Search results for: car sequencing and operator allocation

Authors: Yuli Hou, Jianping Sun, Mengdan Gao, Hui Liu, Ling Qin, Ang Li, Dongfu Li, Yonghong Zhang, Yan Zhao

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Background and Aims: Interferon-induced transmembrane protein 3 (IFITM3) is a component of ISG (Interferon-Stimulated Gene) family. IFITM3 has been recognized as a key signal molecule regulating cell growth in some tumors. However, the function of IFITM3 rs12252-CC genotype in the hepatocellular carcinoma (HCC) remains unknown to author’s best knowledge. A cohort study was employed to clarify the relationship between IFITM3 rs12252-CC genotype and HCC progression, and cellular experiments were used to investigate the correlation of function of IFITM3 and the progress of HCC. Methods: 336 candidates were enrolled in study, including 156 with HBV related HCC and 180 with chronic Hepatitis B infections or liver cirrhosis. Polymerase chain reaction (PCR) was employed to determine the gene polymorphism of IFITM3. The functions of IFITM3 were detected in PLC/PRF/5 cell with different treated:LV-IFITM3 transfected with lentivirus to knockdown the expression of IFITM3 and LV-NC transfected with empty lentivirus as negative control. The IFITM3 expression, proliferation and migration were detected by Quantitative reverse transcription polymerase chain reaction (qRT-PCR), QuantiGene Plex 2.0 assay, western blotting, immunohistochemistry, Cell Counting Kit(CCK)-8 and wound healing respectively. Six samples (three infected with empty lentiviral as control; three infected with LV-IFITM3 vector lentiviral as experimental group ) of PLC/PRF/5 were sequenced at BGI (Beijing Genomics Institute, Shenzhen,China) using RNA-seq technology to identify the IFITM3-related signaling pathways and chose PI3K/AKT pathway as related signaling to verify. Results: The patients with HCC had a significantly higher proportion of IFITM3 rs12252-CC compared with the patients with chronic HBV infection or liver cirrhosis. The distribution of CC genotype in HCC patients with low differentiation was significantly higher than that in those with high differentiation. Patients with CC genotype found with bigger tumor size, higher percentage of vascular thrombosis, higher distribution of low differentiation and higher 5-year relapse rate than those with CT/TT genotypes. The expression of IFITM3 was higher in HCC tissues than adjacent normal tissues, and the level of IFITM3 was higher in HCC tissues with low differentiation and metastatic than high/medium differentiation and without metastatic. Higher RNA level of IFITM3 was found in CC genotype than TT genotype. In PLC/PRF/5 cell with knockdown, the ability of cell proliferation and migration was inhibited. Analysis RNA sequencing and verification of RT-PCR found out the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/AKT/mTOR) pathway was associated with knockdown IFITM3.With the inhibition of IFITM3, the expression of PI3K/AKT/mTOR signaling pathway was blocked and the expression of vimentin was decreased. Conclusions: IFITM3 rs12252-CC with the higher expression plays a vital role in the progress of HCC by regulating HCC cell proliferation and migration. These effects are associated with PI3K/AKT/mTOR signaling pathway.

Keywords: IFITM3, interferon-induced transmembrane protein 3, HCC, hepatocellular carcinoma, PI3K/ AKT/mTOR, phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin

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Authors: H. I. Obadiah, S. N. Wieser, E. A. Omudu, B. O. Atu, O. Byanet, L. Schnittger, M. Florin-Christensen

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Sarcocystis sp. are Apicomplexan protozoan parasites with a life cycle that involves a predator and a prey as final and intermediate hosts, respectively. In tissues of the intermediate hosts, the parasites produce sarcocysts that vary in size and morphology according to the species. When a suitable predator ingests sarcocyst-containing meat, the parasites are released in the intestine and undergo sexual reproduction producing infective sporocysts, which are excreted with the feces into the environment. The cycle is closed when a prey ingests sporocyst-contaminated water or pasture; the parasites gain access to the circulation, and eventually invade tissues and reproduce asexually yielding sarcocysts. Pig farming is a common practice in Nigeria as well as in many countries around the world. In addition to its importance as protein source, pork is also a source of several pathogens relevant to humans. In the case of Sarcocystis, three species have been described both in domestic and wild pigs, namely, S. miescheriana, S. porcifelis and S. suihominis. Humans can act both as final and aberrant intermediate hosts of S. suihominis, after ingesting undercooked sarcocyst-infested pork. Infections are usually asymptomatic but can be associated with inappetence, nausea, vomiting and diarrhea, or with muscle pain, fever, eosinophilia and bronchospasm, in humans acting as final or intermediate hosts, respectively. Moreover, excretion of infective forms with human feces leads to further dissemination of the infection. In this study, macroscopic sarcocysts of white color, oval shape and a size range of approximately 3-5 mm were observed in the skeletal muscle of a slaughtered pig in an abattoir in Makurdi, Benue State, Nigeria, destined to human consumption. Sarcocysts were excised and washed in distilled water, and genomic DNA was extracted using a commercial kit. The near-complete length of the 18S rRNA gene was analyzed after PCR amplification of two overlapping fragments, each of which were submitted to direct sequencing. In addition, the mitochondrial cytochrome oxidase (cox-1) gene was PCR-amplified and directly sequenced. Two phylogenetic trees containing the obtained sequences along with available relevant 18S rRNA and cox-1 sequences were constructed by neighbor joining after alignment, using the corresponding sequences of Toxoplasma gondii as outgroup. The results showed in both cases that the analyzed sequences grouped with S. suihominis with high bootstrap value, confirming the identity of this macroscopic sarcocyst-forming parasite as S. suihominis. To the best of our knowledge, these results represent the first demonstration of this parasite in pigs of Nigeria and the largest sarcocysts described so far for S. suihominis. The close proximity between pigs and humans in pig farms, and the frequent poor sanitary conditions in human dwellings strongly suggest that the parasite undergoes the sexual stages of its life cycle in humans as final hosts. These findings provide an important reference for the examination and control of Sarcocystis species in pigs of Nigeria.

Keywords: nigeria, pork, sarcocystis suihominis, zoonotic parasite

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Authors: Beihai Xiong, Dengke Hua, Wouter Hendriks, Wilbert Pellikaan

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To improve the energy status of dairy cows in the early lactation, lots of jobs have been done on adjusting the starch to fiber ratio in the diet. As a complex ecosystem, the rumen contains a large population of microorganisms which plays a crucial role in feed degradation. Further study on the microbiota alterations and metabolic changes under different dietary energy sources is essential and valuable to better understand the function of the ruminal microorganisms and thereby to optimize the rumen function and enlarge feed efficiency. The present study will focus on the effects of two glucogenic diets (G: ground corn and corn silage; S: steam-flaked corn and corn silage) and a lipogenic diet (L: sugar beet pulp and alfalfa silage) on rumen fermentation, gas production, the ruminal microbiota and metabolome, and also their correlations in vitro. The gas production was recorded consistently, and the gas volume and producing rate at times 6, 12, 24, 48 h were calculated separately. The fermentation end-products were measured after fermenting for 48 h. The ruminal bacteria and archaea communities were determined by 16S RNA sequencing technique, the metabolome profile was tested through LC-MS methods. Compared to the diet G and S, the L diet had a lower dry matter digestibility, propionate production, and ammonia-nitrogen concentration. The two glucogenic diets performed worse in controlling methane and lactic acid production compared to the L diet. The S diet produced the greatest cumulative gas volume at any time points during incubation compared to the G and L diet. The metabolic analysis revealed that the lipid digestion was up-regulated by the diet L than other diets. On the subclass level, most metabolites belonging to the fatty acids and conjugates were higher, but most metabolites belonging to the amino acid, peptides, and analogs were lower in diet L than others. Differences in rumen fermentation characteristics were associated with (or resulting from) changes in the relative abundance of bacterial and archaeal genera. Most highly abundant bacteria were stable or slightly influenced by diets, while several amylolytic and cellulolytic bacteria were sensitive to the dietary changes. The L diet had a significantly higher number of cellulolytic bacteria, including the genera of Ruminococcus, Butyrivibrio, Eubacterium, Lachnospira, unclassified Lachnospiraceae, and unclassified Ruminococcaceae. The relative abundances of amylolytic bacteria genera including Selenomonas_1, Ruminobacter, and Succinivibrionaceae_UCG-002 were higher in diet G and S. These affected bacteria was also proved to have high associations with certain metabolites. The Selenomonas_1 and Succinivibrionaceae_UCG-002 may contribute to the higher propionate production in the diet G and S through enhancing the succinate pathway. The results indicated that the two glucogenic diets had a greater extent of gas production, a higher dry matter digestibility, and produced more propionate than diet L. The steam-flaked corn did not show a better performance on fermentation end-products than ground corn. This study has offered a deeper understanding of ruminal microbial functions which could assistant the improvement in rumen functions and thereby in the ruminant production.

Keywords: gas production, metabolome, microbiota, rumen fermentation

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Authors: László L. Lippai

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In addition to the fact that mental health bears great significance to a particular individual, it can also be regarded as an organizational, community and societal resource. Within the Szeged Health Promotion Research Group, we conducted mental health surveys on two levels: The inhabitants of a medium-sized Hungarian town and students of a Hungarian university with a relatively big headcount were requested to participate in surveys whose goals were to define local government priorities and organization-level health promotion programmes, respectively. To facilitate professional decision-making, we defined three, pragmatically relevant, groups of the target population: the mentally healthy, the vulnerable and the endangered. In order to determine which group a person actually belongs to, we designed a simple and quick measurement tool, which could even be utilised as a smoothing method, the Mental State Questionnaire validity of the above three categories was verified by analysis of variance against psychological quality of life variables. We demonstrate the pragmatic significance of our method via the analyses of the scores of our two mental health surveys. On town level, during our representative survey in Hódmezővásárhely (N=1839), we found that 38.7% of the participants was mentally healthy, 35.3% was vulnerable, while 16.3% was considered as endangered. We were able to identify groups that were in a dramatic state in terms of mental health. For example, such a group consisted of men aged 45 to 64 with only primary education qualification and the ratios of the mentally healthy, vulnerable and endangered were 4.5, 45.5 and 50%, respectively. It was also astonishing to see to what a little extent qualification prevailed as a protective factor in the case of women. Based on our data, the female group aged 18 to 44 with primary education—of whom 20.3% was mentally healthy, 42.4% vulnerable and 37.3% was endangered—as well as the female group aged 45 to 64 with university or college degree—of whom 25% was mentally healthy, 51.3 vulnerable and 23.8% endangered—are to be handled as priority intervention target groups in a similarly difficult position. On organizational level, our survey involving the students of the University of Szeged, N=1565, provided data to prepare a strategy of mental health promotion for a university with a headcount exceeding 20,000. When developing an organizational strategy, it was important to gather information to estimate the proportions of target groups in which mental health promotion methods; for example, life management skills development, detection, psychological consultancy, psychotherapy, would be applied. Our scores show that 46.8% of the student participants were mentally healthy, 42.1% were vulnerable and 11.1% were endangered. These data convey relevant information as to the allocation of organizational resources within a university with a considerable headcount. In conclusion, The Mental State Questionnaire, as a valid smoothing method, is adequate to describe a community in a plain and informative way in the terms of mental health. The application of the method can promote the preparation, design and implementation of mental health promotion interventions. 

