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The Corona Virus Disease (COVID-19) has a huge impact on all of humanity, and people's disregard for COVID-19 regulations has sped up the disease's spread. Our study uses a state-of-the-art object detection model like YOLOv4 (You Only Look Once, version 4), a very effective tool, on real-time 25fps, 1920 X 1080 video data streamed live by a camera-mounted Unmanned Aerial Vehicle (UAV) quad-copter to observe proper maintenance of social distance in an area of 35m range in this study. The model has demonstrated remarkable efficacy in identifying and quantifying instances of social distancing, with an accuracy of 82% and little latency. It has been able to work efficiently with real-time streaming at 25–30 ms. Our model is based on CSPDarkNet-53, which was trained on the MS COCO dataset for image classification. It includes additional layers to capture feature maps from different phases. Additionally, the model's neck is made up of PANet, which is used to aggregate the parameters from various CSPDarkNet-53 layers. The CSPDarkNet-53's 53 convolutional layers are followed by 53 more layers in the model head, for a total of 106 completely convolutional layers in the design. This architecture is further integrated with YOLOv3, resulting in the YOLOv4 model, which will be used by our detection model. Furthermore, to differentiate humans The aforementioned method was used to evaluate drone footage and count social distance violations in real time. Our findings show that our model was reliable and successful at detecting social distance violations in real-time with an average accuracy of 82%. [ABSTRACT FROM AUTHOR]

Deep neural networks (DNNs), the integration of neural networks (NNs) and deep learning (DL), have proven highly efficient in executing numerous complex tasks, such as data and image classification. Because the multilayer in a nonlinearly separable data structure is not transparent, it is critical to develop a specific data classification model from a new and unexpected dataset. In this paper, we propose a novel approach using the concepts of DNN and decision tree (DT) for classifying nonlinear data. We first developed a decision tree-based neural network (DTBNN) model. Next, we extend our model to a decision tree-based deep neural network (DTBDNN), in which the multiple hidden layers in DNN are utilized. Using DNN, the DTBDNN model achieved higher accuracy compared to the related and relevant approaches. Our proposal achieves the optimal trainable weights and bias to build an efficient model for nonlinear data classification by combining the benefits of DT and NN. By conducting in-depth performance evaluations, we demonstrate the effectiveness and feasibility of the proposal by achieving good accuracy over different datasets. [ABSTRACT FROM AUTHOR]

Dietary fibres can exert beneficial anti-inflammatory effects through microbially fermented short-chain fatty acid metabolites1,2, although the immunoregulatory roles of most fibre diets and their microbiota-derived metabolites remain poorly defined. Here, using microbial sequencing and untargeted metabolomics, we show that a diet of inulin fibre alters the composition of the mouse microbiota and the levels of microbiota-derived metabolites, notably bile acids. This metabolomic shift is associated with type 2 inflammation in the intestine and lungs, characterized by IL-33 production, activation of group 2 innate lymphoid cells and eosinophilia. Delivery of cholic acid mimics inulin-induced type 2 inflammation, whereas deletion of the bile acid receptor farnesoid X receptor diminishes the effects of inulin. The effects of inulin are microbiota dependent and were reproduced in mice colonized with human-derived microbiota. Furthermore, genetic deletion of a bile-acid-metabolizing enzyme in one bacterial species abolishes the ability of inulin to trigger type 2 inflammation. Finally, we demonstrate that inulin enhances allergen- and helminth-induced type 2 inflammation. Taken together, these data reveal that dietary inulin fibre triggers microbiota-derived cholic acid and type 2 inflammation at barrier surfaces with implications for understanding the pathophysiology of allergic inflammation, tissue protection and host defence.Dietary inulin fibre alters the composition and metabolism of gut microbiota, resulting in elevated levels of bile acids that subsequently trigger mucosal type 2 inflammation characterized by eosinophilia, with clinical implications for allergy and anti-helminth defence. [ABSTRACT FROM AUTHOR]

