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BACKGROUND: Prospective genetic evaluation of patients at this referral research hospital presents clinical research challenges. OBJECTIVES: This study sought not only a single-gene explanation for participants' immune-related presentations, but viewed each participant holistically, with the potential to have multiple genetic contributions to their immune phenotype and other heritable comorbidities relevant to their presentation and health. METHODS: This study developed a program integrating exome sequencing, chromosomal microarray, phenotyping, results return with genetic counseling, and reanalysis in 1505 individuals from 1000 families with suspected or known inborn errors of immunity. RESULTS: Probands were 50.8% female, 71.5% were ≥18 years, and had diverse immune presentations. Overall, 327 of 1000 probands (32.7%) received 361 molecular diagnoses. These included 17 probands with diagnostic copy number variants, 32 probands with secondary findings, and 31 probands with multiple molecular diagnoses. Reanalysis added 22 molecular diagnoses, predominantly due to new disease-gene associations (9 of 22, 40.9%). One-quarter of the molecular diagnoses (92 of 361) did not involve immune-associated genes. Molecular diagnosis was correlated with younger age, male sex, and a higher number of organ systems involved. This program also facilitated the discovery of new gene-disease associations such as SASH3-related immunodeficiency. A review of treatment options and ClinGen actionability curations suggest that at least 251 of 361 of these molecular diagnoses (69.5%) could translate into ≥1 management option. CONCLUSIONS: This program contributes to our understanding of the diagnostic and clinical utility whole exome analysis on a large scale.
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Exoma , Testes Genéticos , Exoma/genética , Feminino , Testes Genéticos/métodos , Genômica , Humanos , Masculino , Fenótipo , Estudos ProspectivosRESUMO
A simple generic method for enhancing extracellular protein yields in engineered bacteria is still lacking. Here, we demonstrated that phage-encoded holin can be used to export proteins to the extracellular medium in both Gram-negative Escherichia coli and -positive Lactococcus lactis. When a putative holin gene LLNZ_RS10380 annotated in the genome of L. lactis NZ9000 (hol380) was recombinantly expressed in E. coli BL21(DE3), the Hol380 oligomerized up to hexamer in the cytoplasmic membrane, yielding membrane pore to allow the passage of cytosolic ß-galatosidase (116 kDa), whose extracellular production reached 54.59 U/µl, accounting for 76.37% of the total activity. However, the overexpressed Hol380 could not release cytosolic proteins across the membrane in L. lactis NZ9000, but increased the secretory production of staphylococcal nuclease to 2.55-fold and fimbrial adhesin FaeG to 2.40-fold compared with those guided by signal peptide Usp45 alone. By using a combination of proteomics and transcriptional level analysis, we found that overexpression of the Hol380 raised the accumulation of Ffh and YidC involved in the signal recognition particle pathway in L. lactis, suggesting an alternative road participating in protein secretion. This study proposed a new approach by expressing holin in bacterial cell factories to export target proteins of economic or medical interest.
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Proteínas de Escherichia coli , Lactococcus lactis , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Lactococcus lactis/genética , Lactococcus lactis/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismoRESUMO
In the era of the Internet of Things and Artificial Intelligence, the Wi-Fi fingerprinting-based indoor positioning system (IPS) has been recognized as the most promising IPS for various applications. Fingerprinting-based algorithms critically rely on a fingerprint database built from machine learning methods. However, currently methods are based on single-feature Received Signal Strength (RSS), which is extremely unstable in performance in terms of precision and robustness. The reason for this is that single feature machines cannot capture the complete channel characteristics and are susceptible to interference. The objective of this paper is to exploit the Time of Arrival (TOA) feature and propose a heterogeneous features fusion model to enhance the precision and robustness of indoor positioning. Several challenges are addressed: (1) machine learning models based on heterogeneous features, (2) the optimization of algorithms for high precision and robustness, and (3) computational complexity. This paper provides several heterogeneous features fusion-based localization models. Their effectiveness and efficiency are thoroughly compared with state-of-the-art methods.
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In signal array processing, high-resolution direction-of-arrival (DOA) estimation algorithms work well on the assumption that the system models are perfect. However, in practicality, there are imperfect system models in which sensor gain-and-phase errors are considered. In this paper, we propose a novel framework that can effectively solve direction-of-arrival estimation tasks in the presence of sensor gain-and-phase errors. In contrast to existing approaches based on phase retrieval, our method eliminates gain errors by using the compensated covariance matrix. Meanwhile, we propose a data preprocessing method by taking only one column of the compensated covariance matrix without losing any magnitude information. Additionally, the phase retrieval problem is formed by the proposed data preprocessing method. Furthermore, the phase retrieval problem is solved by the recently proposed sparse feasible point pursuit algorithm, and DOA estimates are obtained. To prevent the model from ambiguities, we employ the known DOA to place reference sources. Numerical results show that the proposed scheme achieves better performance compared to state-of-the-art approaches.