Keywords: health promotion, mental health promotion, mental state questionnaire, psychological well-being

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Authors: Alla Ovsyannikova, Oksana Rymar, Elena Shakhtshneider, Mikhail Voevoda

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In Siberia endocrinologists increasingly noted young patients with the course of diabetes mellitus differing from 1 and 2 types. Therefore we did a molecular genetic study for this group of patients to verify the monogenic forms of diabetes mellitus in them and researched the characteristics of this pathology. When confirming the monogenic form of diabetes, we performed a correction therapy for many patients (transfer from insulin to tablets), prevented specific complications, examined relatives and diagnosed their diabetes at the preclinical stage, revealed phenotypic characteristics of the pathology which led to the high significance of this work. Materials and Methods: We observed 5 patients (4 families). We diagnosed MODY (Maturity Onset Diabetes of the Young) during the molecular genetic testing (direct automatic sequencing). All patients had a full clinical examination, blood samples for biochemical research, determination of C-peptide and TSH, antibodies to b-cells, microalbuminuria, abdominal ultrasound, heart and thyroid ultrasound, examination of ophthalmologist. Results: We diagnosed 3 rare types of MODY: two women had MODY8, one man – MODY6 and man and his mother - MODY12. Patients with types 8 and 12 had clinical features. Age of onset hyperglycemia ranged from 26 to 34 years. In a patient with MODY6 fasting hyperglycemia was detected during a routine examination. Clinical symptoms, complications were not diagnosed. The patient observes a diet. In the first patient MODY8 was detected during first pregnancy, she had itchy skin and mostly postprandial hyperglycemia. Upon examination we determined glycated hemoglobin 7.5%, retinopathy, non-proliferative stage, peripheral neuropathy. She uses a basic bolus insulin therapy. The second patient with MODY8 also had clinical manifestations of hyperglycemia (pruritus, thirst), postprandial hyperglycemia and diabetic nephropathy, a stage of microalbuminuria. The patient was diagnosed autoimmune thyroiditis. She used inhibitors of DPP-4. The patient with MODY12 had an aggressive course. In the detection of hyperglycemia he had complaints of visual impairment, intense headaches, leg cramps. The patient had a history of childhood convulsive seizures of non-epileptic genesis, without organic pathology, which themselves were stopped at the age of 12 years. When we diagnosed diabetes a patient was 28 years, he had hypertriglyceridemia, atherosclerotic plaque in the carotid artery, proliferative retinopathy (lacerocoagulation). Diabetes and early myocardial infarction were observed in three cases in family. We prescribe therapy with sulfonylureas and SGLT-2 inhibitors with a positive effect. At the patient's mother diabetes began at a later age (30 years) and a less aggressive course was observed. She also has hypertriglyceridemia and uses oral hypoglycemic drugs. Conclusions: 1) When young patients with hyperglycemia have extrapancreatic pathologies and diabetic complications with a short duration of diabetes we can assume they have one of type of MODY diabetes. 2) In patients with monogenic forms of diabetes mellitus, the clinical manifestations of hyperglycemia in each succeeding generation are revealed at an earlier age. Research had increased our knowledge of the monogenic forms of diabetes. The reported study was supported by RSCF, research project No. 14-15-00496-P.

Keywords: diabetes mellitus, MODY diabetes, monogenic forms, young patients

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Authors: Huseyin Sinan Gunesli

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Process Safety Management is a disciplined framework for managing the integrity of systems and processes that handle hazardous substances. It relies on good design principles, well-implemented automation systems, and operating and maintenance practices. Alarm Management Systems play a critically important role in the safe and efficient operation of modern industrial plants. In that respect, Alarm Management is one of the critical factors feeding the safe operations of the plants in the manner of applying effective process safety principles. Trans Anatolian Natural Gas Pipeline (TANAP) is part of the Southern Gas Corridor, which extends from the Caspian Sea to Italy. TANAP transports Natural Gas from the Shah Deniz gas field of Azerbaijan, and possibly from other neighboring countries, to Turkey and through Trans Adriatic Pipeline (TAP) Pipeline to Europe. TANAP plays a crucial role in maintaining Energy Security for the region and Europe. In that respect, the application of Process Safety principles is vital to deliver safe, reliable and efficient Natural Gas delivery to Shippers both in the region and Europe. Effective Alarm Management is one of those Process Safety principles which feeds safe operations of the TANAP pipeline. Alarm Philosophy was designed and implemented in TANAP Pipeline according to the relevant standards. However, it is essential to manage the alarms received in the control room effectively to maintain safe operations. In that respect, TANAP has commenced Alarm Management & Rationalization program as of February 2022 after transferring to Plateau Regime, reaching the design parameters. While Alarm Rationalization started, there were more than circa 2300 alarms received per hour from one of the compressor stations. After applying alarm management principles such as reviewing and removal of bad actors, standing, stale, chattering, fleeting alarms, comprehensive review and revision of alarm set points through a change management principle, conducting alarm audits/design verification and etc., it has been achieved to reduce down to circa 40 alarms per hour. After the successful implementation of alarm management principles as specified above, the number of alarms has been reduced to industry standards. That significantly improved operator vigilance to focus on mainly important and critical alarms to avoid any excursion beyond safe operating limits leading to any potential process safety events. Following the ‟What Gets Measured, Gets Managed” principle, TANAP has identified key Performance Indicators (KPIs) to manage Process Safety principles effectively, where Alarm Management has formed one of the key parameters of those KPIs. However, review and analysis of the alarms were performed manually. Without utilizing Alarm Management Software, achieving full compliance with international standards is almost infeasible. In that respect, TANAP has started using one of the industry-wide known Alarm Management Applications to maintain full review and analysis of alarms and define actions as required. That actually significantly empowered TANAP’s process safety principles in terms of Alarm Management.

Keywords: process safety principles, energy security, natural gas pipeline operations, alarm rationalization, alarm management, alarm management application

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Authors: W. Staiano, S. Marcora

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Introduction: It has been proposed that acute negative effect of mental fatigue (MF) could potentially become a training stimulus for the brain (Brain endurance training (BET)) to adapt and improve its ability to attenuate MF states during sport competitions. Purpose: The aim of this study was to test the efficacy of 6 weeks of BET on cognitive and cycling tests in a group of well-trained subjects. We hypothesised that combination of BET and standard physical training (SPT) would increase cognitive capacity and cycling performance by reducing rating of perceived exertion (RPE) and increase resilience to fatigue more than SPT alone. Methods: In a randomized controlled trial design, 26 well trained participants, after a familiarization session, cycled to exhaustion (TTE) at 80% peak power output (PPO) and, after 90 min rest, at 65% PPO, before and after random allocation to a 6 week BET or active placebo control. Cognitive performance was measured using 30 min of STROOP coloured task performed before cycling performance. During the training, BET group performed a series of cognitive tasks for a total of 30 sessions (5 sessions per week) with duration increasing from 30 to 60 min per session. Placebo engaged in a breathing relaxation training. Both groups were monitored for physical training and were naïve to the purpose of the study. Physiological and perceptual parameters of heart rate, lactate (LA) and RPE were recorded during cycling performances, while subjective workload (NASA TLX scale) was measured during the training. Results: Group (BET vs. Placebo) x Test (Pre-test vs. Post-test) mixed model ANOVA’s revealed significant interaction for performance at 80% PPO (p = .038) or 65% PPO (p = .011). In both tests, groups improved their TTE performance; however, BET group improved significantly more compared to placebo. No significant differences were found for heart rate during the TTE cycling tests. LA did not change significantly at rest in both groups. However, at completion of 65% TTE, it was significantly higher (p = 0.043) in the placebo condition compared to BET. RPE measured at ISO-time in BET was significantly lower (80% PPO, p = 0.041; 65% PPO p= 0.021) compared to placebo. Cognitive results in the STROOP task showed that reaction time in both groups decreased at post-test. However, BET decreased significantly (p = 0.01) more compared to placebo despite no differences accuracy. During training sessions, participants in the BET showed, through NASA TLX questionnaires, constantly significantly higher (p < 0.01) mental demand rates compared to placebo. No significant differences were found for physical demand. Conclusion: The results of this study provide evidences that combining BET and SPT seems to be more effective than SPT alone in increasing cognitive and cycling performance in well trained endurance participants. The cognitive overload produced during the 6-week training of BET can induce a reduction in perception of effort at a specific power, and thus improving cycling performance. Moreover, it provides evidence that including neurocognitive interventions will benefit athletes by increasing their mental resilience, without affecting their physical training load and routine.