An artificial intelligence-based system was developed to efficiently predict settling velocity (SV) using a large dataset comprised of 2726 samples. The ranges of particle size and fluid viscosity were 0.212 − 98.59 mm and 0.02 − 92800 mPa.s, respectively. Properties of particle and fluid were fed to a model as the inputs to obtain SV as the output. Six machine learning algorithms were tested for the prediction. The random forest (RF) performed better than other algorithms with a coefficient of determination of 0.98 and a mean square error of 0.0027. A simple decision support system was developed using the RF model. The current study demonstrates the complete methodology of modeling SV with ML. [ABSTRACT FROM AUTHOR]

X-ray luminescence tomography (XLT) detects X-ray scintillators contrast agents using a focused or collimated X-ray beam to provide high spatial resolution excitation through thick tissue. The approach requires bright nanophosphors that are either synthesized or purchased. However, currently available commercial nanophosphors are mostly composed of a polydisperse mixture of several micro- to nano-sized particles that are unsuitable for biomedical imaging applications because of their size and aggregated form. Here, we demonstrate a fast and robust method to obtain uniform nano to submicron phosphor particles from a commercial source of polydisperse Eu- and Tb-doped Gd2O2S particles by separating the smaller particles present using gravitational and centrifugal sedimentation. In contrast to ball milling for 15–60 min, which drastically degraded the particles' brightness while reducing their size, our sedimentation method enabled the extraction of comparatively bright nanophosphors (≈100–300 nm in size) with a luminescence intensity of ≈10–20% of the several micron particles in the sample. Moreover, if scale up for higher yielding is required, the sedimentation process can be accelerated using fixed-angle and/or swinging bucket rotating centrifugation. Finally, after separation and characterization, nano and submicron phosphors were suspended and imaged through 5 mm thick porcine tissue using our in-house-built scanning X-ray induced luminescence chemical imaging (XELCI) system. [ABSTRACT FROM AUTHOR]

Cholinergic ILC2s support helminth clearance: The activity of group 2 innate lymphoid cells (ILC2s) is modulated by the action of neuropeptides and small-molecule neurotransmitters. To investigate whether ILC2s are a nonneuronal source of the neurotransmitter acetylcholine (ACh) that could influence potential neuro-immune cross-talk, Chu et al. used a green fluorescent protein (GFP) reporter mouse to analyze ILC2 expression of the ACh-synthesizing enzyme choline acetyltransferase (ChAT) following infection with helminth parasites or exposure to allergens or proinflammatory cytokines. ILC2 expression of the ChAT-GFP reporter was strongly induced in vivo after helminth infection and after exposure to Alternaria or injection of the cytokines IL-33 and IL-25. ACh stimulation of ILC2s increased their production of type 2 cytokines and enhanced helminth expulsion from the small intestine. The cholinergic potential of ILC2s revealed in this study provides further evidence of the complex interplay between the nervous system and innate lymphocytes. Group 2 innate lymphoid cells (ILC2s) reside in multiple tissues, including lymphoid organs and barrier surfaces, and secrete type 2 cytokines including interleukin-5 (IL-5), IL-9, and IL-13. These cells participate in multiple physiological processes including allergic inflammation, tissue repair, metabolic homeostasis, and host defense against helminth infections. Recent studies indicate that neurotransmitters and neuropeptides can play an important role in regulating ILC2 responses; however, the mechanisms that underlie these processes in vivo remain incompletely defined. Here, we identify that activated ILC2s up-regulate choline acetyltransferase (ChAT)—the enzyme responsible for the biosynthesis of acetylcholine (ACh)—after infection with the helminth parasite Nippostrongylus brasiliensis or treatment with alarmins or cytokines including IL-25, IL-33, and thymic stromal lymphopoietin (TSLP). ILC2s also express acetylcholine receptors (AChRs), and ACh administration promotes ILC2 cytokine production and elicits expulsion of helminth infection. In accordance with this, ChAT deficiency in ILC2s leads to defective ILC2 responses and impaired immunity against helminth infection. Together, these results reveal a previously unrecognized role of the ChAT-ACh pathway in promoting type 2 innate immunity to helminth infection. [ABSTRACT FROM AUTHOR]

