Study on Distribution of Four Pseudomonas Species in Living Environment Using Multiplex PCR

Purpose: The genus Pseudomonas is a ubiquitous microorganism frequently detected from immunocompromised patients. The inherent resistance to numerous antimicrobial agents contributes to the opportunistic character of this pathogen exhaustive monitoring of this pathogen is considered of critical importance to public health organizations. The reliable identification method able to distinguish genetic close Pseudomonas species is needed, because these organisms are difficult to differentiate by phenotypic or biochemical methods. The purpose of the present study was to design species-specific primers in order to identify and detect four Pseudomonas species which are frequently detected from the human oral cavities, and to investigate the distribution of these organisms in the living environment using a multiplex PCR. Methods: Polymerase chain reaction (PCR) primers were designed based on partial sequences of the rpoD gene of four Pseudomonas species. Swab samples were collected from fifty washstands, and the distribution of Pseudomonas species was investigated using a conventional PCR at genus level and a multiplex PCR at species level. Results: Multiplex PCR method developed in this study was able to distinguish four Pseudomonas species clearly. The genus Pseudomonas was detected from all samples (100%), whereas P. putida, P, aeruginosa, P. stutzeri and P. fluorescens were detected at 44%, 8%, 4% and 2% in fifty swab samples, respectively. Conclusion: Our developed one-step multiplex PCR method is accurate, specific, cost-effective, time-saving, and works without requiring DNA extraction. It was indicated that washstands were the uninhabitable environment for P. putida, P, aeruginosa, P. stutzeri and P. fluorescens.

A simple and efficient CRISPR/Cas9 system permits ultra-multiplex genome editing in plants

The development and maturation of the CRISPR/Cas genome editing system provides a valuable tool for plant functional genomics and genetic improvement.Currently available genome-editing tools have a limited number of targets,restricting their application in genetic research.In this study,we developed a novel CRISPR/Cas9 plant ultra-multiplex genome editing system consisting of two template vectors,eight donor vectors,four destination vectors,and one primer-design software package.By combining the advantages of Golden Gate cloning to assemble multiple repetitive fragments and Gateway recombination to assemble large fragments and by changing the structure of the amplicons used to assemble sg RNA expression cassettes,the plant ultra-multiplex genome editing system can assemble a single binary vector targeting more than 40 genomic loci.A rice knockout vector containing 49 sg RNA expression cassettes was assembled and a high co-editing efficiency was observed.This plant ultra-multiplex genome editing system advances synthetic biology and plant genetic engineering.

Multi-dimensional multiplexing optical secret sharing framework with cascaded liquid crystal holograms

Secret sharing is a promising technology for information encryption by splitting the secret information into different shares.However,the traditional scheme suffers from information leakage in decryption process since the amount of available information channels is limited.Herein,we propose and demonstrate an optical secret sharing framework based on the multi-dimensional multiplexing liquid crystal(LC)holograms.The LC holograms are used as spatially separated shares to carry secret images.The polarization of the incident light and the distance between different shares are served as secret keys,which can significantly improve the information security and capacity.Besides,the decryption condition is also restricted by the applied external voltage due to the variant diffraction efficiency,which further increases the information security.In implementation,an artificial neural network(ANN)model is developed to carefully design the phase distribution of each LC hologram.With the advantage of high security,high capacity and simple configuration,our optical secret sharing framework has great potentials in optical encryption and dynamic holographic display.

Multiplex Rapid Test with Acceptable Diagnosis Performance as a Solution to Increase Diagnosis of Hepatitis B and C Viruses in Pregnant Women in an Area of High Prevalence of Both Hepatitis Viruses Associated with HIV

