ARTICLE | doi:10.20944/preprints201910.0217.v1
Subject: Engineering, Biomedical & Chemical Engineering Keywords: aptamer: acoustophoresis; microfluidics; gram-negative bacteria
Online: 18 October 2019 (11:41:41 CEST)
Here, we report a simple and effective method for separation of gram-negative bacteria using aptamer-modified microbeads and acoustophoresis. As acoustophoresis allows for simultaneous washing and size-dependent separation in continuous flow mode, we efficiently obtained gram-negative bacteria that showed high affinity without any additional washing steps. The proposed device has a simple and efficient channel design, utilizing a long, square-shaped microchannel that shows excellent separation performance in terms of the purity, recovery, and concentration factor. Microbeads (10 µm) coated with the GN6 aptamer can specifically bind gram-negative bacteria. Using acoustophoresis, gram-negative bacteria-bound microbeads and other unbound/contaminants can be separated by size with high purity and recovery. The device demonstrated excellent separation performance, with high recovery (up to 98%), high purity (up to 99%), and a high volume rate (500 µL/min), and a concentration factor of up to 20×. The acoustophoresis microfluidic device also showed binding affinity to multiple strains of gram-negative bacteria, but not to gram-positive bacteria. This study presents a new paradigm for early diagnosis of bacterial infectious diseases. In addition to detecting living bacteria or bacteria-derived biomarkers, this protocol can be extended to monitoring the contamination of water resources, and may aid quick responses to bioterrorism and pathogenic bacterial infections.
ARTICLE | doi:10.20944/preprints202011.0626.v1
Subject: Keywords: Isolation; Identification; gram positive; gram negative; tests staining; microbio metabolic activities
Online: 25 November 2020 (08:36:32 CET)
AbstractBaraton University dairy farm is an environment that attracts a microbiologist to inquire the composition of bacteria that exist there in. The knowledge of bacteria has in time and again amazed the life scientist community that have invested to acquire more information in this microbiology world.The study engages fundamental tests such as gram stain, endospore stain, and assays for specific microbial activities & enzymes, susceptibility on disinfectant and antibiotic, utilization of specific substrate and culture characteristics. The two organisms (gram negative and positive) tested positive for sucrose & lactose fermentation, Indole & Methly red, catalase & Oxidase, were both facultative and motile. On contrary, gram positive bacteria had spores and had a gamma haemolysis on Blood Agar, while gram negative bacteria haemolysed beta haemolysis. To draw a conclusion on the identity of the two organisms is that, the gram positive is a Bacillus ........ while gram negative is Escherichia coli.
REVIEW | doi:10.20944/preprints201910.0255.v1
Subject: Life Sciences, Microbiology Keywords: antimicrobial resistance; biofilm; efflux pump inhibitors; antibiotic potentiation; eskapee pathogens; gram-negative bacteria
Online: 22 October 2019 (10:22:56 CEST)
Antibiotic resistance represents a significant threat to the modern healthcare provision. The ESKAPEE pathogens, in particular, have proven to be especially challenging to treat, due to their intrinsic and acquired ability to rapidly develop resistance mechanisms in response to environmental threats. The development of biofilm has been characterised as an essential contributing factor towards antimicrobial-resistance and tolerance. Several studies have implicated the involvement of efflux pumps in antibiotic resistance, both directly, via drug extrusion and indirectly, through the formation of biofilm. As a result, the underlying mechanism of these pumps has attracted considerable interest due to the potential of targeting these protein structures and developing novel adjunct therapies. Subsequent investigations have revealed the ability of efflux pump-inhibitors (EPIs) to block drug-extrusion and disrupt biofilm formation, thereby, potentiating antibiotics and reversing resistance of pathogen towards them. This review will discuss the potential of EPIs as a possible solution to antimicrobial resistance, examining different challenges to the design of these compounds, with an emphasis on Gram-negative ESKAPEE pathogens.
