Ebola Virus Epidemiology, Transmission, and Evolution during Seven Months in Sierra Leone.

The 2013-2015 Ebola virus disease (EVD) epidemic is caused by the Makona variant of Ebola virus (EBOV). Early in the epidemic, genome sequencing provided insights into virus evolution and transmission and offered important information for outbreak response. Here, we analyze sequences from 232 patients sampled over 7 months in Sierra Leone, along with 86 previously released genomes from earlier in the epidemic. We confirm sustained human-to-human transmission within Sierra Leone and find no evidence for import or export of EBOV across national borders after its initial introduction. Using high-depth replicate sequencing, we observe both host-to-host transmission and recurrent emergence of intrahost genetic variants. We trace the increasing impact of purifying selection in suppressing the accumulation of nonsynonymous mutations over time. Finally, we note changes in the mucin-like domain of EBOV glycoprotein that merit further investigation. These findings clarify the movement of EBOV within the region and describe viral evolution during prolonged human-to-human transmission.

Extracellular vesicle autoantibodies.

Autoantibodies are immunoglobulin proteins produced by autoreactive B cells responding to self-antigens. Extracellular vesicles (EVs) are membranous structures released by almost all types of cells and extensively distributed in various biological fluids. Studies have indicated that EVs loaded with self-antigens not only play important roles in antigen presentation and autoantibody production but can also form functional immune complexes with autoantibodies (termed EV autoantibodies). While numerous papers have summarized the production and function of pathogenic autoantibodies in diseases, especially autoimmune diseases, reviews on EV autoantibodies are rare. In this review, we outline the existing knowledge about EVs, autoantibodies, and EV antigens, highlighting the formation of EV autoantibodies and their functions in autoimmune diseases and cancers. In conclusion, EV autoantibodies may be involved in the occurrence of disease(s) and also serve as potential non-invasive markers that could help in the diagnosis and/or prognosis of disease. Additional studies designed to define in more detail the molecular characteristics of EV autoantibodies and their contribution to disease are recommended.

Result of a Pilot External Quality Assessment Scheme for Clinical Diagnosis of Inherited Metabolic Disorders in China.

BACKGROUND: We aimed to evaluate the diagnostic capabilities of Chinese laboratories for inherited metabolic disorders (IMDs) using gas chromatography-mass spectrometry (GC-MS) on urine samples. Meanwhile, based on the result of the pilot external quality assessment (EQA) scheme, we hope to establish a standardized and reliable procedure for future EQA practice. METHODS: We recruited laboratories that participated in the EQA of quantitative analysis of urinary organic acids with GC-MS before joining the surveys. In each survey, a set of five real urine samples was distributed to each participant. The participants should analyze the sample by GC-MS and report the "analytical result", "the most likely diagnosis", and "recommendation for further tests" to the NCCL before the deadline. RESULTS: A total of 21 laboratories participated in the scheme. The pass rates were 94.4% in 2020 and 89.5% in 2021. For all eight IMDs tested, the analytical proficiency rates ranged from 84.7% - 100%, and the interpretational performance rate ranged from 88.2% - 97.0%. The performance on hyperphenylalaninemia (HPA), 3-methylcrotonyl-CoA carboxylase deficiency (MCCD), and ethylmalonic encephalopathy (EE) samples were not satisfactory. CONCLUSIONS: In general, the participants of this pilot EQA scheme are equipped with the basic capability for qualitative organic acid analysis and interpretation of the results. Limited by the small size of laboratories and samples involved, this activity could not fully reflect the state of clinical practice of Chinese laboratories. NCCL will improve the EQA scheme and implement more EQA activities in the future.

Results of a Pilot External Quality Assessment Scheme for Genetic Testing of Newborns with Spinal Muscular Atrophy.

BACKGROUND: This study aims to evaluate the ability of laboratories to perform spinal muscular atrophy (SMA) genetic testing in newborns based on dried blood spot (DBS) samples, and to provide reference data and advance preparation for establishing the pilot external quality assessment (EQA) scheme for SMA genetic testing of newborns in China. METHODS: The pilot EQA scheme contents and evaluation principles of this project were designed by National Center for Clinical Laboratories (NCCL), National Health Commission. Two surveys were carried out in 2022, and 5 batches of blood spots were submitted to the participating laboratory each time. All participating laboratories conducted testing upon receiving samples, and test results were submitted to NCCL within the specified date. RESULTS: The return rates were 75.0% (21/28) and 95.2% (20/21) in the first and second surveys, respectively. The total return rate of the two examinations was 83.7% (41/49). Nineteen laboratories (19/21, 90.5%) had a full score passing on the first survey, while in the second survey twenty laboratories (20/20, 100%) scored full. CONCLUSIONS: This pilot EQA survey provides a preliminary understanding of the capability of SMA genetic testing for newborns across laboratories in China. A few laboratories had technical or operational problems in testing. It is, therefore, of importance to strengthen laboratory management and to improve testing capacity for the establishment of a national EQA scheme for newborn SMA genetic testing.

