Search results for: anthropogenic markers

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

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

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

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Authors: Miguel A. Figueroa, José A. Lara-Ramos, Miguel A. Mueses

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Pharmaceutical residues are a section of emerging contaminants of anthropogenic origin that are present in a myriad of waters with which human beings interact daily and are starting to affect the ecosystem directly. Conventional waste-water treatment systems are not capable of degrading these pharmaceutical effluents because their designs cannot handle the intermediate products and biological effects occurring during its treatment. That is why it is necessary to hybridize conventional waste-water systems with non-conventional processes. In the specific case of an ozonation process, its efficiency highly depends on a perfect dispersion of ozone, long times of interaction of the gas-liquid phases and the size of the ozone bubbles formed through-out the reaction system. In order to increase the efficiency of these parameters, the use of a modified flotation cell has been proposed recently as a reactive system, which is used at an industrial level to facilitate the suspension of particles and spreading gas bubbles through the reactor volume at a high rate. The objective of the present work is the development of a mathematical model that can closely predict the kinetic rates of reactions taking place in the flotation cell at an experimental scale by means of identifying proper reaction mechanisms that take into account the modified chemical and hydrodynamic factors in the FeO-catalyzed Ozonation of Diclofenac aqueous solutions in a flotation cell. The methodology is comprised of three steps: an experimental phase where a modified flotation cell reactor is used to analyze the effects of ozone concentration and loading catalyst over the degradation of Diclofenac aqueous solutions. The performance is evaluated through an index of utilized ozone, which relates the amount of ozone supplied to the system per milligram of degraded pollutant. Next, a theoretical phase where the reaction mechanisms taking place during the experiments must be identified and proposed that details the multiple direct and indirect reactions the system goes through. Finally, a kinetic model is obtained that can mathematically represent the reaction mechanisms with adjustable parameters that can be fitted to the experimental results and give the model a proper physical meaning. The expected results are a robust reaction rate law that can simulate the improved results of Diclofenac mineralization on water using the modified flotation cell reactor. By means of this methodology, the following results were obtained: A robust reaction pathways mechanism showcasing the intermediates, free-radicals and products of the reaction, Optimal values of reaction rate constants that simulated Hatta numbers lower than 3 for the system modeled, degradation percentages of 100%, TOC (Total organic carbon) removal percentage of 69.9 only requiring an optimal value of FeO catalyst of 0.3 g/L. These results showed that a flotation cell could be used as a reactor in ozonation, catalytic ozonation and photocatalytic ozonation processes, since it produces high reaction rate constants and reduces mass transfer limitations (Ha > 3) by producing microbubbles and maintaining a good catalyst distribution.

Keywords: advanced oxidation technologies, iron oxide, emergent contaminants, AOTS intensification

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

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

Keywords: arterial stiffness, exercise, inflammation, pedometer

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

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

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

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

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

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

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

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

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

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Authors: C. Villamizar, M. Serrato

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

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

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Authors: Rama Umesh Pandey, Jyoti Yadav

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Flooding is one of the waterborne disasters that causes extensive destruction in urban areas. Developing countries are at a higher risk of such damage and more than half of the global flooding events take place in Asian countries including India. Urban flooding is more of a human-induced disaster rather than natural. This is highly influenced by the anthropogenic factors, besides metrological and hydrological causes. Unplanned urbanization and poor management of cities enhance the impact manifold and cause huge loss of life and property in urban areas. It is an irony that urban areas have been facing water scarcity in summers and flooding during monsoon. This paper is an attempt to highlight the factors responsible for flooding in a city especially from an urban planning perspective and to suggest mitigating measures through spatial planning interventions. Analysis has been done in two stages; first is to assess the impacts of previous flooding events and second to analyze the factors responsible for flooding at macro and micro level in cities. Bhopal, a city in Central India having nearly two million population, has been selected for the study. The city has been experiencing flooding during heavy rains in monsoon. The factors responsible for urban flooding were identified through literature review as well as various case studies from different cities across the world and India. The factors thus identified were analyzed for both macro and micro level influences. For macro level, the previous flooding events that have caused huge destructions were analyzed and the most affected areas in Bhopal city were identified. Since the identified area was falling within the catchment of a drain so the catchment area was delineated for the study. The factors analyzed were: rainfall pattern to calculate the return period using Weibull’s formula; imperviousness through mapping in ArcGIS; runoff discharge by using Rational method. The catchment was divided into micro watersheds and the micro watershed having maximum impervious surfaces was selected to analyze the coverage and effect of physical infrastructure such as: storm water management; sewerage system; solid waste management practices. The area was further analyzed to assess the extent of violation of ‘building byelaws’ and ‘development control regulations’ and encroachment over the natural water streams. Through analysis, the study has revealed that the main issues have been: lack of sewerage system; inadequate storm water drains; inefficient solid waste management in the study area; violation of building byelaws through extending building structures ether on to the drain or on the road; encroachments by slum dwellers along or on to the drain reducing the width and capacity of the drain. Other factors include faulty culvert’s design resulting in back water effect. Roads are at higher level than the plinth of houses which creates submersion of their ground floors. The study recommends spatial planning interventions for mitigating urban flooding and strategies for management of excess rain water during monsoon season. Recommendations have also been made for efficient land use management to mitigate water logging in areas vulnerable to flooding.

Keywords: mitigating strategies, spatial planning interventions, urban flooding, violation of development control regulations

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Authors: A. S. Vidyashree, C. M. Kalleshwaraswamy, R. Asokan, H. M. Mahadevaswamy

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Termites are very vital ecological thespians in tropical ecosystem, having been designated as “ecosystem engineers”, due to their significant role in providing soil ecosystem services. Despite their importance, our understanding of a number of their basic biological processes in termites is extremely limited. Developing a better understanding of termite biology is closely dependent upon consistent species identification. At present, identification of termites is relied on soldier castes. But for many species, soldier caste is not reported, that creates confusion in identification. The use of molecular markers may be helpful in estimating phylogenetic relatedness between the termite species and estimating genetic differentiation among local populations within each species. To understand this, termites samples were collected from various places of Western Ghats covering four states namely Karnataka, Kerala, Tamil Nadu, Maharashtra during 2013-15. Termite samples were identified based on their morphological characteristics, molecular characteristics, or both. Survey on the termite fauna in Karnataka, Kerala, Maharashtra and Tamil Nadu indicated the presence of a 16 species belongs to 4 subfamilies under two families viz., Rhinotermitidae and Termitidae. Termititidae was the dominant family which was belonging to 4 genera and four subfamilies viz., Macrotermitinae, Amitermitinae, Nasutitermitinae and Termitinae. Amitermitinae had three species namely, Microcerotermes fletcheri, M. pakistanicus and Speculitermes sinhalensis. Macrotermitinae had the highest number of species belonging two genera, namely Microtermes and Odontotermes. Microtermes genus was with only one species i.e., Microtermes obesi. The genus Odontotermes was represented by the highest number of species (07), namely, O. obesus was the dominant (41 per cent) and the most widely distributed species in Karnataka, Karala, Maharashtra and Tamil nadu followed by O. feae (19 per cent), O.assmuthi (11 per cent) and others like O. bellahunisensis O. horni O. redemanni, O. yadevi. Nasutitermitinae was represented by two genera namely Nasutitermes anamalaiensis and Trinervitermes biformis. Termitinae subfamily was represented by Labiocapritermes distortus. Rhinotermitidae was represented by single subfamily Heterotermetinae. In Heterotermetinae, two species namely Heterotermes balwanthi and H. malabaricus were recorded. Genetic relationship among termites collected from various locations of Western Ghats of India was characterized based on mitochondrial DNA sequences (12S, 16S, and COII). Sequence analysis and divergence among the species was assessed. These results suggest that the use of both molecular and morphological approaches is crucial in ensuring accurate species identification. Efforts were made to understand their evolution and to address the ambiguities in morphological taxonomy. The implication of the study in revising the taxonomy of Indian termites, their characterization and molecular comparisons between the sequences are discussed.