Background and Objective: HIV, hepatitis B virus (HBV) and hepatitis C virus (HCV) are very widespread in the world, however, less than 20% of the people affected are diagnosed and treated. This study aimed to determine the prevalence of HIV, HCV and HBV co-infections in pregnant women at Bangui Community University Hospital and the cost of screening. Methods: A cross-sectional study involving consenting pregnant women who came for antenatal care was performed. HIV, HCV antibodies and HBV antigens were detected using Exacto Triplex? HIV/HCV/HBsAg rapid test, cross-validated by ELISA tests. Sociodemographic and professional data, the modes of transmission and prevention of HIV and both hepatitis viruses were collected in a standard sheet and analyzed using the Epi-Info software version 7. Results: Pregnant women aged 15 to 24 were the most affected (45.3%);high school girls (46.0%), and pregnant women living in cohabitation (65.3%) were the most represented. Twenty-five (16.7%) worked in the formal sector, 12.7% were unemployed housewives and the remainder in the informal sector. The prevalence of HIV, HBV, and HCV viruses was 11.8%, 21.9% and 22.2%, respectively. The prevalence of co-infections was 8.6% for HIV-HBV, 10.2% for HIV-HCV, 14.7% for HBV-HCV and 6.5% for HIV-HBV-HCV. All positive results and 10% of negative results by the rapid test were confirmed by ELISA tests. The serology of the three viruses costs 39,000 FCFA (60 Euros) by ELISA compared to 10,000 FCFA (15.00 Euros) with Exacto Triplex? HIV/HCV/AgHBs (BioSynex, Strasbourg, France). Conclusion: The low level of education and awareness of hepatitis are barriers to development and indicate the importance of improving the literacy rate of women in the Central African Republic (CAR). Likewise, the high prevalence of the three viruses shows the need for the urgent establishment of a national program to combat viral hepatitis in the CAR.

Semi-implantable device based on multiplexed microfilament electrode cluster for continuous monitoring of physiological ions

Modern medicine is increasingly interested in advanced sensors to detect and analyze biochemical indicators.Ion sensors based on potentiometric methods are a promising platform for monitoring physiological ions in biological subjects.Current semi-implantable devices are mainly based on single-parameter detection.Miniaturized semi-implantable electrodes for multiparameter sensing have more restrictions on the electrode size due to biocompatibility considerations,but reducing the electrode surface area could potentially limit electrode sensitivity.This study developed a semi-implantable device system comprising a multiplexed microfilament electrode cluster(MMEC)and a printed circuit board for real-time monitoring of intra-tissue K^(+),Ca^(2+),and Na^(+)concentrations.The electrode surface area was less important for the potentiometric sensing mechanism,suggesting the feasibility of using a tiny fiber-like electrode for potentiometric sensing.The MMEC device exhibited a broad linear response(K^(+):2–32 mmol/L;Ca^(2+):0.5–4 mmol/L;Na^(+):10–160 mmol/L),high sensitivity(about 20–45 mV/decade),temporal stability(>2weeks),and good selectivity(>80%)for the above ions.In vitro detection and in vivo subcutaneous and brain experiment results showed that the MMEC system exhibits good multi-ion monitoring performance in several complex environments.This work provides a platform for the continuous real-time monitoring of ion fluctuations in different situations and has implications for developing smart sensors to monitor human health.

Real-Time 4-Mode MDM Transmission Using Commercial 400G OTN Transceivers and All-Fiber Mode Multiplexers

Weakly-coupled mode division multiplexing(MDM)technique is considered a promising candidate to enhance the capacity of an optical transmission system,in which mode multiplexers/demultiplexers(MMUX/MDEMUX)with low insertion loss and modal crosstalk are the key components.In this paper,a low-modal-crosstalk 4-mode MMUX/MDEMUX for the weakly-coupled triple-ring-core few-mode fiber(TRC-FMF)is designed and fabricated with side-polishing processing.The measurement results show that a pair of MMUX/MDEMUX and 25 km weakly-coupled TRC-FMF MDM link achieve low modal crosstalk of lower than−17.5 dB and insertion loss of lower than 11.56 dB for all the four modes.Based on the TRC-FMF and all-fiber MMUX/MDEMUX,an experiment for 25 km real-time 4-mode 3-λwavelength division multiplexing(WDM)-MDM transmission is conducted using commercial 400G optical transport network(OTN)transceivers.The experimental results prove weakly-coupled MDM techniques facilitate a smooth upgrade of the optical transmission system.