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The indoor localization method based on the Received Signal Strength (RSS) fingerprint is widely used for its high positioning accuracy and low cost. However, the propagation behavior of radio signals in an indoor environment is complicated and always leads to the existence of outliers and noises that deviate from a normal RSS value in the database. The fingerprint database containing outliers and noises will severely degrade the performance of an indoor localization system. In this paper, an approach to reconstruct the fingerprint database is proposed with the purpose of mitigating the influences of outliers. More specifically, by exploiting the spatial and temporal correlations of RSS data, the database can be transformed into a low-rank matrix. Therefore, the RPCA (Robust Principle Component Analysis) technique can be applied to recover the low-rank matrix from a noisy matrix. In addition, we propose an improved RPCA model which takes advantage of the prior knowledge of a singular value and could remove outliers and structured noise simultaneously. The experimental results show that the proposed method can eliminate outliers and structured noise efficiently.
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A heteroatom-rich 3D noninterpenetrating metal-organic framework (MOF) Cd-EDDA constructed from an ethylene glycol ether bridging tetracarboxylate ligand H4 EDDA (5,5'-(ethane-1,2-diylbis(oxy))diisophthalic acid) shows good chemical resistance to both acidic and alkaline solutions with a pH ranging from 2.0 to 12.2. There is a corresponding ratiometric luminescence response to pH from 2.0 to 11.5, and the sensing mechanism is also discussed through ion chromatography and molecular force field-based calculations. Importantly, the probe can easily be regenerated simply by modulating the pH of the solution, thus being the first example of a regenerable MOF-based ratiometric luminescent probe for pH.
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Lactococcus lactis and Streptococcus thermophilus are considered as ideal chassis of engineered probiotics, while food-grade genetic tools are limited in those strains. Here, a Zn2+-controlled gene expression (ZICE) system was identified in the genome of S. thermophilus CGMCC7.179, including a transcriptional regulator sczAst and a promoter region of cation transporter czcD (PczcDst). Specific binding of the SczAst to the palindromic sequences in PczcDst was demonstrated by EMSA analysis, suggesting the regulation role of SczAst on PczcDst. To evaluate their possibility to control gene expression in vivo, the sczAst-PczcDst was employed to drive the expression of green fluorescence protein (GFP) gene in L. lactis NZ9000 and S. thermophilus CGMCC7.179, respectively. Both of the transformants could express GFP under Zn2+ induction, while no fluorescence without Zn2+ addition. For optimal conditions, Zn2+ was used at a final concentration of 0.8 mM in L. lactis and 0.16 mM in S. thermophilus at OD600 close to 0.4, and omitting yeast extract powder in the medium unexpectedly improved GFP expression level by 2.2-fold. With the help of the ZICE system, engineered L. lactis and S. thermophilus strains were constructed to secret cytokine interleukin-10 (IL-10) with immunogenicity, and the IL-10 content in the supernatant of the engineered L. lactis was 59.37 % of that under the nisin controlled expression system. This study provided a tightly controlled expression system by the food-grade inducer Zn2+, having potential in development of engineered probiotics.
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The effect of atmospheric pressure plasma jet (APPJ) with different discharge power (0, 400, 600, and 800 W) on the structure and physicochemical properties of wheat starch were evaluated in this study. After APPJ treatments, significant declines in peak viscosity, breakdown viscosity, and final viscosity of wheat starch pasting parameters were observed with increase of plasma treatment power. Being treated with discharge power of 800 W, the PV and BD value of wheat starch paste significantly dropped to 2,578 and 331 cP, respectively. Apparently, APPJ could raise the solubility of wheat starch, while reduce the swelling capacity, and also lower the G' and Gâ³ value of wheat starch gel. Roughness and apparent scratch was observed on the surface of the treated wheat starch granules. Although APPJ treatment did not alter wheat starch's crystallization type, it abated the relative crystallinity. APPJ treatment might be useful in producing modified wheat starch with lower viscosity and higher solubility.
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Pea protein is one of plant proteins with high nutritional value, but its lower solubility and poor emulsifying properties limit its application in food industry. Based on wet-heating glycosylation of pea protein and inulin, effects of discharge power of atmospheric pressure plasma jet (APPJ) on structure, solubility, and emulsifying ability of pea protein-inulin glycosylation conjugate were explored. Results indicated that the APPJ discharge power did not affect the primary structure of pea protein. However, changes in secondary and spatial structure of pea protein were observed. When APPJ discharge power was 600 W, the solubility of glycosylation conjugate was 75.0% and the emulsifying stability index was 98.9 min, which increased by 14.85 and 21.95% than that of only glycosylation sample, respectively. These findings could provide technical support for APPJ treatment combination with glycosylation to enhance the physicochemical properties of plant-based proteins.