Keywords: isoptera, mitochondrial DNA sequences, rhinotermitidae, termitidae, Western ghats

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

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

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

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

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

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

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

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

Keywords: biomaterials, bone regeneration, biocompatibility, inflammation

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Authors: A. Kolendo

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Pacific Islands are one of the most vulnerable areas to climate change. Sea level rise and accelerating storm surge continuously threaten the communities' habitats on low-lying atolls. With scientific predictions of encroaching tides on their land, the Islanders have been informed about the need for future relocation planning. However, some communities oppose such retreat strategies through the reasoning that comprehends current climatic changes through the lenses of the biblical ark of Noah. This parable states God's promise never to flood the Earth again and never deprive people of their land and habitats. Several interpretations of this parable emerged in Oceania, prompting either climate action or denial. Resistance to relocation planning expressed through Christian thoughts led religion to be perceived as a barrier to dialogue between the Islanders and scientists. Since climate change concerns natural processes, the attitudes towards environmental stewardship prompt the communities' responses to it; some Christian teachings indicate humanity's responsibility over the environment, whereas others ascertain the people's dominion, which prompts resistance and sometimes denial. With church denominations and their various environmental standpoints, competing responses to climate change emerged in Oceania. Before miss-ionization, traditional knowledge had guided the environmental sphere, influencing current Christian teachings. Each atoll characterizes a distinctive manner of traditional knowledge; however, the unique relationship with nature unites all islands. The interconnectedness between the land, sea and people indicates the integrity between the communities and their environments. Such a factor influences the comprehension of Noah's story in the context of climate change that threatens their habitats. Pacific Islanders experience climate change through the slow disappearance of their homelands. However, the Western world perceives it as a global issue that will affect the population in the long-term perspective. Therefore, the Islanders seek to comprehend this global phenomenon in a local context that reads climate change as the Great Deluge. Accordingly, the safety measures that this parable promotes compensate for the danger of climate change. The rainbow covenant gives hope in God's promise never to flood the Earth again. At the same time, Noah's survival relates to the Islanders' current situation. Since these communities have the lowest carbon emissions rate, their contribution to anthropogenic climate change is scarce. Therefore, the lack of environmental sin would contextualize them as contemporary Noah with the ultimate survival of sea level rise. This study aims to defy religion constituting a barrier through secondary data analysis from a risk compensation perspective. Instead, religion is portrayed as a source of knowledge that enables comprehension of the communities' situation. By demonstrating that the Pacific Islanders utilize Noah's story as a vessel for coping with the danger of climate change, the study argues that religion provides safety measures that compensate for the future projections of land's disappearance. The purpose is to build a bridge between religious communities and scientific bodies and ultimately bring an understanding of two diverse perspectives. By addressing the practical challenges of interdisciplinary research with faith-based systems, this study uplifts the voices of communities and portrays their experiences expressed through Christian thoughts.

Keywords: Christianity, climate change, existential threat, Pacific Islands, story of Noah

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Authors: Eva Filová, Jana Daňková, Věra Sovková, Matej Daniel

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Titanium alloys are biocompatible metals that are widely used in clinical practice as load bearing implants. The chemical modification may influence cell adhesion, proliferation, and differentiation as well as stiffness of the material. The aim of the study was to evaluate the adhesion, growth and differentiation of pig mesenchymal stem cells on the novel beta titanium alloy Ti36Nb6Ta compared to standard medical titanium alloy Ti6Al4V. Discs of Ti36Nb6Ta and Ti6Al4V alloy were sterilized by ethanol, put in 48-well plates, and seeded by pig mesenchymal stem cells at the density of 60×103/cm2 and cultured in Minimum essential medium (Sigma) supplemented with 10% fetal bovine serum and penicillin/streptomycin. Cell viability was evaluated using MTS assay (CellTiter 96® AQueous One Solution Cell Proliferation Assay;Promega), cell proliferation using Quant-iT™ ds DNA Assay Kit (Life Technologies). Cells were stained immunohistochemically using monoclonal antibody beta-actin, and secondary antibody conjugated with AlexaFluor®488 and subsequently the spread area of cells was measured. Cell differentiation was evaluated by alkaline phosphatase assay using p-nitrophenyl phosphate (pNPP) as a substrate; the reaction was stopped by NaOH, and the absorbance was measured at 405 nm. Osteocalcin, specific bone marker was stained immunohistochemically and subsequently visualized using confocal microscopy; the fluorescence intensity was analyzed and quantified. Moreover, gene expression of osteogenic markers osteocalcin and type I collagen was evaluated by real-time reverse transcription-PCR (qRT-PCR). For statistical evaluation, One-way ANOVA followed by Student-Newman-Keuls Method was used. For qRT-PCR, the nonparametric Kruskal-Wallis Test and Dunn's Multiple Comparison Test were used. The absorbance in MTS assay was significantly higher on titanium alloy Ti6Al4V compared to beta titanium alloy Ti36Nb6Ta on days 7 and 14. Mesenchymal stem cells were well spread on both alloys, but no difference in spread area was found. No differences in alkaline phosphatase assay, fluorescence intensity of osteocalcin as well as the expression of type I collagen, and osteocalcin genes were observed. Higher expression of type I collagen compared to osteocalcin was observed for cells on both alloys. Both beta titanium alloy Ti36Nb6Ta and titanium alloy Ti6Al4V Ti36Nb6Ta supported mesenchymal stem cellsˈ adhesion, proliferation and osteogenic differentiation. Novel beta titanium alloys Ti36Nb6Ta is a promising material for bone implantation. The project was supported by the Czech Science Foundation: grant No. 16-14758S, the Grant Agency of the Charles University, grant No. 1246314 and by the Ministry of Education, Youth and Sports NPU I: LO1309.