Thalassemias are emerging as a global public health concern. Due to remarkable success in the reduction of childhood mortality by controlling infectious diseases in developing countries, thalassemias are likely to be a major public health concern in the coming decades in South Asia. Despite the fact that Bangladesh lies in the world's thalassemia belt, the information on different aspects (epidemiology, clinical course, mortality, complications and treatment outcomes) of thalassemias is lacking. In this comprehensive review, the aim is to to depict the epidemiological aspects of thalassemias, mutation profile and current treatment and management practices in the country by sharing the experience of dealing with 1178 cases over 2009-2014 time periods in a specialized thalassemia treatment centre. We have also discussed the preventative strategies of thalassemias from the context of Bangladesh which could be effective for other developing countries. [ABSTRACT FROM AUTHOR]

The existing IEEE and 3GPP standards have laid the foundation for integrating cellular and WiFi network to deliver a seamless experience for the end-users when roaming across multiple access networks. However, in recent studies, the issue of making roaming decision and intelligently selecting the most preferable Point of Service to optimize network resource and improve end user’s experience has not been considered properly. In this paper, we propose a novel cellular and WiFi roaming decision and AP selection scheme based on state of the art, 3GPP TS24.312 and IEEE 802.11u, k standards. Our proposed scheme assists the mobile nodes to decide the right timing to make roaming decision and select preferable point of service based on the operator’s policies and real-time network condition. We also introduce our simulation model of a heterogeneous network with cellular and WiFi interworking as well as 3GPP ANDSF, TS24.312. It is a complete end-to-end system model from application to physical layer with considering user’s mobility and realistic traffic model. The proposed scheme outperformed the conventional WiFi selection scheme in terms of dynamically steering mobile node’s data traffic from macrocell to available Access Points. The proposed scheme increased the utilization and balanced the traffic load of access points and improved user’s experienced throughput. [ABSTRACT FROM AUTHOR]

Advanced metering infrastructure (AMI), as the totality of systems and networks to measure, collect, store, analyze, and use energy usage data, is supposed to be the core component in smart grid. In AMI, there are numerous challenges among which cyber security is a major one that needs to be addressed with priority. The information centric networking (ICN) is a promising architecture for the future Internet that disseminates content based on named data instead of named hosts. The congestion control and self-security can enable more scalable, secure, collaborative, and pervasive networking, these make the ICN a potential network architecture for smart grid. This paper aims to apply the ICN approach on AMI system, which we termed as information centric AMI (ICN-AMI). To the best of our knowledge, this is the first attempt to distribute contents (or requests for contents) based on ICN in AMI system. Moreover, a simulation-based performance evaluation is employed to evaluate the effectiveness of the proposed ICN-AMI approach in traffic control for developing AMI system in smart grid. In addition, we proposed a novel key management scheme (KMS) for a large number of smart meters in this system to ensure confidentiality, integrality, and authentication. To validate the scheme, the security analysis, comparisons are done to demonstrate that the proposed information centric KMS (ICN-KMS) is possible and a promising solution for ICN-AMI system. [ABSTRACT FROM PUBLISHER]

In this paper, we present a novel low power medium access control protocol for wireless sensor networks (WSNs). The proposed protocol, EP-MAC (Efficient MAC with Parallel Transmission) achieves high energy efficiency and high packet delivery ratio under different traffic load. EP-MAC protocol is basically based on the Time Division Multiple Access (TDMA) approach. The power of Carrier Sense Multiple Access (CSMA) is used in order to offset the fundamental problems that the stand-alone TDMA method suffers from, i.e., problems such as lack of scalability, adaptability to varying situations, etc. The novel idea behind the EP-MAC is that it uses the parallel transmission concept with the TDMA link scheduling. EP-MAC uses the methods for the transmission power adjustment, i.e., uses the minimum level power necessary to reach the intended neighbor within a specified bit error rate [BER] target. This reduces energy consumption, as well as further enhances the scope of parallel transmission of the protocol. The simulation studies support the theoretical results, and validate the efficiency of our proposed EP-MAC protocol. [ABSTRACT FROM AUTHOR]

