Screening, genotyping and haematological analysis of glucose-6-phosphate dehydrogenase deficiency in the blood donors of Wuxi City, China.

BACKGROUND AND OBJECTIVES: To investigate the prevalence, genotype and haematological characteristics of glucose-6-phosphate dehydrogenase (G6PD) deficiency in the blood donor population of Wuxi area (Jiangsu Province, China) and to assess the impact of their red blood cell (RBC) units on clinical transfusion. MATERIALS AND METHODS: We conducted genotyping and large-scale screening for G6PD enzyme activity in the blood donors of Wuxi City. In addition, we assessed the haematological parameters of G6PD-deficient and non-deficient blood donors, and investigated the adverse transfusion reactions in patients transfused with G6PD-deficient blood. RESULTS: We investigated 17,113 blood donors, among whom 44 (0.26%) were tested positive for G6PD deficiency. We identified 40 G6PD gene variants, among which c.1388G>A, c.1376G>T, c.1024C>T and c.95A>G were common. In addition, we identified two novel G6PD gene variants, c.1312G>A and c.1316G>A. The G6PD-deficient and non-deficient blood samples showed a significant difference in the RBC, mean corpuscular volume (MCV), mean corpuscular Hb (MCH), RBC distribution width, total bilirubin (TBIL), direct bilirubin (DBIL) and indirect bilirubin (IBIL) values. However, the two samples showed no significant difference in the haemolysis rate at the end of the storage period. Finally, transfusion with G6PD-deficient RBC units did not lead to any adverse transfusion reactions. CONCLUSION: The positive rate of G6PD deficiency in the blood donor population of Wuxi City is 0.26%, and the genetic variants identified in this population are consistent with the common genetic variants observed in the Chinese population. Blood centres can establish a database on G6PD-deficient blood donors and mark their RBC units to avoid their use for special clinical patients.

SlBBX20 attenuates JA signalling and regulates resistance to Botrytis cinerea by inhibiting SlMED25 in tomato.

Jasmonic acid (JA) plays an important role in regulating plant growth and defence responses. Here, we show that a transcription factor that belongs to the B-box (BBX) family named SlBBX20 regulates resistance to Botrytis cinerea in tomato by modulating JA signalling. The response to JA was significantly suppressed when SlBBX20 was overexpressed in tomato. By contrast, the JA response was enhanced in SlBBX20 knockout lines. RNA sequencing analysis provided more evidence that SlBBX20 modulates the expression of genes that are involved in JA signalling. We found that SlBBX20 interacts with SlMED25, a subunit of the Mediator transcriptional co-activator complex, and prevents the accumulation of the SlMED25 protein and transcription of JA-responsive genes. JA contributes to the defence response against necrotrophic pathogens. Knocking out SlBBX20 or overexpressing SlMED25 enhanced tomato resistance to B. cinerea. The resistance was impaired when SlBBX20 was overexpressed in plants that also overexpressed SlMED25. These data show that SlBBX20 attenuates JA signalling by regulating SlMED25. Interestingly, in addition to developing enhanced resistance to B. cinerea, SlBBX20-KO plants also produced higher fruit yields. SlBBX20 is a potential target gene for efforts that aim to develop elite crop varieties using gene editing technologies.

SlTCP24 and SlTCP29 synergistically regulate compound leaf development through interacting with SlAS2 and activating transcription of SlCKX2 in tomato.

The complexity of compound leaves results primarily from the leaflet initiation and arrangement during leaf development. However, the molecular mechanism underlying compound leaf development remains a central research question. SlTCP24 and SlTCP29, two plant-specific transcription factors with the conserved TCP motif, are shown here to synergistically regulate compound leaf development in tomato. When both of them were knocked out simultaneously, the number of leaflets significantly increased, and the shape of the leaves became more complex. SlTCP24 and SlTCP29 could form both homodimers and heterodimers, and such dimerization was impeded by the leaf polarity regulator SlAS2, which interacted with SlTCP24 and SlTCP29. SlTCP24 and SlTCP29 could bind to the TCP-binding cis-element of the SlCKX2 promoter and activate its transcription. Transgenic plants with SlTCP24 and SlTCP29 double-gene knockout had a lowered transcript level of SlCKX2 and an elevated level of cytokinin. This work led to the identification of two key regulators of tomato compound leaf development and their targeted genes involved in cytokinin metabolic pathway. A model of regulation of compound leaf development was proposed based on observations of this study.