Subject: Biology, Anatomy & Morphology Keywords: Wild Edible Plants; antimicrobial effect; Mediterranean plant; Gram+ bacteria; Gram- bacteria; extraction protocols; bioactive compounds; essential oils
Online: 21 July 2021 (10:05:22 CEST)
Introduction. Bacterial resistance to antibiotics is estimated to cause the major number of deaths by 2050 if we do not find strategies to slow down the rise of drug resistance . Reviews on Mediterranean Wild Edible Plants (MWEPs) with antimicrobial properties are scarce in the mean databases (Pubmed, Scopus and WoS). Hence, we proceeded to a new review of the studies on MWEPs. Methods. We used Wild Edible Plant and Antimicrobial as keywords. We only included the Mediterranean plants, and studies in non-Mediterranean countries, but for plants growing in Mediterranean basin. Exclusion criteria were the document type, studies not concerning plant, plants not edible, not antimicrobial properties, or totally out of topic. Results. Finally, the number of studies reviewed, starting from one hundred and ninety-two, was thirty-eight (19,8%), concerning the antimicrobial properties of seventy-four MWEPs species, be-longing to twenty-five Families. Fifty-seven (77%) out of seventy-four species, proved to be antimicrobial with a stringent threshold selection. Conclusions. The studies are still very heterogeneous. We still know too little about MWEPs properties, but what we already know seriously recommends continuing.
ARTICLE | doi:10.20944/preprints202010.0468.v1
Subject: Social Sciences, Accounting Keywords: Gram-Charlier series; DCC; DECO; backtesting; cryptocurrencies
Online: 22 October 2020 (21:50:55 CEST)
The semi-nonparametric (SNP) modeling of the return distribution has been proved to be a flexible and accurate methodology for portfolio risk management that allows two-step estimation of the dynamic conditional correlation (DCC) matrix. For this SNP-DCC model, we propose a stepwise procedure to compute pairwise conditional correlations under bivariate marginal SNP distributions, overcoming the curse of dimensionality. The procedure is compared to the assumption of Dynamic Equicorrelation (DECO), which is a parsimonious model when correlations among the assets are not significantly different but requires joint estimation of the multivariate SNP model. The risk assessment of both methodologies is tested for a portfolio on cryptocurrencies by implementing backtesting techniques and for different risk measures: Value-at-Risk, Expected Shortfall and Median Shortfall. The results support our proposal showing that the SNP-DCC model has better performance for a smaller confidence level than the SNP-DECO model, although both models perform similarly for higher confidence levels.
ARTICLE | doi:10.20944/preprints202208.0511.v1
Subject: Biology, Plant Sciences Keywords: Zingiber cassumunar Roxb.; essential oils; microencapsulated products; antibacterial activity; agricultural pathogens; foodborne pathogens; gram-positive bacteria; gram-negative bacteria
Online: 30 August 2022 (07:02:54 CEST)
Zingiber cassumunar Roxb. is a powerful medicinal plant that has been used as traditional medicine to cure respiratory problems, pain, and inflammation in China, Indonesia, Thailand and other Asian countries by using the crude extracts. The objective of this research is to identify phytochemical composition of Z. cassumunar Roxb. and to analyze antibacterial activity of crude extract, purified compounds, and their microencapsulated products of Rhizome Z. cassumunar Roxb. Identification of phytochemical composition in crude extract of rhizome Z. cassumunar Roxb. was achieved by chromatography-mass spectrophotometer. The major phytochemical composition in crude extract of Z. cassumunar Roxb. is essential oils, including terpinen4-ol (37.7%), β-pinene (20.8%), and (E)-1-(3,4-dimethoxyphenyl)but-1-ene (13.3%). Crude extract of Z. cassumunar Roxb. was purified with silica gel flash column chromatography, resulting two purified compounds. The antibacterial activity of crude extract, purified compounds, and their microencapsulated products of Rhizome Z. cassumunar Roxb. were evaluated against agricultural and foodborne pathogens by using disc agar diffusion and broth microdilution techniques. All of the samples studied (crude extracts, purified compounds, and microencapsulated of Z. cassumunar Roxb.) were effective against all the bacteria. Based on the results of the disc-diffusion assay suggested that amongst the samples studied, purified compounds (compound 1 and 2) and microencapsulated purified compounds (compound 1 and 2) exhibited more effective against all the bacteria compared to the crude extracts. Antibacterial activity of the rhizome of Z. cassumunar Roxb. was contributed mainly by the essential oils components as the active compounds. Gram-negative bacteria (X. oryzae, X. translucens, Pseudomonas spp, E. coli, and S. typhimurium) appeared to the most resistant to the crude extracts, purified compounds, and microencapsulated of Z. cassumunar Roxb. compared to the gram-positive bacteria (S. aureus, B. cereus, and L. monocytogenes). Microencapsulated of the tested samples (crude extract, purified compound 1, and purified compound 2) of the rhizome Z. cassumunar Roxb. exhibited high antibacterial activity with no significantly different with the tested samples without microencapsulation. These results suggest potential antibacterial properties of Z. cassumunar Roxb., which useful for agricultural plant health, food preservation, natural therapies, and pharmaceuticals.