Background: Human genetic factors such as blood group antigens may affect the severity of infectious diseases. Presence of specific ABO and Lewis blood group antigens has been shown previously to be associated with the risk of different enteric infections. The aim of this study was to determine the relationship of the Lewis blood group antigens with susceptibility to cholera, as well as severity of disease and immune responses to infection. Methodology: We determined Lewis and ABO blood groups of a cohort of patients infected by Vibrio cholerae O1, their household contacts, and healthy controls, and analyzed the risk of symptomatic infection, severity of disease if infected and immune response following infection. Principal Findings: We found that more individuals with cholera expressed the Le(a+b−) phenotype than the asymptomatic household contacts (OR 1.91, 95% CI 1.03–3.56) or healthy controls (OR 1.90, 95% CI 1.13–3.21), as has been seen previously for the risk of symptomatic ETEC infection. Le(a–b+) individuals were less susceptible to cholera and if infected, required less intravenous fluid replacement in hospital, suggesting that this blood group may be associated with protection against V. cholerae O1. Individuals with Le(a–b−) blood group phenotype who had symptomatic cholera had a longer duration of diarrhea and required higher volumes of intravenous fluid replacement. In addition, individuals with Le(a–b−) phenotype also had lessened plasma IgA responses to V. cholerae O1 lipopolysaccharide on day 7 after infection compared to individuals in the other two Lewis blood group phenotypes. Conclusion: Individuals with Lewis blood type Le(a+b−) are more susceptible and Le(a–b+) are less susceptible to V. cholerae O1 associated symptomatic disease. Presence of this histo-blood group antigen may be included in evaluating the risk for cholera in a population, as well as in vaccine efficacy studies, as is currently being done for the ABO blood group antigens. Author Summary: Cholera remains a severe diarrheal disease, capable of causing extensive outbreaks and high mortality. Blood group is one of the genetic factors determining predisposition to disease, including infectious diseases. Expression of different Lewis or ABO blood group types has been shown to be associated with risk of different enteric infections. For example, individuals of blood group O have a higher risk of severe illness due to V. cholerae compared to those with non-blood group O antigens. In this study, we have determined the relationship of the Lewis blood group antigen phenotypes with the risk of symptomatic cholera as well as the severity of disease and immune responses following infection. We show that individuals expressing the Le(a+b−) phenotype were more susceptible to symptomatic cholera, while Le(a–b+) expressing individuals were less susceptible. Individuals with the Le(a–b−) blood group had a longer duration of diarrhea when infected, required more intravenous fluid replacement, and had lower plasma IgA antibody responses to V. cholerae LPS on day 7 following infection. We conclude that there is an association between the Lewis blood group and the risk of cholera, and that this risk may affect the outcome of infection as well as possibly the efficacy of vaccination. [ABSTRACT FROM AUTHOR]

