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Introduction: Reflective thinking offers learners insight and encourages deeper understanding by leveraging past experiences. This study explores the impact of reflective writing, a self-assessment tool, on undergraduate medical students. The focus is on training students using author-specific reflection rubrics based on Moon's model. Methods: A mixed-methods study involving 32 volunteered students undertaking an interactive 3-hour session on reflective thinking and writing (RT&W). 19 students submitted reflections, which were self-graded by students and two faculties independently. The perceptions of students were gathered through questionnaires and focus group discussions. The analysis was done using the mean, inter-class correlational coefficient, and thematic analysis. Results: Inter-rater reliability and inter-class correlation coefficient for reflective writing rubric scores was 63.2%, i.e. below the acceptable threshold. Cronbach's Alpha for the learner perception questionnaire was 0.90. The outcome of the student's perception questionnaire recognized the value of reflective writing in terms of professional skills enhancement (4.83±0.39) and improvement after feedback (4.17±0.72). However, satisfaction with overall training was comparatively lower (2.5±0.52). Focus group discussions revealed six themes. Conclusion: Reflective writing enhances the learning outcomes, deepens understanding, and refines judgment. The author-specific reflection rubric, though reliable, warrants empirical validation with a larger and more diverse participant pool. Undergraduate programs should prioritize mastery of reflection and meta-cognitive learning approaches to optimize educational outcomes.
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Ricinoleic acid (RA) from castor oil was employed in biotransformation of peach-flavoured γ-decalactone (GDL), using a Candida parapsilosis strain (MTCC13027) which was isolated from waste of pineapple crown base. Using four variables-pH, cell density, amount of RA, and temperature-the biotransformation parameters were optimized using RSM and BBD. Under optimized conditions (pH 6, 10â¯% of microbial cells, 10â¯g/L RA at 28°C), the conversion was maximum and resulted to 80â¯% (+)-GDL (4.4â¯g/L/120â¯h) yield in shake flask (500â¯mL). Furthermore, optimization was achieved by adjusting the aeration and agitation parameters in a 3â¯L bioreactor, which were then replicated in a 10â¯L bioreactor to accurately determine the amount of (+)-GDL. In bioreactor condition, 4.7â¯g/L (>85â¯%) of (+)-GDL is produced with 20â¯% and 40â¯% dissolved oxygen (1.0 vvm) at 150â¯rpm in 72â¯h and 66â¯h, respectively. Further, a new Al-Mg-Ca-Si composite column-chromatography method is developed to purify enantiospecific (+)-GDL (99.9â¯%). This (+)-GDL is 100â¯% nature-identical as validated through 14C-radio-carbon dating. Thorough chemical investigation of enantiospecific (+)-GDL is authenticated for its use as flavour. This bioflavour has been developed through a cost-effective biotechnological process in response to the demand from the food industry on commercial scale.
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Reatores Biológicos , Candida parapsilosis , Óleo de Rícino , Lactonas , Ácidos Ricinoleicos , Ácidos Ricinoleicos/metabolismo , Ácidos Ricinoleicos/química , Reatores Biológicos/microbiologia , Óleo de Rícino/química , Óleo de Rícino/metabolismo , Candida parapsilosis/metabolismo , Lactonas/metabolismo , Lactonas/química , Aromatizantes/metabolismo , Aromatizantes/química , BiotransformaçãoRESUMO
Background: Multimorbidity is a group of conditions, it has significant impact on the population as a whole, resulting in lower quality of life, higher mortality, frequent use of medical services, and consequently higher healthcare costs. The objective of this study is to document the prevalence of common multimorbidity and its associated risk factors among population of Mechinagar Municipality. Methods: Community-based cross-sectional study was conducted where selected multimorbidity were assessed in selected areas of Mechinagar municipality of Jhapa District . Systematic random sampling technique was used to select 590 adult participants from three pre-defined pocket areas. Pre-designed semi-structured multimorbidity assessment questionnaire for primary care (MAQ-PC)was used to assess prevalence of multimorbidity. Multiple logistic regression was conducted to identify the strongest determinants of multimorbidity. Results: The prevalence of multimorbidity was 22.4%.Hypertension, Diabetes mellitus and COPD was seen in 39.2%, 7.8.% and 4.4% of the participants respectively . Participants with advancing age i. e. 40-49yrs were 12.62 times (AOR) more likely to have multimorbidity compared to their counterparts who were 20-29yrs old( p=<0.01,CI3.01-15.28) after adjusting for occupation, physical activity and family history of kidney disease. Working individuals, Physical inactivity and positive family history of kidney disease were the strongest determinates of multimorbidity. Conclusions: The study revealed that participants with increasing age, working individuals, physical inactivity and family history of kidney disease were more vulnerable of having multimorbidity. The findings of our study indicate need of intervention strategies and community-based health promotion programs in reducing burden of chronic disease among adult population.