Keywords: beta titanium, cell growth, mesenchymal stem cells, titanium alloy, implant

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Authors: Eleni Akritopoulou, Katerina Topouzoglou

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The last 20 years marine debris has been identified as one of the main marine pollution sources caused by anthropogenic activities. Plastics has reached the farthest marine areas of the planet affecting all marine trophic levels including the, recently discovered, amphipoda Eurythenes plasticus inhabiting Mariana Trench to large cetaceans, marine reptiles and sea birds causing immunodeficiency disorders, deteriorating health and death overtime. For the time period 2016-20, in the framework of the national initiative ‘Keep Aegean Blue”, All for Blue team has been collecting marine debris (coastline and underwater) following a modified in situ MEDSEALITTER monitoring protocol from eight Greek islands. After collection, marine debris was weighted, sorted and categorised according to material; plastic (PL), glass (G), metal (M), wood (W), rubber (R), cloth (CL), paper (P), mixed (MX). The goal of the project included the documentation of marine debris sources, human trends, waste management and public marine environmental awareness. Waste management was focused on plastics recycling and utilisation into daily useful products. This research is focused on the island of Mykonos due to its continuous touristic activity and lack of scientific information. In overall, a field work area of 1.832.856 m2 was cleaned up yielding 5092 kg of marine debris. The preliminary results indicated PL as main source of marine debris (62,8%) followed by M (15,5%), GL (13,2%) and MX (2,8%). Main items found were fishing tools (lines, nets), disposable cutlery, cups and straws, cigarette butts, flip flops and other items like plastic boat compartments. In collaboration with a local company for plastic management and the Circular Economy and Eco Innovation Institute (Sweden), all plastic debris was recycled. Granulation process was applied transforming plastic into building materials used for refugees’ houses, litter bins bought by municipalities and schools and, other items like shower components. In terms of volunteering and attendance in public awareness seminars, there was a raise of interest by 63% from different age ranges and professions. Regardless, the research being fairly new for Mykonos island and logistics issues potentially affected systemic sampling, it appeared that plastic debris is the main littering source attributed, possibly to the intense touristic activity of the island all year around. However, marine environmental awareness activities were pointed out to be an effective tool in forming public perception against marine debris and, alter the daily habits of local society. Since the beginning of this project, three new local environmental teams were formed against marine pollution supported by the local authorities and stakeholders. The continuous need and request for the production of items made by recycled marine debris appeared to be beneficial socio-economically to the local community and actions are taken to expand the project nationally. Finally, as an ongoing project and whilst, new scientific information is collected, further funding and research is needed.

Keywords: Greece, marine debris, marine environmental awareness, Mykonos island, plastics debris, plastic granulation, recycled plastic, tourism, waste management

Procedia PDF Downloads 108

Authors: Monika Trzaskowska, Dorota Zielinska, Anna Lepecka, Katarzyna Neffe-Skocinska, Beata Bilska, Marzena Tomaszewska, Danuta Kolozyn-Krajewska

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Microbiological quality and food safety are important food characteristics. Regulation (EU) No 1169/2011 of the European Parliament and of the Council on the provision of food information to consumers introduces the obligation to provide information on the 'use-by' date or the date of minimum durability (DMD). The second term is the date until which the properly stored or transported foodstuff retains its physical, chemical, microbiological and organoleptic properties. The date should be preceded by 'best before'. It is used for durable products, e.g., pasta. In relation to reducing food losses, the question may be asked whether products with the date of minimum durability currently declared retain quality and safety beyond this. The aim of the study was to assess the sensory quality and microbiological safety of selected cereal products, i.e., pasta and millet after DMD. The scope of the study was to determine the markers of microbiological quality, i.e., the total viable count (TVC), the number of bacteria from the Enterobacteriaceae family and the number of yeast and mold (TYMC) on the last day of DMD and after 1 and 3 months of storage. In addition, the presence of Salmonella and Listeria monocytogenes was examined on the last day of DMD. The sensory quality of products was assessed by quantitative descriptive analysis (QDA), the intensity of 14 differentiators and overall quality were defined and determined. In the tested samples of millet and pasta, no pathogenic bacteria Salmonella and Listeria monocytogenes were found. The value of the distinguishing features of selected quality and microbiological safety indicators on the last DMD day was in the range of about 3-1 log cfu/g. This demonstrates the good microbiological quality of the tested food. Comparing the products, a higher number of microorganisms was found in the samples of millet. After 3 months of storage, TVC decreased in millet, while in pasta, it was found to increase in value. In both products, the number of bacteria from the Enterobacretiaceae family decreased. In contrast, the number of TYMCs increased in samples of millet, and in pasta decreased. The intensity of sensory characteristic in the studied period varied. It remained at a similar level or increased. Millet was found to increase the intensity and flavor of 'cooked porridge' 3 months after DMD. Similarly, in the pasta, the smell and taste of 'cooked pasta' was more intense. To sum up, the researched products on the last day of the minimum durability date were characterized by very good microbiological and sensory quality, which was maintained for 3 months after this date. Based on these results, the date of minimum durability of tested products could be extended. The publication was financed on the basis of an agreement with the National Center for Research and Development No. Gospostrateg 1/385753/1/NCBR/2018 for the implementation and financing of the project under the strategic research and development program 'social and economic development of Poland in the conditions of globalizing markets – GOSPOSTRATEG - acronym PROM'.

Keywords: date of minimum durability, food losses, food quality and safety, millet, pasta

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Authors: Aoxue Yang, Weili Tian, Haikun Liu

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Brain tumor stem cells (BTSCs) are resistant to therapy and give rise to recurrent tumors. These rare and elusive cells are likely to disseminate during cancer progression, and some may enter dormancy, remaining viable but not increasing. The identification of dormant BTSCs is thus necessary to design effective therapies for glioblastoma (GBM) patients. Glucocorticoids (GCs) are used to treat GBM-associated edema. However, glucocorticoids participate in the physiological response to psychosocial stress, linked to poor cancer prognosis. This raises concern that glucocorticoids affect the tumor and BTSCs. Identifying markers specifically expressed by brain tumor stem cells (BTSCs) may enable specific therapies that spare their regular tissue-resident counterparts. By ribosome profiling analysis, we have identified that glycerol-3-phosphate dehydrogenase 1 (GPD1) is expressed by dormant BTSCs but not by NSCs. Through different stress-induced experiments in vitro, we found that only dexamethasone (DEXA) can significantly increase the expression of GPD1 in NSCs. Adversely, mifepristone (MIFE) which is classified as glucocorticoid receptors antagonists, could decrease GPD1 protein level and weaken the proliferation and stemness in BTSCs. Furthermore, DEXA can induce GPD1 expression in tumor-bearing mice brains and shorten animal survival, whereas MIFE has a distinct adverse effect that prolonged mice lifespan. Knocking out GR in NSC can block the upregulation of GPD1 inducing by DEXA, and we find the specific sequences on GPD1 promotor combined with GR, thus improving the efficiency of GPD1 transcription from CHIP-Seq. Moreover, GR and GPD1 are highly co-stained on GBM sections obtained from patients and mice. All these findings confirmed that GR could regulate GPD1 and loss of GPD1 Impairs Multiple Pathways Important for BTSCs Maintenance GPD1 is also a critical enzyme regulating glycolysis and lipid synthesis. We observed that DEXA and MIFE could change the metabolic profiles of BTSCs by regulating GPD1 to shift the transition of cell dormancy. Our transcriptome and lipidomics analysis demonstrated that cell cycle signaling and phosphoglycerides synthesis pathways contributed a lot to the inhibition of GPD1 caused by MIFE. In conclusion, our findings raise concern that treatment of GBM with GCs may compromise the efficacy of chemotherapy and contribute to BTSC dormancy. Inhibition of GR can dramatically reduce GPD1 and extend the survival duration of GBM-bearing mice. The molecular link between GPD1 and GR may give us an attractive therapeutic target for glioblastoma.