CASE REPORT | doi:10.20944/preprints201912.0170.v1
Subject: Medicine & Pharmacology, Dermatology Keywords: sphingomonas; gram-negative bacterial infection; skin disease; bacterial
Online: 12 December 2019 (10:01:35 CET)
Sphingomonas paucimobilis is an opportunist pathogen bacillus gram-negative aerobic with a rare occurrence. We present a case in an immunocompetent man successfully treated by surgical debridement, purulent drainage and with an associated course of antibiotics. A large necrotic infection, approximately 5 cm x 3 cm, in a 74-year-old man was identified. Empirical antibiotic therapy with ciprofloxacin 400mg EV 12/12 hours, associated with clindamycin 600mg EV 6/6 hours and pain control was done through dipyrone 1gr, tramadol 400 mg. Deep venous thrombosis was prevented through the prescription of enoxaparin 40mg subcutaneous once a day during hospitalization. The case was well illustrated with pictures throughout treatment. Complete healing was achieved after 90 days. Herein, we present a case of cutaneous contamination. The presented case is the third cutaneous contamination case reported in the literature and the first reported case in the Amazonia region in Brazil.
REVIEW | doi:10.20944/preprints202101.0517.v1
Subject: Life Sciences, Biochemistry Keywords: Gram-negative bacterial; Metabolite transporter; pathways; metabolic engineering approaches
Online: 25 January 2021 (15:49:13 CET)
Metabolite transport across cellular membranes is a key feature of living organisms. Specialized proteins or protein complexes mediate such transport processes and are accessible to genetic engineering approaches. This special issue will reflect on the different transport between cellular compartments. It will provide an overview about different transporter classes and metabolic engineering approaches in different organisms ranging from Bacteria to Eukarya. The transport mechanisms energetics of these compounds were thought little of, and most consideration was given to the designing of metabolic pathways. Gram-negative microbes discharge a wide scope of proteins whose capacities incorporate biogenesis of organelles, for example, pilli and flagella, supplement securing, destructiveness, and efflux of medications and different poisons. Six particular emission frameworks have been appeared to intervene protein send out through the inward and external layers of Gram-negative microbes. These pathways are exceptionally monitored all through the Gram-negative bacterial species. In Gram-positive microscopic organisms, discharged proteins are usually moved across the single layer by the Sec pathway or the two-arginine (Tat) pathway. Recently efforts focused on transporter and metabolite engineering, visualize the improvement and enhancement of microbial cell to pass such metabolites these metabolite carrier proteins are inserted into the inner membrane of mitochondria across the inner membrane, specialized carrier proteins are necessary that mediate the transport into and out of the mitochondrial matrix. Two main strategies for metabolic engineering in plants: the introduction of genes encoding new enzymes and the use of TFs controlling specialized metabolic pathways
ARTICLE | doi:10.20944/preprints202111.0018.v1
Subject: Life Sciences, Immunology Keywords: Newcastle Disease Virus (NDV); NDV ZG1999HDS; NDV La Sota; TLT (Human macrophage cell line; Immunomodulation; Anti - Gram Positive bacteria; Anti - Gram - Negative bacteria; activity
Online: 1 November 2021 (13:01:38 CET)
ABSTRACT. The immunotherapies, as a modern therapeutic approach, get an attention because of theirs’ promise to treat a large number of different medical disorders. Immunomodulation effects of low titres (10 HA/ml) of NDV (Newcastle Disease Virus) ZG1999HDS or La Sota were tested on TLT (Human macrophage cell line) bound to PBMC (Peripheral Blood Mononuclear Cells). During the immunomodulation, the amount of NO, H2O2, lysozym and induced antibacterial activity against Gram - positive bacteria (Staphylococcus aureus, MRSA, Streptococcus pyogenes, Streptococcus agalactiae and Streptococcus mutants) and against Gram - negative bacteria (Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis and Acinetobacter baumanii) were analysed. In addition, the cytokine secretion, IL-1α, IL-2, IL-4, GM-CSF, TNF-α, IFN-α and IFN-α were evaluated. Firstly, the TLT cells are activated through the NDV ZG1999HDS or La Sota binding, followed by the NO “burst” and H2O2 and lysozyme level increase. Secondly, after the binding to the TLT cells and interaction with the PBMCs, the decrease of GM-CSF, and an increase of TNF – α and IFN – γ were found. Simultaneously, the decrease of pro – inflammatory cytokine IFN-α and the differentially increase of IL-1α, IL-2 and IL-4 were recorded. During the induction of the antibacterial response, against Gram - positive bacteria (Staphylococcus aureus, MRSA, Streptococcus pyogenes, Streptococcus agalactiae and Streptococcus mutants) the effect was one third higher with NDV ZG1999HDS compared to La Sota. Antibacterial response against Gram - negative bacteria (Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis and Acinetobacter baumanii) was not so clear. In general, NDV ZG1999HDS or La Sota activated TLT cells, further bound to PBMC; the ZG1999HDS is stronger immunomodulator than La Sota.