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Purpose: Identifying the etiology of acute ischemic stroke (AIS) before endovascular treatment (EVT) is important but challenging. In CT perfusion imaging processed by perfusion software, we observed a phenomenon called patchy profile sign (PPS), that is, the hypoperfusion morphology in RAPID software is a discontinuous sheet pattern. This phenomenon is predominantly observed in patients diagnosed with intracranial atherosclerotic stenosis (ICAS). The study intends to assess whether the PPS can be used to differentiate ICAS from intracranial embolism. Method: Patients with AIS due to M1 segment occlusion of the MCA who underwent mechanical thrombectomy were retrospectively enrolled. The receiver operating characteristic (ROC) curve analysis was performed to assess the value of PPS in predicting ICAS. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy of the PPS for prediction of ICAS were calculated. Results: A total of 51 patients were included in the study. The PPS was observed in 10 of 19 (52.6%) patients with ICAS, and in 2 of 32 (6.3%) patients with intracranial embolism (p < 0.001). Interobserver agreement for identifying PPS was excellent (κ = 0.944). The sensitivity, specificity, PPV, NPV, and accuracy of the PPS for predicting ICAS were 52.6, 93.8, 83.3, 76.9, and 78.4%, respectively. Conclusion: The PPS on RAPID software is an imaging marker with high specificity for ICAS. Larger sample sizes are imperative to validate the findings.
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Purpose: Though copy number variants (CNVs) have been suggested to play a significant role in inborn errors of immunity (IEI), the precise nature of this role remains largely unexplored. We sought to determine the diagnostic contribution of CNVs using genome-wide chromosomal microarray analysis (CMA) in children with IEI. Methods: We performed exome sequencing (ES) and CMA for 332 unrelated pediatric probands referred for evaluation of IEI. The analysis included primary, secondary, and incidental findings. Results: Of the 332 probands, 134 (40.4%) received molecular diagnoses. Of these, 116/134 (86.6%) were diagnosed by ES alone. An additional 15/134 (11.2%) were diagnosed by CMA alone, including two likely de novo changes. Three (2.2%) participants had diagnostic molecular findings from both ES and CMA, including two compound heterozygotes and one participant with two distinct diagnoses. Half of the participants with CMA contribution to diagnosis had CNVs in at least one non-immune gene, highlighting the clinical complexity of these cases. Overall, CMA contributed to 18/134 diagnoses (13.4%), increasing the overall diagnostic yield by 15.5% beyond ES alone. Conclusion: Pairing ES and CMA can provide a comprehensive evaluation to clarify the complex factors that contribute to both immune and non-immune phenotypes. Such a combined approach to genetic testing helps untangle complex phenotypes, not only by clarifying the differential diagnosis, but in some cases by identifying multiple diagnoses contributing to the overall clinical presentation.
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Cromossomos , Testes Genéticos , Humanos , Criança , Sequenciamento do Exoma , Análise em Microsséries , FenótipoRESUMO
In this study, an experiment was designed to optimize the synthesis of seleno-Capparis spionosa L. polysaccharide (Se-CSPS) by response surface methodology. Three independent variables (reaction time, reaction temperature and ratio of Na(2)SeO(3) to CSPS) were tested. Furthermore, the thermal stability, particle size, shape and cytotoxic activity of Se-CSPS in vitro were investigated. The optimum reaction conditions were obtained shown as follows: reaction time 7.5 h, reaction temperature 71 °C, and ratio of Na(2)SeO(3) to CSPS 0.9 g/g. Under these conditions, the Se content in Se-CSPS reached 5.547 mg/g, which was close to the predicted value (5.518 mg/g) by the model. The thermal stability, particle size and shape of Se-CSPS were significantly different from those of CSPS. Additionally, a MTT assay indicated that the Se-CSPS could inhibit the proliferation of human gastric cancer SGC-7901 cells in a dose-dependent manner.
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Capparis/química , Proliferação de Células/efeitos dos fármacos , Citotoxinas , Polissacarídeos , Selênio , Linhagem Celular Tumoral , Citotoxinas/síntese química , Citotoxinas/química , Citotoxinas/farmacologia , Relação Dose-Resposta a Droga , Humanos , Polissacarídeos/síntese química , Polissacarídeos/química , Polissacarídeos/farmacologia , Selênio/química , Selênio/farmacologiaRESUMO
Capparis spionosa L. is a traditional medicinal plant in China and central Asia. In this study, an experiment was designed to investigate the optimization of the extraction of anti-tumor polysaccharides from the fruit of Capparis spionosa L. (CSPS) by response surface methodology (RSM). Four independent variables (extraction temperature, extraction time, ratio of water to sample and extraction cycles) were explored. Meanwhile, the in vivo anti-tumor activity of CSPS was investigated. The results showed that the experimental data could be fitted to a second-order polynomial equation using multiple regression analysis. The optimum extraction conditions were as follows: extraction temperature 92 °C, extraction time 140 min, ratio of water to sample 26 mL/g, and three extraction cycle. Under these conditions, the yield of polysaccharides reached 13.01%, which was comparable to the predicted yield (12.94%, p > 0.05). This indicated that the model was adequate for the extraction process. Additionally, CSPS could prolong the survival time of H22 bearing mice in vivo. The anti-tumor activities of CSPS were dose-dependent.