Establishment of a visual gene knockout system based on CRISPR/Cas9 for the rare actinomycete Nonomuraea gerenzanensis.

OBJECTIVES: To develop a modified CRISPR/Cas9 system with the ß-glucuronidase (GusA) reporter and a dual sgRNA cassette for Nonomuraea gerenzanensis (N. gerenzanensis). RESULTS: With the aid of a visual GusA reporter, the complicated and tedious process of cloning and gene identification could be abandoned entirely in the genetic editing of N. gerenzanensis. Moreover, introducing a dual sgRNA cassette into the CRISPR/Cas9 system significantly improved gene deletion efficiency compared to the single sgRNA element. Furthermore, the length of the homologous flanking sequences set to the lowest value of 500 bp in this system could still reach the relatively higher conjugation transfer frequency. CONCLUSIONS: The enhanced CRISPR/Cas9 system could efficiently perform genetic manipulation on the rare actinomycete N. gerenzanensis.

Asymmetrical cross-modal influence on neural encoding of auditory and visual features in natural scenes.

Natural scenes contain multi-modal information, which is integrated to form a coherent perception. Previous studies have demonstrated that cross-modal information can modulate neural encoding of low-level sensory features. These studies, however, mostly focus on the processing of single sensory events or rhythmic sensory sequences. Here, we investigate how the neural encoding of basic auditory and visual features is modulated by cross-modal information when the participants watch movie clips primarily composed of non-rhythmic events. We presented audiovisual congruent and audiovisual incongruent movie clips, and since attention can modulate cross-modal interactions, we separately analyzed high- and low-arousal movie clips. We recorded neural responses using electroencephalography (EEG), and employed the temporal response function (TRF) to quantify the neural encoding of auditory and visual features. The neural encoding of sound envelope is enhanced in the audiovisual congruent condition than the incongruent condition, but this effect is only significant for high-arousal movie clips. In contrast, audiovisual congruency does not significantly modulate the neural encoding of visual features, e.g., luminance or visual motion. In summary, our findings demonstrate asymmetrical cross-modal interactions during the processing of natural scenes that lack rhythmicity: Congruent visual information enhances low-level auditory processing, while congruent auditory information does not significantly modulate low-level visual processing.

Fresh hetero-metallic compound as luminescent sensor for detection of CS2 and inhibitory activity on children diabetes.

The combination of the dicarboxylate ligand of 2-aminoterephthalic acid (H2L), Cd(NO3)2·4H2O, and KNO3 under hydrothermal condition formed a fresh hetero-metallic coordination polymer, namely, [CdK2(L)2(H2O)4]n (1). The compound exhibited a high luminescence intensity at normal environment as well as high sensitivity and selectivity in the measurement of CS2. The effect of the fresh compound on childhood diabetes was investigated. A blood glucose monitor was utilized to assess the blood glucose level of the body after the addition of the compound. Moreover, the relative expression level of the insulin recipient on hepatocytes was assessed by real time RT-PCR.

Integration of metabolic pathway manipulation and promoter engineering for the fine-tuned biosynthesis of malic acid in Bacillus coagulans.

Bacillus coagulans, a thermophilic facultative anaerobe, is a favorable chassis strain for the biosynthesis of desired products. In this study, B. coagulans was converted into an efficient malic acid producer by metabolic engineering and promoter engineering. Promoter mapping revealed that the endogenous promoter Pldh was a tandem promoter. Accordingly, a promoter library was developed, covering a wide range of relative transcription efficiencies with small increments. A reductive tricarboxylic acid pathway was established in B. coagulans by introducing the genes encoding pyruvate carboxylase (pyc), malate dehydrogenase (mdh), and phosphoenolpyruvate carboxykinase (pckA). Five promoters of various strengths within the library were screened to fine-tune the expression of pyc to improve the biosynthesis of malic acid. In addition, genes involved in the competitive metabolic pathways were deleted to focus the substrate and energy flux toward malic acid. Dual-phase fed-batch fermentation was performed to increase the biomass of the strain, further improving the titer of malic acid to 25.5 g/L, with a conversion rate of 0.3 g/g glucose. Our study is a pioneer research using promoter engineering and genetically modified B. coagulans for the biosynthesis of malic acid, providing an effective approach for the industrialized production of desired products using B. coagulans.

Purification and characterization of a cold-adapted κ-carrageenase from Pseudoalteromonas sp. ZDY3.