Background: Salmonella enterica serotype Typhi is the cause of typhoid fever. It is a human-restricted pathogen, and few data exist on S. Typhi gene expression in humans. Methodology/Principal Findings: We applied an RNA capture and amplification technique, Selective Capture of Transcribed Sequences (SCOTS), and microarray hybridization to identify S. Typhi transcripts expressed in the blood of five humans infected with S. Typhi in Bangladesh. In total, we detected the expression of mRNAs for 2,046 S. Typhi genes (44% of the S. Typhi genome) in human blood; expression of 912 genes was detected in all 5 patients, and expression of 1,100 genes was detected in 4 or more patients. Identified transcripts were associated with the virulence-associated PhoP regulon, Salmonella pathogenicity islands, the use of alternative carbon and energy sources, synthesis and transport of iron, thiamine, and biotin, and resistance to antimicrobial peptides and oxidative stress. The most highly represented group were genes currently annotated as encoding proteins designated as hypothetical, unknown, or unclassified. Of the 2,046 detected transcripts, 1,320 (29% of the S. Typhi genome) had significantly different levels of detection in human blood compared to in vitro cultures; detection of 141 transcripts was significantly different in all 5 patients, and detection of 331 transcripts varied in at least 4 patients. These mRNAs encode proteins of unknown function, those involved in energy metabolism, transport and binding, cell envelope, cellular processes, and pathogenesis. We confirmed increased expression of a subset of identified mRNAs by quantitative-PCR. Conclusions/Significance: We report the first characterization of bacterial transcriptional profiles in the blood of patients with typhoid fever. S. Typhi is an important global pathogen whose restricted host range has greatly inhibited laboratory studies. Our results suggest that S. Typhi uses a largely uncharacterized genetic repertoire to survive within cells and utilize alternate energy sources during infection. Author Summary: Salmonella enterica serotype Typhi is the cause of typhoid fever and infects over 21 million cases and causes 200,000 deaths each year. S. Typhi only infects humans and this has greatly limited studies of S. Typhi pathogenesis. To study bacterial gene expression in human hosts, we used Selective Capture of Transcribed Sequences (SCOTS) and array hybridization to identify S. Typhi mRNAs expressed in the blood of 5 patients with S. Typhi infection. In total, we detected the expression of 2,046 S. Typhi genes (44% of the S. Typhi genome) in human blood; of these, 1,320 (29% of the S. Typhi genome) had significantly different levels of detection in human blood compared to in vitro cultures. Our results provide insight into S. Typhi pathogenesis, identifying both previously described and novel interactions occurring between host and microbe during the natural course of human infection. Further study of these genes, especially those of unknown function, may further our understanding of S. Typhi pathogenesis and aid in vaccine, diagnostic, and/or drug target development. [ABSTRACT FROM AUTHOR]

Background: Salmonella enterica serotype Typhi is the cause of typhoid fever. It is a human-restricted pathogen, and few data exist on S. Typhi gene expression in humans. Methodology/Principal Findings: We applied an RNA capture and amplification technique, Selective Capture of Transcribed Sequences (SCOTS), and microarray hybridization to identify S. Typhi transcripts expressed in the blood of five humans infected with S. Typhi in Bangladesh. In total, we detected the expression of mRNAs for 2,046 S. Typhi genes (44% of the S. Typhi genome) in human blood; expression of 912 genes was detected in all 5 patients, and expression of 1,100 genes was detected in 4 or more patients. Identified transcripts were associated with the virulence-associated PhoP regulon, Salmonella pathogenicity islands, the use of alternative carbon and energy sources, synthesis and transport of iron, thiamine, and biotin, and resistance to antimicrobial peptides and oxidative stress. The most highly represented group were genes currently annotated as encoding proteins designated as hypothetical, unknown, or unclassified. Of the 2,046 detected transcripts, 1,320 (29% of the S. Typhi genome) had significantly different levels of detection in human blood compared to in vitro cultures; detection of 141 transcripts was significantly different in all 5 patients, and detection of 331 transcripts varied in at least 4 patients. These mRNAs encode proteins of unknown function, those involved in energy metabolism, transport and binding, cell envelope, cellular processes, and pathogenesis. We confirmed increased expression of a subset of identified mRNAs by quantitative-PCR. Conclusions/Significance: We report the first characterization of bacterial transcriptional profiles in the blood of patients with typhoid fever. S. Typhi is an important global pathogen whose restricted host range has greatly inhibited laboratory studies. Our results suggest that S. Typhi uses a largely uncharacterized genetic repertoire to survive within cells and utilize alternate energy sources during infection. [ABSTRACT FROM AUTHOR]