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Although the functional role of circular RNA (circRNA) interaction with microRNAs and proteins has been studied extensively, circRNA interactions with the protein-coding mRNAs in intact cells remain largely unknown. Here, by employing AMT-mediated proximity ligation of RNA-RNA duplexes followed by circRNA enrichment and deep sequencing, we report a novel Cross-Linking Poly(A) Pulldown RNase R Sequencing (CLiPPR-seq) technology which identified hundreds of mRNA-interacting circRNAs in three different cell types, including ßTC6, C2C12 and HeLa cells. Furthermore, CLiPP-seq without RNase R treatment was also performed to identify the mRNA expression in these cells. BLAST analysis of circRNAs in CLiPPR-seq sample with the mRNAs in CLiPP-seq samples determined their potential complementary sequences for circRNA-mRNA interaction. Pulldown of circRNAs and poly(A) RNAs confirmed the direct interaction of circRNAs with target mRNAs. Silencing of mRNA-interacting circRNAs led to the altered expression of target mRNAs in ßTC6 cells, suggesting the role of direct interaction of circRNAs with mRNAs in gene expression regulation. CLiPPR-seq thus represents a novel method for illuminating the myriad of uncharacterized circRNA-mRNA hybrids that may regulate gene expression.
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Técnicas Genéticas , MicroRNAs , RNA Circular , Análise de Sequência de RNA , Humanos , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica , Células HeLa , MicroRNAs/genética , MicroRNAs/metabolismo , RNA Circular/genética , RNA Circular/isolamento & purificação , RNA Circular/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Análise de Sequência de RNA/métodosRESUMO
The intriguing network of antibody-antigen (Ab-Ag) interactions is highly governed by environmental perturbations and the nature of biomolecular interaction. Protein-protein interactions (PPIs) have potential applications in developing protein-adsorption-based sensors and nano-scale materials. Therefore, characterizing PPIs in the presence of a nanomaterial at the molecular level becomes imperative. The present work involves the investigation of antiferritin-ferritin (Ab-Ag) protein interactions under the influence of tungsten disulfide quantum dots (WS2 QDs). Isothermal calorimetry and contact angle measurements validated the strong influence of WS2 QDs on Ab-Ag interactions. The interfacial signatures of nano-bio-interactions were evaluated using sum frequency generation vibration spectroscopy (SFG-VS) at the air-water interface. Our SFG results reveal a variation in the tilt angle of methyl groups by â¼12° ± 2° for the Ab-Ag system in the presence of WS2 QDs. The results illustrated an enhanced ordering of water molecules in the presence of QDs, which underpins the active role of interfacial water molecules during nano-bio-interactions. We have also witnessed a differential impact of QDs on Ab-Ag by raising the concentration of the Ab-Ag combination, which showcased an increased inter-molecular interaction among the Ab and Ag molecules and a minimal influence on the methyl tilt angle. These findings suggest the formation of stronger and ordered Ab-Ag complexes upon introducing WS2 QDs in the aqueous medium and signify the potentiality of WS2 QDs relevant to protein-based sensing assays.