κ-Carrageenase (EC3.2.1.83) is a class of glycoside hydrolase, which can be used for hydrolysis of κ-carrageenan to κ-carrageenan oligosaccharides. In this study, a bacterium, identified as Pseudoalteromonas sp. ZDY3 isolated from rotten algae, was capable to degrade κ-carrageenan. The κ-carrageenase produced by Pseudoalteromonas sp. ZDY3 was purified to homogeneity and named as CgkZDY3. The accurate molecular mass of CgkZDY3 was determined through LC-HRMS, and a posttranslational removal of C-terminal end of the protein was discovered. CgkZDY3 had strict hydrolysis specificity to κ-carrageenan, the values of Km and kcat/Km of CgkZDY3 were 3.67 mg mL-1 and 53.0 mL mg-1 s-1, respectively. CgkZDY3 was a cold-adapted κ-carrageenase with excellent storage stability of both the temperature below 35 °C and a wide pH range, and was an endo-type κ-carrageenase with high hydrolysis rate, oligosaccharides with different degrees of polymerization can be obtained by controlling the hydrolysis time, and the final products were κ-neocarrabiose and κ-neocarratetraose. These properties are of great significance for production of κ-carrageenan oligosaccharides with different polymerization degrees under process control.

Photoredox-catalyzed redox-neutral difluoroalkylation to construct perfluoroketones with difluoroenoxysilanes.

A mild and facile approach to construct various perfluoroketones via photo-catalyzed difluoroalkylation of difluoroenoxysilanes is developed. The reaction includes a strategy of combination of two fluorine-containing functional groups, which confers the reaction with characteristics like high efficiency, mild conditions, and broad scope. A variety of fluoroalkyl halides including perfluoroalkyl iodides, bromo difluoro esters and amides can be employed as radical precursors. Control experiments indicate that a single-electron transfer pathway may be involved in the reaction.

Functional daidzein enhances the anticancer effect of topotecan and reverses BCRP-mediated drug resistance in breast cancer.

Topotecan (TPT), a semisynthetic derivative of camptothecin, has been used in cancer chemotherapy, but side effects and drug resistance limit its clinical application. Daidzein (DAI), a natural isoflavone and bioactive food component widely existing in fruits, nuts, soybeans and soy-based products, is a type of phytoestrogen. Combination treatment with DAI and TPT showed a strong synergistic effect on tumor cells, with a 0.10˜0.66 combined index, by increasing TPT inhibition on Topo Ⅰ, resulting in more cells arresting at the G2/M phase and inducing more cells to undergo apoptosis. In addition, the resistance of MCF7/ADR cells to TPT was reversed (the resistance index decreased from 7.17 to 0.77) by inhibiting the expression of ERα and BCRP to increase TPT accumulation intracellularly. Moreover, the combination of DAI and TPT showed a stronger inhibitory effect (P < 0.01) on tumor growth in both MCF7 and MCF7/ADR xenograft models than the 9 mg/kg TPT monotherapy group. Our results may provide a reasonable, new approach to develop safe and efficient nutrition components from foods for breast cancer combination treatment.

A molecular level study of the phase transition process of hydrogen-bonding UCST polymers.

Most of the existing studies have focused on the development of the lower critical solution temperature (LCST) behavior of thermoresponsive polymers, whereas research on the upper critical solution temperature (UCST) behavior from a molecular point of view is extremely rare. In this work, a molecular level study of the UCST thermal transition of poly-N-acryloylglycinamide (PNAGA)-based polymers in water and in a water/methanol mixture (χm = 6.9 mol%) was performed. Turbidimetric measurements revealed the increased UCST and enlarged hysteresis of the thermal transition of the polymers with the addition of methanol. Fourier transform infrared (FTIR) spectral analysis indicated that the increased hydrophilicity among the C-H groups along the polymer chains upon heating induced the UCST-type transition of the polymers in water, while the hydrogen bonding transformation predominated the thermal transition in the mixture. Moreover, in combination with two-dimensional correlation spectroscopy (2Dcos), the structures with different types of inter/intramolecular hydrogen bonds formed among the NAGA units have been distinguished and the transformation of the amide groups in PNAGA-based polymers from C[double bond, length as m-dash]OD-N hydrogen bonds to C[double bond, length as m-dash]OD2O ones during the UCST-type transition upon heating was captured.

Switching between Polymer Architectures with Distinct Thermoresponses.