Keywords: cognitive training, perception of effort, endurance performance, neuro-performance

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Authors: Kaomud Tyagi, Rajasree Chakraborty, Shantanu Kundu, Devkant Singha, Kailash Chandra, Vikas Kumar

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The melon thrips, Thrips palmi is a serious pest of a wide range of agriculture crops and also act as vectors for plant viruses (genus Tospovirus, family Bunyaviridae). More molecular data on this species is required to understand the cryptic speciation and evolutionary affiliations. Mitochondrial genomes have been widely used in phylogenetic and evolutionary studies in insect. So far, mitogenomes of five thrips species (Anaphothrips obscurus, Frankliniella intonsa, Frankliniella occidentalis, Scirtothrips dorsalis and Thrips imaginis) is available in the GenBank database. In this study, we sequenced the first complete mitogenome T. palmi and compared it with available thrips mitogenomes. We assembled the mitogenome from the whole genome sequencing data generated using Illumina Hiseq2500. Annotation was performed using MITOS web-server to estimate the location of protein coding genes (PCGs), transfer RNA (tRNAs), ribosomal RNAs (rRNAs) and their secondary structures. The boundaries of PCGs and rRNAs was confirmed manually in NCBI. Phylogenetic analyses were performed using the 13 PCGs data using maximum likelihood (ML) in PAUP, and Bayesian inference (BI) in MrBayes 3.2. The complete mitogenome of T. palmi was 15,333 base pairs (bp), which was greater than the genomes of A. obscurus (14,890bp), F. intonsa (15,215 bp), F. occidentalis (14,889 bp) and S. dorsalis South Asia strain (SA1) (14,283 bp), but smaller than the genomes of T. imaginis (15,407 bp) and S. dorsalis East Asia strain (EA1) (15,343bp). Like in other thrips species, the mitochondrial genome of T. palmi was represented by 37 genes, including 13 PCGs, large and small ribosomal RNA (rrnL and rrnS) genes, 22 transfer RNA (tRNAs) genes (with one extra gene for trn-Serine) and two A+T-rich control regions (CR1 and CR2). Thirty one genes were observed on heavy (H) strand and six genes on the light (L) strand. The six tRNA genes (trnG,trnK, trnY, trnW, trnF, and trnH) were found to be conserved in all thrips species mitogenomes in their locations relative to a protein-coding or rRNA gene upstream or downstream. The gene arrangements of T. palmi is very close to T. imaginis except the rearrangements in tRNAs genes: trnR (arginine), and trnE (glutamic acid) were found to be located between cox3 and CR2 in T. imaginis which were translocated between atp6 and CR1 in T. palmi; trnL1 (Leucine) and trnS1(Serine) were located between atp6 and CR1 in T. imaginis which were translocated between cox3 and CR2 in T. palmi. The location of CR1 upstream of nad5 gene was suggested to be ancestral condition of the thrips species in subfamily Thripinae, was also observed in T. palmi. Both the Maximum likelihood (ML) and Bayesian Inference (BI) phylogenetic trees generated resulted in similar topologies. The T. palmi was clustered with T. imaginis. We concluded that more molecular data on the diverse thrips species from different hierarchical level is needed, to understand the phylogenetic and evolutionary relationships among them.

Keywords: thrips, comparative mitogenomics, gene rearrangements, phylogenetic analysis

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Authors: Okolie Pius Ifeanyi, Emerenini Emilymary Chima

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Aims: The study investigated the diversity and identities of Lactic Acid Bacteria (LAB) isolated from different fresh fruits using Molecular Nested PCR analysis and the efficacy of cell free supernatants from Lactic Acid Bacteria (LAB) isolated from fresh fruits for in vitro control of some tomato pathogens. Study Design: Nested PCR approach was used in this study employing universal 16S rRNA gene primers in the first round PCR and LAB specific Primers in the second round PCR with the view of generating specific Nested PCR products for the LAB diversity present in the samples. The inhibitory potentials of supernatant obtained from LAB isolates of fruits origin that were molecularly characterized were investigated against some tomato phytopathogens using agar-well method with the view to develop biological agents for some tomato disease causing organisms. Methodology: Gram positive, catalase negative strains of LAB were isolated from fresh fruits on Man Rogosa and Sharpe agar (Lab M) using streaking method. Isolates obtained were molecularly characterized by means of genomic DNA extraction kit (Norgen Biotek, Canada) method. Standard methods were used for Nested Polymerase Chain Reaction (PCR) amplification targeting the 16S rRNA gene using universal 16S rRNA gene and LAB specific primers, agarose gel electrophoresis, purification and sequencing of generated Nested PCR products (Macrogen Inc., USA). The partial sequences obtained were identified by blasting in the non-redundant nucleotide database of National Center for Biotechnology Information (NCBI). The antimicrobial activities of characterized LAB against some tomato phytopathogenic bacteria which include (Xanthomonas campestries, Erwinia caratovora, and Pseudomonas syringae) were obtained by using the agar well diffusion method. Results: The partial sequences obtained were deposited in the database of National Centre for Biotechnology Information (NCBI). Isolates were identified based upon the sequences as Weissella cibaria (4, 18.18%), Weissella confusa (3, 13.64%), Leuconostoc paramensenteroides (1, 4.55%), Lactobacillus plantarum (8, 36.36%), Lactobacillus paraplantarum (1, 4.55%) and Lactobacillus pentosus (1, 4.55%). The cell free supernatants of LAB from fresh fruits origin (Weissella cibaria, Weissella confusa, Leuconostoc paramensenteroides, Lactobacillus plantarum, Lactobacillus paraplantarum and Lactobacillus pentosus) can inhibits these bacteria by creating clear zones of inhibition around the wells containing cell free supernatants of the above mentioned strains of lactic acid bacteria. Conclusion: This study shows that potentially LAB can be quickly characterized by molecular methods to specie level by nested PCR analysis of the bacteria isolate genomic DNA using universal 16S rRNA primers and LAB specific primer. Tomato disease causing organisms can be most likely biologically controlled by using extracts from LAB. This finding will reduce the potential hazard from the use of chemical herbicides on plant.

Keywords: nested pcr, molecular characterization, 16s rRNA gene, lactic acid bacteria

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Authors: Andreas Kerstan, Darren Robey, Wesam Alvan, David Troiani

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Over the last 3.5 years, Quantum Cascade Lasers (QCL) technology has become increasingly important in infrared (IR) microscopy. The advantages over fourier transform infrared (FTIR) are that large areas of a few square centimeters can be measured in minutes and that the light intensive QCL makes it possible to obtain spectra with excellent S/N, even with just one scan. A firmly established solution of the laser direct infrared imaging (LDIR) 8700 is the analysis of microplastics. The presence of microplastics in the environment, drinking water, and food chains is gaining significant public interest. To study their presence, rapid and reliable characterization of microplastic particles is essential. Significant technical hurdles in microplastic analysis stem from the sheer number of particles to be analyzed in each sample. Total particle counts of several thousand are common in environmental samples, while well-treated bottled drinking water may contain relatively few. While visual microscopy has been used extensively, it is prone to operator error and bias and is limited to particles larger than 300 µm. As a result, vibrational spectroscopic techniques such as Raman and FTIR microscopy have become more popular, however, they are time-consuming. There is a demand for rapid and highly automated techniques to measure particle count size and provide high-quality polymer identification. Analysis directly on the filter that often forms the last stage in sample preparation is highly desirable as, by removing a sample preparation step it can both improve laboratory efficiency and decrease opportunities for error. Recent advances in infrared micro-spectroscopy combining a QCL with scanning optics have created a new paradigm, LDIR. It offers improved speed of analysis as well as high levels of automation. Its mode of operation, however, requires an IR reflective background, and this has, to date, limited the ability to perform direct “on-filter” analysis. This study explores the potential to combine the filter with an infrared reflective surface filter. By combining an IR reflective material or coating on a filter membrane with advanced image analysis and detection algorithms, it is demonstrated that such filters can indeed be used in this way. Vibrational spectroscopic techniques play a vital role in the investigation and understanding of microplastics in the environment and food chain. While vibrational spectroscopy is widely deployed, improvements and novel innovations in these techniques that can increase the speed of analysis and ease of use can provide pathways to higher testing rates and, hence, improved understanding of the impacts of microplastics in the environment. Due to its capability to measure large areas in minutes, its speed, degree of automation and excellent S/N, the LDIR could also implemented for various other samples like food adulteration, coatings, laminates, fabrics, textiles and tissues. This presentation will highlight a few of them and focus on the benefits of the LDIR vs classical techniques.

Keywords: QCL, automation, microplastics, tissues, infrared, speed

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Authors: Ricardo Godoy, Herbert Venthur, Hector Jimenez, Andres Quiroz, Ana Mutis

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In agriculture, grape production has great economic importance at global level, considering that in 2013 it reached 7.4 million hectares (ha) covered by plantations of this fruit worldwide. Chile is the number one exporter in the world with 800,000 tons. However, these values have been threatened by the attack of the grapevine moth, Lobesia botrana (Denis & Schiffermuller) (Lepidoptera: Tortricidae), since its detection in 2008. Nowadays, the use of semiochemicals, in particular the major component of the sex pheromone, (E,Z)-7.9-dodecadienil acetate, are part of mating disruption methods to control L. botrana. How insect pests can recognize these molecules, is being part of huge efforts to deorphanize their olfactory mechanism at molecular level. Thus, an interesting group of proteins has been identified in the antennae of insects, where odorant-binding proteins (OBPs) are known by transporting molecules to odorant receptors (ORs) and a co-receptor (ORCO) causing a behavioral change in the insect. Other proteins such as chemosensory proteins (CSPs), ionotropic receptors (IRs), odorant degrading enzymes (ODEs) and sensory neuron membrane proteins (SNMPs) seem to be involved, but few studies have been performed so far. The above has led to an increasing interest in insect communication at a molecular level, which has contributed to both a better understanding of the olfaction process and the design of new pest management strategies. To date, it has been reported that the ORs can detect one or a small group of odorants in a specific way. Therefore, the objective of this study is the identification of genes that encode these ORs using the antennal transcriptome of L. botrana. Total RNA was extracted for females and males of L. botrana, and the antennal transcriptome sequenced by Next Generation Sequencing service using an Illumina HiSeq2500 platform with 50 million reads per sample. Unigenes were assembled using Trinity v2.4.0 package and transcript abundance was obtained using edgeR. Genes were identified using BLASTN and BLASTX locally installed in a Unix system and based on our own Tortricidae database. Those Unigenes related to ORs were characterized using ORFfinder and protein Blastp server. Finally, a phylogenetic analysis was performed with the candidate amino acid sequences for LbotORs including amino acid sequences of other moths ORs, such as Bombyx mori, Cydia pomonella, among others. Our findings suggest 61 genes encoding ORs and one gene encoding an ORCO in both sexes, where the greatest difference was found in the OR6 because of the transcript abundance according to the value of FPKM in females and males was 1.48 versus 324.00. In addition, according to phylogenetic analysis OR6 is closely related to OR1 in Cydia pomonella and OR6, OR7 in Epiphyas postvittana, which have been described as pheromonal receptors (PRs). These results represent the first evidence of ORs present in the antennae of L. botrana and a suitable starting point for further functional studies with selected ORs, such as OR6, which is potentially related to pheromonal recognition.

