Search results for: cell-free protein expression

Authors: Marie Lesly Fontin, Audalbert Bien-Aimé, Didier Marlier, Yves Beckers

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Fecal digestibility of mature plantain leaves was determined in castrated Creolegoatsin order to better assess them. Five diets made from plantain leaves were used in an in vivo digestibility study on 20 castrated Creole goats over three periods using a completely random design in order to assess their apparent fecal digestibility (Dg). These diets consisted of sun-dried leaves (DL), sun-dried urea treated leaves (DUTL, 5kg of urea per 100kg of raw product ensilaged during 90 days with 60 kg of water), sun-dried leaves + hoopvine (Trichostigma octandrum, L)(DLH, DL: 61.4% + Hoopvine: 38.6%), sun-dried leaves + urea (DLU, DL: 98.2%+ U: 1.8%), and fresh leaves. (FL).0.5% of salt diluted with water was added to diets before distribution to the goats. A mineral lick block was available for each goat in its digestibility cage. During each period, diets were distributed to meet the maintenance needs of the goats for 21 days, including 14 days of adaptation and 7 days of measurement. Offered and refused diets and feces were weighed every day, and samples were taken for laboratory analysis. Results showed that the urea treatment increasedCP (Crude Protein) content of DL by 44% (from 10.4% for DL to 15.0% for DUTL) and decreased their NDF (Neutral Detergent Fiber) content (55.5% to 52.4%). Large amounts of refused feed (around 40%) were observed in goats fed with FL, DLU, and DL diets, for which no significant difference was observed for DM (Dry Matter) intakes (40.3; 36.6 and 35.1g/kg0.75 respectively) (p>0.05). DM intakes of DUTL (59.9 g/kg0.75) were significantly (p<0.05) greater than DLH (50.2 g/kg0.75). DM Dg of DL was very low (29.2%). However, supplementation with hoopvine and urea treatment resulted in a significant increase of DM Dg (40.3% and 42.1%, respectively), but the addition of urea (DLU) had no effect on it. FL showed a DM Dg similar to DHL and DUTL diets (39.0%). OM (Organic Matter)Dg was higher for the DUTL diet (45.1%), followed by DLH (40.9%), then by DLU and FL (32.9% and 40.7% respectively) and finally by DL (29.8%). CP Dg was higher for the FL diet (65.7%) and lower for the DL diet (39.9%). NDF Dg was also increased with urea treatment (54.8% for DUTL) and with the addition of hoopvine (41.4%) compared to the DL diet (31.0% for DLH). In conclusion, urea treatment and complementation with hoopvine of plantain leaves are the best treatments among those tested for increasing the nutritive value of this foragein the castrated Creole goats.

Keywords: apparent fecal digestibility, nitrogen supplements, plantain leaves, urea treatment

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Authors: Israel B. Bitton

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“Memory” is associated with various phenomena, from physical to mental, personal to collective and historical to cultural. As part of a broader exploration of memory studies in philosophy and science (slated for academic publication October 2021), this specific study employs analytical methods of cognitive psychology and philosophy of memory to theorize that A) the primary human will (drive) is to significance, in that every human action and expression can be rooted in a most primal desire to be cosmically significant (however that is individually perceived); and B) that the will to significance manifests as the will to memorability, an innate desire to be remembered by others after death. In support of these broad claims, a review of various popular culture “touchpoints”—historic and contemporary records spanning literature, film and television, traditional news media, and social media—is presented to demonstrate how this very theory is repeatedly and commonly expressed (and has been for a long time) by many popular public figures as well as “everyday people.” Though developed before COVID, the crisis only increased the theory’s relevance: so many people were forced to die alone, leaving them and their loved ones to face even greater existential angst than what ordinarily accompanies death since the usual expectations for one’s “final moments” were shattered. To underscore this issue of, and response to, what can be considered a sociocultural “memory gap,” this study concludes with a summary of several projects launched by journalists at the height of the pandemic to document the memorable human stories behind COVID’s tragic warped speed death toll that, when analyzed through the lens of Viktor E. Frankl’s psychoanalytical perspective on “existential meaning,” shows how countless individuals were robbed of the last wills and testaments to their self-significance and memorability typically afforded to the dying and the aggrieved. The resulting insight ought to inform how government and public health officials determine what is truly “non-essential” to human health, physical and mental, at times of crisis.

Keywords: cognitive psychology, covid, neuroscience, philosophy of memory

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Authors: Aldé Belgard Tchicaya Loemba, Baraka Kichonge, Thomas Kivevele, Juma Rajabu Selemani

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Plants used for medicinal purposes are extremely perishable, owing to moisture-enhanced enzymatic and microorganism activity, climate change, and improper handling and storage. Experiments have shown that drying the medicinal plant without affecting the active nutrients and controlling the moisture content as much as possible can extend its shelf life. Different traditional and modern drying techniques for preserving medicinal plants have been developed, with some still being improved in Sub-Saharan Africa. However, many of these methods fail to address the most common issues encountered when drying medicinal plants, such as nutrient loss, long drying times, and a limited capacity to dry during the evening or cloudy hours. Heat pump drying is an alternate drying method that results in no nutritional loss. Furthermore, combining a heat pump dryer with a solar energy storage system appears to be a viable option for all-weather drying without affecting the nutritional values of dried products. In this study, a solar-assisted heat pump dryer integrated with thermal energy storage is developed for drying moringa leaves. The study also discusses the performance analysis of the developed dryer as well as the proximate analysis of the dried moringa leaves. All experiments were conducted from 11 a.m. to 4 p.m. to assess the dryer's performance in “daytime mode”. Experiment results show that the drying time was significantly reduced, and the dryer demonstrated high performance in preserving all of the nutrients. In 5 hours of the drying process, the moisture content was reduced from 75.7 to 3.3%. The average COP value was 3.36, confirming the dryer's low energy consumption. The findings also revealed that after drying, the content of protein, carbohydrates, fats, fiber, and ash greatly increased.

Keywords: heat pump dryer, efficiency, moringa leaves, proximate analysis

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Authors: Nemat Sokhandan Bashir

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Threats from viruses to agricultural crops could be even larger than the losses caused by the other pathogens because, in many cases, the viral infection is latent but crucial from an epidemic point of view. Wild vegetation can be a source of many viruses that eventually find their destiny in crop plants. Although often asymptomatic in wild plants due to adaptation, they can potentially cause serious losses in crops. Therefore, exploring viruses in wild vegetation is very important. Recently, omics have been quite useful for exploring plant viruses from various plant sources, especially wild vegetation. For instance, we have discovered viruses such as Ambrossia asymptomatic virus I (AAV-1) through the application of metagenomics from Oklahoma Prairie Reserve. Accordingly, extracts from randomly-sampled plants are subjected to high speed and ultracentrifugation to separated virus-like particles (VLP), then nucleic acids in the form of DNA or RNA are extracted from such VLPs by treatment with phenol—chloroform and subsequent precipitation by ethanol. The nucleic acid preparations are separately treated with RNAse or DNAse in order to determine the genome component of VLPs. In the case of RNAs, the complementary cDNAs are synthesized before submitting to DNA sequencing. However, for VLPs with DNA contents, the procedure would be relatively straightforward without making cDNA. Because the length of the nucleic acid content of VPLs can be different, various strategies are employed to achieve sequencing. Techniques similar to so-called "chromosome walking" may be used to achieve sequences of long segments. When the nucleotide sequence data were obtained, they were subjected to BLAST analysis to determine the most related previously reported virus sequences. In one case, we determined that the novel virus was AAV-l because the sequence comparison and analysis revealed that the reads were the closest to the Indian citrus ringspot virus (ICRSV). AAV—l had an RNA genome with 7408 nucleotides in length and contained six open reading frames (ORFs). Based on phylogenies inferred from the replicase and coat protein ORFs of the virus, it was placed in the genus Mandarivirus.