Keywords: cancer stem cell, dormancy, glioblastoma, glycerol-3-phosphate dehydrogenase 1, glucocorticoid receptor, dexamethasone, RNA-sequencing, phosphoglycerides

Procedia PDF Downloads 128

Authors: Ildiko Fekete-Kertesz, Jade Chaker, Sylvain Berthelot, Viktoria Feigl, Monika Molnar, Lidia Favier

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There has been growing concern about emerging micropollutants in recent years, because of the possible environmental and health risk posed by these substances, which are released into the environment as a consequence of anthropogenic activities. Among them pharmaceuticals are currently not considered under water quality regulations; however, their potential effect on the environment have become more frequent in recent years. Due to the fact that these compounds can be detected in natural water matrices, it can be concluded, that the currently applied water treatment processes are not efficient enough for their effective elimination. To date, advanced oxidation processes (AOPs) are considered as highly competitive water treatment technologies for the removal of those organic micropollutants not treatable by conventional techniques due to their high chemical stability and/or low biodegradability. AOPs such as (photo)chemical oxidation and heterogeneous photocatalysis have proven their potential in degrading harmful organic compounds from aqueous matrices. However, some of these technologies generate reaction by-products, which can even be more toxic to aquatic organisms than the parent compounds. Thus, target compound removal does not necessarily result in the removal of toxicity. Therefore, to evaluate process efficiency the determination of the toxicity and ecotoxicity of the reaction intermediates is crucial to estimate the environmental risk of such techniques. In this context, the present study investigates the effectiveness of TiO2 assisted photodegradation for the removal of emerging water contaminants. Two drugs named losartan (used in high blood pressure medication) and levetiracetam (used to treat epilepsy) were considered in this work. The photocatalytic reactions were carried out with a commercial catalyst usually employed in photocatalysis. Moreover, the toxicity of the by-products generated during the process was assessed with various ecotoxicological methods applying aquatic test organisms from different trophic levels. A series of experiments were performed to evaluate the toxicity of untreated and treated solutions applying the Aliivibrio fischeri bioluminescence inhibition test, the Tetrahymena pyriformis proliferation inhibition test, the Daphnia magna lethality and immobilization tests and the Lemna minor growth inhibition test. The applied ecotoxicological methodology indicated sensitively the toxic effects of the treated and untreated water samples, hence the applied test battery is suitable for the ecotoxicological characterization of TiO2 based photocatalytic water treatment technologies and the indication of the formation of toxic by-products from the parent chemical compounds. Obtained results clearly showed that the TiO2 assisted photodegradation was more efficient in the elimination of losartan than levetiracetam. It was also observed that the treated levetiracetam solutions had more severe effect on the applied test organisms. A possible explanation would be the production of levetiracetam by-products, which are more toxic than the parent compound. The increased toxicity and the risk of formation of toxic metabolites represent one possible limitation to the implementation of photocatalytic treatment using TiO2 for the removal of losartan and levetiracetam. Our results proved that, the battery of ecotoxicity tests used in this work can be a promising investigation tool for the environmental risk assessment of photocatalytic processes.

Keywords: aquatic micropollutants, ecotoxicology, nano titanium dioxide, photocatalysis, water treatment

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Authors: Aaron Derner

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Unlike the histories of France, the UK, or even Spain, the Russian colonial past often dissolves before the seemingly more salient Cold War figurations or Soviet dissolution. The obvious explanation behind Caucasian states’ roles—that of Russian-propped governments obeying the whims of their patron—is but the latest instance of such oversight. Where the results of colonial social and cultural interactions are indelibly stamped across France, Algeria, and every other former (and current) French holding, so to are the Muscovite and Russian colonial ambitions embedded within the modern politics and cultures of both Russia and the Caucasus. Russian colonial artefacts are enhanced and perhaps granted an additional social explanatory edge over those of the ‘typical’ colonizers, by the cyclical adoration for and noisy rejection of European cultural markers over the centuries, along with the somewhat unusual composition of the Cossacks: Russia’s main agents of colonialization within the Caucasian frontier. The story of Russia and Chechnya, of all the Caucasus, is of the manufacture of social and individual identity through “modes of subjectification” inherent within the region’s colonial history and driven by the triangular interactions between three main groups: the Cossacks, the Caucasian Mountain Tribes, and the Russian Metropol. Together, interactions between these social groups worked to shape and transform the lifestyles and institutional pathologies that constitute the Russian and Chechen states and the politics between them. At the core of this (Western) state-building is the simultaneous and seemingly contradictory desire to be more Western and emulate Western cultural and political practices while also desperately grasping for a uniquely Russian identity. This sits somewhat ironically against the backdrop that Russia hosted a frontier-based settler society and had established that distinctly European feature of settler colonialism early in its history—arguably establishing a claim to being the most “colonial” of the colonial powers. There is no doubt that these forces worked to shape contemporary Russian political and social identity—apparent in the mythic popularity of the Cossack in Russian literature, politics, and academic discourse. What needs to be expanded from the current narrative, however, is that beyond the Cossack identity’s attractiveness on the grounds of its tones of freedom and resistance to unjust authority, the identity is rooted in the imperial ambitions and colonial experiences of the Russian state, and is, therefore, a direct marker of domination and subjectification. Adding an unusual dimension to this not-uncommon cultural progression, the Russian state needed to colonize both the Caucases and the Russian Cossacks, appropriating them in much the same way they appropriated the Circassian mountain tribes. The focus of this paper is not to tell yet another story of how one culture entered an area to overpower another but how a ‘powerful,’ ‘modern,’ ‘Western(ish)’ culture was profoundly and continually changed through its contact with a group of tribal ‘savages’ and ‘braves.’

Keywords: Russia, chechnya, subjectification, caucasus, cossacks, Ukraine

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Authors: Alebiosu Olugbenga Shadrak, Victor Victoria

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Honey bees exhibit preferences in their foraging behaviour on pollen and nectar for food and honey production, respectively. Melissopalynology is the study of pollen in honey and other honey products. Several work have been conducted on the palynological studies of honeys from the southern parts of Nigeria but with relatively scant records from the Northern region of the country. This present study aimed at revealing the favourably visited plants by honey bees, Apis melifera var. adansonii, at some apiaries in Northern Nigeria, as well as determining the quality of honeys produced. Honeys were harvested and collected from four apiaries of the region, namely: Sarkin Dawa missionary bee farm, Taraba State; Eleeshuwa Bee Farm, Keffi, Nassarawa State, Bulus Beekeeper Apiaries, Kagarko, Kaduna State and Mai Gwava Bee Farm, Kano State. These honeys were acetolysed for palynological microscopic analysis and subjected to standard treatment methods for the determination of their proximate composition and sugar profiling. Fresh anthers of two dominantly represented plants in the honeys were then collected for the quantification of their pollen protein contents, using the micro-kjeldhal procedure. A total of 30 pollen types were identified in the four honeys, and some of them were common to the honeys. A classification method for expressing pollen frequency class was employed: Senna cf. siamea, Terminalia cf. catappa, Mangifera indica, Parinari curatelifolia, Vitellaria paradoxa, Elaeis guineensis, Parkia biglobosa, Phyllantus muellerianus and Berlina Grandiflora, as “Frequent” (16-45%); while the others are either Rare (3-15%) or Sporadic (less than 3 %). Pollen protein levels of the two abundantly represented plants, Senna siamea (15.90mg/ml) and Terminalia catappa (17.33mg/ml) were found to be considerably lower. The biochemical analyses revealed varying amounts of proximate composition, non-reducing sugar and total sugar levels in the honeys. The results of this study indicate that pollen and nectar of the “Frequent” plants were preferentially foraged by honeybees in the apiaries. The estimated pollen protein contents of Senna same and Terminalia catappa were considerably lower and not likely to have influenced their favourable visitation by honeybees. However, a relatively higher representation of Senna cf. siamea in the pollen spectrum might have resulted from its characteristic brightly coloured and well scented flowers, aiding greater entomophily. Terminalia catappa, Mangifera indica, Elaeis guineensis, Vitellaria paradoxa, and Parkia biglobosa are typical food crops; hence they probably attracted the honeybees owing to the rich nutritional values of their fruits and seeds. Another possible reason for a greater entomophily of the favourably visited plants are certain nutritional constituents of their pollen and nectar, which were not investigated in this study. The nutritional composition of the honeys was observed to fall within the safe limits of international norms, as prescribed by Codex Alimentarius Commission, thus they are good honeys for human consumption. It is therefore imperative to adopt strategic conservation steps in ensuring that these favourably visited plants are protected from indiscriminate anthropogenic activities and also encourage apiarists in the country to establish their bee farms more proximally to the plants for optimal honey yield.