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Pontos Quânticos , Compostos de Tungstênio , Pontos Quânticos/química , Água/química , Sulfetos/químicaRESUMO
The pre-existing theory of pre-mRNA splicing into linear mature RNA was questioned with the introduction of circular RNAs (circRNAs). Hundreds of studies using high throughput RNA-sequencing (RNA-seq) techniques and novel computational programs reported the abundant and ubiquitous expression of circRNAs originating by pre-mRNA backsplicing. CircRNAs are mostly involved in gene expression by regulating functions of interacting microRNAs (miRNAs) and RNA-binding proteins (RBPs) or translating into functional polypeptides. Although all circRNA annotation tools identify circRNAs based on the backsplice junction (BSJ) sequences, only a few identify the internal sequences of circRNAs. However, the full-length sequence of circRNAs from RNA-seq data could be error-prone due to its similarity with the counterpart linear RNA. Since circRNA function depends on the mature sequence, validation of the mature sequence is the prerequisite for their further characterization. In this chapter, we discuss the validation of circRNA BSJ sequence by RT-PCR using divergent primer followed by Sanger sequencing. Furthermore, we describe the circRNA-rolling circle amplification (circRNA-RCA; circRNA enrichment by RNase R treatment, full-length cDNA synthesis, rolling circle PCR amplification using full-length primers, and Sanger sequencing of the PCR product) to validate the mature splice sequence of circRNAs. This chapter highlights the basic guidelines for designing divergent and full-length primers for PCR amplification and Sanger sequencing to validate circRNA sequences.
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Circular (circ)RNAs have emerged as novel regulators of gene expression through various mechanisms. However, most publications focus on functional circRNAs regulating target gene expression by interacting with micro (mi)RNAs and acting as competing endogenous RNAs (ceRNAs). Although the theory of miRNA sponging by ceRNAs suggests the inhibition of miRNA activity, many studies are biased toward the selection of miRNAs showing a reverse expression pattern compared with circRNA expression. Although several computational tools and molecular assays have been used to predict and validate the interaction of miRNAs with circRNAs, the actual validation of functional in vivo interactions needs careful consideration of molecular experiments with specific controls. As extensive research is being performed on circRNA, many questions arise on the functional significance of circRNA-miRNA interactions. We hope the critical discussion on the criteria for selecting circRNA-miRNA pairs for functional analysis and providing standard methods for validating circRNA-miRNA interactions will advance our understanding of circRNAs as novel gene regulators. This article is categorized under: Regulatory RNAs/RNAi/Riboswitches > Regulatory RNAs Translation > Regulation RNA Methods > RNA Analyses in Cells.
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Flavour molecules are generated now-a-days through microbial fermentation on a commercial scale. γ-Decalactone (GDL) is an important molecule due to its long-lasting flavouring impact as buttery, coconut and peach-type. In the current study, 33 microorganisms were isolated from different fruit sources, and their screening for target GDL production was performed. Using DNA sequencing, two potential strains yielding good amounts of GDL were identified from pineapple and strawberry fruits. The identified strains were Metschnikowia vanudenii (OP954735) and Candida parapsilosis (OP954733), and further optimized by Taguchi method. The effectiveness of lactone production is influenced by the rate of microbial growth under various operating conditions. The factors such as substrate concentration, pH, temperature, cell density and rotation (rpm) with 3 levels were applied for the GDL production using M. vanudenii (OP954735) and C. parapsilosis (OP954733) strains. The results revealed that the highest molar conversion of GDL was 24.69% (115.7â¯mg/g quantitative yield) and 52.69% (272.0â¯mg/g quantitative yield) at the optimal conditions using SB-62 and PA-19 strains, respectively. The two novel strains are reported for the first time for production of γ-decalactone and overall, this study opens up the possibility of using Taguchi design for large scale up process development for producing food flavours utilising environmentally friendly natural strains.