Keywords: wild, plant, novel, metagenomics

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Authors: Mohammad Hossein Habibi, Atena Sadat Hosseini

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Colorectal cancer is the third leading cause of cancer-related death in the world. Colorectal cancer screening, early detection, and treatment programs could benefit from the most up-to-date information on the disease's burden, given the present worldwide trend of increasing colorectal cancer incidence. Tumor recurrence and resistance are exacerbated by the presence of chemotherapy-resistant cancer stem cells that can generate rapidly proliferating tumor cells. In addition, tumor cells can evolve chemoresistance through adaptation mechanisms. In this work, we used in silico analysis to select suitable GEO datasets. In this study, we compared slow-growing cancer stem cells with high-growth colorectal cancer-derived cancer stem cells. We then evaluated the signal pathways, transcription factors, and kinases associated with these two types of cancer stem cells. A total of 980 upregulated genes and 870 downregulated genes were clustered. MAPK signaling pathway, AGE-RAGE signaling pathway in diabetic complications, Fc gamma R-mediated phagocytosis, and Steroid biosynthesis signaling pathways were observed in upregulated genes. Also, caffeine metabolism, amino sugar and nucleotide sugar metabolism, TNF signaling pathway, and cytosolic DNA-sensing pathway were involved in downregulated genes. In the next step, we evaluated the best transcription factors and kinases in two types of cancer stem cells. In this regard, NR2F2, ZEB2, HEY1, and HDGF as transcription factors and PRDM5, SMAD, CBP, and KDM2B as critical kinases in upregulated genes. On the other hand, IRF1, SPDEF, NCOA1, and STAT1 transcription factors and CTNNB1 and CDH7 kinases were regulated low expression genes. Using bioinformatics analysis in the present study, we conducted an in-depth study of colorectal cancer stem cells at low and high growth rates so that we could take further steps to detect and even target these cells. Naturally, more additional tests are needed in this direction.

Keywords: colorectal cancer, bioinformatics analysis, transcription factor, kinases, cancer stem cells

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

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

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

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Authors: Suveen Kumar

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Oral cancer (OC) is the sixth most death causing cancer in world which includes tumour of lips, floor of the mouth, tongue, palate, cheeks, sinuses, throat, etc. Conventionally, the techniques used for OC detection are toluidine blue staining, biopsy, liquid-based cytology, visual attachments, etc., however these are limited by their highly invasive nature, low sensitivity, time consumption, sophisticated instrument handling, sample processing and high cost. Therefore, we developed biosensing chips for non-invasive detection of OC via CYFRA-21-1 biomarker. CYFRA-21-1 (molecular weight: 40 kDa) is secreted in saliva of OC patients which is a non-invasive biological fluid with a cut-off value of 3.8 ng mL-1, above which the subjects will be suffering from oral cancer. Therefore, in first work, 3-aminopropyl triethoxy silane (APTES) functionalized zirconia (ZrO2) nanoparticles (APTES/nZrO2) were used to successfully detect CYFRA-21-1 in a linear detection range (LDR) of 2-16 ng mL-1 with sensitivity of 2.2 µA mL ng-1. Successively, APTES/nZrO2-RGO was employed to prevent agglomeration of ZrO2 by providing high surface area reduced graphene oxide (RGO) support and much wider LDR (2-22 ng mL-1) was obtained with remarkable limit of detection (LOD) as 0.12 ng mL-1. Further, APTES/nY2O3/ITO platform was used for oral cancer bioseneor development. The developed biosensor (BSA/anti-CYFRA-21-1/APTES/nY2O3/ITO) have wider LDR (0.01-50 ng mL-1) with remarkable limit of detection (LOD) as 0.01 ng mL-1. To improve the sensitivity of the biosensing platform, nanocomposite of yattria stabilized nanostructured zirconia-reduced graphene oxide (nYZR) based biosensor has been developed. The developed biosensing chip having ability to detect CYFRA-21-1 biomolecules in the range of 0.01-50 ng mL-1, LOD of 7.2 pg mL-1 with sensitivity of 200 µA mL ng-1. Further, the applicability of the fabricated biosensing chips were also checked through real sample (saliva) analysis of OC patients and the obtained results showed good correlation with the standard protein detection enzyme linked immunosorbent assay (ELISA) technique.

Keywords: non-invasive, oral cancer, nanomaterials, biosensor, biochip

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Authors: Kamsiah Jaarin, Yusof Kamisah, Faizah Othman Nurul Akmal Muhammad, Zakiah Jubri, Qodriyah Mohd Saad, Srijit Das

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Forty (40) normotensive adult male Sprague-Dawley rats aged three months weighing 180-200 g were divided into 4 groups with 10 rats per group: (1) normotensive control; (2) hypertensive rats; (3) hypertensive rats treated with Nigella sativa (2.5 ml/kg/day); and (4) hypertensive rats treated with nicardipine (5 mg/kg/day). After acclimatization, the hypertensive rats of the group 2, 3 and 4 were induced to be hypertensive by giving NW–nitro-L-arginine methyl ester (L-NAME; 30 mg/kg/day) in their drinking water for consecutive 7 days. After one week, rats in the group 3 were given a daily oral dose of 2.5 ml/kg/day of Nigella sativa (NS) by oral gavage. Rats in the group 4 were given nicardipine (5 mg/kg/day) via oral gavages. All rats in this study received L-NAME continuously throughout the treatment duration. The blood pressure will be measured pre-treatment and weekly for 8 weeks using power lab. Blood was taken before and at the end of study for measurement of nitric oxide. At the end of 8 weeks, the rats are sacrificed and descending thoracic aorta was disserted for measurement of vascular reactivity, and intracellular adhesion molecules (ICAM-1) and vascular cell adhesion molecules (VCAM-1). Nigella sativa reduced both systolic and diastolic BP compared to control and L-name group. The BP lowering effect of NS was comparable to nicardipine a calcium antagonist. The blood pressure lowering effect of NS was accompanied with an increasing relaxation response to nitroprusside and acetylcholine and reducing vasoconstriction response to epinephrine. L-NAME and nicardipine on the other hand, reduced plasma nitric oxide concentration. In contrast, NS increased NO concentration. However, Nigella sativa had no significant effect on aortic VCAM- 1 and ICAM-1 expression. In conclusion; Nigella sativa oil reduces both systolic and diastolic blood pressure in L-NAME treated rats. The antihypertensive effect of NS was comparable to nicardipine. The BP lowering effect may be mediated via stimulating nitric oxide release from vascular endothelium.