Keywords: honeybees, melissopalynology, preferentially foraged, nutritional, bee farms, proximally

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Authors: Charlotte Entwistle, Ryan Boyd

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Recent developments in information retrieval techniques and natural language processing have allowed for greater exploration of psychological and social processes. Linguistic analysis methods for understanding behaviour have provided useful insights within the field of mental health. One area within mental health that has received little attention though, is borderline personality disorder (BPD). BPD is a common mental health disorder characterised by instability of interpersonal relationships, self-image and affect. It also manifests through maladaptive behaviours, such as impulsivity and self-harm. Examination of language patterns associated with BPD could allow for a greater understanding of the disorder and its links to maladaptive thoughts and behaviours. Language analysis methods could also be used in a predictive way, such as by identifying indicators of BPD or predicting maladaptive thoughts, emotions and behaviours. Additionally, associations that are uncovered between language and maladaptive thoughts and behaviours could then be applied at a more general level. This study explores linguistic characteristics of BPD, and their links to maladaptive thoughts and behaviours, through the analysis of social media data. Data were collected from a large corpus of posts from the publicly available social media platform Reddit, namely, from the ‘r/BPD’ subreddit whereby people identify as having BPD. Data were collected using the Python Reddit API Wrapper and included all users which had posted within the BPD subreddit. All posts were manually inspected to ensure that they were not posted by someone who clearly did not have BPD, such as people posting about a loved one with BPD. These users were then tracked across all other subreddits of which they had posted in and data from these subreddits were also collected. Additionally, data were collected from a random control group of Reddit users. Disorder-relevant behaviours, such as self-harming or aggression-related behaviours, outlined within Reddit posts were coded to by expert raters. All posts and comments were aggregated by user and split by subreddit. Language data were then analysed using the Linguistic Inquiry and Word Count (LIWC) 2015 software. LIWC is a text analysis program that identifies and categorises words based on linguistic and paralinguistic dimensions, psychological constructs and personal concern categories. Statistical analyses of linguistic features could then be conducted. Findings revealed distinct linguistic features associated with BPD, based on Reddit posts, which differentiated these users from a control group. Language patterns were also found to be associated with the occurrence of maladaptive thoughts and behaviours. Thus, this study demonstrates that there are indeed linguistic markers of BPD present on social media. It also implies that language could be predictive of maladaptive thoughts and behaviours associated with BPD. These findings are of importance as they suggest potential for clinical interventions to be provided based on the language of people with BPD to try to reduce the likelihood of maladaptive thoughts and behaviours occurring. For example, by social media tracking or engaging people with BPD in expressive writing therapy. Overall, this study has provided a greater understanding of the disorder and how it manifests through language and behaviour.

Keywords: behaviour analysis, borderline personality disorder, natural language processing, social media data

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Authors: Almudena Espin Perez, Tyler Risom, Carl Pelz, Isabel English, Robert M. Angelo, Rosalie Sears, Andrew J. Gentles

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Pancreatic ductal adenocarcinoma (PDAC) is one of the most deadly malignancies. The role of the tumor microenvironment (TME) is gaining significant attention in cancer research. Despite ongoing efforts, the nature of the interactions between tumors, immune cells, and stromal cells remains poorly understood. The cell-intrinsic properties that govern cell lineage plasticity in PDAC and extrinsic influences of immune populations require technically challenging approaches due to the inherently heterogeneous nature of PDAC. Understanding the cell lineage plasticity of PDAC will improve the development of novel strategies that could be translated to the clinic. Members of the team have demonstrated that the acquisition of ductal to neuroendocrine lineage plasticity in PDAC confers therapeutic resistance and is a biomarker of poor outcomes in patients. Our approach combines computational methods for deconvolving bulk transcriptomic cancer data using CIBERSORTx and high-throughput single-cell imaging using Multiplexed Ion Beam Imaging (MIBI) to study lineage plasticity in PDAC and its relationship to the infiltrating immune system. The CIBERSORTx algorithm uses signature matrices from immune cells and stroma from sorted and single-cell data in order to 1) infer the fractions of different immune cell types and stromal cells in bulked gene expression data and 2) impute a representative transcriptome profile for each cell type. We studied a unique set of 300 genomically well-characterized primary PDAC samples with rich clinical annotation. We deconvolved the PDAC transcriptome profiles using CIBERSORTx, leveraging publicly available single-cell RNA-seq data from normal pancreatic tissue and PDAC to estimate cell type proportions in PDAC, and digitally reconstruct cell-specific transcriptional profiles from our study dataset. We built signature matrices and optimized by simulations and comparison to ground truth data. We identified cell-type-specific transcriptional programs that contribute to cancer cell lineage plasticity, especially in the ductal compartment. We also studied cell differentiation hierarchies using CytoTRACE and predict cell lineage trajectories for acinar and ductal cells that we believe are pinpointing relevant information on PDAC progression. Collaborators (Angelo lab, Stanford University) has led the development of the Multiplexed Ion Beam Imaging (MIBI) platform for spatial proteomics. We will use in the very near future MIBI from tissue microarray of 40 PDAC samples to understand the spatial relationship between cancer cell lineage plasticity and stromal cells focused on infiltrating immune cells, using the relevant markers of PDAC plasticity identified from the RNA-seq analysis.