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Lactonas , Leveduras , Leveduras/genética , Leveduras/metabolismo , Lactonas/química , BiotransformaçãoRESUMO
Salinity is a significant concern in crop production, causing severe losses in agricultural yields. Ocimum sanctum, also known as Holy Basil, is an important ancient medicinal plant used in the Indian traditional system of medicine. The present study explores the use of 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase-producing strains of plant-growth-promoting bacteria (PGPB) namely Str-8 (Halomonas desiderata), Sd-6 (Brevibacterium halotolerans), Fd-2 (Achromobacter xylosoxidans), Art-7 (Burkholderia cepacia), and Ldr-2 (Bacillus subtilis), and T. harzianum (Th), possessing multi-functional properties like growth promotion, stress alleviation, and for enhancing O. sanctum yield under salt stress. The results showed that co-inoculation of Th and PGPBs enhanced plant height and fresh herb weight by 3.78-17.65% and 7.86-58.76%, respectively; highest being in Th + Fd-2 and Th + Art-7 compared to positive control plants. The doubly inoculated plants showed increased pigments, phenol, flavonoids, protein, sugar, relative water content, and nutrient uptake (Nitrogen and Phosphorous) as compared to monocultures and untreated positive control plants. In addition, co-inoculation in plants resulted in lower Na+, MDA, H2O2, CAT, APX activities, and also lower ACC accumulation (49.75 to 72.38% compared to non-treated salt- stressed plant) in O. sanctum, which probably played a significant role in minimizing the deleterious effects of salinity. Finally, multifactorial analysis showed that co-inoculation of Th and PGPBs improved O. sanctum growth, its physiological activities, and alleviated salt stress compared to single inoculated and positive control plants. These microbial consortia were evaluated for the first time on O. sanctum under salt stress. Therefore, the microbial consortia application could be employed to boost crop productivity in poor, marginalized and stressed agricultural fields. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-023-01328-2.
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BACKGROUND: To motivate students toward the path of lifelong learning, it is important to train them for a self-directed learning (SDL) approach, where they identify the need of learning the content as well as look forward to attaining the learning outcome, independently. The level of readiness for SDL will allow the learner to be self-disciplined, self-organized, an effective team builder and communicator, a self-assessor, and a self-reflector, thus being a self-learner with the ability to accept and give constructive feedback. The aim of the study was to develop, validate, and implement the SDL readiness scale for health professional students. MATERIALS AND METHOD: The readiness scale of 43 items was used in four sub-titles (awareness, learning strategies, and style, motivation, team building) was developed using the Delphi method with 12 experts and was implemented for the medical students at Karamsad, Gujarat as a cross-sectional survey after a pilot trial scale from May 2021 to September 2021, using mean, the standard deviation for each item, and arrived sub-titles. The ANOVA test was used to find differences in readiness scores as per different years of the medical program. RESULT: The result demonstrates that the maximum score was obtained for the first-year medical student (149.89 ± 24.72), which dropped in year 2 (136.35 ± 32.26) but increased by the final year (147.67 ± 56.66), although not as high as the initial joining year. However, a statistically significant difference per gender was identified for a few items of the scale [(items 24 (P < 0.034), 26 (P < 0.0005), 37 (P < 0.035), and 40 (P < 0.013)]. The logistic regression analysis showed no statistical significance for the DSVS-self-directed learning readiness scale (SDLRS) score and demographic variables. CONCLUSION: The outcome of the study strongly suggests training/sensitization sessions for students to highlight the importance of a SDL approach in a digital millennium. Moreover, a longitudinal follow-up needs to be conducted for the readiness score of the students based on the developed scale and subsequent training sessions need to be organized for students as well as faculty for better outcomes for the students toward SDL sessions.