Keywords: Nigella sativa, ICAM, VCAM, blood pressure, vascular reactivity

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Authors: Rabia Göçmen, Gülşah Kanbur, Sinan Sefa Parlat

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In this study, in the breeding Japanese quails (coturnix coturnix japonica), it was aimed to study the effects of cellular insulin receptor stimulation on the performance, some blood parameters, and hatchability features. In the study, a total of 84 breeding quails were used, which are in 6 weeks age, and whose 24 are male and 60 female. In the trial, rations which contain 2900 kcal/kg metabolic energy; crude protein of 20%, and water whose pH is calibrated to 7.45 were administered as ad-libitum, to the animals, as metformin source, metformin-HCl was used and as chrome resource, Chromium Picolinate. Trial groups were formed as control group (basal ration), metformin group (basal ration, added metformin at the level of fodder of 20 mg/kg), and chromium picolinate group (basal ration, added fodder of 1500 ppb Cr. When regarded to the results of performance at the end of trial, it is seen that live weight gain, fodder consumption, egg weight, fodder evaluation coefficient, and egg production were affected at the significant level (p < 0.05). When the results are evaluated in terms of incubation features at the end of trial, it was identified that incubation yield and hatchability are not affected by the treatments but in the groups, in which metformin and chromium picolinate are added to ration, that fertility rose at the significant level compared to control group (p < 0,05). According to the results of blood parameters and hormone at the end of the trial, while the level of plasma glucose level was not affected by treatments (p > 0.05), with the addition of metformin and chromium picolinate to ration, plasma, total control, cholesterol, HDL, LDL, and triglyceride levels were significantly affected from insulin receptor stimulators added to ration (p<0,05). Hormone level of Plasma T3 and T4 were also affected at the significant level from insulin receptor stimulators added to ration (p < 0,05).

Keywords: cholesterol, chromium picolinate, hormone, metformin, performance, quail

Procedia PDF Downloads 187

Authors: Geevika J. Ganegama Arachchi, Steve H. Flint, Lynn McIntyre, Cristina D. Cruz, Beatrice M. Dias-Wanigasekera, Craig Billington, J. Andrew Hudson, Anthony N. Mutukumira

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In seafood processing plants, Listeriamonocytogenes(L. monocytogenes)likely exists in a metabolically stressed state due to the nutrient-deficient environment, processing treatments such as heating, curing, drying, and freezing, and exposure to detergents and disinfectants. Stressed L. monocytogenes cells have been shown to be as pathogenic as unstressed cells. This study investigated lytic efficacy of (LiMN4L, LiMN4p, and LiMN17) which were previouslycharacterized as virulent against physiologically stressed cells of three seafood borne L. monocytogenesstrains (19CO9, 19DO3, and 19EO3).Physiologically compromised cells ofL. monocytogenesstrains were prepared by aging cultures in TrypticaseSoy Broth at 15±1°C for 72 h; heat injuringcultures at 54±1 - 55±1°C for 40 - 60 min;salt-stressing cultures in Milli-Q water were incubated at 25±1°C in darkness for three weeks; and incubating cultures in 9% (w/v) NaCl at 15±1°C for 72 h. Low concentrations of physiologically compromised cells of three L. monocytogenesstrainswere challenged in vitrowith high titre of three phages in separate experiments using Fish Broth medium (aqueous fish extract) at 15 °C in order to mimic the environment of seafood processing plant. Each phage, when present at ≈9 log10 PFU/ml, reduced late exponential phase cells of L. monocytogenes suspended in fish protein broth at ≈2-3 log10 CFU/ml to a non-detectable level (< 10 CFU/ml). Each phage, when present at ≈8.5 log10 PFU/ml, reduced both heat-injured cells present at 2.5-3.6 log10 CFU/ml and starved cells that were showed coccoid shape, present at ≈2-3 log10 CFU/ml to < 10 CFU/ml after 30 min. Phages also reduced salt-stressed cellspresent at ≈3 log10 CFU/ml by > 2 log10. L. monocytogenes (≈8 log10 CFU/ml) were reduced to below the detection limit (1 CFU/ml) by the three successive phage infections over 16 h, indicating that emergence of spontaneous phage resistance was infrequent. The three virulent phages showed high decontamination potential for physiologically stressed L. monocytogenes strains from seafood processing environments.

Keywords: physiologically stressed L. monocytogenes, heat injured, seafood processing environment, virulent phage

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Authors: Jovana Petrovic, Biljana Pajin, Ivana Loncarevic, Aleksandar Fistes, Antun Jozinivic, Durdica Ackar, Drago Subaric

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Recovery of food wastes and their conversion to economically viable products will play a vital role for the management strategies in the years to come. Apple pomace may be considered as wastes, but they contain considerable amounts of high value reusable materials. Apple pomace, the by-product of apple juice and cider production, is a good source of fibre, particularly insoluble one. The remaining apple pulp contains 12% dry residue, which is half dietary fibre. Another remarkable aspect is its richness in polyphenols, components with antioxidant activity. Apple pomace could be an interesting alternative source for fibre and polyphenols in extruded corn meals. The extruded corn meals with the addition of finely ground apple pomace were prepared (the ratio of corn meal: apple pomace was 85:15 and 70:30). Characterization of the extrudates in terms of determining the chemical composition and colour was performed. The color of samples was measured by MINOLTA Chroma Meter CR-400 (Minolta Co., Ltd., Osaka, Japan) using D 65 lighting, a 2º standard observer angle and an 8-mm aperture in the measuring head. The following CIELab color coordinates were determined: L* – lightness, a* – redness to greenness and b* – yellowness to blueness. Protein content decreased significantly from 7.91% to 5.19% with increase in pomace from 0% to 30%, while total fibre content increase from 3.39% to 16.62%. The apple pomace addition produced extrudates with a significantly lower L* value and significantly higher a* value. This study has been fully supported by the Provincial Secretariat for High Education and Scientific Research of the Government of Autonomous Province of Vojvodina, Republic of Serbia, project 142-451-2483/2017 and the Ministry of Science and Technological Development of the Republic of Serbia (Project no. 31014).