Background: Human genetic factors such as blood group antigens may affect the severity of infectious diseases. Presence of specific ABO and Lewis blood group antigens has been shown previously to be associated with the risk of different enteric infections. The aim of this study was to determine the relationship of the Lewis blood group antigens with susceptibility to cholera, as well as severity of disease and immune responses to infection. Methodology: We determined Lewis and ABO blood groups of a cohort of patients infected by Vibrio cholerae O1, their household contacts, and healthy controls, and analyzed the risk of symptomatic infection, severity of disease if infected and immune response following infection. Principal Findings: We found that more individuals with cholera expressed the Le(a+b-) phenotype than the asymptomatic household contacts (OR 1.91, 95% CI 1.03-3.56) or healthy controls (OR 1.90, 95% CI 1.13-3.21), as has been seen previously for the risk of symptomatic ETEC infection. Le(a-b+) individuals were less susceptible to cholera and if infected, required less intravenous fluid replacement in hospital, suggesting that this blood group may be associated with protection against V. cholerae O1. Individuals with Le(a-b-) blood group phenotype who had symptomatic cholera had a longer duration of diarrhea and required higher volumes of intravenous fluid replacement. In addition, individuals with Le(a-b-) phenotype also had lessened plasma IgA responses to V. cholerae O1 lipopolysaccharide on day 7 after infection compared to individuals in the other two Lewis blood group phenotypes. Conclusion: Individuals with Lewis blood type Le(a+b-) are more susceptible and Le(a-b+) are less susceptible to V. cholerae O1 associated symptomatic disease. Presence of this histo-blood group antigen may be included in evaluating the risk for cholera in a population, as well as in vaccine efficacy studies, as is currently being done for the ABO blood group antigens. [ABSTRACT FROM AUTHOR]

Background: Salmonella enterica serotype Typhi is a human-restricted intracellular pathogen and the cause of typhoid fever. Cellular immune responses are required to control and clear Salmonella infection. Despite this, there are limited data on cellular immune responses in humans infected with wild type S. Typhi. Methodology/Principal Findings: For this work, we used an automated approach to purify a subset of S. Typhi proteins identified in previous antibody-based immuno-affinity screens and antigens known to be expressed in vivo, including StaF-putative fimbrial protein-STY0202, StbB-fimbrial chaperone-STY0372, CsgF-involved in curli production-STY1177, CsgD- putative regulatory protein-STY1179, OppA-periplasmic oligopeptide binding protein precursor-STY1304, PagC-outer membrane invasion protein-STY1878, and conserved hypothetical protein-STY2195; we also generated and analyzed a crude membrane preparation of S. Typhi (MP). In comparison to samples collected from uninfected Bangladeshi and North American participants, we detected significant interferon-γ responses in PBMCs stimulated with MP, StaF, StbB, CsgF, CsgD, OppA, STY2195, and PagC in patients bacteremic with S. Typhi in Bangladesh. The majority of interferon-γ expressing T cells were CD4 cells, although CD8 responses also occurred. We also assessed cellular proliferation responses in bacteremic patients, and confirmed increased responses in infected individuals to MP, StaF, STY2195, and PagC in convalescent compared to acute phase samples and compared to controls. StaF is a fimbrial protein homologous to E. coli YadK, and contains a Pfam motif thought to be involved in cellular adhesion. PagC is expressed in vivo under the control of the virulence-associated PhoP-regulon required for intra-macrophage survival of Salmonella. STY2195 is a conserved hypothetical protein of unknown function. Conclusion/Significance: This is the first analysis of cellular immune responses to purified S. Typhi antigens in patients with typhoid fever. These results indicate that patients generate significant CD4 and CD8 interferon-γ responses to specific S. Typhi antigens during typhoid fever, and that these responses are elevated at the time of clinical presentation. These observations suggest that an interferon-γ based detection system could be used to diagnose individuals with typhoid fever during the acute stage of illness. Author Summary: Salmonella enterica serotype Typhi infection is a significant global public health problem and the cause of typhoid fever. Salmonella are intracellular pathogens, and cellular immune responses are required to control and clear Salmonella infections. Despite this, there are limited data on cellular immune responses during wild type S. Typhi infection in humans. Here we report the assessment of cellular immune responses in humans with S. Typhi bacteremia through a screening approach that permitted us to evaluate interferon-γ and proliferation responses to a number of S. Typhi antigens. We detected significant interferon-γ CD4 and CD8 responses, as well as proliferative responses, to a number of recombinantly purified S. Typhi proteins as well as membrane preparation in infected patients. Antigen-specific interferon-γ responses were present at the time of clinical presentation in patients and absent in healthy controls. These observations could assist in the development of interferon-γ-based diagnostic assays for typhoid fever. [ABSTRACT FROM AUTHOR]