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INTRODUCTION: Lumbar spondylosis (LS) is a degenerative disorder of the lumbar spine. Despite substantial research efforts, no gold-standard treatment for LS has been identified. The efficacy of individualized homeopathic medicines (IHMs) in LS has remained under-researched. In this study, the efficacy of IHMs was compared with identical-looking placebos in the treatment of low back pain associated with LS. METHODS: A double-blind, randomized (1:1), placebo-controlled trial was conducted at the National Institute of Homoeopathy, West Bengal, India. Patients were randomized to receive IHMs or placebos, along with standardized concomitant care for both the groups. The Oswestry low back pain and disability questionnaire (ODQ) was the primary outcome; the Roland-Morris questionnaire (RMQ) and the short form of the McGill pain questionnaire (SF-MPQ) were the secondary outcomes. Each was measured at baseline and every month for 3 months. The intention-to-treat (ITT) sample was analyzed to detect any inter-group differences using two-way repeated measures analysis of variance models overall and by unpaired t-tests at different time points. RESULTS: Enrolment was stopped prematurely because of time restrictions; 55 patients were randomized (verum: 28; control: 27); 49 were analyzed by ITT (verum: 26; control: 23). Inter-group differences in ODQ (F 1, 47 = 0.001, p = 0.977), RMQ (F 1, 47 = 0.190, p = 0.665) and SF-MPQ total score (F 1, 47 = 3.183, p = 0.081) at 3 months were not statistically significant. SF-MPQ total score after 2 months (p = 0.030) revealed inter-group statistical significance, favoring IHMs against placebos. Some of the SF-MPQ sub-scales at different time points were also statistically significant: e.g., the SF-MPQ average pain score after 2 months (p = 0.002) and 3 months (p = 0.007). Rhus toxicodendron, Sulphur and Pulsatilla nigricans were the most frequently indicated medicines. CONCLUSION: Owing to failure in detecting a statistically significant effect for the primary outcome and in recruiting a sufficient number of participants, our trial remained inconclusive. TRIAL REGISTRATION: CTRI/2019/11/021918.
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Homeopatia , Dor Lombar , Espondilose , Humanos , Dor Lombar/tratamento farmacológico , Resultado do Tratamento , Método Duplo-CegoRESUMO
The enantiomeric pure and natural (+)-Lactones (C ≤ 14) with aromas obtained from fruits and milk are considered flavoring compounds. The flavoring value is related to the lactones' ring size and chain length, which blend in varying concentrations to produce different stone-fruit flavors. The nature-identical and enantiomeric pure (+)-lactones are only produced through whole-cell biotransformation of yeast. The industrially important γ-decalactone and δ-decalactone are produced by a four-step aerobic-oxidation of ricinoleic acid (RA) following the lactonization mechanism. Recently, metabolic engineering strategies have opened up new possibilities for increasing productivity. Another strategy for increasing yield is to immobilize the RA and remove lactones from the broth regularly. Besides flavor impact, γ-, δ-, ε-, ω-lactones of the carbon chain (C8-C12), the macro-lactones and their derivatives are vital in pharmaceuticals and healthcare. These analogues are isolated from natural sources or commercially produced via biotransformation and chemical synthesis processes for medicinal use or as active pharmaceutical ingredients. The various approaches to biotransformation have been discussed in this review to generate more prospects from a commercial point of view. Finally, this work will be regarded as a magical brick capable of containing both traditional and genetic engineering technology while contributing to a wide range of commercial applications.
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Lactonas , Engenharia Metabólica , Lactonas/química , Lactonas/metabolismo , Biotransformação , Oxirredução , Saccharomyces cerevisiae/metabolismoRESUMO
INTRODUCTION: This study was conducted with the objective to analyze the out-of-pocket (OOP) healthcare expenditure and catastrophic healthcare expenditure (CHE) on chronic non-communicable diseases (CNCD) among the elderly population, and the association of CHE on CNCD with associated factors among the same population. MATERIALS AND METHODS: We collected data from the elderly population of Dharan Sub-metropolitan city of the Eastern Nepal via door-to-door survey and face-to-face interview. The ten wards out of twenty were chosen by lottery method, and the equal proportion out of 280 samples was purposively chosen from each of ten wards (28 participants from each selected ward). The data were entered in Microsoft Excel 2019 v16.0 and statistical analysis was performed by using statistical package for social sciences, IBM SPSS® v21. The chi-square test was used to test the group differences. Multivariable logistic regression was used to determine independent factors associated with CHE (all variables with P < 0.20), and adjusted odds ratios (AOR) were calculated at 95% confidence interval (CI). RESULTS: The median household, food and health expenditures were 95325 (72112.50-126262.50), 45000 (33000-60000) and 2100 (885.00-6107.50) NPR respectively. The proportion of the participants with CHE was 14.6%. The single living participants had 3.4 times higher odds of catastrophic health expenditure (AOR = 3.4, 95% CI = 1.2-9.6, P-value = 0.022) than those who are married. Similarly, those who had cancer had 0.1 times lower odds of CHE (AOR = 0.1, 95% CI = 0.0-0.2, P-value = <0.001) than those without cancer. CONCLUSION: The elder population had significant financial health shocks due to chronic health ailments. There should be the provision of mandatory health insurance programmes for elderly to cut down the catastrophic healthcare expenditure. Similarly, there should be the provision of exemption scheme for vulnerable elderly who are more likely to face catastrophic expenditure from all available health facilities.