Keywords: apple pomace, chemical composition, colour, extruded corn snack products, food waste recovery

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Authors: Tugce Duygu Koksal

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In recent years, given the fact that the acts of terrorism and the threat of the latter are taking place without any border and distinction, it has led the states to deal with the terrorism as a priority issue. More recently, as seen in different countries during state of emergency, the adoption of anti-terrorism measures motivated by the sole need of the prevention of terrorism targets directly the fundamental rights of individuals. Therefore, a contribution to the understanding of the value of the principle of legality is becoming more and more important nowadays. This paper aims to reflect the probable effects of the adoption of anti-terrorism measures regardless of the principle of legality, on the fundamental rights. In this respect, this paper will first discuss the margin of appreciation of the national authorities by countering terrorism, and then, the importance of the respect of the legality of the anti-terrorism measures will be examined in the light of actual examples. Indeed, one of the major findings of this study is the fact that the anti-terrorism laws and measures were taken in this framework must be subject to close scrutiny in democracies, which adopted the principle of the rule of law and respect human rights. Although the state's margin of appreciation in the field of counter-terrorism is broad, these measures which are based on the legitimate aim of a democracies’ legitimate right to protect itself against the activities of terrorist organizations should have the legal basis and be strictly required by the exigencies of the fight against terrorism. While combating terrorism, the legal basis shall only be achieved if the legal consequences of an individuals’ actions related to terrorism shall be clear and foreseeable by the individuals of a society. On the other hand, particularly during the state of emergency, the ambiguity of the law might be used to include a wide range of actions under acts of terrorism. This is becoming more dangerous where freedom of expression, freedom of the press, freedom of association and the right to information is in the substance of these actions. Disregarding the principle of legality is susceptible to create a chilling effect on the exercise of human rights, and therefore, the fight against terrorism can be transformed into a repressive regime on opponents. As a result, the efforts to counter terrorism of the national authorities irrespective of the principle of legality are susceptible to cause a transformation of the rule of law to a state of law which cannot be appreciated in a democratic society.

Keywords: anti-terrorism measures, chilling effect, predictability, the principle of legality, state of emergency

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Authors: Sonal Tuljaram Kame

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Growth and development is based on proper diet. A balanced diet contains all the nutrients in required quantum. Although physical growth is completed by young adulthood, the body tissues remain in a dynamic state with catabolism slightly exceeding anabolism, resulting in a net decrease in the number of cells. After the years of adolescence which cause upheavals in the life of the person, the individual struggle to emerge as an adult who know who he is and what his goals are. During this period nutrients are needed for maintaining the health and energy is required for physical functions and physical activities. The nutritional requirement in young adulthood differs from other periods of life. Iron is needed for haemoglobin synthesis and necessitates by the considerable examination of blood volume. Young adult girls need to ensure adequate intake of iron as they loose 0.5 mg/day by way of menstruation. This is complete awareness about nutritional and health on the other side there is widespread ignorance about nutrition and health among young adult girls. The young adult girls who are aware about nutrition and health seem to be very conscious about nutritional intake and health. Figure consciousness and fear of obesity leads to self imposed intake of nutrients. It may result in various health problems. The study was planned to investigate nutrient intake, find relation between nutritional intake, clinical examination score and health status of young adult girls. The present study is based on the data collected from 120 young adult girls studying in four different competitive exams coaching academies in Akola city of Maharashtra. It was found that nutritional intake of these young adult girls was below the recommended level, nutritional knowledge level and nutritional intake are associated attributes, calories, calcium and protein intake is positively correlated with clinical examination and health status. It was concluded that well planned nutritional counseling for the young adult girls can help prevent nutritional deficiency diseases and disorders which may lead to anaemic condition in young adult girls. Girls need to be educated on intake of iron and vitamin B12.

Keywords: nutritional intake, health status, young adult girls, correlation studies

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Authors: Patricia Portugal Marques de Carvalho Lourenco

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A three-phase methodological multi-lingual path was designed, constructed and carried out using the 2020 Chinese Internet Drama Series We Are All Alone as a case study. Phase one, the backbone of the research, comprised of secondary data analysis, providing the structure on which the next two phases would be built on. Phase one incorporated a Google Scholar and a Baidu Index analysis, Star Network Influence Index and Mydramalist.com top two drama reviews, along with an article written about the drama and scrutiny of Chinese related blogs and websites. Phase two was field research elaborated across Latin Europe, and phase three was social media focused, having into account that perceptions are going to be memory conditioned based on past ideas recall. Overall, research has shown the poor cultural expression of Chinese entertainment in Latin Europe and demonstrated the inexistence of Chinese content in French, Italian, Portuguese and Spanish Business to Consumer retailers; a reflection of their low significance in Latin European markets and the short-life cycle of entertainment products in general, bubble-gum, disposable goods without a mid to long-term effect in consumers lives. The process of conducting comprehensive international research was complex and time-consuming, with data not always available in Mandarin, the researcher’s linguistic deficiency, limited Chinese Cultural Knowledge and cultural equivalence. Despite steps being taken to minimize the international proposed research, theoretical limitations concurrent to Latin Europe and China still occurred. Data accuracy was disputable; sampling, data collection/analysis methods are heterogeneous; ascertaining data requirements and the method of analysis to achieve a construct equivalence was challenging and morose to operationalize. Secondary data was also not often readily available in Mandarin; yet, in spite of the array of limitations, research was done, and results were produced.

Keywords: research methodologies, international research, primary data, secondary data, research limitations, online dramas, china, latin europe

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Authors: Muhammad Naeem, Juho Lee, Jin-Wook Yoo

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In this study, we hypothesized that prolonged gastrointestinal transit at the inflamed colon conferred by a pH-triggered mucoadhesive smart nanoparticulate drug delivery system aids in achieving selective and sustained levels of the drug within the inflamed colon for the treatment of ulcerative colitis. We developed budesonide-loaded pH-sensitive charge-reversal solid lipid nanoparticles (SLNs) using a hot homogenization method. Polyetylenimine (PEI) was used to render SLNs cationic (PEI-SLNs). Eudragit S100 (ES) was coated on PEI-SLNs for pH-trigger charge-reversal SLNs (ES-PEI-SLNs). Therapeutic potential of the prepared SNLs formulation was evaluated in ulcerative colitis in mice. The transmission electron microscopy, zeta size and zeta potential data showed the successful formation of SLNs formulations. SLNs and PEI-SLNs showed burst drug release in acidic pH condition mimicking stomach and early small intestine environment which limiting their application as oral delivery systems. However, ES-PEI-SLNs prevented a burst drug release in acidic pH conditions and showed sustained release at a colonic pH. Most importantly, the surface charge of ES-PEI-SLNs switched from negative to positive in colonic conditions by pH-triggered removal of ES coating and accumulated selectively in inflamed colon. Furthermore, a charge reversal ES-PEI-SLNs showed a superior mitigation of dextran sulfate sodium (DSS)-induced acute colitis in mice as compared to SLNs and PEI-SLNs treated groups. Moreover, histopathological analysis of distal colon sections stained with hematoxylin/eosin and E-cadherin immunostaining revealed attenuated inflammation in an ES-PEI-SLNs-treated group. We also found that ES-PEI-SLNs markedly reduced the myeloperoxidase level and expression of TNF-alpha in colon tissue. Our results suggest that the pH-triggered charge reversal SLNs presented in this study would be a promising approach for ulcerative colitis therapy.