Development of a multiplex polymerase chain reaction assay for detection of hepatitis C virus,hepatitis B virus,and human immunodeficiency virus 1

BACKGROUND Hepatitis C virus(HCV),hepatitis B virus(HBV),and human immunodeficiency virus 1(HIV-1)are the most epidemic blood-borne viruses,posing threats to human health and causing economic losses to nations for combating the infection transmission.The diagnostic methodologies that depend on the detection of viral nucleic acids are much more expensive,but they are more accurate than sero-logical testing.AIM To develop a rapid,cost-effective,and accurate diagnostic multiplex polymerase chain reaction(PCR)assay for simultaneous detection of HCV,HBV,and HIV-1.METHODS The design of the proposed PCR assay targets the amplification of a short conserved region featured with a distinguishable melting profile and electro-phoretic molecular weight inside each viral genome.Therefore,this diagnostic method will be appropriate for application in both conventional(combined with electrophoresis)and real-time PCR facilities.Confirmatory in silico investigations were conducted to prove the capability of the approached PCR assay to detect variants of each virus.Then,Egyptian isolates of each virus were subjected to the wet lab examination using the given diagnostic assay.RESULTS The in silico investigations confirmed that the PCR primers can match many viral variants in a multiplex PCR assay.The wet lab experiment proved the efficiency of the assay in distinguishing each viral type through high-resolution melting analysis.Compared to related published assays,the proposed assay in the current study is more sensitive and competitive with many expensive PCR assays.CONCLUSION This study provides a simple,cost-effective,and sensitive diagnostic PCR assay facilitating the detection of the most epidemic blood-borne viruses;this makes the proposed assay promising to be substitutive for the mistakable and cheap serological-based assays.

Time-sequential color code division multiplexing holographic display with metasurface

Color metasurface holograms are powerful and versatile platforms for modulating the amplitude,phase,polarization,and other properties of light at multiple operating wavelengths.However,the current color metasurface holography can only realize static manipulation.In this study,we propose and demonstrate a multiplexing metasurface technique combined with multiwavelength code-division multiplexing(CDM)to realize dynamic manipulation.Multicolor code references are utilized to record information within a single metasurface and increase the information capacity and security for anticracks.A total of 48 monochrome images consisting of pure color characters and multilevel color video frames were reconstructed in dual polarization channels of the birefringent metasurface to exhibit high information density,and a video was displayed via sequential illumination of the corresponding code patterns to verify the ability of dynamic manipulation.Our approach demonstrates significant application potential in optical data storage,optical encryption,multiwavelengthversatile diffractive optical elements,and stimulated emission depletion microscopy.

Flat multifunctional liquid crystal elements through multi-dimensional information multiplexing

Flat optical elements have attracted enormous attentions and act as promising candidates for the next generation of optical components.As one of the most outstanding representatives,liquid crystal(LC)has been widely applied in flat panel display industries and inspires the wavefront modulation with the development of LC alignment techniques.However,most LC elements perform only one type of optical manipulation and are difficult to realize the multifunctionality and light integration.Here,flat multifunctional liquid crystal elements(FMLCEs),merely composed of anisotropic LC molecules with space-variant orientations,are presented for multichannel information manipulation by means of polarization,space and wavelength multiplexing.Specifically,benefiting from the unique light response with the change of the incident polarization,observation plane,and working wavelength,a series of FMLCEs are demonstrated to achieve distinct near-and far-field display functions.The proposed strategy takes full advantage of basic optical parameters as the decrypted keys to improve the information capacity and security,and we expect it to find potential applications in information encryption,optical anti-counterfeiting,virtual/augmented reality,etc.

Planar peristrophic multiplexing metasurfaces

As a promising counterpart of two-dimensional metamaterials,metasurfaces enable to arbitrarily control the wavefront of light at subwavelength scale and hold promise for planar holography and applicable multiplexing devices.Nevertheless,the degrees of freedom(DoF)to orthogonally multiplex data have been almost exhausted.Compared with state-of-theart methods that extensively employ the orthogonal basis such as wavelength,polarization or orbital angular momentum,we propose an unprecedented method of peristrophic multiplexing by combining the spatial frequency orthogonality with the subwavelength detour phase principle.The orthogonal relationship between the spatial frequency of incident light and the locally shifted building blocks of metasurfaces can be regarded as an additional DoF.We experimentally demonstrate the viability of the multiplexed holograms.Moreover,this newly-explored orthogonality is compatible with conventional DoFs.Our findings will contribute to the development of multiplexing metasurfaces and provide a novel solution to nanophotonics,such as large-capacity chip-scale devices and highly integrated communication.