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Gastos em Saúde , Doenças não Transmissíveis , Idoso , Humanos , Doença Catastrófica , Estudos Transversais , Nepal , Doenças não Transmissíveis/epidemiologiaRESUMO
Consistently emerging variants and the life-threatening consequences of SARS-CoV-2 have prompted worldwide concern about human health, necessitating rapid and accurate point-of-care diagnostics to limit the spread of COVID-19. Still, However, the availability of such diagnostics for COVID-19 remains a major rate-limiting factor in containing the outbreaks. Apart from the conventional reverse transcription polymerase chain reaction, loop-mediated isothermal amplification-based (LAMP) assays have emerged as rapid and efficient systems to detect COVID-19. The present study aims to develop RT-LAMP-based assay system for detecting multiple targets in N, ORF1ab, E, and S genes of the SARS-CoV-2 genome, where the end-products were quantified using spectrophotometry, paper-based lateral-flow devices, and electrochemical sensors. The spectrophotometric method shows a LOD of 10 agµL-1 for N, ORF1ab, E genes and 100 agµL-1 for S gene in SARS-CoV-2. The developed lateral-flow devices showed an LOD of 10 agµL-1 for all four gene targets in SARS-CoV-2. An electrochemical sensor developed for N-gene showed an LOD and E-strip sensitivity of log 1.79 ± 0.427 pgµL-1 and log 0.067 µA/pg µL-1/mm2, respectively. The developed assay systems were validated with the clinical samples from COVID-19 outbreaks in 2020 and 2021. This multigene target approach can effectively detect emerging COVID-19 variants using combination of various analytical techniques at testing facilities and in point-of-care settings.
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COVID-19 , SARS-CoV-2 , Humanos , SARS-CoV-2/genética , COVID-19/diagnóstico , Sensibilidade e Especificidade , Técnicas de Amplificação de Ácido Nucleico/métodos , Técnicas de Diagnóstico Molecular/métodos , RNA Viral/genéticaRESUMO
Circular RNAs (circRNAs) are a novel class of covalently closed RNA molecules that recently emerged as a critical regulator of gene expression in development and diseases. Recent research has highlighted the importance of novel circRNAs in the biosynthesis and secretion of insulin from ß-cells of pancreatic islets. However, all circRNAs expressed in pancreatic islets or ß-cells are not readily available in the database. In this study, we analyzed publicly available RNA-sequencing datasets of the pancreatic islets to catalog all circRNAs expressed in pancreatic islets to construct the PanCircBase (https://www.pancircbase.net/) database that provides the following resources: 1) pancreatic islet circRNA annotation details (genomic position, host gene, exon information, splice length, sequence, other database IDs, cross-species conservation), 2) divergent primers for PCR analysis of circRNAs, 3) siRNAs for silencing of target circRNAs, 4) miRNAs associated with circRNAs, 5) possible protein-coding circRNAs and their polypeptides. In summary, this is a comprehensive online resource for exploring circRNA expression and its possible function in pancreatic ß-cells.