Keywords: antennal transcriptome, lobesia botrana, odorant receptors (ORs), phylogenetic analysis

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Authors: Tumer Orhun Aykut, Robin Michael Crucitti-Thoo, Agnieszka Rudak, Iwona Jasser

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Due to eutrophication connected to the growing human population, intensive agriculture, industrialization, and reinforcement of global warming, freshwater resources are changing negatively in every region of the World. This change also concerns the replacement of native species by invasive ones that can spread in many ways. Biological invasions are a developing problem to ecosystem continuity and their presence is mostly common in freshwater bodies. The occurrence and potential invasion of the species depends on associations between abiotic and biotic variables. Due to climate change, many species can extend their range from low to high latitudes and differ in their geographic ranges. In addition, the hydrological issues strongly influence the physicochemical parameters and biological processes, especially the growth rates of species and bloom formation of Cyanobacteria. Among tropical invasive species noted in temperate Europe, Raphidiopsis raciborskii, Chrysosporum bergii, and Sphaerospermopsis aphanizomenoides are considered a serious threat. R. raciborskii being the most important one as it is already known as a highly invasive species in almost all around the World, is a freshwater, planktonic, filamentous, potentially toxic, and nitrogen-fixing Cyanobacteria. This study aimed to investigate the presence of invasive cyanobacterial species in temperate lakes in Northeastern Poland, reveal the composition of phytoplankton communities, determine the effect of environmental variables, and identify the toxic properties of invasive Cyanobacteria and other phytoplankton groups. Our study was conducted in twenty-five lakes in August 2023. The lakes represent a geographical gradient from central Poland to the Northeast and have different depths, sizes, and trophic statuses. According to performed analyses, the presence of R. raciborskii was recorded in five lakes: Szczęśliwickie (Warsaw), Mikołajskie, Rekąty, Sztynorckie (Masurian Lakeland), and further East, in Pobondzie (Suwałki Lakeland). On the other hand, C. bergii was found in three lakes: Rekąty (Masurian Lakeland), Żabinki, and Pobondzie (Suwałki Lakeland), while S. aphanizomenoides only in Pobondzie (Suwałki Lakeland). Maximum phytoplankton diversity was found in Lake Rekąty, a small and shallow lake mentioned above. The highest phytoplankton biomass was detected in highly eutrophic Lake Suskie, followed by Lake Sztynorckie. In this last lake, which is also strongly eutrophic, the highest biomass of R. raciborskii was found. Cyanophyceae had the highest biovolume and was followed by Chlorophyceae in the entire study. Numerous environmental parameters, including nutrients, were studied, and their relationships with the invasive species and the whole phytoplankton community will be presented. In addition, toxic properties of environmental DNA results from each lake will also be shown. In conclusion, investigated invasive cyanobacterial species were found in a few Northeastern Polish temperate lakes, but the number of individuals was quite low, so the biomass was quite low. It has been observed that the structure of phytoplankton changed based on lakes and environmental parameters.

Keywords: biological invasion, cyanobacteria, cyanotoxins, phytoplankton ecology, sanger sequencing

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Authors: Arsyam Mawardi, Sony Suhandono, Azzania Fibriani, Fifi Fitriyah Masduki

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Malaria is an infectious disease caused by Plasmodium sp. This disease has a high prevalence in Indonesia, especially in Jayapura. The vaccine that is currently being developed has not been effective in overcoming malaria. This is due to the high polymorphism in the Plasmodium genome especially in areas that encode Plasmodium surface proteins. Merozoite Surface Protein 1 (MSP1) Plasmodium falciparum is a surface protein that plays a role in the invasion process in human erythrocytes through the interaction of Glycophorin A protein receptors and sialic acid in erythrocytes with Reticulocyte Binding Proteins (RBP) and Duffy Adhesion Protein (DAP) ligands in merozoites. MSP1 can be targeted to be a specific antigen and predicted epitope area which will be used for the development of diagnostic and malaria vaccine therapy. MSP1 consists of 17 blocks, each block is dimorphic, and has been marked as the K1 and MAD20 alleles. Exceptions only in block 2, because it has 3 alleles, among others K1, MAD20 and RO33. These polymorphisms cause allelic variations and implicate the severity of patients infected P. falciparum. In addition, polymorphism of MSP1 in Jayapura isolates has not been reported so it is interesting to be further identified and projected as a specific antigen. Therefore, in this study, we analyzed the allele polymorphism as well as detected the MSP1 epitope antigen candidate on block 2 P. falciparum. Clinical samples of selected malaria patients followed the consecutive sampling method, examining malaria parasites with blood preparations on glass objects observed through a microscope. Plasmodium DNA was isolated from the blood of malarial positive patients. The block 2 MSP1 gene was amplified using PCR method and cloned using the pGEM-T easy vector then transformed to TOP'10 E.coli. Positive colonies selection was performed with blue-white screening. The existence of target DNA was confirmed by PCR colonies and DNA sequencing methods. Furthermore, DNA sequence analysis was done through alignment and formation of a phylogenetic tree using MEGA 6 software and insilico analysis using IEDB software to predict epitope candidate for P. falciparum. A total of 15 patient samples have been isolated from Plasmodium DNA. PCR amplification results show the target gene size about ± 1049 bp. The results of MSP1 nucleotide alignment analysis reveal that block 2 MSP1 genes derived from the sample of malarial patients were distributed in four different allele family groups, K1 (7), MAD20 (1), RO33 (0) and MSP1_Jayapura (10) alleles. The most commonly appears of the detected allele is MSP1_Jayapura single allele. There was no significant association between sex variables, age, the density of parasitemia and alel variation (Mann Whitney, U > 0.05), while symptomatic signs have a significant difference as a trigger of detectable allele variation (U < 0.05). In this research, insilico study shows that there is a new epitope antigen candidate from the MSP1_Jayapura allele and it is predicted to be recognized by B cells with 17 amino acid lengths in the amino acid sequence 187 to 203.

Keywords: epitope candidate, insilico analysis, MSP1 P. falciparum, polymorphism

Procedia PDF Downloads 165

Authors: Eric Bang

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Interactions between receptors and ligands are crucial for many essential biological processes, including neurotransmission and metabolism. Ligand-receptor analyses that examine cell behavior and interactions often utilize cell type-specific RNA expressions from single-cell RNA sequencing (scRNA-seq) data. Using CellPhoneDB, a public repository consisting of ligands, receptors, and ligand-receptor interactions, the cell-cell interactions were explored in a specific scRNA-seq dataset from kidney tissue and portrayed the results with dot plots and heat maps. Depending on the type of cell, each ligand-receptor pair was aligned with the interacting cell type and calculated the positori probabilities of these associations, with corresponding P values reflecting average expression values between the triads and their significance. Using single-cell data (sample kidney cell references), genes in the dataset were cross-referenced with ones in the existing CellPhoneDB dataset. For example, a gene such as Pleiotrophin (PTN) present in the single-cell data also needed to be present in the CellPhoneDB dataset. Using the single-cell transcriptomics data via slide-seq and reference data, the CellPhoneDB program defines cell types and plots them in different formats, with the two main ones being dot plots and heat map plots. The dot plot displays derived measures of the cell to cell interaction scores and p values. For the dot plot, each row shows a ligand-receptor pair, and each column shows the two interacting cell types. CellPhoneDB defines interactions and interaction levels from the gene expression level, so since the p-value is on a -log10 scale, the larger dots represent more significant interactions. By performing an interaction analysis, a significant interaction was discovered for myeloid and T-cell ligand-receptor pairs, including those between Secreted Phosphoprotein 1 (SPP1) and Fibronectin 1 (FN1), which is consistent with previous findings. It was proposed that an effective protocol would involve a filtration step where cell types would be filtered out, depending on which ligand-receptor pair is activated in that part of the tissue, as well as the incorporation of the CellPhoneDB data in a streamlined workflow pipeline. The filtration step would be in the form of a Python script that expedites the manual process necessary for dataset filtration. Being in Python allows it to be integrated with the CellPhoneDB dataset for future workflow analysis. The manual process involves filtering cell types based on what ligand/receptor pair is activated in kidney cells. One limitation of this would be the fact that some pairings are activated in multiple cells at a time, so the manual manipulation of the data is reflected prior to analysis. Using the filtration script, accurate sorting is incorporated into the CellPhoneDB database rather than waiting until the output is produced and then subsequently applying spatial data. It was envisioned that this would reveal wherein the cell various ligands and receptors are interacting with different cell types, allowing for easier identification of which cells are being impacted and why, for the purpose of disease treatment. The hope is this new computational method utilizing spatially explicit ligand-receptor association data can be used to uncover previously unknown specific interactions within kidney tissue.