Performance of phase-matching quantum key distribution based on wavelength division multiplexing technology

Quantum key distribution(QKD) generates information-theoretical secure keys between two parties based on the physical laws of quantum mechanics. The phase-matching(PM) QKD protocol allows the key rate to break the quantum channel secret key capacity limit without quantum repeaters, and the security of the protocol is demonstrated by using equivalent entanglement. In this paper, the wavelength division multiplexing(WDM) technique is applied to the PM-QKD protocol considering the effect of crosstalk noise on the secret key rate. The performance of PM-QKD protocol based on WDM with the influence of adjacent classical channels and Raman scattering is analyzed by numerical simulations to maximize the total secret key rate of the QKD, providing a reference for future implementations of QKD based on WDM techniques.

Impact of individual behavior adoption heterogeneity on epidemic transmission in multiplex networks

In recent years, the impact of information diffusion and individual behavior adoption patterns on epidemic transmission in complex networks has received significant attention. In the immunization behavior adoption process, different individuals often make behavioral decisions in different ways, and it is of good practical importance to study the influence of individual heterogeneity on the behavior adoption process. In this paper, we propose a three-layer coupled model to analyze the process of co-evolution of official information diffusion, immunization behavior adoption and epidemic transmission in multiplex networks, focusing on individual heterogeneity in behavior adoption patterns. Specifically, we investigate the impact of the credibility of social media and the risk sensitivity of the population on behavior adoption in further study of the effect of heterogeneity of behavior adoption on epidemic transmission. Then we use the microscopic Markov chain approach to describe the dynamic process and capture the evolution of the epidemic threshold. Finally, we conduct extensive simulations to prove our findings. Our results suggest that enhancing the credibility of social media can raise the epidemic transmission threshold, making it effective at controlling epidemic transmission during the dynamic process. In addition, improving an individuals' risk sensitivity, and thus their taking effective protective measures, can also reduce the number of infected individuals and delay the epidemic outbreak. Our study explores the role of individual heterogeneity in behavior adoption in real networks, more clearly models the effect of the credibility of social media and risk sensitivity of the population on the epidemic transmission dynamic, and provides a useful reference for managers to formulate epidemic control and prevention policies.

Study on Distribution of Acinetobacter baumannii Complex in Dental Hospital Using Multiplex PCR

Purpose: In recent years, multidrug-resistant Acinetobacter baumannii has appeared and caused outbreaks of hospital infections all over the world. Close monitoring of this pathogen and other A. baumanii complex species is considered of critical importance to public health organizations. The reliable identification method able to distinguish A. baumannii from genetically close Acinetobacter species is needed, because these species are unable to be differentiated by phenotypic or biochemical methods. The purpose of the present study was to design species-specific primers in order to identify and detect A. baumanii complex species, and Acinetobacter lwoffii which is frequently detected from the human specimens, and to investigate the distribution of these organisms in dental hospital using a multiplex PCR. Methods: Polymerase chain reaction (PCR) primers were designed based on partial sequences of the 16S ribosomal RNA (16S rRNA) gene and DNA gyrase subunit B (gyrB) of each species of A. baumanii complex species. Swab samples were collected from ten dental spittoon units in dental hospital, and the distribution of A. baumanii complex species was investigated using a multiplex PCR. Results: These primers were able to distinguish each species of A. baumanii complex species clearly. A. baumanii and A. calcoaceticus were detected at 20.0% and 10.0% in ten swab samples, respectively. On the other hand, A. nosocomialis, A. lowffii, and A. pittii were detected from no sample. Conclusion: Our developed one-step multiplex PCR method is accurate, specific, cost-effective, time-saving, and worked without requiring DNA extraction.

