Identification of variants in 94 Chinese patients with hereditary spherocytosis by next-generation sequencing.

Hereditary spherocytosis (HS) is the most common type of hereditary erythrocyte membrane disease and has varied phenotypic features and genetic patterns. We herein performed a retrospective study of 94 patients with HS and aimed to investigate the genetic variations and genotype-phenotype correlations using targeted next-generation sequencing. In 79/94 (84%) patients, 83 HS variants including 67 novel variants were identified. Pathogenic variants of SPTB, ANK1, SLC4A1, SPTA1, and EPB42 were found in 32/79(41%), 22/79(28%), 15/79 (19%), 8/79 (9%), and 3/79 (4%) of the patients respectively, revealing that SPTB is the most frequently mutated HS gene in Eastern China. Most SPTB and ANK1 gene variations were nonsense and frameshift variations. Missense variants were the main variant type of SLC4A1, SPTA1, and EPB42 genes. Interestingly, one SPTA1 variant (p. Arg1757Cys) showed an autosomal dominant inheritance pattern and one EPB42 variant (p. Gln377His) was apparent as a hotspot variation. Furthermore, genotype-phenotype analysis was performed among the five mutated gene groups. Besides the finding that patients with the SLC4A1 variant had the highest mean corpuscular hemoglobin levels, no clear correlations between genotype and phenotype were observed.

Single-short-cavity dual-comb fiber laser with over 120†kHz repetition rate difference based on polarization multiplexing.

An all-fiber single-short-cavity dual-comb laser with a high repetition rate of up to 500 MHz and a high repetition rate difference of over 120 kHz was demonstrated. The laser setup exploits high birefringence of a polarization-maintaining gain fiber to generate asynchronous combs based on the polarization-multiplexing method. By adopting short-linear-cavity and all-birefringent configuration, a repetition rate difference several orders of magnitude larger than that of a previous work was achieved. The soliton dual-comb showed good mutual coherence and stability, which reveals the potential to enhance the acquisition rate and accuracy of dual-comb measurement systems.

Fist Detection of Hb Suqian[ß42(CD1) Phe→Leu, HBB:c.129T > A] in Han Chinese.

We have identified a variant on the ß-globin gene in a Chinese female. Sequencing of the HBB gene revealed a Phe→Leu substitution at codon 42[ß42(CD1) Phe→Leu, HBB:c.129T > A] which has been named Hb Suqian for where the proband was born.

The molecular characteristics, clinical manifestations, and risk factors of hypervirulent Klebsiella pneumoniae infections in a large teaching hospital in southwest China.

Hypervirulent Klebsiella pneumoniae (hvKp) has been globally disseminated recently, especially in Asia. The purpose of this study was to identify the molecular characteristics, clinical manifestations, and clinical risk factors of hvKp infections among patients in a large teaching hospital. A retrospective study was conducted in 123 patients infected with K. pneumoniae at the Affiliated Hospital of Southwest Medical University (Luzhou, China) from October 2016 to November 2018. An isolate that positive for both PCR amplification of aerobactin gene and Galleria mellonella infection model was defined as hvKp. Overall, 43.1% (53/123) of K. pneumoniae isolates were hvKp. String tests were performed on all isolates, and MLSTs of all hvKp were conducted. The K1 ST23 isolates were the dominant clone of hvKp (35.8%). Univariate analysis revealed the following risk factors for hvKp: hepatic abscess (OR = 41.818 [95% CI, 5.379-335.086]), bacteremia (OR = 19.94 [95% CI, 5.565-71.446]), metastatic spread (OR = 19.938 [95% CI, 6.344-62.654]), CRP (OR = 1.008 [95% CI, 1.001-1.015]), nitroimidazole treatment (OR = 7.907 [95% CI, 1.652-37.843]), diabetes (OR = 3.067 [95% CI, 1.38-6.817]), and admission to positive culture interval (OR = 3.636 [95% CI, 1.524-8.678]). Moreover, Multivariate analysis implicated hepatic abscess (OR = 74.332 [95% CI, 3.121-1769.588]), bacteremia (OR = 28.388 [95% CI, 3.039-264.200]), and metastatic spread (OR = 19.391 [95% CI, 3.633-103.498]) as independent risk factors for hvKp infections. Thirteen of twenty-one tested antibiotics were founded resistant to non-hvKp, which is significantly greater than hvKp. Importantly, the ESBL-hvKp and MDR-hvKp were responsible for 7.5% and 15.1% in the hvKp group, respectively.