Ultrastretchable and Self-Healing Conductors with Double Dynamic Network for Omni-Healable Capacitive Strain Sensors.

Skin-mountable capacitive-type strain sensors with great linearity and low hysteresis provide inspiration for the interactions between human and machine. For practicality, high sensing performance, large stretchability, and self-healing are demanded but limited by stretchable electrode and dielectric and interfacial compatibility. Here, we demonstrate an extremely stretchable and self-healing conductor via both hard and soft tactics that combine conductive nanowire assemblies with double dynamic network based on π-π attractions and Ag-S coordination bonds. The obtained conductor outperforms the reported stretchable conductors by delivering an elongation of 3250%, resistance change of 223% at 2000% strain, high durability, and multiresponsive self-healability. Especially, this conductor accommodates large strain of 1500% at extremely knotted and twisted deformations. By sandwiching hydrogel conductors with a newly developed dielectric, ultrahigh stretchability and omni-healability are simultaneously achieved for the first time for a capacitive strain sensor inspired by metal-thiolate coordination chemistry, showing great potentials in wearable electronics and soft robotics.

Abelson tyrosine kinase controls BCR signalling and B-cell differentiation by promoting B-cell metabolism.

As a nonreceptor tyrosine kinase, Abelson tyrosine kinase (c-Abl) was first studied in chronic myelogenous leukaemia, and its role in lymphocytes has been well characterised. c-Abl is involved in B-cell development and CD19-associated B-cell antigen receptor (BCR) signalling. Although c-Abl regulates different metabolic pathways, the role of c-Abl is still unknown in B-cell metabolism. In this study, B-cell-specific c-Abl knockout (KO) mice (Mb1Cre+/- c-Ablfl/fl ) were used to investigate how c-Abl regulates B-cell metabolism and BCR signalling. We found that the levels of activation positive BCR signalling proximal molecules, phosphorylated spleen tyrosine kinase (pSYK) and phosphorylated Bruton tyrosine kinase (pBTK), were decreased, while the level of key negative regulator, phosphorylated SH2-containing inositol phosphatase 1 (pSHIP1), was increased in Mb1Cre+/- c-Ablfl/fl mice. Furthermore, we found c-Abl deficiency weakened the B-cell spreading, formation of BCR signalosomes, and the polymerisation of actin during BCR activation, and also impaired the differentiation of germinal center (GC) B-cells both in quiescent condition and after immunisation. Moreover, B-cell mitochondrial respiration and the expression of B-cell metabolism-regulating molecules were downregulated in c-Abl deficiency mice. Overall, c-Abl, which involved in actin remodelling and B-cell metabolism, positively regulates BCR signalling and promotes GC differentiation.

Ag nanocubes monolayer-modified PDMS as flexible SERS substrates for pesticides sensing.

A new approach is presented to fabricate flexible surface-enhanced Raman scattering (SERS) substrate of Ag nanocubes monolayer-modified polydimethylsiloxane (Ag NCs/PDMS) through a powerful three-phase interface self-assemble method. The morphologies and crystal structures were characterized by scanning electron microscopy and X-ray diffraction. The self-assembled Ag NCs/PDMS substrate exhibited high SERS activity and good signal homogeneity, which was successfully used for quantitative detection of thiram; the detection limit reached 10 ng/mL, and the linear range is 10-1000 ng/mL. Furthermore, the flexible SERS substrates were successfully employed to detect thiram residues on factual apple samples, and trace amount (1 ng/cm2) of thiram residues was detected on apple peels. The excellent SERS detection ability of self-assembled Ag NCs/PDMS substrate indicated that it will play an important role in pesticide detection in the future.

Core-shell architectured NH2-UiO-66@ZIF-8/multi-walled carbon nanotubes nanocomposite-based sensitive electrochemical sensor towards simultaneous determination of Pb2+ and Cu2.