Demonstration of a manufacturable SOT-MRAM multiplexer array towards industrial applications

We have successfully demonstrated a 1 Kb spin-orbit torque(SOT)magnetic random-access memory(MRAM)multiplexer(MUX)array with remarkable performance.The 1 Kb MUX array exhibits an in-die function yield of over 99.6%.Additionally,it provides a sufficient readout window,with a TMR/RP_sigma%value of 21.4.Moreover,the SOT magnetic tunnel junctions(MTJs)in the array show write error rates as low as 10^(-6)without any ballooning effects or back-hopping behaviors,ensuring the write stability and reliability.This array achieves write operations in 20 ns and 1.2 V for an industrial-level temperature range from-40 to 125℃.Overall,the demonstrated array shows competitive specifications compared to the state-of-the-art works.Our work paves the way for the industrial-scale production of SOT-MRAM,moving this technology beyond R&D and towards widespread adoption.

Multiplex PCR for Identification and Detection of Cassava Mosaic Begomoviruses in Togo

Cassava mosaic disease (CMD) caused by Cassava Mosaic Begomoviruses (CMBs) is one of the most devastating crop diseases and a major constraint for cassava production. In order to ensure surveillance for epidemic prevention, low-cost diagnostic tools are appropriate for large-scale testing of cassava viruses. Multiplex PCR diagnosis is one approach that can reduce diagnostic costs and delays. A multiplex PCR approach was developed for simultaneous detection of African cassava mosaic virus (ACMV), East African Cassava Mosaic Virus and East African cassava mosaic Cameroon virus (EACMV/CM) in Togo CMD-infected cassava leaves. Three primers pairs were used to target their respective viruses in a single tube PCR. Multiplex PCR detected ACMV, EACMV and EACMV/CM in plant DNA extracts prepared from cassava leaves infected with CMB. The primers amplified 783 bp specific to ACMV, 650 bp specific to EACMV and 560 bp specific to EACMCV/CM in both uniplex and multiplex formats. Multiplex PCR is an excellent tool for the effective control of cassava diseases. .

Multiplex network infomax:Multiplex network embedding via information fusion

For networking of big data applications,an essential issue is how to represent networks in vector space for further mining and analysis tasks,e.g.,node classification,clustering,link prediction,and visualization.Most existing studies on this subject mainly concentrate on monoplex networks considering a single type of relation among nodes.However,numerous real-world networks are naturally composed of multiple layers with different relation types;such a network is called a multiplex network.The majority of existing multiplex network embedding methods either overlook node attributes,resort to node labels for training,or underutilize underlying information shared across multiple layers.In this paper,we propose Multiplex Network Infomax(MNI),an unsupervised embedding framework to represent information of multiple layers into a unified embedding space.To be more specific,we aim to maximize the mutual information between the unified embedding and node embeddings of each layer.On the basis of this framework,we present an unsupervised network embedding method for attributed multiplex networks.Experimental results show that our method achieves competitive performance on not only node-related tasks,such as node classification,clustering,and similarity search,but also a typical edge-related task,i.e.,link prediction,at times even outperforming relevant supervised methods,despite that MNI is fully unsupervised.

One Step Multiplex PCR for Identification of Candida haemulonii complex and Candia auris

Purpose: Recently, Candida haemulonii complex (Candida haemulonii, Candida duobushaemulonii and Candida haemulonii var. vulnera) and two genetically close species (Candida pseudohaemulonii and Candida auris) have emerged as an opportunistic fungal pathogen associated with various infectious diseases of humans, and most of those isolates have displayed antifungal resistance. Because it is difficult to differentiate these microorganisms by a current technique, unfortunately, it is important to establish a method for identifying them accurately. The purpose of the present study was to design species-specific primers in order to identify and detect C. auris, C. pseudohaemulonii, and C. haemulonii complex, i.e. , C. haemulonii, C. duobushaemulonii and C. haemulonii var. vulnera using a multiplex PCR. Methods: Polymerase chain reaction (PCR) primers were designed based on partial sequences of the 26S rRNA, 18S rRNA, and RPB1 genes and ITS region of five Candida species. Results: The multiplex PCR method developed in this study was able to distinguish five Candida species clearly. Conclusion: Our developed one-step multiplex PCR method is accurate, specific, cost-effective, time-saving, and works without requiring DNA extraction.