Keywords: solid lipid nanoparticles, stimuli-triggered charge-reversal, ulcerative colitis, methacrylate copolymer, budesonide

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Authors: Olufemi M. Alabi, Foluke A. Aderemi, Adebayo A. Adewumi, Banwo O. Alabi

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General well-being of animals is of paramount interest in some developed countries and of global importance hence the shift onto alternative housing systems for egg-type chickens as replacement for conventional battery cage system. However, there is paucity of information on the effect of this shift on physiological status of the hens to judge their health via the blood profile. Therefore, investigation was carried out on two strains of hen kept in three different housing systems in humid tropics to evaluate changes in their blood parameters. 108, 17-weeks old super black (SBL) hens and 108, 17-weeks old super brown (SBR) hens were randomly allotted to three different intensive systems Partitioned Conventional Cage (PCC), Extended Conventional Cage (ECC) and Deep Litter System (DLS) in a randomized complete block design with 36 hens per housing system, each with three replicates. The experiment lasted 37 weeks during which blood samples were collected at 18th week of age and bi-weekly thereafter for analyses. Parameters measured are packed cell volume (PCV), hemoglobin concentration (Hb), red blood counts (RBC), white blood counts (WBC) and serum metabolites such as total protein (TP), albumin (Alb), globulin (Glb), glucose, cholesterol, urea, bilirubin, serum cortisol while blood indices such as mean corpuscular hemoglobin (MCH), mean cell volume (MCV) and mean corpuscular hemoglobin concentration (MCHC) were calculated. The hematological values of the hens were not significantly (p>0.05) affected by the housing system and strain, so also the serum metabolites except for the serum cortisol which was significantly (p<0.05) affected by the housing system only. Hens housed on PCC had higher values (20.05 ng/ml for SBL and 20.55 ng/ml for SBR) followed by hens on ECC (18.15ng/ml for SBL and 18.38ng/ml for SBL) while hens on DLS had the lowest value (16.50ng/ml for SBL and 16.00ng/ml for SBR) thereby confirming indication of stress with conventionally caged birds. Alternative housing systems can also be adopted for egg-type chickens in the humid tropics from welfare point of view with the results of this work confirming stress among caged hens.

Keywords: blood, housing, humid-tropics, layers

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Authors: Auwal Ibrahim Magashi, Lawan Dan Larai Fagwalawa, Muhammad Bello Ibrahim

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A research was conducted to study genetic variability of some quantitative traits in varieties of cowpea (Vigna unguiculata L. [Walp]) under water stressed from Zaria, Nigeria. Seeds of seven varieties of cowpea (Sampea 1, Sampea 2, IAR1074, Sampea 7, Sampea 8, Sampea 10 and Sampea 12) collected from Institute for Agricultural Research (IAR), Samaru, Zaria were screened for water stressed tolerance. The seeds were then sown in poly bags containing sandy-loam arranged in Completely Randomized Design with three replications for quantitative traits evaluation. The nutritional composition of the seeds obtained from the water stress tolerant varieties of cowpea were analyzed. The result obtained revealed highly significant difference (P ≤ 0.01) in the effects of water stress on the number of wilted and dead plants at 40 days after sowing (DAS) and significant (P ≤ 0.05) 34 DAS. However, sampea 10 has the highest mean performance in terms of number of wilted plants at 34 DAS while sampea 2 and IAR 1074 has the lowest mean performance. However, sampea 7 was found to have the highest mean performance for the number of wilted plants at 40 DAS and sampea 2 is lowest. The result for quantitative traits study indicated highly significant difference (P ≤ 0.01) in the plant height, number of days to 50% flowering, number of days to maturity, number of pods per plant, pod length, number of seeds per plant and 100 seed weight; and significant (P ≤ 0.05) at seedling height and number of branches per plant. Similarly, IAR1074 was found to have high performance in terms of most of the quantitative traits under study. However, sampea 8 has the highest mean performance at nutritional level. It was therefore concluded that, all the seven cowpea genotypes were water stress tolerant and produced considerable yield that contained significant nutrients. It was recommended that IAR1074 should be grown for yield while sampea 8 should be grown for protein supplements.

Keywords: cowpea, genetic variability, quantitative traits, water stress

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Authors: Luiz Nolasco Jr. Rezende, Antonio Villar M. Sá, Claudia Marcia L. Pato

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The increasing urban violence in Brazil involves more and more infractions committed by children and youths. The challenges faced by the institutional environments responsible for the education and resocialization of adolescents in conflict with the law are enormous, especially of those deprived of their liberty. These institutions have an inadequate educational structure. They are characterized by a dirty and unhealthy environment without the minimum basic conditions for their activities, by frequent practices of degradation, humiliation, and the physical and psychological punishment of inmates. This mixed-method study investigated the personal values of adolescents with restriction of freedom in a socio-educational institutional environment aiming to contribute to the development of their morality through an educational process. For that, we used a survey and transdisciplinary play workshops involving thirty-two boys aged between 15 and 19 years old and at least two years out of school. To evaluate the survey the reduced version of the Portrait Questionnaire—PQ21—was used. The workshops happened once a week, lasting 80 minutes each, totaling twelve meetings. By using the game of chess and its metaphors, participants produced texts and engaged in critical brainstorming about their lives. The survey results pointed out that these young people showed a predominance of values of openness to change and self-transcendence, dissatisfaction with one's own reality and surroundings, not considering the consequences of their actions on themselves and others, difficulties in speaking and writing, and desire for changes in their lives. After the pedagogical interventions, these adolescents demonstrated an understanding of the implications of their actions for themselves, for their families, especially for the mothers, with whom they demonstrated stronger bonds. It was possible to observe evidence of improvement in the capacity of linguistic expression, more autonomy and critical vision, including about themselves and their respective contexts. These results demonstrated the educational potential of lively, symbolic, dynamic and playful activities that favor the mediation and identification of these adolescents with their lives, and contribute to the projection of dreams.

Keywords: adolescents arrested, human values, moral development, playful workshops

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

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

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

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Authors: Hacer Kobya Bulut, Kıymet Yeşilçiçek Çalık, Birsel Canan Demirbağ, Hacer Erdöl, Songül Aktaş

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Introduction: As an expression of an emotion which always exists in life, crying is regarded as one of the problematic behaviors of patients by nurses. Towards such patients, nurses may exhibit emotional and behavioral reactions such as feeling helpless, anger, indifferent, defense, and opposition. However crying either meets a need, reduces the tension to cope with problems or helps patient to gain strength. Therefore, nurses must accept that crying is a normal mechanism that reduces emotional tension and should approach a crying patient accordingly. Objective: This study was carried out to evaluate the communicational behaviors of the nurses towards ‘crying patient’. Methods: This descriptive study was conducted with the nurses working at a university hospital in a city in the Eastern Black Sea in June-September 2015. The entire universe was tried to be reached without sampling. 90% of the population was reached and the study was completed with 309 nurses who volunteered to participate in the study. Data were collected through a questionnaire which was prepared reviewing the literature by researchers. Data were evaluated in SPSS analysis program using percentages, numbers and chi-square test with the 95% confidence interval and p <0.05significance level. Findings: The findings showed that the average age of nurses was 31.52 ± 7.96, work experience was 10:09 ± 7.69 and only 22.7% had training about ‘approach to crying patient’ during their education. 97.1% of the nurses often faced with crying patients in their professional lives, 62.8% stated that they faced crying women patients. When they see crying patients, 84.8% of the nurses ‘do not want the patient to cry’, 80.9% wonder ‘why they are crying’, % 79.6 ‘feel uneasiness’,% 79.3 ‘feel sorry’ and 41.4% ‘ feel helpless’. The question ‘Why do you think the patient is crying?’ was answered by 93.5% nurses as ‘they are suffering’, by 86.1% ‘they are helpless’, 80.9% ‘they are sad’, 79.6% ‘they need help’, 54.4% ‘because they feel inadequate,’ and 44.7% ‘they fail to control their crying behavior. ‘How do you approach to your patient when she/he is crying?’ question was answered by 82.5% of nurses as ‘I would console’, 77.3% as ‘I would ask the reason’, 63.1% as ‘I would try to stop her from crying’ all of which are actually inappropriate nursing approaches. However, 92.2% of the nurses stated that ‘I do not judge the crying patient’, ‘87.1% said ‘I allocate time to crying patients’ and 85.8% said ‘ I ask patient whether they want to cry alone’. The study showed that educational background and work experience of the nurses affected the appropriate approach to crying patients (P <0.05). Conclusion: As a result of the study, it was found out that nurses do not want patients to cry, so they exhibit inappropriate approach such as consoling the patients and they have difficulty in approaching crying patients.