Background: Salmonella enterica serotype Typhi is a human-restricted intracellular pathogen and the cause of typhoid fever. Cellular immune responses are required to control and clear Salmonella infection. Despite this, there are limited data on cellular immune responses in humans infected with wild type S. Typhi. Methodology/Principal Findings: For this work, we used an automated approach to purify a subset of S. Typhi proteins identified in previous antibody-based immuno-affinity screens and antigens known to be expressed in vivo, including StaFputative fimbrial protein-STY0202, StbB-fimbrial chaperone-STY0372, CsgF-involved in curli production-STY1177, CsgDputative regulatory protein-STY1179, OppA-periplasmic oligopeptide binding protein precursor-STY1304, PagC-outer membrane invasion protein-STY1878, and conserved hypothetical protein-STY2195; we also generated and analyzed a crude membrane preparation of S. Typhi (MP). In comparison to samples collected from uninfected Bangladeshi and North American participants, we detected significant interferon-c responses in PBMCs stimulated with MP, StaF, StbB, CsgF, CsgD, OppA, STY2195, and PagC in patients bacteremic with S. Typhi in Bangladesh. The majority of interferon-c expressing T cells were CD4 cells, although CD8 responses also occurred. We also assessed cellular proliferation responses in bacteremic patients, and confirmed increased responses in infected individuals to MP, StaF, STY2195, and PagC in convalescent compared to acute phase samples and compared to controls. StaF is a fimbrial protein homologous to E. coli YadK, and contains a Pfam motif thought to be involved in cellular adhesion. PagC is expressed in vivo under the control of the virulence-associated PhoP-regulon required for intra-macrophage survival of Salmonella. STY2195 is a conserved hypothetical protein of unknown function. Conclusion/Significance: This is the first analysis of cellular immune responses to purified S. Typhi antigens in patients with typhoid fever. These results indicate that patients generate significant CD4 and CD8 interferon-c responses to specific S. Typhi antigens during typhoid fever, and that these responses are elevated at the time of clinical presentation. These observations suggest that an interferon-c based detection system could be used to diagnose individuals with typhoid fever during the acute stage of illness. [ABSTRACT FROM AUTHOR]

Based on the genomic sequence data of Escherichia coli K-12 strain, we have constructed a complete set of cloned individual genes encoding Histidine-tagged proteins with or without GFP fused for functional genomic analysis. Each clone encodes a protein of predicted ORF attached by Histidines and seven spacer amino acids at the N-terminal end, and five spacer amino acids and GFP at the C-terminal end. SfiI restriction sites are generated at both the N- and C-terminal boundaries of ORF upon cloning, which enables easy transfer of ORF to other vector systems by cutting with SfiI. Expression of cloned ORF is under the control of an IPTG-inducible promoter, which is strictly repressed by lacIq repressor gene product. The set of cloned ORFs described here should provide unique resources for systematic functional genomic approaches including (i) construction of DNA microarray, (ii) production and purification of proteins, (iii) analysis of protein localization by monitoring GFP fluorescence and (iv) analysis of protein–protein interaction. [ABSTRACT FROM PUBLISHER]

Investigates the role of chaperones in the folding of fusion proteins from Escherichia coli. Production of fusion proteins with predicted molecular masses; Use of luciferases in biotechnology applications; Means of analyzing proteins.