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This manuscript reports the design and fabrication of conducting plastibody based electrochemical sensor for the detection of Salmonella typhimurium. The conductive plastibody was fabricated on an Indium Tin Oxide surface through potentiostatic method (electrodeposition for 400 s), wherein a polymer mix of pyrrole, lactic acid, ammonium chloride, and sodium dodecyl sulfate was employed for the electrodeposition. Various template removal methods were tested and electrochemical cleaning in the MES buffer was found to be the most suitable, which was optimized further. The synthesized plastibody sensors were characterized using electrochemical impedance spectroscopy, contact angle, FTIR spectroscopy and scanning electron microscopy. Amperometry was used as the electrochemical analytical technique for the determination of the analyte in the concentration range of 100 -108 CFU/mL having a limit of detection of 3.42 CFU/mL. Sensor's performance was also compared with the non-imprinted electrode and an imprinting factor of 3.8 was found. The plastibody sensor was tested against other bacteria and coefficient of selectivity was calculated to be 1.0, 10.8, 5.6 and 2.4 towards S. typhi, S. aureus, E. coli and L. monocytogenes respectively. The sensor was also found to be reproducible in nature (RSD 0.11 %) and this generic concept presented herein may be extended for the detection of pathogens in other matrices as well.
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Impressão Molecular , Polímeros , Técnicas Eletroquímicas/métodos , Eletrodos , Escherichia coli , Limite de Detecção , Impressão Molecular/métodos , Polímeros/química , Salmonella typhimurium , Staphylococcus aureusRESUMO
Hepcidin, a key regulator of iron homeostasis, has been implicated in the pathogenesis of various iron-related diseases. Although small interfering RNA (siRNA) are potent to modulate the expression of hepcidin, their bioavailability remains a major issue. The ß-galactopyranoside-conjugated liposomes (GAL-liposome) targeting liver synthesized hepcidin were prepared by thin lipid film hydration method to encapsulate siRNA and the conjugation of ß-galactopyranoside to the lipid nanocarrier was achieved by covalent chemistry. The prepared siRNA loaded GAL-lip were spherical with around 50 nm radius in size as observed by HR-TEM. The zeta potential and polydispersity index of the prepared liposomes were - 19.9 ± 0.96 mV and 0.44 ± 0.05, respectively. The encapsulation efficiency as determined by dialysis bag method was around 91.76 ± 1.74%. The cell viability and cellular uptake analysis was examined in HepG2 cells by MTT assay and flow cytometry, respectively. The stability and cumulative release of siRNA was also assessed. The hepcidin mRNA expression on administration of siRNA loaded GAL-lip was determined in HepG2 cells and in lipopolysaccharide-induced mice model followed by examining itsin vivo biodistribution by fluorescence microscopy. The results suggested thatsiRNA loaded GAL-lip reduced the hepcidin levels, thus, highlighting a novel ligand conjugated ionizable lipid-based nanocarrier for inducing RNA interference.
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Hepcidinas , Lipossomos , Animais , Galactose/metabolismo , Hepcidinas/genética , Hepcidinas/metabolismo , Ferro/metabolismo , Lipídeos , Lipossomos/metabolismo , Fígado/metabolismo , Camundongos , RNA Interferente Pequeno , Distribuição TecidualRESUMO
BACKGROUND: Acne is estimated to affect 9.4% of the global population, making it the 8th most prevalent disease worldwide. Acne vulgaris (AV) is among the diseases that directly affect quality of life. This trial evaluated the efficacy of individualized homeopathic medicines (IHM) against placebo in AV. METHODS: In this double-blind, randomized, placebo-controlled trial conducted at the National Institute of Homoeopathy, India, 126 patients suffering from AV were randomized in a 1:1 ratio to receive either IHM (verum) in centesimal potencies or identical-looking placebo (control). The primary outcome measure was the Global Acne Grading System score; secondary outcomes were the Cardiff Acne Disability Index and Dermatology Life Quality Index questionnaires - all measured at baseline and 3 months after the intervention. Group differences and effect sizes (Cohen's d) were calculated on the intention-to-treat sample. RESULTS: Overall, improvements were greater in the IHM group than placebo, with small to medium effect sizes after 3 months of intervention; however, the inter-group differences were statistically non-significant. Sulphur (17.5%), Natrum muriaticum (15.1%), Calcarea phosphorica (14.3%), Pulsatilla nigricans (10.3%), and Antimonium crudum (7.1%) were the most frequently prescribed medicines; Pulsatilla nigricans, Tuberculinum bovinum and Natrum muriaticum were the most effective of those used. No harms, unintended effects, homeopathic aggravations or any serious adverse events were reported from either group. CONCLUSION: There was non-significant direction of effect favoring homeopathy against placebo in the treatment of AV. TRIAL REGISTRATION: CTRI/2018/11/016248; UTN: U1111-1221-8164.