Antioxidant and antibacterial study of 10 flavonoids revealed rutin as a potential antibiofilm agent in Klebsiella pneumoniae strains isolated from hospitalized patients.

The emergence of multidrug resistance (MDR) and extensive drug resistance (XDR) in Klebsiella pneumoniae strains has posed great threats to conventional antibiotics. Previous studies have shown that plant-derived flavonoids have inhibitory functions against pathogens. However, in K. pneumoniae, the antibacterial activity of different flavonoids against growth and biofilm formation remains a mystery. The aim of the present study was to evaluate the antioxidant abilities of different flavonoids, to screen active ingredients and to identify their inhibitory effects on K. pneumoniae growth and biofilm formation. In total, 10 flavonoids representing 4 major categories were screened and used in this study. The antioxidant capacity of each flavonoid was evaluated through a DPPH (2,2-diphenyl-1-picrylhydrazyl) assay. Rutin showed the highest level of free radical scavenging capacity, followed by kaempferol, luteolin, quercetin, apigenin, hesperidin, sinensetin, naringenin, naringin and 3,5,6,7,8,3',4'-heptamethoxyflavone. The inhibitory effects of rutin and naringin on bacterial growth were also compared. The lowest MICs of rutin were found against K. pneumoniae ATCC700603 (1024 µg/mL) and E. coli ATCC25922 (512 µg/mL). However, the MBICs were not found. Rutin showed strong inhibitory ability against both the growth curve and biofilm production. The expression profiles of 15 biofilm-related genes were analyzed in biofilm cells both with and without rutin treatment. The luxS gene and wabG gene were downregulated significantly by rutin treatment. Correlation analysis showed that mrkA gene expression was positively correlated with biofilm biomass accumulation. Our study indicated that biofilm production is correlated with the expression of several genes rather than one. MrkA gene expression was positively correlated with biofilm biomass accumulation. Our study screened rutin as a potential agent to inhibit K. pneumoniae biofilm formation.

Comparison of MPL-ANN and PLS-DA models for predicting the severity of patients with acute pancreatitis: An exploratory study.

OBJECTIVE: Acute pancreatitis (AP) is a common inflammatory disorder that may develop into severe AP (SAP), resulting in life-threatening complications and even death. The purpose of this study was to explore two different machine learning models of multilayer perception-artificial neural network (MPL-ANN) and partial least squares-discrimination (PLS-DA) to diagnose and predict AP patients' severity. METHODS: The MPL-ANN and PLS-DA models were established using candidate markers from 15 blood routine parameters and five serum biochemical indexes of 133 mild acute pancreatitis (MAP) patients, 167 SAP (including 88 moderately SAP) patients, and 69 healthy controls (HCs). The independent parameters and combined model's diagnostic efficiency in AP severity differentiation were analyzed using the area under the receiver operating characteristic curve (AUC). RESULTS: The neutrophil to lymphocyte ratio (NLR) is the most useful marker in 20 parameters for screening AP patients [AUC = 0.990, 95% confidence interval (CI): 0.984-0.997, sensitivity 94.3%, specificity 98.6%]. The MPL-ANN model based on six optimal parameters exhibited better diagnostic and predict performance (AUC = 0.984, 95% CI: 0.960-1.00, sensitivity 92.7%, specificity 93.3%, accuracy 93.0%) than the PLS-DA model based on five optimal parameters (AUC = 0.912, 95% CI: 0.853-0.971, sensitivity 87.8%, specificity 84.4%, accuracy 84.8%) in discriminating MAP patients from SAP patients. CONCLUSION: The results demonstrated that the MPL-ANN model based on routine blood and serum biochemical indexes provides a reliable and straightforward daily clinical practice tool to predict AP patients' severity.

Identification of a Novel ß-Thalassemia Mutation at Codon 130 (+T) (HBB: c.391insT) in Han Chinese.

ß-Thalassemia (ß-thal) is an inherited blood disorder characterized by reduced or absent synthesis of the ß chains of hemoglobin (Hb). Although more than 900 ß-globin gene mutations around the world have been identified, here we report a novel mutation detected in a Chinese subject of Han ethnicity. This allele develops by insertion of one nucleotide (+T) at codon 130 (HBB: c.391insT) in the third exon of the ß-globin gene. The mutation causes a frameshift that leads to a termination codon at codon 139. The identification of the novel mutation will facilitate future diagnosis of ß-thal and will also be useful the genetic counseling and prenatal diagnosis.

The levels of bioactive ingredients in Citrus aurantium L. at different harvest periods and antioxidant effects on H2 O2 -induced RIN-m5F cells.