Keywords: deconvolution, imaging, microenvironment, PDAC

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Authors: Eashwar V. Somasundaram, Raoul R. Wadhwa, Jacob G. Scott

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The use of radiomics in measuring geometric properties of tumor images such as size, surface area, and volume has been invaluable in assessing cancer diagnosis, treatment, and prognosis. In addition to analyzing geometric properties, radiomics would benefit from measuring topological properties using persistent homology. Intuitively, features uncovered by persistent homology may correlate to tumor structural features. One example is necrotic cavities (corresponding to 2D topological features), which are markers of very aggressive tumors. We develop a data pipeline in R that clusters tumors images based on persistent homology is used to identify meaningful clinical distinctions between tumors and possibly new relationships not captured by established clinical categorizations. A preliminary analysis was performed on 16 Magnetic Resonance Imaging (MRI) breast tissue segments downloaded from the 'Investigation of Serial Studies to Predict Your Therapeutic Response with Imaging and Molecular Analysis' (I-SPY TRIAL or ISPY1) collection in The Cancer Imaging Archive. Each segment represents a patient’s breast tumor prior to treatment. The ISPY1 dataset also provided the estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2) status data. A persistent homology matrix up to 2-dimensional features was calculated for each of the MRI segmentation. Wasserstein distances were then calculated between all pairwise tumor image persistent homology matrices to create a distance matrix for each feature dimension. Since Wasserstein distances were calculated for 0, 1, and 2-dimensional features, three hierarchal clusters were constructed. The adjusted Rand Index was used to see how well the clusters corresponded to the ER/PR/HER2 status of the tumors. Triple-negative cancers (negative status for all three receptors) significantly clustered together in the 2-dimensional features dendrogram (Adjusted Rand Index of .35, p = .031). It is known that having a triple-negative breast tumor is associated with aggressive tumor growth and poor prognosis when compared to non-triple negative breast tumors. The aggressive tumor growth associated with triple-negative tumors may have a unique structure in an MRI segmentation, which persistent homology is able to identify. This preliminary analysis shows promising results in the use of persistent homology on tumor imaging to assess the severity of breast tumors. The next step is to apply this pipeline to other tumor segment images from The Cancer Imaging Archive at different sites such as the lung, kidney, and brain. In addition, whether other clinical parameters, such as overall survival, tumor stage, and tumor genotype data are captured well in persistent homology clusters will be assessed. If analyzing tumor MRI segments using persistent homology consistently identifies clinical relationships, this could enable clinicians to use persistent homology data as a noninvasive way to inform clinical decision making in oncology.

Keywords: cancer biology, oncology, persistent homology, radiomics, topological data analysis, tumor imaging

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Authors: Nikhilesh Singh, Shishir Gaur, Anitha K. Sharma

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Climate change and anthropogenic stress have severely affected ecosystems all over the globe. Indian rivers are under immense pressure, facing challenges like pollution, encroachment, extreme fluctuation in the flow regime, local ignorance and lack of coordination between stakeholders. To counter all these issues a holistic river rejuvenation plan is needed that tests, innovates and implements sustainable solutions in the river space for sustainable river management. Smart Laboratory for Clean Rivers (SLCR) an Indo-Danish collaboration project, provides a living lab setup that brings all the stakeholders (government agencies, academic and industrial partners and locals) together to engage, learn, co-creating and experiment for a clean and sustainable river that last for ages. Just like every mega project requires piloting, SLCR has opted for a small catchment of the Varuna River, located in the Middle Ganga Basin in India. Considering the integrated approach of river rejuvenation, SLCR embraces various techniques and upgrades for rejuvenation. Likely, maintaining flow in the channel in the lean period, Managed Aquifer Recharge (MAR) is a proven technology. In SLCR, Floa-TEM high-resolution lithological data is used in MAR models to have better decision-making for MAR structures nearby of the river to enhance the river aquifer exchanges. Furthermore, the concerns of quality in the river are a big issue. A city like Varanasi which is located in the last stretch of the river, generates almost 260 MLD of domestic waste in the catchment. The existing STP system is working at full efficiency. Instead of installing a new STP for the future, SLCR is upgrading those STPs with an IoT-based system that optimizes according to the nutrient load and energy consumption. SLCR also advocate nature-based solutions like a reed bed for the drains having less flow. In search of micropollutants, SLCR uses fingerprint analysis involves employing advanced techniques like chromatography and mass spectrometry to create unique chemical profiles. However, rejuvenation attempts cannot be possible without involving the entire catchment. A holistic water management plan that includes storm management, water harvesting structure to efficiently manage the flow of water in the catchment and installation of several buffer zones to restrict pollutants entering into the river. Similarly, carbon (emission and sequestration) is also an important parameter for the catchment. By adopting eco-friendly practices, a ripple effect positively influences the catchment's water dynamics and aids in the revival of river systems. SLCR has adopted 4 villages to make them carbon-neutral and water-positive. Moreover, for the 24×7 monitoring of the river and the catchment, robust IoT devices are going to be installed to observe, river and groundwater quality, groundwater level, river discharge and carbon emission in the catchment and ultimately provide fuel for the data analytics. In its completion, SLCR will provide a river restoration manual, which will strategise the detailed plan and way of implementation for stakeholders. Lastly, the entire process is planned in such a way that will be managed by local administrations and stakeholders equipped with capacity-building activity. This holistic approach makes SLCR unique in the field of river rejuvenation.

Keywords: sustainable management, holistic approach, living lab, integrated river management

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Authors: Maia Kharadze, Indira Djaparidze, Maia Vanidze, Aleko Kalandia

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Modern scientific world studies chemical components and antioxidant activity of different kinds of vines according to their breed purity and location. To our knowledge, this kind of research has not been conducted in Georgia yet. The object of our research was to study Chkhaveri vine, which is included in the oldest varieties of the Black Sea basin vine. We have studied different-altitude Chkaveri grapes, juice, and wine (half dry rose-colored produced with European technologies) and their technical markers, qualitative and quantitive composition of their biologically active compounds and their antioxidant activity. We were determining the amount of phenols using Folin-Ciocalteu reagent, Flavonoids, Catechins and Anthocyanins using Spectral method and antioxidant activity using DPPH method. Several compounds were identified using –HPLC-UV-Vis, UPLC-MS methods. Six samples of Chkhaveri species– 5, 300, 360, 380, 400, 780 meter altitudes were taken and analyzed. The sample taken from 360 m altitude is distinguished by its cluster mass (383.6 grams) and high amount of sugar (20.1%). The sample taken from the five-meter altitude is distinguished by having high acidity (0.95%). Unlike other grapes varieties, such concentration of sugar and relatively low levels of citric acid ultimately leads to Chkhaveri wine individuality. Biologically active compounds of Chkhaveri were researched in 2014, 2015, 2016. The amount of total phenols in samples of 2016 fruit varies from 976.7 to 1767.0 mg/kg. Amount of Anthocians is 721.2-1630.2 mg/kg, and the amount of Flavanoids varies from 300.6 to 825.5 mg/kg. Relatively high amount of anthocyanins was found in the Chkhaveri at 780-meter altitude - 1630.2 mg/kg. Accordingly, the amount of Phenols and Flavanoids is high- 1767.9 mg/kg and 825.5 mg/kg. These characteristics are low in samples gathered from 5 meters above sea level, Anthocyanins-721.2 mg/ kg, total Phenols-976.7 mg/ kg, and Flavanoids-300.6 mg/kg. The highest amount of bioactive compounds can be found in the Chkhaveri samples of high altitudes because with rising height environment becomes harsh, the plant has to develop a better immune system using Phenolic compounds. The technology that is used for the production of wine also plays a huge role in the composition of the final product. Optimal techniques of maceration and ageing were worked out. While squeezing Chkhaveri, there are no anthocyanins in the juice. However, the amount of Anthocyanins rises during maceration. After the fermentation of dregs, the amount of anthocyanins is 55%, 521.3 gm/l, total Phenols 80% 1057.7 mg/l and Flavanoids 23.5 mg/l. Antioxidant activity of samples was also determined using the effect of 50% inhibition of the samples. All samples have high antioxidant activity. For instance, in samples at 780 meters above the sea-level antioxidant activity was 53.5%. It is relatively high compared to the sample at 5 m above sea-level with the antioxidant activity of 30.5%. Thus, there is a correlation between the amount Anthocyanins and antioxidant activity. The designated project has been fulfilled by financial support of the Georgia National Science Foundation (Grant AP/96/13, Grant 216816), Any idea in this publication is possessed by the author and may not represent the opinion of the Georgia National Science Foundation.