HscC, a DnaK homolog in Escherichia coli, consists of adenosine triphosphatase (ATPase), substrate-binding and C-terminal domains. Overexpression of HscC markedly inhibits growth of host cell and reduces the σ70-dependent promoter activity presumably by forming a complex with σ70. To identify the region(s) of HscC responsible for growth inhibition and complex formation with σ70, domain swapping experiments were carried out between DnaK and HscC. Thus the chimeric proteins carrying the ATPase domain of HscC and substrate-binding domains of either HscC or DnaK were found to inhibit the growth of the cell, reduce the σ70-dependent promoter activity and form a complex with σ70. These results indicate that the ATPase domain of HscC rather than the substrate-binding domain is important for determining its functional specificity. [Copyright &y& Elsevier]

Abstract Background: HscC, the third member of the Hsp70 family in Escherichia coli , shares 33% identity with the other two homologues, DnaK and HscA, and displays ATPase activity. Genetic and biochemical evidence indicates that the DnaK-DnaJ chaperone system interacts with σ32 and is involved in the negative regulation of the heat shock response. Although HscC is a highly conserved protein in the Hsp70 family, its function is still unknown. Results: We observed that the over-expression of HscC caused severe growth inhibition. To explore this effect, we performed primer extension analysis and a β-galactosidase assay and found that HscC reduced the σ70 -dependent promoter activity. An in vitro transcription assay revealed that HscC inhibited σ70 -dependent transcription. In addition, the co-purification analysis showed that σ70 co-eluted with HscC. Conclusion: These results indicate that HscC forms a complex with σ70 and may function as its negative modulator. [ABSTRACT FROM AUTHOR]

Significance: The ability to detect and localize specific molecules through tissue is important for elucidating the molecular basis of disease and treatment. Unfortunately, most current molecular imaging tools in tissue either lack high spatial resolution (e.g., diffuse optical fluorescence tomography or positron emission tomography) or lack molecular sensitivity (e.g., micro-computed tomography, μCT). X-ray luminescence imaging emerged about 10 years ago to address this issue by combining the molecular sensitivity of optical probes with the high spatial resolution of x-ray imaging through tissue. In particular, x-ray luminescence computed tomography (XLCT) has been demonstrated as a powerful technique for the high-resolution imaging of deeply embedded contrast agents in three dimensions (3D) for small-animal imaging. Aim: To facilitate the translation of XLCT for small-animal imaging, we have designed and built a small-animal dedicated focused x-ray luminescence tomography (FXLT) scanner with a μCT scanner, synthesized bright and biocompatible nanophosphors as contrast agents, and have developed a deep-learning-based reconstruction algorithm. Approach: The proposed FXLT imaging system was designed using computer-aided design software and built according to specifications. NaGdF4 nanophosphors doped with europium or terbium were synthesized with a silica shell for increased biocompatibility and functionalized with biotin. A deep-learning-based XLCT image reconstruction was also developed based on the residual neural network as a data synthesis method of projection views from few-view data to enhance the reconstructed image quality. Results: We have built the FXLT scanner for small-animal imaging based on a rotational gantry. With all major imaging components mounted, the motor controlling the gantry can be used to rotate the system with a high accuracy. The synthesized nanophosphors displayed distinct x-ray luminescence emission, which enables multi-color imaging, and has successfully been bound to streptavidin-coated substrates. Lastly, numerical simulations using the proposed deep-learning-based reconstruction algorithm has demonstrated a clear enhancement in the reconstructed image quality. Conclusions: The designed FXLT scanner, synthesized nanophosphors, and deep-learning-based reconstruction algorithm show great potential for the high-resolution molecular imaging of small animals. [ABSTRACT FROM AUTHOR]