BACKGROUND: Citrus aurantium L. (Aurantii fructus) is a multi-purpose citrus fruit with high medicinal and nutritional value, but currently there are no data that can be used to investigate the appropriate harvest time to obtain high-quality citrus bioactive ingredients from it. RESULTS: Phytochemicals and the levels of the main bioactive ingredients were investigated by ultra high performance liquid chromatography electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-ESI-Q-TOF/MS). The flavanone, polymethoxyflavone, coumarin, synephrine, and limonin content in the citrus fruit was analyzed at different harvest periods, and significant differences, ranging from 0.03 ± 0.01 to 116.26 ± 40.20 g kg-1 (DW), were shown. These compounds were present in higher amounts in June and then decreased gradually, while the biomass accumulation of most of them showed an increasing tendency around harvest time. The H2 O2 -induced RIN-m5F cells model was employed to evaluate their antioxidant capacity. Citrus fruit harvested from June 11 to July 7 possessed an excellent antioxidant capacity by inhibiting the intensity of intracellular reactive oxygen species (ROS) (P < 0.01) and improving superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH) activity (P < 0.01). The chemical composition and antioxidant capacity of citrus leaves, stems, and roots were also evaluated, and these showed great variation compared with other citrus fruits. Multivariate statistical analysis indicated that harvesting time was related closely to the phytochemical contents and antioxidant capacity. CONCLUSION: Citrus fruit can be appropriately harvested from June to early July when the levels of bioactive ingredients and antioxidant activity reach higher values. This research provides practical information for producing high-quality citrus products. © 2020 Society of Chemical Industry.

The role and relationship with efflux pump of biofilm formation in Klebsiella pneumoniae.

This study aimed to identify the role and relationship with efflux pump of biofilm formation in Klebsiella pneumoniae. Sixty-one K. pneumoniae clinical isolates were collected between January and June of 2017 from the affiliated hospital of southwest medical university in Luzhou, China. The minimum inhibitory concentration (MIC) and minimum biofilm eradication concentration (MBEC) were determined using broth microdilution method. Crystal violet (CV) staining and confocal laser scanning microscope (CLSM) were used to monitor biofilm formation. Efflux pump expression was investigated qualitatively and quantitatively by polymerase chain reaction (PCR) and reverse transcriptase quantitative PCR (RT-qPCR). Crystal violet staining was performed to evaluate the effect of efflux pump inhibitor carbonyl cyanide m-chlorophenyl hydrazine (CCCP) on K. pneumoniae biofilms. Our results showed that crystal violet staining and CLSM had good consistency in biofilm detection. Biofilm formation was an independent biological behavior of the strain and measured at 24 h was reasonable. Biofilms up-regulated antimicrobial resistance and expression of efflux pump gene acrA, emrB, oqxA, and qacEΔ1 in K. pneumoniae. CCCP inhibited biofilms but dose-dependent effect was obvious. Altogether, our data demonstrates that biofilm formation, as well as its interaction with efflux pump, promotes antimicrobial resistance in K. pneumoniae.

In-situ growth of high-density ultrafine Ag3PO4 nanoparticles on 3D TiO2 hierarchical spheres for enhanced photocatalytic degradation of organic pollutants.

Silver phosphate (Ag3PO4, APO) has attracted intense attention as a visible-light-driven photocatalyst, but its large-scale application is limited by severe charge recombination and inevitable photo-corrosion. Various rational APO-based heterostructures composed of APO nanoparticles (NPs) and band-matched semiconductor support are designed to address the above issues. Nevertheless, the size, density, stability, and dispersion of APO NPs are critical challenges for the photocatalytic performance of APO-based photocatalysts. Here, three-dimensional (3D) self-assembled TiO2 hierarchical spheres (THS) prepared by a simple one-step hydrothermal method are employed as innovative support, and ultrafine high-density APO NPs with an average size of about 3 nm are successfully deposited and uniformly dispersed throughout THS to form hierarchical THS/APO composites. The novel THS/APO microstructure provides abundant reactive sites for photocatalytic reactions and promotes the photogenerated charge separation and transfer due to the ultrafine size of APO NPs and the TiO2/APO Type-II heterojunction. As a result, the THS/APO composites show significant improvement in photocatalytic activity and stability in methylene blue (MB) degradation. The reaction constant of THS/APO composites far exceeds that of either THS or APO, roughly 16 and 7 times higher than that of THS and APO under full-spectrum light, and 41 and 4 times higher under visible light. Our results strongly suggest new insights into the low-cost, large-scale application of high-efficiency APO-based photocatalyst.