Thermoresponsive linear polymers and their corresponding aggregates or nanogels typically show similar thermoresponsive profiles. In this study, the authors demonstrate reversible chemical switching between linear polymers and their cross-linked nanogels. The linear polymers exhibit sharp thermal transitions typical of common thermoresponsive polymers but the cross-linked nanogels exhibit "linear" thermal transitions over a relatively broad temperature range. The reversible switching between these two different polymer architectures with distinct thermoresponses represents a unique example of how the responsive properties of smart polymers can be significantly manipulated via polymer architecture engineering.

Revealing the distinct thermal transition behavior between PEGA-based linear polymers and their disulfide cross-linked nanogels.

The distinct thermal transition behavior of thermoresponsive block polymers based on poly(ethylene glycol)methyl ether acrylate (PEGA) and their corresponding disulfide-cross-linked nanogels was studied by using FTIR measurements in combination with two-dimensional correlation spectroscopy (2Dcos). Spectral analysis clearly reveals that the sharp thermal transition of the precursor polymer is accompanied by a forced hydration process induced by the formation of hydrogen bonds between the entrapped water molecules and the C[double bond, length as m-dash]O groups, while the nanogel with a relatively continuous thermal transition is related to the existence of various dehydrating states of the C[double bond, length as m-dash]O groups. The C-H groups in the pyridyl disulfide (PDS) units exhibit a distinct change in the thermoresponsive profile of the precursor and the nanogel to show the effect of the polymer architecture on the thermal transition behavior. Additionally, a portion of the poly(N,N-dimethylacrylamide) (PDMA) segments is entrapped in the nanogel core, indicating that the thiol-disulfide exchange reaction occurs rapidly within the nanogels.

Transcriptome Analysis of Tomato Leaf Spot Pathogen Fusarium proliferatum: De novo Assembly, Expression Profiling, and Identification of Candidate Effectors.

Leaf spot disease caused by the fungus Fusarium proliferatum (Matsushima) Nirenberg is a destructive disease of tomato plants in China. Typical symptoms of infected tomato plants are softened and wilted stems and leaves, leading to the eventual death of the entire plant. In this study, we resorted to transcriptional profile analysis to gain insight into the repertoire of effectors involved in F. proliferatum-tomato interactions. A total of 61,544,598 clean reads were de novo assembled to provide a F. proliferatum reference transcriptome. From these, 75,044 unigenes were obtained, with 19.46% of the unigenes being assigned to 276 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, with 22.3% having a homology with genes from F. fujikuroi. A total of 18,075 differentially expressed genes (DEGs) were identified, 720 of which were found to code for secreted proteins. Of these, 184 were identified as candidate effectors, while 79.89% had an upregulated expression. Moreover, 17 genes that were differentially expressed in RNA-seq studies were randomly selected for validation by quantitative real-time polymerase chain reaction (qRT-PCR). The study demonstrates that transcriptome analysis could be an effective method for identifying the repertoire of candidate effectors and may provide an invaluable resource for future functional analyses of F. proliferatum pathogenicity in F. proliferatum and tomato plant-host interactions.

The structure and volume phase transition behavior of poly(N-vinylcaprolactam)-based hybrid microgels containing carbon nanodots.

In this paper, we investigated the internal structure and the volume phase transition (VPT) behavior of poly(N-vinylcaprolactam-co-vinylimidazole)/polymerizable carbon nanodot (P(VCL-co-VIM)/PCND) microgels with different amounts of PCNDs. Our study shows that compared to the pure P(VCL-co-VIM) microgel, the hybrid microgels undergo a two-step VPT as the temperature increases and a core-shell(-corona) structure of the hybrid microgels is formed by copolymerization with PCNDs. A change in the amount of PCNDs has effects on both of the volume phase transition temperature and internal structure of microgels. Moreover, based on FT-IR in combination with perturbation correlation moving window (PCMW) and two-dimensional correlation spectral (2Dcos) analyses, the difference in VPT behavior between the shell and the core (corona) structure of the hybrid microgels at the molecular level is elucidated. The core/shell of the hybrid microgels fixed with hydrophilic PCNDs has a higher transition temperature during heating and a more compact structure due to the additional crosslinkers PCNDs.

STAT3 represents a molecular switch possibly inducing astroglial instead of oligodendroglial differentiation of oligodendroglial progenitor cells in Theiler's murine encephalomyelitis.