Keywords: antioxidants, bioactive content, wine, chkhaveri

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Authors: Stephanie P. Nigro

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The Exhale Breath Hold (EBH) technique presents a promising approach to enhance the precision and efficacy of External Beam Radiation Therapy (EBRT) for abdominal tumours, which include liver, pancreas, kidney, and adrenal glands. These tumours are challenging to treat due to their proximity to organs at risk (OARs) and the significant motion induced by respiration and physiological variations, such as stomach filling. Respiratory motion can cause up to 40mm of displacement in abdominal organs, complicating accurate targeting. While current practices like limiting fasting help reduce motion related to digestive processes, they do not address respiratory motion. 4DCT scans are used to assess this motion, but they require extensive workflow time and expose patients to higher doses of radiation. The EBH technique, which involves holding the breath in an exhale with no air in the lungs, stabilizes internal organ motion, thereby reducing respiratory-induced motion. The primary benefit of EBH is the reduction in treatment volume sizes, specifically the Internal Target Volume (ITV) and Planning Target Volume (PTV), as demonstrated by smaller ITVs when gated in EBH. This reduction also improves the quality of 3D Cone Beam CT (CBCT) images by minimizing respiratory artifacts, facilitating soft tissue matching akin to stereotactic treatments. Patients suitable for EBH must meet criteria including the ability to hold their breath for at least 15 seconds and maintain a consistent breathing pattern. For those who do not qualify, the traditional 4DCT protocol will be used. The implementation involves an EBH planning scan and additional short EBH scans to ensure reproducibility and assist in contouring and volume expansions, with a Free Breathing (FB) scan used for setup purposes. Treatment planning on EBH scans leads to smaller PTVs, though intrafractional and interfractional breath hold variations must be accounted for in margins. The treatment decision process includes performing CBCT in EBH intervals, with careful matching and adjustment based on soft tissue and fiducial markers. Initial studies at two sites will evaluate the necessity of multiple CBCTs, assessing shifts and the benefits of initial versus mid-treatment CBCT. Considerations for successful implementation include thorough patient coaching, staff training, and verification of breath holds, despite potential disadvantages such as longer treatment times and patient exhaustion. Overall, the EBH technique offers significant improvements in the accuracy and quality of abdominal EBRT, paving the way for more effective and safer treatments for patients.

Keywords: abdominal cancers, exhale breath hold, radiation therapy, respiratory motion

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Authors: Shaista Suhail

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As cancer populations is increasing sharply, the incidence of oral squamous cell carcinoma (OSCC) has also been expected to increase. Oral carcinogenesis is a highly complex, multistep process which involves accumulation of genetic alterations that lead to the induction of proteins promoting cell growth (encoded by oncogenes), increased enzymatic (telomerase) activity promoting cancer cell proliferation. The global increase in frequency and mortality, as well as the poor prognosis of oral squamous cell carcinoma, has intensified current research efforts in the field of prevention and early detection of this disease. The advances in the understanding of the molecular basis of oral cancer should help in the identification of new markers. The study of the carcinogenic process of the oral cancer, including continued analysis of new genetic alterations, along with their temporal sequencing during initiation, promotion and progression, will allow us to identify new diagnostic and prognostic factors, which will provide a promising basis for the application of more rational and efficient treatments. Telomerase activity has been readily found in most cancer biopsies, in premalignant lesions or germ cells. Activity of telomerase is generally absent in normal tissues. It is known to be induced upon immortalization or malignant transformation of human cells such as in oral cancer cells. Maintenance of telomeres plays an essential role during transformation of precancer to malignant stage. Mammalian telomeres, a specialized nucleoprotein structures are composed of large conctamers of the guanine-rich sequence 5_-TTAGGG-3_. The roles of telomeres in regulating both stability of genome and replicative immortality seem to contribute in essential ways in cancer initiation and progression. It is concluded that activity of telomerase can be used as a biomarker for diagnosis of malignant oral cancer and a target for inactivation in chemotherapy or gene therapy. Its expression will also prove to be an important diagnostic tool as well as a novel target for cancer therapy. The activation of telomerase may be an important step in tumorgenesis which can be controlled by inactivating its activity during chemotherapy. The expression and activity of telomerase are indispensable for cancer development. There are no drugs which can effect extremely to treat oral cancers. There is a general call for new emerging drugs or methods that are highly effective towards cancer treatment, possess low toxicity, and have a minor environment impact. Some novel natural products also offer opportunities for innovation in drug discovery. Natural compounds isolated from medicinal plants, as rich sources of novel anticancer drugs, have been of increasing interest with some enzyme (telomerase) blockage property. The alarming reports of cancer cases increase the awareness amongst the clinicians and researchers pertaining to investigate newer drug with low toxicity.

Keywords: oral carcinoma, telomere, telomerase, blockage

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Authors: Shikha Rani, Neeta Sehgal, Vipin Kumar Verma, Om Prakash

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Introduction: Fish is an important protein source for humans and has great economic value. Fish cultures are affected due to various anthropogenic activities that lead to bacterial and viral infections. Aeromonas hydrophila is a fish pathogenic bacterium that causes several aquaculture outbreaks throughout the world and leads to huge mortalities. In this study, plants of no commercial value were used to investigate their immunostimulatory, antioxidant, anti-inflammatory, anti-bacterial, and disease resistance potential in fish against Aeromonas hydrophila, through fish feed fortification. Methods: The plant was dried at room temperature in the shade, dissolved in methanol, and analysed for biological compounds through GC-MS/MS. DPPH, FRAP, Phenolic, and flavonoids were estimated following standardized protocols. In silico molecular docking was also performed to validate its broad-spectrum activities based on binding affinity with specific proteins. Fish were divided into four groups (n=6; total 30 in a group): Group 1, non-challenged fish (fed on a non-supplemented diet); Group 2, fish challenged with bacteria (fed on a non-supplemented diet); Group 3 and 4, fish challenged with bacteria (A. hydrophila) and fed on plant supplemented feed at 2.5% and 5%. Blood was collected from the fish on 0, 7th, 14th, 21st, and 28th days. Serum was separated for glutamic-oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), alkaline phosphatase assay (ALP), lysozyme activity assay, superoxide dismutase assay (SOD), lipid peroxidation assay (LPO) and molecular parameters (including cytokine levels) were estimated through ELISA. The phagocytic activity of macrophages from the spleen and head kidney, along with quantitative analysis of immune-related genes, were analysed in different tissue samples. The digestive enzymes (Pepsin, Trypsin, and Chymotrypsin) were also measured to evaluate the effect of plant-supplemented feed on freshwater fish. Results and Discussion: GC-MS/MS analysis of a methanolic extract of plant validated the presence of key compounds having antioxidant, anti-inflammatory, anti-bacterial, anti-inflammatory, and immunomodulatory activities along with disease resistance properties. From biochemical investigations like ABTS, DPPH, and FRAP, the amount of total flavonoids, phenols, and promising binding affinities towards different proteins in molecular docking analysis helped us to realize the potential of this plant that can be used for investigation in the supplemented feed of fish. Measurement liver function tests, ALPs, oxidation-antioxidant enzyme concentrations, and immunoglobulin concentrations in the experimental groups (3 and 4) showed significant improvement as compared to the positive control group. The histopathological evaluation of the liver, spleen, and head kidney supports the biochemical findings. The isolated macrophages from the group fed on supplemented feed showed a higher percentage of phagocytosis and a phagocytic index, indicating an enhanced cell-mediated immune response. Significant improvements in digestive enzymes were also observed in fish fed on supplemented feed, even after weekly challenges with bacteria. Hence, the plant-fortified feed can be recommended as a regular feed to enhance fish immunity and disease resistance against the Aeromonas hydrophila infection after confirmation from the field trial.