Keywords: approach to patient, communication, crying patient, nurse, Turkey

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Authors: Armando A. Rodríguez, Emilio Salceda, Anoland Garateix, André J. Zaharenko, Steve Peigneur, Omar López, Tirso Pons, Michael Richardson, Maylín Díaz, Yasnay Hernández, Ludger Ständker, Jan Tytgat, Enrique Soto

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Acid-sensing ion channels are cation (Na+) channels activated by a pH drop. These proteins belong to the ENaC/degenerin superfamily of sodium channels. ASICs are involved in sensory perception, synaptic plasticity, learning, memory formation, cell migration and proliferation, nociception, and neurodegenerative disorders, among other processes; therefore those molecules that specifically target these channels are of growing pharmacological and biomedical interest. Sea anemones produce a large variety of ion channels peptide toxins; however, those acting on ligand-gated ion channels, such as Glu-gated, Ach-gated ion channels, and acid-sensing ion channels (ASICs), remain barely explored. The peptide PhcrTx1 is the first compound characterized from the sea anemone Phymanthus crucifer, and it constitutes a novel ASIC inhibitor. This peptide was purified by chromatographic techniques and pharmacologically characterized on acid-sensing ion channels of mammalian neurons using patch-clamp techniques. PhcrTx1 inhibited ASIC currents with an IC50 of 100 nM. Edman degradation yielded a sequence of 32 amino acids residues, with a molecular mass of 3477 Da by MALDI-TOF. No similarity to known sea anemone peptides was found in protein databases. The computational analysis of Cys-pattern and secondary structure arrangement suggested that this is a structurally ICK (Inhibitor Cystine Knot)-type peptide, a scaffold that had not been found in sea anemones but in other venomous organisms. These results show that PhcrTx1 represents the first member of a new structural group of sea anemones toxins acting on ASICs. Also, this peptide constitutes a novel template for the development of drugs against pathologies related to ASICs function.

Keywords: animal toxin, inhibitor cystine knot, ion channel, sea anemone

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Authors: S. Kozlovski, S.W. Feigelson, R. Alon

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The b2 integrin ligands ICAM-1 ICAM-2 and the endothelial VLA-4 integrin ligand VCAM-1 are constitutively expressed on different lung vessels and on high endothelial venules (HEVs), the main portal for lymphocyte entry from the blood into lung draining lymph nodes. ICAMs are also ubiquitously expressed by many antigen-presenting leukocytes and have been traditionally suggested as critical for the various antigen-specific immune synapses generated by these distinct leukocytes and specific naïve and effector T cells. Loss of both ICAM-1 and ICAM-2 on the lung vasculature reduces the ability to patrol monocytes and Tregs to patrol the lung vasculature at a steady state. Our new findings suggest, however, that in terms of innate leukocyte trafficking into the lung lamina propria, both constitutively expressed and virus-induced vascular VCAM-1 can functionally compensate for the loss of these ICAMs. In a mouse model for influenza infection, neutrophil and NK cell recruitment and clearance of influenza remained normal in mice deficient in both ICAMs. Strikingly, mice deficient in both ICAMs also mounted normal influenza-specific CD8 proliferation and differentiation. In addition, these mice normally combated secondary influenza infection, indicating that the presence of ICAMs on conventional dendritic cells (cDCs) that present viral antigens are not required for immune synapse formation between these APCs and naïve CD8 T cells as previously suggested. Furthermore, long-lasting humoral responses critical for protection from a secondary homosubtypic influenza infection were also normal in mice deficient in both ICAM-1 and ICAM-2. Collectively, our results suggest that the expression of ICAM-1 and ICAM-2 on lung endothelial and epithelial cells, as well as on DCs and B cells, is not critical for the generation of innate or adaptive anti-viral immunity in the lungs. Our findings also suggest that endothelial VCAM-1 can substitute for the functions of vascular ICAMs in leukocyte trafficking into various lung compartments.

Keywords: emigration, ICAM-1, lymph nodes, VCAM-1

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Authors: Allioua Meryem

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Similar to much of the developing world, Algeria is currently undergoing an epidemiological transition. While mal- and under-nutrition and infectious diseases used to be the main causes of poor health, today there is a higher proportion of chronic, non-communicable diseases (NCDs), including cardiovascular disease, diabetes mellitus, cancer, etc. According to estimates for Algeria from the World Health Organization (WHO), NCDs accounted for 63% of all deaths in 2010. The objective of this study was the assessment of eating habits and anthropometric characteristics in a group of youth aged 15 to 19 years in Tlemcen. This study was conducted on a total effective of 806 youth enrolled in a descriptive cross-sectional study; the classification of nutritional status has been established by international standards IOTF, youth were defined as obese if they had a BMI ≥ 95th percentile, and youth with 85th ≤ BMI ≤ 95th percentile were defined as overweight. Wc is classified by the criteria HD, Wc with moderate risk ≥ 90th percentile and Wc with high risk ≥ 95th percentile. The dietary assessment was based on a 24-hour dietary recall assisted by food records. USDA’S nutrient database for Nutrinux® program was used to analyze dietary intake. Nutrients adequacy ratio was calculated by dividing daily individual intake to dietary recommended intake DRI for each nutrient. 9% of the population was overweight, 3% was obese, 7.5% had abdominal obesity, foods eaten in moderation are chips, cookies, chocolate 1-3 times/day and increased consumption of fried foods in the week, almost half of youth consume sugary drinks more than 3 times per week, we observe a decreased intake of energy, protein (P < 0.001, P = 0.003), SFA (P = 0.018), the NAR of phosphorus, iron, magnesium, vitamin B6, vitamin E, folate, niacin, and thiamin reflecting less consumption of fruit, vegetables, milk, and milk products. Youth surveyed have eating habits at risk of developing obesity and chronic disease.