AIMS: Insufficient oligodendroglial differentiation of oligodendroglial progenitor cells (OPCs) is suggested to be responsible for remyelination failure and astroglial scar formation in Theiler's murine encephalomyelitis (TME). The aim of the present study is to identify molecular key regulators of OPC differentiation in TME, and to dissect their mechanism of action in vitro. METHODS: TME virus (TMEV) infected SJL/J-mice were evaluated by rotarod analysis, histopathology, immunohistology and gene expression microarray analysis. The STAT3 pathway was activated using meteorin and inhibited using STAT3 inhibitor VII in the glial progenitor cell line BO-1 and in primary rat OPCs in vitro. RESULTS: As expected, immunohistology demonstrated progressively decreasing myelin basic protein-positive white matter in TME. In contrast, intralesional NG2-positive OPCs as well as GFAP-positive astrocytes were increased. Gene Set Enrichment Analysis revealed 26 Gene Ontology terms including 'JAK-STAT cascade' to be significantly positively correlated with the density of NG2-positive OPCs. Immunohistology revealed an increased amount of activated, phosphorylated STAT3-expressing astrocytes, OPCs, and microglia/macrophages within the lesions. Meteorin-induced activation of STAT3-signalling in BO-1 cells and primary rat OPCs resulted in an enhanced GFAP and reduced CNPase expression. In contrast, an oppositional result was observed in BO-1 cells treated with STAT3 inhibitor VII. CONCLUSIONS: The STAT3 pathway is a key regulator of OPC-differentiation, suggested to shift their differentiation from an oligodendroglial towards an astrocytic fate, thereby inducing astrogliosis and insufficient remyelination in TME.

Improved wavelength coded optical time domain reflectometry based on the optical switch.

This paper presents an improved wavelength coded time-domain reflectometry based on the 2 × 1 optical switch. In this scheme, in order to improve the signal-noise-ratio (SNR) of the beat signal, the improved system used an optical switch to obtain wavelength-stable, low-noise and narrow optical pulses for probe and reference. Experiments were set up to demonstrate a spatial resolution of 2.5m within a range of 70km and obtain the beat signal with line width narrower than 15 MHz within a range of 50 km in fiber break detection. A system for wavelength-division-multiplexing passive optical network (WDM-PON) monitoring was also constructed to detect the fiber break of different channels by tuning the current applied on the gating section of the distributed Bragg reflector (DBR) laser.

Life cycle assessment of biochar for sustainable agricultural application: A review.

Biochar application is an effective strategy to address Agro-climatic challenges. However, the agro-environmental impacts of different biochar technology models are lacking of systematic summaries and reviews. Therefore, this paper comprehensively reviews recent developments derived from published literature, delving into the economic implications and environmental benefits of three distinct process namely technologies-pyrolysis, gasification, and hydrothermal carbonization. This paper specifically focuses on the agricultural life cycle assessment (LCA) methodology, and the influence of biochar preparation technologies and products on energy consumption and agricultural carbon emissions. LCA analysis shows that process and feedstock pose a predominant role on the properties and production rate of biochar, while gasification technology exhibits excellent economic attributes compared to the other two technologies. Biochar applications in agricultural has the beneficial effect of sequestering carbon and reducing emissions, especially in the area of mitigating the carbon footprint of farmland. However, the complexity of the composition of the prepared feedstock and the mismatch between the biochar properties and the application scenarios are considered as potential sources of risks. Notably, mechanism of carbon sequestration and emission reduction by soil microorganisms and agro-environmental sequestration by biochar application remains unclear, calling for in-depth studies. We review novel aspects that have not been covered by previous reviews by comparing the technical, economic, and environmental benefits of pyrolysis, gasification, and hydrothermal carbonization systematically. Overall, this study will provide a valuable framework to environmental implications of biochar preparation, application, and life cycle assessments.

Linguistic feedback supports rapid adaptation to acoustically degraded speech.

Humans can quickly adapt to recognize acoustically degraded speech, and here we hypothesize that the quick adaptation is enabled by internal linguistic feedback - Listeners use partially recognized sentences to adapt the mapping between acoustic features and phonetic labels. We test this hypothesis by quantifying how quickly humans adapt to degraded speech and analyzing whether the adaptation process can be simulated by adapting an automatic speech recognition (ASR) system based on its own speech recognition results. We consider three types of acoustic degradation, i.e., noise vocoding, time compression, and local time-reversal. The human speech recognition rate can increase by >20% after exposure to just a few acoustically degraded sentences. Critically, the ASR system with internal linguistic feedback can adapt to degraded speech with human-level speed and accuracy. These results suggest that self-supervised learning based on linguistic feedback is a plausible strategy for human adaptation to acoustically degraded speech.