Mild heat stress limited the post-acidification caused by Lactobacillus rhamnosus hsryfm 1301 in fermented milk.

OBJECTIVE: Fermented milk is the optimal vehicle for delivering probiotic bacteria. However, the viable count of probiotic bacteria such as some lactic acid bacteria and the post-acidification of fermented milk are a contradiction. The objective of this study was to restrict the post-acidification of the fermented milk containing living Lactobacillus rhamnosus hsryfm 1301. RESULTS: Mild heat stress treatment (46 °C, 1 h) was chosen to help control the post-acidification caused by L. rhamnosus hsryfm 1301. When fermented milk was produced by single L. rhamnosus hsryfm 1301, the heat stress treatment reduced the post-acidification from 0.39 to 0.11% lactic acid, and the viable cells were maintained above 2.0 × 108 CFU mL-1 during 21 days of storage. Although the post-acidification limitation of heat treatment was relatively weak in fermented milk produced by L. rhamnosus hsryfm 1301 and S. thermophilus grx02 (from 0.26 to 0.10% lactic acid), this treatment was still effective. Furthermore, no whey separation in the fermented milk was caused by the treatment. CONCLUSIONS: Mild heat stress treatment could limit the post-acidification caused by L. rhamnosus hsryfm 1301 by decreasing its metabolism and proliferation. This treatment is a promising strategy to improve the shelf life of probiotic fermented milk.

Isopropanol-promoted DNA extraction by polydopamine functionalized magnetic particles based on metal coordination.

High-quality DNA is an important guarantee to start downstream experiments in many biological and medical research areas. Magnetic particle-based DNA extraction methods from blood mainly depend on electrostatic adsorption in a low-pH environment. However, the strong acidic environment can influence the DNA stability. Herein, a polydopamine-functionalized magnetic particle (PDA@Fe3O4)-based protocol was developed for DNA extraction from whole blood samples. In the protocol, Mg2+ and Ca2+ were utilized to bridge the adsorption of DNA by PDA@Fe3O4 via the metal-mediated coordination. Isopropanol was found to efficiently promote DNA adsorption by triggering the change of the conformation of DNA from B-form to more compact A-form. In 50 % isopropanol solution, the DNA adsorption efficiency was nearly 100 % in the presence of 0.5 mM Ca2+ or 1.5 mM Mg2+. The role of metal ions and isopropanol in DNA adsorption was explored. The protocol averts the strong acidic environment and PCR inhibitors, such as high concentrations of salt or polyethylene glycol. It demonstrates superiority in DNA yield (59.13 ± 3.63 ng µL-1) over the commercial kit (27.33 ± 4.98 ng µL-1) and phenol-chloroform methods (37.90 ± 0.47 ng µL-1). In addition, to simplify the operastion, an automated nucleic acid extraction device was designed and fabricated to extract whole genomic DNA from blood. The feasibility of the device was verified by extracting DNA from cattle and pig blood samples. The extracted DNA was successfully applied to discriminate the beef authenticity by a duplex PCR system. The results demonstrate that the DNA extraction protocol and the automated device have great potential in blood samples.

Smartphone-based colorimetric detection platform using color correction algorithms to reduce external interference.

Smartphone-based digital image colorimetry is a powerful, fast, low-cost approach to detecting target analytes. However, lighting conditions and camera parameters easily affect the detection results, significantly curtailing its applicability in multiple scenarios. In this study, an Android-based mobile application (SMP-CC) is developed, which offers a comprehensive package that includes image acquisition, color correction, and colorimetric analysis functions. Using a custom color card, a built-in algorithm in SMP-CC can minimize the color difference between the standard color block image captured by different smartphones under different lighting conditions and the standard value by an LS171 colorimeter less than 4.36. The algorithm significantly eliminates the impacts of external lighting conditions and differences in cell phone models. Furthermore, the feasibility of SMP-CC was verified by successful colorimetric detection of urine pH, glucose, and protein, demonstrating its potential in smartphone-based digital image colorimetry.

Peroxidase like Zn doped Prussian blue facilitates salinity tolerance in winter wheat through seed dressing.