Keywords: food intake, health, anthropometric characteristics, Algeria

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Authors: Sarah Nasser, Anne Moreau, Alain Hedoux, Romain Jeantet, Guillaume Delaplace

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Background: Spray dryed powders containing some caseins are commonly produced in dairy industry. It is widely admitted that the structure of casein evolves during powder storage, inducing a loss of solubility. However few studies evaluate accurately the destabilization mechanisms at molecular and mesoscopic level, in particular for Native Phospho Casein powder (NPC). Consequently, at the state of the art, it is very difficult to assess which secondary structure change or crosslinks initiate insolubility during storage. To address this issue, controlled ageing conditions have been applied to a NPC powder (which was obtained by spray drying a concentrate containing a higher content of casein (90%), whey protein (8%) and lactose (few %)). Evolution of structure and loss of solubility, with the effects of temperature and time of storage were systematically reported. Methods: FTIR spectroscopy, Raman and Circular Dichroism were used to monitor changes of secondary structure in dry powder and in solution after rehydration. Besides, proteomic tools and electrophoresis have been performed after varying storage conditions for evaluating aggregation and post translational modifications, like lactosylation or phosphorylation. Finally, Tof Sims and MEB were used to follow in parallel evolution of structure in surface and skin formation due to storage. Results + conclusion: These results highlight the important role of storage temperature in the stability of NPC. It is shown that this is not lactosylation at the heart of formation of aggregates, as advanced in others publications This is almost the rise of multitude post translational modifications (chemical cross link), added to disulphide bridges (physical cross link) wich contribute to the destabilisation of structure and aggregation of casein. A relative quantification of each kind of cross link, source of aggregates, is proposed. In addition, it has been proved that migration of lipids and formation of skin in surface during the ageing also explains the evolution of structure casein and thus the alterations of functional properties of NPC powder.

Keywords: casein, cross link, powder, storage

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Authors: Vasilis Andriopoulos, Maria D. Gkioni, Elena Koutra, Savvas G. Mastropetros, Fotini N. Lamari, Sofia Hatziantoniou, Michalis Kornaros

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In the present study, the antioxidant activity of aqueous and methanolic extracts of Chlorella minutissima, Dunaliella salina, Isochrysis galbana, Nannochloropsis oculata and Tisohrysis lutea, as well as the proximate composition and fatty acid profile were evaluated, with the aim to select species suitable for co-production of antioxidants and aquaculture feed. Batch cultivation was performed at 25oC in a modified f/2 medium under continuous illumination and aeration with ambient air. Biomass was collected via centrifugation and extracted first with H2O and subsequently with methanol at two growth phases (early and late stationary). Total phenolic content and antioxidant and reducing activity of the extracts were evaluated. The highest phenolic content was found in the methanolic extract of C. minutissima at the early stationary phase (9.04±0.68 mg Gallic Acid Equivalent g-1 dry weight), and the aqueous extract of D. salina at the late stationary phase (8.78±1.49 mg Gallic Acid Equivalent g-1 Dry weight). Antioxidant activity, measured as 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, and Ferric reducing antioxidant power assay of methanolic extracts were comparable to the literature and correlated to Total phenolic content and Chlorophyll content of the biomass. No such correlation was found in the aqueous extracts. N. oculata and T. lutea were high in protein (39.88±1.72% Dry weight and 43.30±1.33% Dry weight, respectively) and carotenoids (0.64±0.13% and 0.92±0.02%, respectively). Additionally, they presented high eicosapentaenoic acid and docosahexaenoic acid levels (33.74±9.98 mg eicosapentaenoic acid g-1 DW and 31.31±2.92 mg docosahexaenoic acid g-1 dry weight, respectively). N. oculata and T. lutea are promising candidates for the co-production of antioxidants and aquaculture feed, while C. minutissima and D. salina showed promise due to their higher antioxidant content.

Keywords: aquaculture fee, antioxidant activity, fatty acids, microalgae, total phenolic content

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Authors: Adel Alblihy, Reem Ali, Mashael Algethami, Ahmed Shoqafi, Michael S. Toss, Juliette Brownlie, Natalie J. Tatum, Ian Hickson, Paloma Ordonez Moran, Anna Grabowska, Jennie N. Jeyapalan, Nigel P. Mongan, Emad A. Rakha, Srinivasan Madhusudan

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Platinum resistance is a clinical challenge in ovarian cancer. Platinating agents induce DNA damage which activate Mre11 nuclease directed DNA damage signalling and response (DDR). Upregulation of DDR may promote chemotherapy resistance. Here we have comprehensively evaluated Mre11 in epithelial ovarian cancers. In clinical cohort that received platinum- based chemotherapy (n=331), Mre11 protein overexpression was associated with aggressive phenotype and poor progression free survival (PFS) (p=0.002). In the ovarian cancer genome atlas (TCGA) cohort (n=498), Mre11 gene amplification was observed in a subset of serous tumours (5%) which correlated highly with Mre11 mRNA levels (p<0.0001). Altered Mre11 levels was linked with genome wide alterations that can influence platinum sensitivity. At the transcriptomic level (n=1259), Mre11 overexpression was associated with poor PFS (p=0.003). ROC analysis showed an area under the curve (AUC) of 0.642 for response to platinum-based chemotherapy. Pre-clinically, Mre11 depletion by gene knock down or blockade by small molecule inhibitor (Mirin) reversed platinum resistance in ovarian cancer cells and in 3D spheroid models. Importantly, Mre11 inhibition was synthetically lethal in platinum sensitive XRCC1 deficient ovarian cancer cells and 3D-spheroids. Selective cytotoxicity was associated with DNA double strand break (DSB) accumulation, S-phase cell cycle arrest and increased apoptosis. We conclude that pharmaceutical development of Mre11 inhibitors is a viable clinical strategy for platinum sensitization and synthetic lethality in ovarian cancer.

Keywords: MRE11; XRCC1, ovarian cancer, platinum sensitization, synthetic lethality

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Authors: Umar Saeed

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Peptidyl-prolyl cis/trans isomerases Par14 and Par17 in humans play crucial roles in diverse cellular processes, including protein folding, chromatin remodeling, DNA binding, ribosome biogenesis, and cell cycle progression. However, the effects of Par14 and Par17 on viral replication have been explored to a limited extent. We first time discovered their influential roles in promoting Hepatitis B Virus replication. In this study, we observed that in the presence of HBx, either Par14 or Par17 could upregulate HBV replication. However, in the absence of HBx, neither Par14 nor Par17 had any effect on replication. Their mechanism of action involves binding to specific motifs within HBc and HBx proteins. Notably, they target the conserved 133Arg-Pro134 (RP) motif of HBc and the 19RP20-28RP29 motifs of HBx. This interaction is fundamental for the stability of HBx, core particles, and HBc. Par14 and Par17 exhibit versatility by binding both outside and inside core particles, thereby facilitating core particle assembly through their participation in HBc dimer-dimer interactions. NAGE and immunoblotting analyses unveiled the binding of Par14/Par17 to core particles. Co-immunoprecipitation experiments further demonstrated the interaction of Par14/Par17 with core particle assembly-defective and dimer-positive HBc-Y132A. It's essential to emphasize that R133 is the key residue in the HBc RP motif that governs their interaction with Par14/Par17. Chromatin immunoprecipitation conducted on HBV-infected cells elucidated the participation of residues S19 and E46/D74 in Par14 and S44 and E71/D99 in Par17 in the recruitment of 133RP134 motif-containing HBc into cccDNA. Depleting PIN4 in liver cell lines results in a significant reduction in cccDNA levels, pgRNA, sgRNAs, HBc, core particle assembly, and HBV DNA synthesis. Notably, parvulin inhibitors like juglone and PiB have proven to be effective in substantially reducing HBV replication. These inhibitors weaken the interaction between HBV core particles and Par14/Par17, underscoring the dynamic nature of this interaction. It's also worth noting that specific Par14/Par17 inhibitors hold promise as potential therapeutic options for chronic hepatitis B.