Keywords: bioinformatics, Ligands, kidney tissue, receptors, spatial transcriptome

Procedia PDF Downloads 127

Authors: Niklas Ravn-Boess, Nainita Bhowmick, Takamitsu Hattori, Shohei Koide, Christopher Park, Dimitris Placantonakis

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Background: Glioblastoma (GBM) is the most common and deadly primary brain malignancy in adults. Tumor propagation, brain invasion, and resistance to therapy critically depend on GBM stem-like cells (GSCs); however, the mechanisms that regulate GSC self-renewal are incompletely understood. Given the aggressiveness and poor prognosis of GBM, it is imperative to find biomarkers that could also translate into novel drug targets. Along these lines, we have identified a cell surface antigen, CD97 (ADGRE5), an adhesion G protein-coupled receptor (GPCR), that is expressed on GBM cells but is absent from non-neoplastic brain tissue. CD97 has been shown to promote invasiveness, angiogenesis, and migration in several human cancers, but its frequency of expression and functional role in regulating GBM growth and survival, and its potential as a therapeutic target has not been investigated. Design: We assessed CD97 mRNA and protein expression in patient derived GBM samples and cell lines using publicly available RNA-sequencing datasets and flow cytometry, respectively. To assess CD97 function, we generated shRNA lentiviral constructs that target a sequence in the CD97 extracellular domain (ECD). A scrambled shRNA (scr) with no predicted targets in the genome was used as a control. We evaluated CD97 shRNA lentivirally transduced GBM cells for Ki67, Annexin V, and DAPI. We also tested CD97 KD cells for their ability to self-renew using clonogenic tumorsphere formation assays. Further, we utilized synthetic Abs (sAbs) generated against the ECD of CD97 to test for potential antitumor effects using patient-derived GBM cell lines. Results: CD97 mRNA expression was expressed at high levels in all GBM samples available in the TCGA cohort. We found high levels of surface CD97 protein expression in 6/6 patient-derived GBM cell cultures, but not human neural stem cells. Flow cytometry confirmed downregulation of CD97 in CD97 shRNA lentivirally transduced cells. CD97 KD induced a significant reduction in cell growth in 3 independent GBM cell lines representing mesenchymal and proneural subtypes, which was accompanied by reduced (~20%) Ki67 staining and increased (~30%) apoptosis. Incubation of GBM cells with sAbs (20 ug/ ml) against the ECD of CD97 for 3 days induced GSC differentiation, as determined by the expression of GFAP and Tubulin. Using three unique GBM patient derived cultures, we found that CD97 KD attenuated the ability of GBM cells to initiate sphere formation by over 300 fold, consistent with an impairment in GSC self-renewal. Conclusion: Loss of CD97 expression in patient-derived GBM cells markedly decreases proliferation, induces cell death, and reduces tumorsphere formation. sAbs against the ECD of CD97 reduce tumorsphere formation, recapitulating the phenotype of CD97 KD, suggesting that sAbs that inhibit CD97 function exhibit anti-tumor activity. Collectively, these findings indicate that CD97 is necessary for the proliferation and survival of human GBM cells and identify CD97 as a promising therapeutically targetable vulnerability in GBM.

Keywords: adhesion GPCR, CD97, GBM stem cell, glioblastoma

Procedia PDF Downloads 114

Authors: Sogand Barghi

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The integration of artificial intelligence (AI) in accounting practices is reshaping the landscape of financial management. This paper explores the innovative applications of AI in the realm of accounting, emphasizing its transformative impact on efficiency, accuracy, decision-making, and financial insights. By harnessing AI's capabilities in data analysis, pattern recognition, and automation, accounting professionals can redefine their roles, elevate strategic decision-making, and unlock unparalleled value for businesses. This paper delves into AI-driven solutions such as automated data entry, fraud detection, predictive analytics, and intelligent financial reporting, highlighting their potential to revolutionize the accounting profession. Artificial intelligence has swiftly emerged as a game-changer across industries, and accounting is no exception. This paper seeks to illuminate the profound ways in which AI is reshaping accounting practices, transcending conventional boundaries, and propelling the profession toward a new era of efficiency and insight-driven decision-making. One of the most impactful applications of AI in accounting is automation. Tasks that were once labor-intensive and time-consuming, such as data entry and reconciliation, can now be streamlined through AI-driven algorithms. This not only reduces the risk of errors but also allows accountants to allocate their valuable time to more strategic and analytical tasks. AI's ability to analyze vast amounts of data in real time enables it to detect irregularities and anomalies that might go unnoticed by traditional methods. Fraud detection algorithms can continuously monitor financial transactions, flagging any suspicious patterns and thereby bolstering financial security. AI-driven predictive analytics can forecast future financial trends based on historical data and market variables. This empowers organizations to make informed decisions, optimize resource allocation, and develop proactive strategies that enhance profitability and sustainability. Traditional financial reporting often involves extensive manual effort and data manipulation. With AI, reporting becomes more intelligent and intuitive. Automated report generation not only saves time but also ensures accuracy and consistency in financial statements. While the potential benefits of AI in accounting are undeniable, there are challenges to address. Data privacy and security concerns, the need for continuous learning to keep up with evolving AI technologies, and potential biases within algorithms demand careful attention. The convergence of AI and accounting marks a pivotal juncture in the evolution of financial management. By harnessing the capabilities of AI, accounting professionals can transcend routine tasks, becoming strategic advisors and data-driven decision-makers. The applications discussed in this paper underline the transformative power of AI, setting the stage for an accounting landscape that is smarter, more efficient, and more insightful than ever before. The future of accounting is here, and it's driven by artificial intelligence.

Keywords: artificial intelligence, accounting, automation, predictive analytics, financial reporting

Procedia PDF Downloads 52

Authors: Mohamed M. Zaki, Maha M. Hady

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Salmonella spp. have been categorized as the world’s biggest threats to human health and poultry products are mostly incriminated sources. In Egypt, it was found that S. enteritidis and S. typhimurium are the most prevalent ones in poultry farms. It is recommended to eliminate salmonella from living bird by competing for salmonella contamination in feed in order to establish a healthy gut. The Feed acidifiers are the group of feed additives containing low-molecular-weight organic acids and/ or their salts which act as performance promoters by lowering the pH in the gut, optimizes digestion and inhibit bacterial growth. The inclusion of organic acid in pure form nonetheless effective in feed, yet, it is difficult to handle in feed mills as it is corrosive and produce more losses during pelleting process. The current study aimed at to evaluate the impact of incorporation of sodium diformate (SDF) and a commercial acidifier, CA (a mixture of butyric and propionic acids and their ammonium salts) at 0.4% dietary levels on broilers performance and the control S. enteritidis infection. Two hundreds and seventy unsexed cobb chickens were allotted in one of three treatments (90/ group) which were, the control (no acidifier, C- &C+), the 0.4% SDF (SDF- & SDF +) and the 0.4% CA (CA- & CA +) dietary levels for 35 days. Before the allocation of the groups, ten extra birds and a diet sample were bacteriologically examined to ensure negative contamination with salmonella. The birds were raised on deep-litter separated pens and had free access to feed and water all the time. The experimentally formulated diets were kept at 40C. After 24h access to the different dietary treatments, all the birds in the positive groups (n=15/ replicate) were inoculated intra-crop with 0.2 ml of 24 h broth culture of S. entertidis containing 1X 107 organisms while the negative-treated groups were inoculated with the same amount of the negative broth and second inoculation was done at 22 d of age. Colocal swabs were collected individually from all birds 2 h pre-inoculation to assure the absence of salmonella, then 1, 3, 5, 7, 21 days post-inoculation to recover salmonella. Performance parameter (body weight gain and feed efficiency) were calculated. Mortalities were recorded and reisolation of the salmonella was adopted to ensure it was the inoculated ones. The results revealed that the dietary acidification with sodium diformate significantly improved broilers performance and tends to produce heavier birds as compared to the negative control and CA groups. Moreover, the dietary inclusion of both acidifiers at level of 0.4% was able to eliminate mortalities completely at the relevant inoculation time. Regarding the shedding of S. enteritidius in positive groups, the SDF treatment resulted in significant (p<0.05) cessation of the shedding at 3 days post-inoculation compared to 7 days post-inoculation for the CA-group. In conclusion, sodium diformate at 0.4% dietary level in broiler diets has a valuable effect not only on broilers performance but also by eliminating S. enteritidis the main source of salmonella contamination in poultry farms which is feed.

Keywords: acidifier, broilers, Salmonalla spp, sodium diformate

Procedia PDF Downloads 272

Authors: Imen Soltani, Takoua Soltani, Taoufik Aguili

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Metamaterials crossover classic physical boundaries and gives rise to new phenomena and applications in the domain of beam steering and shaping. Where electromagnetic near and far field manipulations were achieved in an accurate manner. In this sense, 3D imaging is one of the beneficiaries and in particular Denis Gabor’s invention: holography. But, the major difficulty here is the lack of a suitable recording medium. So some enhancements were essential, where the 2D version of bulk metamaterials have been introduced the so-called metasurface. This new class of interfaces simplifies the problem of recording medium with the capability of tuning the phase, amplitude, and polarization at a given frequency. In order to achieve an intelligible wavefront control, the electromagnetic properties of the metasurface should be optimized by means of solving Maxwell’s equations. In this context, integral methods are emerging as an important method to study electromagnetic from microwave to optical frequencies. The method of moment presents an accurate solution to reduce the problem of dimensions by writing its boundary conditions in the form of integral equations. But solving this kind of equations tends to be more complicated and time-consuming as the structural complexity increases. Here, the use of equivalent circuit’s method exhibits the most scalable experience to develop an integral method formulation. In fact, for allaying the resolution of Maxwell’s equations, the method of Generalised Equivalent Circuit was proposed to convey the resolution from the domain of integral equations to the domain of equivalent circuits. In point of fact, this technique consists in creating an electric image of the studied structure using discontinuity plan paradigm and taken into account its environment. So that, the electromagnetic state of the discontinuity plan is described by generalised test functions which are modelled by virtual sources not storing energy. The environmental effects are included by the use of an impedance or admittance operator. Here, we propose a tunable metasurface composed of graphene-based elements which combine the advantages of reflectarrays concept and graphene as a pillar constituent element at Terahertz frequencies. The metasurface’s building block consists of a thin gold film, a dielectric spacer SiO₂ and graphene patch antenna. Our electromagnetic analysis is based on the method of moment combined with generalised equivalent circuit (MoM-GEC). We begin by restricting our attention to study the effects of varying graphene’s chemical potential on the unit cell input impedance. So, it was found that the variation of complex conductivity of graphene allows controlling the phase and amplitude of the reflection coefficient at each element of the array. From the results obtained here, we were able to determine that the phase modulation is realized by adjusting graphene’s complex conductivity. This modulation is a viable solution compared to tunning the phase by varying the antenna length because it offers a full 2π reflection phase control.