ARTICLE | doi:10.20944/preprints202211.0296.v1
Subject: Chemistry, Medicinal Chemistry Keywords: antimicrobial resistance; broad-spectrum antibiotics; antibacterial drug discovery; gram-negative bacteria; pyrrolobenzodiazepines
Online: 16 November 2022 (04:20:55 CET)
It is urgent to find new antibiotic classes to replenish the empty development pipeline of antibiotics. Recently, pyrrolobenzodiazepines (PBDs) with a C8-linked aliphatic-heterocycle have been identified as a new broad spectrum antibiotic class with activity against Gram-negative bacteria. The active imine moiety of the reported lead pyrrolobenzodiazepine compounds was replaced with amide to obtain the non-DNA binding and non-cytotoxic dilactam analogues to further understand the structure activity relationship and improve the safety potential of this class. The synthesized compounds were tested against panels of multidrug resistant Gram-positive and Gram-negative bacteria, including WHO priority pathogens. Minimum inhibitory concentrations for the dilactam analogues ranged from 4 – 32 mg/L for MDR Gram-positive bacteria, compared to 0.03 to 2 mg/L for the corresponding imine analogues while they were found to be inactive against MDR Gram-negative bacteria, with an MIC >32 mg/L, compared to an MIC of 0.5 to 32 mg/L. A molecular modelling study suggests the lack of imine functionality also affects the interaction of PBDs with DNA gyrase. This study suggests the presence of N10-C11 imine moiety is crucial for broad spectrum activity of pyrrolobenzodiazepines.
ARTICLE | doi:10.20944/preprints202208.0044.v1
Subject: Engineering, Other Keywords: Electro Hydrostatic Actuator; Fusion Convolutional Neural Networks; Particle Swarm Optimization; Gram Angle Difference Field
Online: 2 August 2022 (07:45:42 CEST)
Contrapose the highly integrated, multiple types of faults and complex working conditions of aircraft Electro Hydrostatic Actuator (EHA), to effectively identify its typical faults, we propose a fault diagnosis method based on the fusion convolutional neural networks (FCNN). First, the aircraft EHA fault data is encoded by GADF to obtain the fault feature images. Then we build an FCNN model that integrates the 1DCNN and 2DCNN, where the original 1D fault data is the input of the 1DCNN model, and the feature images obtained by GADF transformation are used as the input of 2DCNN. Multiple convolution and pooling operations are performed on each of these inputs to extract the features, next these feature vectors are spliced in the convergence layer, and the fully connected layers and the Softmax layers are finally used to attain the classification of aircraft EHA faults. Furthermore, the multi-strategy hybrid particle swarm optimization (MSPSO) algorithm is applied to optimize the FCNN to obtain a better combination of FCNN hyperparameters; MSPSO incorporates various strategies, including an initialization strategy based on homogenization and randomization, and an adaptive inertia weighting strategy, etc. The experimental result indicates that the FCNN model optimized by MSPSO achieves an accuracy of 96.86% for identifying typical faults of the aircraft EHA, respectively higher than the 1DCNN and the 2DCNN about 16.5% and 5.7%. Additionally, the FCNN model improved by MSPSO has a higher accuracy rate when compared to PSO.