Salt stress severely limits the growth and yield of wheat in saline-alkali soil. While nanozymes have shown promise in mitigating abiotic stress by scavenging reactive oxygen species (ROS) in plants, their application in alleviating salt stress for wheat is still limited. This study synthesized a highly active nanozyme catalyst known as ZnPB (Zn-modified Prussian blue) to improve the yield and quality of wheat in saline soil. According to the Michaelis-Menten equation, ZnPB demonstrates exceptional peroxidase-like enzymatic activity, thereby mitigating oxidative damage caused by salt stress. Additionally, studies have shown that the ZnPB nanozyme is capable of regulating intracellular Na+ efflux and K+ retention in wheat, resulting in a decrease in proline and soluble protein levels while maintaining the integrity of macromolecules within the cell. Consequently, field experiments demonstrated that the ZnPB nanozyme increased winter wheat yield by 12.15 %, while also significantly enhancing its nutritional quality. This research offers a promising approach to improving the salinity tolerance of wheat, while also providing insights into its practical application.

Data-Driven Engineering of Phages with Tunable Capsule Tropism for Klebsiella pneumoniae.

Klebsiella pneumoniae, a major clinical pathogen known for causing severe infections, is attracting heightened attention due to its escalating antibiotic resistance. Phages are emerging as a promising alternative to antibiotics; however, their specificity to particular hosts often restricts their use. In this study, a collection of 114 phages is obtained and subjected to analysis against 238 clinical K. pneumoniae strains, revealing a spectrum of lytic behaviors. A correlation between putative tail protein clusters and lysis patterns leads to the discovery of six receptor-binding protein (RBP) clusters that determine host capsule tropism. Significantly, RBPs with cross-capsular lysis capabilities are identified. The newly-identified RBPs provide a toolbox for customizing phages to target diverse capsular types. Building on the toolbox, the engineered phages with altered RBPs successfully shifted and broadened their host capsule tropism, setting the stage for tunable phage that offer a precise and flexible solution to combat K. pneumoniae infections.

Autophagy plays an antiviral defence role against tomato spotted wilt orthotospovirus and is counteracted by viral effector NSs.

Autophagy, an intracellular degradation process, has emerged as a crucial innate immune response against various plant pathogens, including viruses. Tomato spotted wilt orthotospovirus (TSWV) is a highly destructive plant pathogen that infects over 1000 plant species and poses a significant threat to global food security. However, the role of autophagy in defence against the TSWV pathogen, and whether the virus counteracts this defence, remains unknown. In this study, we report that autophagy plays an important role in antiviral defence against TSWV infection; however, this autophagy-mediated defence is counteracted by the viral effector NSs. Transcriptome profiling revealed the up-regulation of autophagy-related genes (ATGs) upon TSWV infection. Blocking autophagy induction by chemical treatment or knockout/down of ATG5/ATG7 significantly enhanced TSWV accumulation. Notably, the TSWV nucleocapsid (N) protein, a major component of the viral replication unit, strongly induced autophagy. However, the TSWV nonstructural protein NSs was able to effectively suppress N-induced autophagy in a dose-dependent manner. Further investigation revealed that NSs inhibited ATG6-mediated autophagy induction. These findings provide new insights into the defence role of autophagy against TSWV, a representative segmented negative-strand RNA virus, as well as the tospoviral pathogen counterdefence mechanism.

QRS detection in single-lead, telehealth electrocardiogram signals: Benchmarking open-source algorithms.

BACKGROUND AND OBJECTIVES: A key step in electrocardiogram (ECG) analysis is the detection of QRS complexes, particularly for arrhythmia detection. Telehealth ECGs present a new challenge for automated analysis as they are noisier than traditional clinical ECGs. The aim of this study was to identify the best-performing open-source QRS detector for use with telehealth ECGs. METHODS: The performance of 18 open-source QRS detectors was assessed on six datasets. These included four datasets of ECGs collected under supervision, and two datasets of telehealth ECGs collected without clinical supervision. The telehealth ECGs, consisting of single-lead ECGs recorded between the hands, included a novel dataset of 479 ECGs collected in the SAFER study of screening for atrial fibrillation (AF). Performance was assessed against manual annotations. RESULTS: A total of 12 QRS detectors performed well on ECGs collected under clinical supervision (F1 score ≥0.96). However, fewer performed well on telehealth ECGs: five performed well on the TELE ECG Database; six performed well on high-quality SAFER data; and performance was poorer on low-quality SAFER data (three QRS detectors achieved F1 of 0.78-0.84). The presence of AF had little impact on performance. CONCLUSIONS: The Neurokit and University of New South Wales QRS detectors performed best in this study. These performed sufficiently well on high-quality telehealth ECGs, but not on low-quality ECGs. This demonstrates the need to handle low-quality ECGs appropriately to ensure only ECGs which can be accurately analysed are used for clinical decision making.