lamina, petiole, and internodes). Explants were cultured for 1 month on an MS media with different concentrations of 6-Benzylaminopurine (BAP) and 1-Naphthaleneacetic acid (NAA) (No hormones, 1 mg⁻¹L BAP only, 3 mg⁻¹L NAA only, 1 mg⁻¹L BAP and 3 mg⁻¹L NAA, and 3 mg⁻¹L BAP and 1 mg⁻¹L NAA) to produce a callus. Samples were then treated with Agrobacterium tumefaciens at an OD600 of 0.6-0.8 to insert the transgene GFP for 30 minutes, co-cultivated for 72 hours, and selected on the same media type they were cultured on with added 7.5 mg⁻¹L of Hygromycin for 1 week before GFP visualization under a UV dissecting scope. Percentage of GFP expressing calli as well as the average number of fluorescing GFP units per callus were recorded and results were evaluated through an ANOVA test (α = 0.05). The WT1 internode-derived calli on media with 3 mg-1L NAA+1 mg⁻¹L BAP and mg⁻¹L BAP alone produced a significantly higher percentage of GFP expressing calli than each other group (19.1% and 19.4%, respectively). Additionally, The WT1 internode group cultured with 3 mg⁻¹L NAA+1 mg⁻¹L BAP produced an average of 2.89 GFP units per callus while the group cultivated with 1 mg⁻¹L BAP produced an average of 0.84 GFP units per callus. In conclusion, genotype, explant choice, and hormones all play a significant role in increasing successful transformation in willows. Future studies to produce whole callus GFP expression and subsequent plantlet regeneration are necessary for a complete willow transformation protocol.

Keywords: agrobacterium, callus, Salix, tissue culture

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Authors: Sajewicz-Krukowska Joanna, Domańska-Blicharz Katarzyna, Tarasiuk Karolina, Marzec-Kotarska Barbara

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Astroviral infections pose a significant problem in the poultry industry, leading to multiple adverse effects such as decreased egg production, breeding disorders, poor weight gain, and even increased mortality. Commonly observed chicken astrovirus (CAstV) was recently reported to be responsible for "white chicks syndrome" associated with increased embryo/chick mortality. The CAstV-mediated pathogenesis in chicken occurs due to complex interactions between the infectious pathogen and the immune system. Many aspects of CAstV-chicken interactions remain unclear, and there is no information available regarding gene expression changes in the chicken's spleen in response to CAstV infection. We aimed to investigate the molecular background triggered by CAstV infection. Ten 21-day-old SPF White Leghorn chickens were divided into two groups of 5 birds each. One group was inoculated with CAstV, and the other was used as the negative control. On 4th dpi, spleen samples were collected and immediately frozen at -70°C for RNA isolation. We analysed transcriptional profiles of the chickens' spleens at the 4th day following infection using RNA-seq to establish differentially expressed genes (DEGs). The RNA-seq findings were verified by quantitative real-time PCR (qRT-PCR). A total of 31959 transcripts were identified in response to CAstV infection. Eventually 45 DEGs (p-value<0.05; Log2Foldchange>1)were recognized in the spleen after CAstV infection (26 upregulated DEGs and 19 downregulated DEGs). qRT-PCR performed on 4 genes (IFIT5, OASL, RASD1, DDX60) confirmed RNAseq results. Top differentially expressed genes belonged to novel putative IFN-induced CAstV restriction factors. Most of the DEGs were associated with RIG-I–like signalling pathway or, more generally, with an innate antiviral response(upregulated: BLEC3, CMPK2, IFIT5, OASL, DDX60, IFI6, and downregulated: SPIK5, SELENOP, HSPA2, TMEM158, RASD1, YWHAB). The study provided a global analysis of host transcriptional changes that occur during CAstV infection in vivo and proved the cell cycle in the spleen and immune signalling in chickens were predominantly affected upon CAstV infection.

Keywords: chicken astrovirus, CastV, RNA-seq, transcriptome, spleen

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