Keywords: graphene, method of moment combined with generalised equivalent circuit, reconfigurable metasurface, reflectarray, terahertz domain

Procedia PDF Downloads 166

Authors: Julia Kapr, Andrea Rossi, Haribaskar Ramachandran, Marius Pollet, Ilka Egger, Selina Dangeleit, Katharina Koch, Jean Krutmann, Ellen Fritsche

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It is widely acknowledged that animal models do not always represent human disease. Especially human brain development is difficult to model in animals due to a variety of structural and functional species-specificities. This causes significant discrepancies between predicted and apparent drug efficacies in clinical trials and their subsequent failure. Emerging alternatives based on 3D in vitro approaches, such as human brain spheres or organoids, may in the future reduce and ultimately replace animal models. Here, we present a human induced pluripotent stem cell (hiPSC)-based 3D neural in a vitro disease model for the Cockayne Syndrome B (CSB). CSB is a rare hereditary disease and is accompanied by severe neurologic defects, such as microcephaly, ataxia and intellectual disability, with currently no treatment options. Therefore, the aim of this study is to investigate the molecular and cellular defects found in neural hiPSC-derived CSB models. Understanding the underlying pathology of CSB enables the development of treatment options. The two CSB models used in this study comprise a patient-derived hiPSC line and its isogenic control as well as a CSB-deficient cell line based on a healthy hiPSC line (IMR90-4) background thereby excluding genetic background-related effects. Neurally induced and differentiated brain sphere cultures were characterized via RNA Sequencing, western blot (WB), immunocytochemistry (ICC) and multielectrode arrays (MEAs). CSB-deficiency leads to an altered gene expression of markers for autophagy, focal adhesion and neural network formation. Cell migration was significantly reduced and electrical activity was significantly increased in the disease cell lines. These data hint that the cellular pathologies is possibly underlying CSB. By induction of autophagy, the migration phenotype could be partially rescued, suggesting a crucial role of disturbed autophagy in defective neural migration of the disease lines. Altered autophagy may also lead to inefficient mitophagy. Accordingly, disease cell lines were shown to have a lower mitochondrial base activity and a higher susceptibility to mitochondrial stress induced by rotenone. Since mitochondria play an important role in neurotransmitter cycling, we suggest that defective mitochondria may lead to altered electrical activity in the disease cell lines. Failure to clear the defective mitochondria by mitophagy and thus missing initiation cues for new mitochondrial production could potentiate this problem. With our data, we aim at establishing a disease adverse outcome pathway (AOP), thereby adding to the in-depth understanding of this multi-faced disorder and subsequently contributing to alternative drug development.

Keywords: autophagy, disease modeling, in vitro, pluripotent stem cells

Procedia PDF Downloads 108

Authors: Nisha Sharma, Mili Kaur, Ashutosh Halder, Seema Kaushal, Manoj Kumar, Manish Jain

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Purpose: To investigate microRNAs (miRNA) as an epigenomic etiological factor in idiopathic non-obstructive azoospermia (NOA). In order to achieve the same, an association was seen between occupational exposure to radiation, thermal, and chemical factors and idiopathic cases of non-obstructive azoospermia, and later, testicular differential miRNA expression profiling was done in exposure group NOA cases. Method: It is a prospective study in which 200 apparent idiopathic male factor infertility cases, who have been advised to undergo testicular fine needle aspiration (FNA) evaluation, are recruited. A detailed occupational history was taken to understand the possible type of exposure due to the nature and duration of work. A total of 26 patients were excluded upon XY-FISH and Yq microdeletion tests due to the presence of genetic causes of infertility, 6 hypospermatogeneis (HS), six Sertoli cell-only syndrome (SCOS), and six normospermatogeneis patients testicular FNA samples were used for RNA isolation followed by small RNA sequencing and nCounter miRNA expression analysis. Differential miRNA expression profile of HS and SCOS patients was done. A web-based tool, miRNet, was used to predict the interacting compounds or chemicals using the shortlisted miRNAs with high fold change. The major limitation encountered in this study was the insufficient quantity of testicular FNA sample used for total RNA isolation, which resulted in a low yield and RNA integrity number (RIN) value. Therefore, the number of RNA samples admissible for differential miRNA expression analysis was very small in comparison to the total number of patients recruited. Results: Differential expression analysis revealed 69 down-regulated and 40 up-regulated miRNAs in HS and 66 down-regulated and 33 up-regulated miRNAs in SCOS in comparison to normospermatogenesis controls. The miRNA interaction analysis using the miRNet tool showed that the differential expression profiles of HS and SCOS patients were associated with arsenic trioxide, bisphenol-A, calcium sulphate, lithium, and cadmium. These compounds are reproductive toxins and might be responsible for miRNA-mediated epigenetic deregulation leading to NOA. The association between occupational risk factor exposure and the non-exposure group of NOA patients was not statistically significant, with ꭓ2 (3, N= 178) = 6.70, p= 0.082. The association between individual exposure groups (radiation, thermal, and chemical) and various sub-types of NOA is also not significant, with ꭓ2 (9, N= 178) = 15.06, p= 0.089. Functional analysis of HS and SCOS patients' miRNA profiles revealed some important miR-family members in terms of male fertility. The miR-181 family plays a role in the differentiation of spermatogonia and spermatocytes, as well as the transcriptional regulation of haploid germ cells. The miR-34 family is expressed in spermatocytes and round spermatids and is involved in the regulation of SSCs differentiation. Conclusion: The reproductive toxins might adopt the miRNA-mediated mechanism of disease development in idiopathic cases of NOA. Chemical compound induced; miRNA-mediated epigenetic deregulation can give a future perspective on the etiopathogenesis of the disease.

Keywords: microRNA, non-obstructive azoospermia (NOA), occupational exposure, hypospermatogenesis (HS), Sertoli cell only syndrome (SCOS)

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Authors: Alexis D'Agostino, Amanda Pomeroy

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Background: Resource availability is essential to effective implementation of national nutrition policies. To this end, the SPRING Project has collected and analyzed budget data from government ministries in Uganda, international donors, and other nutrition implementers to provide data for the first time on what funding is actually allocated to implement nutrition activities named in the national nutrition plan. Methodology: USAID’s SPRING Project used the Uganda Nutrition Action Plan (UNAP) as the starting point for budget analysis. Thorough desk reviews of public budgets from government, donors, and NGOs were mapped to activities named in the UNAP and validated by key informants (KIs) across the stakeholder groups. By relying on nationally-recognized and locally-created documents, SPRING provided a familiar basis for discussions to increase credibility and local ownership of findings. Among other things, the KIs validated the amount, source, and type (specific or sensitive) of funding. When only high-level budget data were available, KIs provided rough estimates of the percentage of allocations that were actually nutrition-relevant, allowing creation of confidence intervals around some funding estimates. Results: After validating data and narrowing in on estimates of funding to nutrition-relevant programming, researchers applied a formula to estimate overall nutrition allocations. In line with guidance by the SUN Movement and its three-step process, nutrition-specific funding was counted at 100% of its allocation amount, while nutrition sensitive funding was counted at 25%. The vast majority of nutrition funding in Uganda is off-budget, with over 90 percent of all nutrition funding is provided outside of the government system. Overall allocations are split nearly evenly between nutrition-specific and –sensitive activities. In FY 2013/14, the two-year study’s baseline year, on- and off-budget funding for nutrition was estimated to be around 60 million USD. While the 60 million USD allocations compare favorably to the 66 million USD estimate of the cost of the UNAP, not all activities are sufficiently funded. Those activities with a focus on behavior change were the most underfunded. In addition, accompanying qualitative research suggested that donor funding for nutrition activities may shift government funding into other areas of work, making it difficult to estimate the sustainability of current nutrition investments.Conclusions: Beyond providing figures, these estimates can be used together with the qualitative results of the study to explain how and why these amounts were allocated for particular activities and not others, examine the negotiation process that occurred, and suggest options for improving the flow of finances to UNAP activities for the remainder of the policy tenure. By the end of the PBN study, several years of nutrition budget estimates will be available to compare changes in funding over time. Halfway through SPRING’s work, there is evidence that country stakeholders have begun to feel ownership over the ultimate findings and some ministries are requesting increased technical assistance in nutrition budgeting. Ultimately, these data can be used within organization to advocate for more and improved nutrition funding and to improve targeting of nutrition allocations.

Keywords: budget, nutrition, financing, scale-up

Procedia PDF Downloads 423

Authors: Javier E. García-Gallo, Nelson J. Fonseca-Ruiz, John F. Duitama-Munoz

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Sepsis is a syndrome that occurs with physiological and biochemical abnormalities induced by severe infection and carries a high mortality and morbidity, therefore the severity of its condition must be interpreted quickly. After patient admission in an intensive care unit (ICU), it is necessary to synthesize the large volume of information that is collected from patients in a value that represents the severity of their condition. Traditional severity of illness scores seeks to be applicable to all patient populations, and usually assess in-hospital mortality. However, the use of machine learning techniques and the data of a population that shares a common characteristic could lead to the development of customized mortality prediction scores with better performance. This study presents the development of a score for the one-year mortality prediction of the patients that are admitted to an ICU with a sepsis diagnosis. 5650 ICU admissions extracted from the MIMICIII database were evaluated, divided into two groups: 70% to develop the score and 30% to validate it. Comorbidities, demographics and clinical information of the first 24 hours after the ICU admission were used to develop a mortality prediction score. LASSO (least absolute shrinkage and selection operator) and SGB (Stochastic Gradient Boosting) variable importance methodologies were used to select the set of variables that make up the developed score; each of this variables was dichotomized and a cut-off point that divides the population into two groups with different mean mortalities was found; if the patient is in the group that presents a higher mortality a one is assigned to the particular variable, otherwise a zero is assigned. These binary variables are used in a logistic regression (LR) model, and its coefficients were rounded to the nearest integer. The resulting integers are the point values that make up the score when multiplied with each binary variables and summed. The one-year mortality probability was estimated using the score as the only variable in a LR model. Predictive power of the score, was evaluated using the 1695 admissions of the validation subset obtaining an area under the receiver operating characteristic curve of 0.7528, which outperforms the results obtained with Sequential Organ Failure Assessment (SOFA), Oxford Acute Severity of Illness Score (OASIS) and Simplified Acute Physiology Score II (SAPSII) scores on the same validation subset. Observed and predicted mortality rates within estimated probabilities deciles were compared graphically and found to be similar, indicating that the risk estimate obtained with the score is close to the observed mortality, it is also observed that the number of events (deaths) is indeed increasing as the outcome go from the decile with the lowest probabilities to the decile with the highest probabilities. Sepsis is a syndrome that carries a high mortality, 43.3% for the patients included in this study; therefore, tools that help clinicians to quickly and accurately predict a worse prognosis are needed. This work demonstrates the importance of customization of mortality prediction scores since the developed score provides better performance than traditional scoring systems.