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Acne Vulgar , Homeopatia , Materia Medica , Humanos , Qualidade de Vida , Materia Medica/uso terapêutico , Método Duplo-Cego , Acne Vulgar/tratamento farmacológico , Resultado do TratamentoRESUMO
This study investigates the static and fatigue behavior of porous and conventional anterior cervical cages. Porous structure titanium anterior cervical cages were manufactured using direct selective laser sintering technique. Four different types of cervical cages were designed and manufactured, among which three designs consist of porous structure (type 1, type 2, and type 3) and manufactured using metal 3D printing. Remaining one design (type 4) was manufactured using conventional machining and did not consist any porous structure. All types of manufactured cages were tested in compression under static and fatigue loading conditions as per ASTM F2077 standard. Static and fatigue subsidence testing was performed using ASTM F2267 standard. Static compression testing results of type 1 and type 4 cages reported higher yield load when compared to the type 2 and type 3 cages. Static subsidence testing results reported almost 11% less subsidence rate for additively manufactured cages than the conventional cages. Fatigue subsidence testing results showed that type 2 and type 3 cages can withstood approximately 21% higher number of cycles before subsidence as compare to the type 1 and type 4 cages. During fatigue testing, all the cages design survived 5 million cycles at the 3000 N loading. For 6000 N and 8000 N, loading rate type 2 and type 3 cages showed lower fatigue life when compared to other cages design. Since fatigue life of type 2 and type 3 cage designs were reported lower than other cages design, it is concluded that the performance of the additively manufactured porous cages can be significantly varied based upon the cage design features.
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Fusão Vertebral , Teste de Materiais , Porosidade , Desenho de Prótese , Fusão Vertebral/métodos , Titânio , Suporte de CargaRESUMO
The last decade has seen a robust increase in various types of novel RNA molecules and their complexity in gene regulation. RNA molecules play a critical role in cellular events by interacting with other biomolecules, including protein, DNA, and RNA. It has been established that RNA-RNA interactions play a critical role in several biological processes by regulating the biogenesis and function of RNA molecules. Interestingly, RNA-RNA interactions regulate the biogenesis of diverse RNA molecules, including mRNAs, microRNAs, tRNAs, and circRNAs, through splicing or backsplicing. Structured RNAs like rRNA, tRNA, and snRNAs achieve their functional conformation by intramolecular RNA-RNA interactions. In addition, functional consequences of many intermolecular RNA-RNA interactions have been extensively studied in the regulation of gene expression. Hence, it is essential to understand the mechanism and functions of RNA-RNA interactions in eukaryotes. Conventionally, RNA-RNA interactions have been identified through diverse biochemical methods for decades. The advent of high-throughput RNA-sequencing technologies has revolutionized the identification of global RNA-RNA interactome in cells and their importance in RNA structure and function in gene expression regulation. Although these technologies revealed tens of thousands of intramolecular and intermolecular RNA-RNA interactions, we further look forward to future unbiased and quantitative high-throughput technologies for detecting transcriptome-wide RNA-RNA interactions. With the ability to detect RNA-RNA interactome, we expect that future studies will reveal the higher-order structures of RNA molecules and multi-RNA hybrids impacting human health and diseases. This article is categorized under: RNA Methods > RNA Analyses In Vitro and In Silico RNA Structure and Dynamics > Influence of RNA Structure in Biological Systems.