Keywords: Par14Par17, HBx, HBc, cccDNA, HBV

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Authors: P. Saha, R. Ganguly, N. K .Singha, A. Pich

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Microgel, a smart class of material, has drawn attention in the past few years due to its response to external stimuli like temperature, pH, and ionic strength of the solution. Among them, one type of polymer becomes soluble, and the other becomes insoluble in water upon heating displaying upper critical solution temperature (UCST) (e.g., polysulfobetaine, PSB) and lower critical solution temperature (LCST) (e.g., poly(N-vinylcaprolactam, PVCL)) respectively. Polyzwitterions, electrically neutral polymers are biocompatible, biodegradable, and non-cytotoxic in nature, and presence of zwitterionic pendant group in the main backbone makes them stable against temperature and pH variations and strong hydration capability in salt solution promotes them to be used as interfacial bio-adhesion resistance material. Majority of zwitterionic microgels have been synthesized in mini- emulsion technique using free radical polymerization approach. Here, a new route to synthesize dual thermo-responsive PVCL microgels decorated with appreciable amount of zwitterionic PSB chains was developed by a purely water-based surfactant-free reversible addition–fragmentation chain transfer (RAFT) precipitation polymerization. PSB macro-RAFTs having different molecular weights were synthesized and utilized for surface-grafting with PVCL microgels varying the macro-RAFT concentration using N,N′-methylenebis(acrylamide) (BIS) as cross-linker. Increasing the PSB concentration in the PVCL microgels resulted in a linear increase in UCST but decrease in hydrodynamic radius due to strong intrachain coulombic attraction forces acting between the opposite charges present in the zwitterionic groups. Anti- fouling effect was observed on addition of BSA protein solution on the microgel-coated membrane surfaces as studied by fluorescence spectrophotoscopy.

Keywords: microgels, polyzwitterions, upper critical solution temperature-lower critical solution temperature, UCST-LCST, ionic crosslinking

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Authors: Nadia Naseem, Farwa Batool, Nasir Mehmood, AbdulHannan Nagi

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Background: The incidence and mortality of breast cancer vary significantly in geographically distinct populations. In Pakistan, breast cancer has shown an increase in incidence in young females and is characterized by more aggressive behavior. The tumor suppressor TP53 gene is a crucial genetic factor that plays a significant role in breast carcinogenesis. This study investigated the TP53 mutations in molecular subtypes of both nodes negative and positive breast cancer in young Pakistani patients. Material and Methods: p53, Estrogen Receptor (ER), Progesterone Receptor (PR), Her-2 neu and Ki 67 expressions were analyzed immunohistochemically in a series of 75 node negative (A) and 75 node positive (B) young (aged: 19-40 years) breast cancer patients diagnosed between 2014 to 2017 at two leading hospitals of Punjab, Pakistan. Tumor tissue specimens and peripheral blood samples were examined for TP53 mutations by direct sequencing of the gene (exons 4-9). The relation of TP53 mutations to these markers and clinicopathological data was investigated. Results: Mean age of the patients was 32.4 + 9.1 SD. Invasive breast carcinoma was the most frequent histological variant (A=92%, B=94.6%). Grade 3 carcinoma was the commonest grade (A=72%, B=81.3%). Triple negative cases (ER-, PR-, Her-2) formed most of the molecular subtypes (A=44%, B=50.6%). A total of 17.2% (A: 6.6%, B: 10.6%) patients showed TP53 mutations. Mutations were significantly more frequent in triple negative cases (A: 74.8%, B: 62.2%) compared to HER2-positive patients (P < 0.0001). In the multivariate analysis of the whole patient group, the independent prognosticator were triple negative cases (P=0.021), TP53 overexpression by IHC (P=0.001) and advanced-stage disease (P=0.007). No statistically significant correlation between TP53 mutations and clinicopathological parameters was found (P < 0.05). Conclusions: It is concluded that TP53 mutations are infrequently present in breast carcinoma of young Pakistani population and there was no significant correlation between p53 mutation and early onset disease. Immunohistochemically detected TP53 expression in our resource-constrained to set up can be beneficial in predicting mutations at the younger age in our population.

Keywords: immunohistochemistry (IHC), invasive breast carcinoma (IBC), Pakistan, TP53

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Authors: Mpho Mokoatle, Darlington Mapiye, James Mashiyane, Stephanie Muller, Gciniwe Dlamini

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Great advances in high-throughput sequencing technologies have resulted in availability of huge amounts of sequencing data in public and private repositories, enabling a holistic understanding of complex biological phenomena. Sequence data are used for a wide range of applications such as gene annotations, expression studies, personalized treatment and precision medicine. However, this rapid growth in sequence data poses a great challenge which calls for novel data processing and analytic methods, as well as huge computing resources. In this work, a machine and statistical learning approach for DNA sequence classification based on $k$-mer representation of sequence data is proposed. The approach is tested using whole genome sequences of Mycobacterium tuberculosis (MTB) isolates to (i) reduce the size of genomic sequence data, (ii) identify an optimum size of k-mers and utilize it to build classification models, (iii) predict the phenotype from whole genome sequence data of a given bacterial isolate, and (iv) demonstrate computing challenges associated with the analysis of whole genome sequence data in producing interpretable and explainable insights. The classification models were trained on 104 whole genome sequences of MTB isoloates. Cluster analysis showed that k-mers maybe used to discriminate phenotypes and the discrimination becomes more concise as the size of k-mers increase. The best performing classification model had a k-mer size of 10 (longest k-mer) an accuracy, recall, precision, specificity, and Matthews Correlation coeffient of 72.0%, 80.5%, 80.5%, 63.6%, and 0.4 respectively. This study provides a comprehensive approach for resampling whole genome sequencing data, objectively selecting a k-mer size, and performing classification for phenotype prediction. The analysis also highlights the importance of increasing the k-mer size to produce more biological explainable results, which brings to the fore the interplay that exists amongst accuracy, computing resources and explainability of classification results. However, the analysis provides a new way to elucidate genetic information from genomic data, and identify phenotype relationships which are important especially in explaining complex biological mechanisms.

Keywords: AWD-LSTM, bootstrapping, k-mers, next generation sequencing

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