Keywords: intensive care, logistic regression model, mortality prediction, sepsis, severity of illness, stochastic gradient boosting

Procedia PDF Downloads 201

Authors: Paul Moses Ndegwa, Teresia Kabucho, Lucy Wanjiru, Esther Wanjiru, Brian Githaiga, Jecinta Wambui

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Health outcome has improved in the country since 2013 following the adoption of the new constitution in Kenya with devolved governance with administration and health planning functions transferred to county governments. 2018-2022 development agenda prioritized universal healthcare coverage, food security, and nutrition, however, the emergence of Covid-19 and the increase of non-communicable diseases pose a challenge and constrain in an already overwhelmed health system. A study was conducted July-November 2021 to establish key challenges in achieving universal healthcare coverage within the county and best practices for improved non-communicable disease control. 14 health workers ranging from nurses, doctors, public health officers, clinical officers, and pharmaceutical technologists were purposely engaged to provide critical information through questionnaires by a trained duo observing ethical procedures on confidentiality. Data analysis. Communicable diseases are major causes of morbidity and mortality. Non-communicable diseases contribute to approximately 39% of deaths. More than 45% of the population does not have access to safe drinking water. Study noted geographic inequality with respect to distribution and use of health resources including competing non-health priorities. 56% of health workers are nurses, 13% clinical officers, 7% doctors, 9%public health workers, 2% are pharmaceutical technologists. Poor-quality data limits the validity of disease-burdened estimates and research activities. Risk factors include unsafe water, sanitation, hand washing, unsafe sex, and malnutrition. Key challenge in achieving universal healthcare coverage is the rise in the relative contribution of non-communicable diseases. Improve targeted disease control with effective and equitable resource allocation. Develop high infectious disease control mechanisms. Improvement of quality data for decision making. Strengthen electronic data-capture systems. Increase investments in the health workforce to improve health service provision and achievement of universal health coverage. Create a favorable environment to retain health workers. Fill in staffing gaps resulting in shortages of doctors (7%). Develop a multi-sectional approach to health workforce planning and management. Need to invest in mechanisms that generate contextual evidence on current and future health workforce needs. Ensure retention of qualified, skilled, and motivated health workforce. Deliver integrated people-centered health services.

Keywords: multi-sectional approach, equity, people-centered, health workforce retention

Procedia PDF Downloads 87

Authors: Harpal Singh, Sakshi Batra

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The widespread and easy accesses to multimedia contents and possibility to make numerous copies without loss of significant fidelity have roused the requirement of digital rights management. Thus this problem can be effectively solved by Digital watermarking technology. This is a concept of embedding some sort of data or special pattern (watermark) in the multimedia content; this information will later prove ownership in case of a dispute, trace the marked document’s dissemination, identify a misappropriating person or simply inform user about the rights-holder. The primary motive of digital watermarking is to embed the data imperceptibly and robustly in the host information. Extensive counts of watermarking techniques have been developed to embed copyright marks or data in digital images, video, audio and other multimedia objects. With the development of digital video-based innovations, copyright dilemma for the multimedia industry increases. Video watermarking had been proposed in recent years to serve the issue of illicit copying and allocation of videos. It is the process of embedding copyright information in video bit streams. Practically video watermarking schemes have to address some serious challenges as compared to image watermarking schemes like real-time requirements in the video broadcasting, large volume of inherently redundant data between frames, the unbalance between the motion and motionless regions etc. and they are particularly vulnerable to attacks, for example, frame swapping, statistical analysis, rotation, noise, median and crop attacks. In this paper, an effective, robust and imperceptible video watermarking algorithm is proposed based on hybridization of powerful mathematical transforms; Fractional Fourier Transform (FrFT), Discrete Wavelet transforms (DWT) and Singular Value Decomposition (SVD) using redundant wavelet. This scheme utilizes various transforms for embedding watermarks on different layers by using Hybrid systems. For this purpose, the video frames are portioned into layers (RGB) and the watermark is being embedded in two forms in the video frames using SVD portioning of the watermark, and DWT sub-band decomposition of host video, to facilitate copyright safeguard as well as reliability. The FrFT orders are used as the encryption key that allows the watermarking method to be more robust against various attacks. The fidelity of the scheme is enhanced by introducing key generation and wavelet based key embedding watermarking scheme. Thus, for watermark embedding and extraction, same key is required. Therefore the key must be shared between the owner and the verifier via some safe network. This paper demonstrates the performance by considering different qualitative metrics namely Peak Signal to Noise ratio, Structure similarity index and correlation values and also apply some attacks to prove the robustness. The Experimental results are presented to demonstrate that the proposed scheme can withstand a variety of video processing attacks as well as imperceptibility.

Keywords: discrete wavelet transform, robustness, video watermarking, watermark

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Authors: Kyoungrean Kim

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Importance of deep-sea mineral resources is dramatically increasing due to the depletion of land mineral resources corresponding to increasing human’s economic activities. Korea has acquired exclusive exploration licenses at four areas which are the Clarion-Clipperton Fracture Zone in the Pacific Ocean (2002), Tonga (2008), Fiji (2011) and Indian Ocean (2014). The preparation for commercial mining of Nautilus minerals (Canada) and Lockheed martin minerals (USA) is expected by 2020. The London Protocol 1996 (LP) under International Maritime Organization (IMO) and International Seabed Authority (ISA) will set environmental guidelines for deep-sea mining until 2020, to protect marine environment. In this research, the applicability of washing/extraction treatment for the remediation of deep-sea mining tailings was mainly evaluated in order to present preliminary data to develop practical remediation technology in near future. Polymetallic nodule samples were collected at the Clarion-Clipperton Fracture Zone in the Pacific Ocean, then stored at room temperature. Samples were pulverized by using jaw crusher and ball mill then, classified into 3 particle sizes (> 63 µm, 63-20 µm, < 20 µm) by using vibratory sieve shakers (Analysette 3 Pro, Fritsch, Germany) with 63 µm and 20 µm sieve. Only the particle size 63-20 µm was used as the samples for investigation considering the lower limit of ore dressing process which is tens to 100 µm. Rhamnolipid and sodium alginate as biosurfactant and aluminum sulfate which are mainly used as flocculant were used as environmentally friendly additives. Samples were adjusted to 2% liquid with deionized water then mixed with various concentrations of additives. The mixture was stirred with a magnetic bar during specific reaction times and then the liquid phase was separated by a centrifugal separator (Thermo Fisher Scientific, USA) under 4,000 rpm for 1 h. The separated liquid was filtered with a syringe and acrylic-based filter (0.45 µm). The extracted heavy metals in the filtered liquid were then determined using a UV-Vis spectrometer (DR-5000, Hach, USA) and a heat block (DBR 200, Hach, USA) followed by US EPA methods (8506, 8009, 10217 and 10220). Polymetallic nodule was mainly composed of manganese (27%), iron (8%), nickel (1.4%), cupper (1.3 %), cobalt (1.3%) and molybdenum (0.04%). Based on remediation standards of various countries, Nickel (Ni), Copper (Cu), Cadmium (Cd) and Zinc (Zn) were selected as primary target materials. Throughout this research, the use of rhamnolipid was shown to be an effective approach for removing heavy metals in samples originated from manganese nodules. Sodium alginate might also be one of the effective additives for the remediation of deep-sea mining tailings such as polymetallic nodules. Compare to the use of rhamnolipid and sodium alginate, aluminum sulfate was more effective additive at short reaction time within 4 h. Based on these results, sequencing particle separation, selective extraction/washing, advanced filtration of liquid phase, water treatment without dewatering and solidification/stabilization may be considered as candidate technologies for the remediation of deep-sea mining tailings.

Keywords: deep-sea mining tailings, heavy metals, remediation, extraction, additives

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Authors: Rumbidzai Sipiwe Zimvumi

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The study aimed to identify the top management team attributes for ambidexterity in small and medium-sized enterprises by utilizing the upper echelons theory. The conventional opinion holds that an organization's ability to pursue both exploitative and explorative innovation methods at the same time is reflected in its ambidexterity. Top-level managers are critical to this matrix because they forecast and explain strategic choices that guarantee success by improving organizational performance. Since the focus of the study was on the unique characteristics of TMTs that can facilitate ambidexterity, the primary goal was to comprehend how TMTs in SMEs can better manage ambidexterity. The study used document analysis to collect information on ambidexterity and TMT traits. Finding, choosing, assessing, and synthesizing data from peer-reviewed publications allowed for the review and evaluation of papers. The fact that SMEs will perform better if they can achieve a balance between exploration and exploitation cannot be overstated. Unfortunately, exploitation is the main priority for most SMEs. The results showed that some of the noteworthy TMT traits that support ambidexterity in SMEs are age diversity, shared responsibility, leadership impact, psychological safety, and self-confidence. It has been shown that most SMEs confront significant obstacles in recruiting people, including formalizing their management and assembling executive teams with seniority. Small and medium-sized enterprises (SMEs) are often held by families or people who neglect to keep their personal lives apart from the firm, which eliminates the opportunity for management and staff to take the initiative. This helps to explain why exploitative strategies, which preserve present success, are used rather than explorative strategies, which open new economic opportunities and dimensions. It is evident that psychological safety deteriorates, and creativity is hindered in the process. The study makes the case that TMTs who are motivated to become ambidextrous can exist. According to the report, small- and medium-sized business owners should value the opinions of all parties involved and provide their managers and regular staff the freedom to think creatively and in a safe environment. TMTs who experience psychological safety are more likely to be inventive, creative, and productive. A team's collective perception that it is acceptable to take chances, voice opinions and concerns, ask questions, and own up to mistakes without fear of unfavorable outcomes is known as team psychological safety. Thus, traits like age diversity, leadership influence, learning agility, psychological safety, and self-assurance are critical to the success of SMEs. As a solution to ensuring ambidexterity is attained, the study suggests a clear separation of ownership and control, the adoption of technology to stimulate creativity, team spirit and excitement, shared accountability, and good management of diversity. Among the suggestions for the SME's success are resource allocation and important collaborations.