Identification of Mixtures of Two Types of Body Fluids Using the Multiplex Methylation System and Random Forest Models

Objective Body fluid mixtures are complex biological samples that frequently occur in crime scenes,and can provide important clues for criminal case analysis.DNA methylation assay has been applied in the identification of human body fluids,and has exhibited excellent performance in predicting single-source body fluids.The present study aims to develop a methylation SNaPshot multiplex system for body fluid identification,and accurately predict the mixture samples.In addition,the value of DNA methylation in the prediction of body fluid mixtures was further explored.Methods In the present study,420 samples of body fluid mixtures and 250 samples of single body fluids were tested using an optimized multiplex methylation system.Each kind of body fluid sample presented the specific methylation profiles of the 10 markers.Results Significant differences in methylation levels were observed between the mixtures and single body fluids.For all kinds of mixtures,the Spearman’s correlation analysis revealed a significantly strong correlation between the methylation levels and component proportions(1:20,1:10,1:5,1:1,5:1,10:1 and 20:1).Two random forest classification models were trained for the prediction of mixture types and the prediction of the mixture proportion of 2 components,based on the methylation levels of 10 markers.For the mixture prediction,Model-1 presented outstanding prediction accuracy,which reached up to 99.3%in 427 training samples,and had a remarkable accuracy of 100%in 243 independent test samples.For the mixture proportion prediction,Model-2 demonstrated an excellent accuracy of 98.8%in 252 training samples,and 98.2%in 168 independent test samples.The total prediction accuracy reached 99.3%for body fluid mixtures and 98.6%for the mixture proportions.Conclusion These results indicate the excellent capability and powerful value of the multiplex methylation system in the identification of forensic body fluid mixtures.

Research on Weighted Directed Dynamic Multiplexing Network of World Grain Trade Based on Improved MLP Framework

As the main food source for humans, the global movement of the three major grains significantly impacts human survival and development. To investigate the evolution of the world cereal trade network and its development trend, a weighted directed dynamic multiplexed network was established using historical data on cereal trade, cereal import dependency ratio, and arable land per capita. Inspired by the MLP framework, we redefined the weight determination method for computing layer weights and edge weights of the target layer, modified the CN, RA, AA, and PA indicators, and proposed the node similarity indicator for weighted directed networks. The AUC metric, which measures the accuracy of the algorithm, has also been improved in order to finally obtain the link prediction results for the grain trading network. The prediction results were processed, such as web-based presentation and community partition. It was found that the number of generalized trade agreements does not have a decisive impact on inter-country cereal trade. The former large grain exporters continue to play an important role in this trade network. In the future, the world trade in cereals will develop in the direction of more frequent intercontinental trade and gradually weaken the intracontinental cereal trade.

CoLM^(2)S:Contrastive self‐supervised learning on attributed multiplex graph network with multi‐scale information

Contrastive self‐supervised representation learning on attributed graph networks with Graph Neural Networks has attracted considerable research interest recently.However,there are still two challenges.First,most of the real‐word system are multiple relations,where entities are linked by different types of relations,and each relation is a view of the graph network.Second,the rich multi‐scale information(structure‐level and feature‐level)of the graph network can be seen as self‐supervised signals,which are not fully exploited.A novel contrastive self‐supervised representation learning framework on attributed multiplex graph networks with multi‐scale(named CoLM^(2)S)information is presented in this study.It mainly contains two components:intra‐relation contrast learning and interrelation contrastive learning.Specifically,the contrastive self‐supervised representation learning framework on attributed single‐layer graph networks with multi‐scale information(CoLMS)framework with the graph convolutional network as encoder to capture the intra‐relation information with multi‐scale structure‐level and feature‐level selfsupervised signals is introduced first.The structure‐level information includes the edge structure and sub‐graph structure,and the feature‐level information represents the output of different graph convolutional layer.Second,according to the consensus assumption among inter‐relations,the CoLM^(2)S framework is proposed to jointly learn various graph relations in attributed multiplex graph network to achieve global consensus node embedding.The proposed method can fully distil the graph information.Extensive experiments on unsupervised node clustering and graph visualisation tasks demonstrate the effectiveness of our methods,and it outperforms existing competitive baselines.