Keywords: immunostimulation, antipathogen, plant fortified feed, macrophages, GC-MS/MS, in silico molecular docking

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Authors: Wei Xiao, Gangzhi Cai, Xingliang Qin, Hongyan Ren, Zaidong Hua, Zhe Zhu, Hongwei Xiao, Ximin Zheng, Jie Yao, Yanzhen Bi

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Myostatin (MSTN) is the master regulator of double muscling phenotype with overgrown muscle and decreased fatness in animals, but its action mode to regulate fat deposition remains to be elucidated. In this study a swin-specific pathway through which MSTN acts to regulate the fat deposition was deciphered. Deep sequenincing of the mRNA and miRNA of fat tissues of MSTN knockout (KO) and wildtype (WT) pigs discovered the positive correlation of myocyte enhancer factor 2C (MEF2C) and fat-inhibiting miR222 expression, and the inverse correlation of miR222 and stearoyl-CoA desaturase 5 (SCD5) expression. SCD5 is rodent-absent and expressed only in pig, sheep and cattle. Fatty acid spectrum of fat tissues revealed a lower percentage of oleoyl-CoA (18:1) and palmitoleyl CoA (16:1) in MSTN KO pigs, which are the catalyzing products of SCD5-mediated desaturation of steroyl CoA (18:0) and palmitoyl CoA (16:0). Blood metrics demonstrated a 45% decline of triglyceride (TG) content in MSTN KO pigs. In light of these observations we hypothesized that MSTN might act through MEF2C/miR222/SCD5 pathway to regulate desaturation of fatty acid as well as triglyceride synthesis in pigs. To this end, real-time PCR and Western blotting were carried out to detect the expression of the three genes stated above. These experiments showed that MEF2C expression was up-regulated by nearly 2-fold, miR222 up-regulated by nearly 3-fold and SCD5 down-regulated by nearly 50% in MSTN KO pigs. These data were consistent with the expression change in deep sequencing analysis. Dual luciferase reporter was then used to confirm the regulation of MEF2C upon the promoter of miR222. Ecotopic expression of MEF2C in preadipocyte cells enhanced miR222 expression by 3.48-fold. CHIP-PCR identified a putative binding site of MEF2C on -2077 to -2066 region of miR222 promoter. Electrophoretic mobility shift assay (EMSA) demonstrated the interaction of MEF2C and miR222 promoter in vitro. These data indicated that MEF2C transcriptionally regulates the expression of miR222. Next, the regulation of miR222 on SCD5 mRNA as well as its physiological consequences were examined. Dual luciferase reporter testing revealed the translational inhibition of miR222 upon the 3´ UTR (untranslated region) of SCD5 in preadipocyte cells. Transfection of miR222 mimics and inhibitors resulted in the down-regulation and up-regulation of SCD5 in preadipocyte cells respectively, consistent with the results from reporter testing. RNA interference of SCD5 in preadipocyte cells caused 26.2% reduction of TG, in agreement with the results of TG content in MSTN KO pigs. In summary, the results above supported the existence of a molecular pathway that MSTN signals through MEF2C/miR222/SCD5 to regulate the fat deposition in pigs. This swine-specific pathway offers potential molecular markers for the development and breeding of a new pig line with optimised fatty acid composition. This would benefit human health by decreasing the takeup of saturated fatty acid.

Keywords: fat deposition, MEF2C, miR222, myostatin, SCD5, pig

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Authors: Mustafa M. Donma, Orkide Donma

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Cardiovascular pathology is one of the expected consequences of excessive fat gain. The role of zinc in thyroid hormone metabolism is an important matter. The concentrations of both thyroid stimulating hormone (TSH) and zinc are subject to variation in obese individuals. Zinc exhibits protective effects on cardiovascular health and is inversely correlated with cardiovascular markers in childhood obesity. The association between subclinical hypothyroidism (SCHT) and metabolic disorders is under investigation due to its clinical importance. Underactive thyroid gland causes high TSH levels. Subclinical hypothyroidism is defined as the elevated serum TSH levels in the presence of normal free thyroxin (T4) concentrations. The aim of this study was to evaluate the associations between TSH levels and zinc concentrations in morbid obese (MO) children exhibiting SCHT. The possibility of using the probable association between these parameters was also evaluated for the discrimination of metabolic syndrome positive (MetS+) and metabolic syndrome negative (MetS-) groups. Forty-two children were present in each group. Informed consent forms were obtained. Institutional Ethics Committee approved the study protocol. Tables prepared by World Health Organization were used for the definition of MO children. Children, whose age- and sex-dependent body mass index percentile values were above 99, were defined as MO. Children with at least two MetS components were included in MOMetS+ group. Elevated systolic/diastolic blood pressure values, increased fasting blood glucose, triglycerides (TRG)/decreased high density lipoprotein-cholesterol (HDL-C) concentrations in addition to central obesity were listed as MetS components. Anthropometric measures were recorded. Routine biochemical analyses were performed. Thirteen and fifteen children had SCHT in MOMetS- and MOMetS+ groups, respectively. Statistical analyses were performed. p<0.05 was accepted as statistically significant. In MOMetS- and MOMetS+ groups, TSH levels were 4.1±2.9 mU/L and 4.6±3.1 mU/L, respectively. Corresponding values for SCHT cases in these groups were 7.3±3.1 mU/L and 8.0±2.7 mU/L. Free T4 levels were within normal limits. Zinc concentrations were negatively correlated with TSH levels in both groups. The significant negative correlation calculated in MOMetS+ group (r= -0.909; p<0.001) was much stronger than that found in MOMetS- group (r= -0.706; p<0.05). This strong correlation (r= -0.909; p<0.001) calculated for cases with SCHT in MOMetS+ group was much lower (r= -0.793; p<0.001) when all MOMetS+ cases were considered. Zinc is closely related to T4 and TSH therefore, it participates in thyroid hormone metabolism. Since thyroid hormones are required for zinc absorption, hypothyroidism can lead to zinc deficiency. The presence of strong correlations between TSH and zinc in SCHT cases found in both MOMetS- and MOMetS+ groups pointed out that MO children were under the threat of cardiovascular pathologies. The detection of the much stronger correlation in MOMetS+ group in comparison with the correlation found in MOMetS- group was the indicator of greater cardiovascular risk due to the presence of MetS. In MOMetS+ group, correlation in SCHT cases found higher than correlation calculated for all cases confirmed much higher cardiovascular risk due to the contribution of SCHT.

Keywords: cardiovascular risk, children, morbid obesity, subclinical hypothyroidism, zinc

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