Keywords: navigating, ambidexterity, top management team, small and medium enterprises

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Authors: Ch. Sridevi, S. P. Shailender Kumar, B. Gurudayal, A. Chalapathi Rao, K. Koteswara Rao, P. Srinivasulu

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System reliability and system availability evaluation plays a crucial role in ensuring the seamless operation of complex satellite data reception system with consistent performance for longer periods. This paper presents a novel approach for the same using a case study on one of the antenna systems at satellite data reception ground station in India. The methodology involves analyzing system's components, their failure rates, system's architecture, generation of logical reliability block diagram model and estimating the reliability of the system using the component level mean time between failures considering exponential distribution to derive a baseline estimate of the system's reliability. The model is then validated with collected system level field failure data from the operational satellite data reception systems that includes failure occurred, failure time, criticality of the failure and repair times by using statistical techniques like median rank, regression and Weibull analysis to extract meaningful insights regarding failure patterns and practical reliability of the system and to assess the accuracy of the developed reliability model. The study mainly focused on identification of critical units within the system, which are prone to failures and have a significant impact on overall performance and brought out a reliability model of the identified critical unit. This model takes into account the interdependencies among system components and their impact on overall system reliability and provides valuable insights into the performance of the system to understand the Improvement or degradation of the system over a period of time and will be the vital input to arrive at the optimized design for future development. It also provides a plug and play framework to understand the effect on performance of the system in case of any up gradations or new designs of the unit. It helps in effective planning and formulating contingency plans to address potential system failures, ensuring the continuity of operations. Furthermore, to instill confidence in system users, the duration for which the system can operate continuously with the desired level of 3 sigma reliability was estimated that turned out to be a vital input to maintenance plan. System availability and station availability was also assessed by considering scenarios of clash and non-clash to determine the overall system performance and potential bottlenecks. Overall, this paper establishes a comprehensive methodology for reliability and availability analysis of complex satellite data reception systems. The results derived from this approach facilitate effective planning contingency measures, and provide users with confidence in system performance and enables decision-makers to make informed choices about system maintenance, upgrades and replacements. It also aids in identifying critical units and assessing system availability in various scenarios and helps in minimizing downtime and optimizing resource allocation.

Keywords: exponential distribution, reliability modeling, reliability block diagram, satellite data reception system, system availability, weibull analysis

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Authors: Sachin Mulmi Shrestha, Xin Fang, Hui Ye, Lihua Ren, Qinghua Ji, Ruihua Shi

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Background: Esophageal squamous cell carcinoma (ESCC) is a tumor arising from esophageal epithelial cells and is one of the major disease subtype in Asian countries, including China. Esophageal cancer is the 7th highest incidence based on the 2020 data of GLOBOCAN. The pathogenesis of cancer is still not well understood as many molecular and genetic basis of esophageal carcinogenesis has yet to be clearly elucidated. Circular RNAs are RNA molecules that are formed by back-splicing covalently joined 3′- and 5′-endsrather than canonical splicing, and recent data suggest circular RNAs could sponge miRNAs and are enriched with functional miRNA binding sites. Hence, we studied the mechanism of circular RNA, its biological function, and the relationship between microRNA in the carcinogenesis of ESCC. Methods: 4 pairs of normal and esophageal cancer tissues were collected in Zhongda hospital, affiliated to Southeast University, and high-throughput RNA sequencing was done. The result revealed that circ_0032746 was upregulated, and thus we selected circ_0032746 for further study. The backsplice junction of circRNA was validated by sanger sequence, and stability was determined by RNASE R assay. The binding site of circRNA and microRNA was predicted by circinteractome,mirandaand RNAhybrid database. Furthermore, circRNA was silenced by siRNA and then by lentivirus. The regulatory axis of circ0032746/miR4270 was validated by shRNA, mimic, and inhibitor transfection. Then, in vitro experiments were performed to assess the role of circ0032746 on proliferation (CCK-8 assay and colon formation assay), migration and invasion (Transewell assay), and apoptosis of ESCC. Results: The upregulated circ0032746 was validated in 9 pairs of tissues and 5 types of cell lines by qPCR, which showed high expression and was statistically significant (P<0.005) ). Upregulated circ0032746 was silenced by shRNA, which showed significant knockdown in KYSE 30 and TE-1 cell lines expression compared to control. Nuclear and cytoplasmic mRNA fraction experiment displayed the cytoplasmic location of circ0032746. The sponging of miR4270 was validated by co-transfection of sh-circ0032746 and mimic or inhibitor. Transfection with mimic showed the decreased expression of circ_0032746, whereas inhibitor inhibited the result. In vitro experiments showed that silencing of circ_0032746 inhibited the proliferation, migration, and invasion compared to the negative control group. The apoptosis was seen higher in a knockdown group than in the control group. Furthermore, 11 common mircoRNA target mRNAs were predicted by Targetscan, MirTarbase, and miRanda database, which may further play role in the pathogenesis. Conclusion: Our results showed that novel circ_0032746 is upregulated in ESCC and plays role in itsoncogenicity. Silencing of circ_0032746 inhibits the proliferation and migration of ESCC whereas increases the apoptosis of cancer cells. Hence, circ0032746 acts as an oncogene in ESCC by sponging with miR4270 and could be a potential biomarker in the diagnosis of ESCC in the future.

Keywords: circRNA, esophageal squamous cell carcinoma, microRNA, upregulated

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Authors: Hale Kirer Silva Lecuna, Beyza Aydin

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Health is one of the main components of human capital and sustainable development, and it is very important for economic growth. Health economics, which is an indisputable part of the science of economics, has five stages in general. These are health and development, financing of health services, economic regulation in the health, allocation of resources and efficiency of health services. A well-developed and efficient health sector plays a major role by increasing the level of development of countries. The most crucial pillars of the health sector are the hospitals that are divided into public and private. The main purpose of the hospitals is to provide more efficient services. Therefore the aim is to meet patients’ satisfaction by increasing the service quality. Health-related studies in Turkey date back to the Ottoman and Seljuk Empires. In the near past, Turkey applied 'Health Sector Transformation Programs' under different titles between 2003 and 2010. Our aim in this paper is to measure how effective these transformation programs are for the health sector, to see how much they can increase the efficiency of hospitals over the years, to see the return of investments, to make comments and suggestions on the results, and to provide a new reference for the literature. Within this framework, the public and private hospitals in Balıkesir, Bilecik, Bursa, Çanakkale, Edirne, Istanbul, Kirklareli, Kocaeli, Sakarya, Tekirdağ, Yalova will be examined by using Data Envelopment Analysis (DEA) for the years between 2000 and 2019. DEA is a linear programming-based technique, which gives relatively good results in multivariate studies. DEA basically estimates an efficiency frontier and make a comparison. Constant returns to scale and variable returns to scale are two most commonly used DEA methods. Both models are divided into two as input and output-oriented. To analyze the data, the number of personnel, number of specialist physicians, number of practitioners, number of beds, number of examinations will be used as input variables; and the number of surgeries, in-patient ratio, and crude mortality rate as output variables. 11 hospitals belonging to the Marmara region were included in the study. It is seen that these hospitals worked effectively only in 7 provinces (Balıkesir, Bilecik, Bursa, Edirne, İstanbul, Kırklareli, Yalova) for the year 2001 when no transformation program was implemented. After the transformation program was implemented, for example, in 2014 and 2016, 10 hospitals (Balıkesir, Bilecik, Bursa, Çanakkale, Edirne, İstanbul, Kocaeli, Kırklareli, Tekirdağ, Yalova) were found to be effective. In 2015, ineffective results were observed for Sakarya, Tekirdağ and Yalova. However, since these values are closer to 1 after the transformation program, we can say that the transformation program has positive effects. For Sakarya alone, no effective results have been achieved in any year. When we look at the results in general, it shows that the transformation program has a positive effect on the effectiveness of hospitals.

Keywords: data envelopment analysis, efficiency, health sector, Marmara region

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Authors: Meiling Yu, Nadia Rouatbi, Khuloud T. Al-Jamal

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Background: Treatment of neurological disorders with modern medical and surgical approaches remains difficult. Gene therapy, allowing the delivery of genetic materials that encodes potential therapeutic molecules, represents an attractive option. The treatment of brain diseases with gene therapy requires the gene-editing tool to be delivered efficiently to the central nervous system. In this study, we explored the efficiency of different delivery routes, namely intravenous (i.v.), intra-cranial (i.c.), and intra-nasal (i.n.), to deliver stable nucleic acid-lipid particles (SNALPs) containing gene-editing tools namely Cas9 mRNA and sgRNA encoding for GFP as a reporter protein. We hypothesise that SNALPs can reach the brain and perform gene-editing to different extents depending on the administration route. Intranasal administration (i.n.) offers an attractive and non-invasive way to access the brain circumventing the blood–brain barrier. Successful delivery of gene-editing tools to the brain offers a great opportunity for therapeutic target validation and nucleic acids therapeutics delivery to improve treatment options for a range of neurodegenerative diseases. In this study, we utilised Rosa26-Cas9 knock-in mice, expressing GFP, to study brain distribution and gene-editing efficiency of SNALPs after i.v.; i.c. and i.n. routes of administration. Methods: Single guide RNA (sgRNA) against GFP has been designed and validated by in vitro nuclease assay. SNALPs were formulated and characterised using dynamic light scattering. The encapsulation efficiency of nucleic acids (NA) was measured by RiboGreen™ assay. SNALPs were incubated in serum to assess their ability to protect NA from degradation. Rosa26-Cas9 knock-in mice were i.v., i.n., or i.c. administered with SNALPs to test in vivo gene-editing (GFP knockout) efficiency. SNALPs were given as three doses of 0.64 mg/kg sgGFP following i.v. and i.n. or a single dose of 0.25 mg/kg sgGFP following i.c.. knockout efficiency was assessed after seven days using Sanger Sequencing and Inference of CRISPR Edits (ICE) analysis. In vivo, the biodistribution of DiR labelled SNALPs (SNALPs-DiR) was assessed at 24h post-administration using IVIS Lumina Series III. Results: Serum-stable SNALPs produced were 130-140 nm in diameter with ~90% nucleic acid loading efficiency. SNALPs could reach and stay in the brain for up to 24h following i.v.; i.n. and i.c. administration. Decreasing GFP expression (around 50% after i.v. and i.c. and 20% following i.n.) was confirmed by optical imaging. Despite the small number of mice used, ICE analysis confirmed GFP knockout in mice brains. Additional studies are currently taking place to increase mice numbers. Conclusion: Results confirmed efficient gene knockout achieved by SNALPs in Rosa26-Cas9 knock-in mice expressing GFP following different routes of administrations in the following order i.v.= i.c.> i.n. Each of the administration routes has its pros and cons. The next stages of the project involve assessing gene-editing efficiency in wild-type mice and replacing GFP as a model target with therapeutic target genes implicated in Motor Neuron Disease pathology.

Keywords: CRISPR, nanoparticles, brain diseases, administration routes

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