Amino functionalized zirconium-based metal-organic framework (NH2-UiO-66) and zinc-based zeolitic imidazolate framework (ZIF-8) were integrated to develop a core-shell architectured hybrid material (NH2-UiO-66@ZIF-8, NU66@Z8). The morphology and structure evolutions of core-shell NU6@Z8 were investigated by FE-SEM, XRD, FTIR, and XPS. The NU66@Z8 combined with carboxylated multi-walled carbon nanotubes (CMWCNT) was deposited on a glassy carbon electrode (GCE) for fabricating an electrochemical platform towards detecting Pb2+ and Cu2+. The NU66@Z8/CMWCNT/GCE revealed significantly improved electrochemical performance for determination of Pb2+ and Cu2+ compared with the individual components, which can be attributed to the strong adsorption capacity, unique core-shell structure, and large electrochemical active surface area of NU66@Z8/CMWCNT. Under the optimal conditions, the developed sensor exhibited excellent sensing capability with a low limit of detection (Pb2+,1 nM; Cu2+, 10 nM) and a wide determination range (Pb2+,0.003-70 µM; Cu2+, 0.03-50 µM). The sensor showed high selectivity towards common interfering ions and good repeatability. The real sample recoveries of proposed sensor were in the range 95.0-103% for Pb2+ (RSD ≤ 5.3%) and 94.2-106% for Cu2+ (RSD ≤ 5.9%), suggesting that the NU66@Z8/CMWCNT is suitable for examining trace heavy metals in natural environment.

Genome-wide identification of sucrose nonfermenting-1-related protein kinase (SnRK) genes in barley and RNA-seq analyses of their expression in response to abscisic acid treatment.

BACKGROUND: Sucrose nonfermenting-1 (SNF1)-related protein kinases (SnRKs) play important roles in regulating metabolism and stress responses in plants, providing a conduit for crosstalk between metabolic and stress signalling, in some cases involving the stress hormone, abscisic acid (ABA). The burgeoning and divergence of the plant gene family has led to the evolution of three subfamilies, SnRK1, SnRK2 and SnRK3, of which SnRK2 and SnRK3 are unique to plants. Therefore, the study of SnRKs in crops may lead to the development of strategies for breeding crop varieties that are more resilient under stress conditions. In the present study, we describe the SnRK gene family of barley (Hordeum vulgare), the widespread cultivation of which can be attributed to its good adaptation to different environments. RESULTS: The barley HvSnRK gene family was elucidated in its entirety from publicly-available genome data and found to comprise 50 genes. Phylogenetic analyses assigned six of the genes to the HvSnRK1 subfamily, 10 to HvSnRK2 and 34 to HvSnRK3. The search was validated by applying it to Arabidopsis (Arabidopsis thaliana) and rice (Oryza sativa) genome data, identifying 50 SnRK genes in rice (four OsSnRK1, 11 OsSnRK2 and 35 OsSnRK3) and 39 in Arabidopsis (three AtSnRK1, 10 AtSnRK2 and 26 AtSnRK3). Specific motifs were identified in the encoded barley proteins, and multiple putative regulatory elements were found in the gene promoters, with light-regulated elements (LRE), ABA response elements (ABRE) and methyl jasmonate response elements (MeJa) the most common. RNA-seq analysis showed that many of the HvSnRK genes responded to ABA, some positively, some negatively and some with complex time-dependent responses. CONCLUSIONS: The barley HvSnRK gene family is large, comprising 50 members, subdivided into HvSnRK1 (6 members), HvSnRK2 (10 members) and HvSnRK3 (34 members), showing differential positive and negative responses to ABA.

CircJARID2 Regulates Hypoxia-Induced Injury in H9c2 Cells by Affecting miR-9-5p-Mediated BNIP3.

ABSTRACT: Myocardial infarction (MI) is a common cardiovascular disease, and many circular RNAs (circRNAs) have been found to participate in the pathological process. This study was to research circRNA jumonji and AT-rich interaction domain containing 2 (circJARID2) in MI. MI cell model was established by hypoxia treatment in H9c2 cells. CircJARID2 and microRNA-9-5p (miR-9-5p) levels were examined using real-time polymerase chain reaction. Cell viability detection was performed by Cell Counting Kit-8 (CCK-8) and 5-ethynyl-2'-deoxyuridine (Edu) assays. Cell apoptosis was evaluated by flow cytometry and caspase-3 activity assay. Apoptotic markers and B-cell lymphoma-2 (Bcl-2) interacting protein 3 (BNIP3) were quantified by western blot. Inflammatory cytokines were determined via enzyme-linked immunosorbent assay. The genic interaction was analyzed through dual-luciferase reporter and RNA immunoprecipitation assays. Hypoxia induced the upregulation of circJARID2 expression in H9c2 cells. The hypoxia-induced cell viability inhibition, apoptosis promotion, and inflammatory response were all counterbalanced by knockdown of circJARID2. CircJARID2 interacted with miR-9-5p, and its function in regulating the hypoxia-induced cell injury was also dependent on targeting miR-9-5p. BNIP3 acted as a target gene of miR-9-5p, and circJARID2 had positive effect on BNIP3 expression by binding to miR-9-5p. MiR-9-5p played a protective role for H9c2 cells against the hypoxia-induced injury via targeting BNIP3. CircJARID2 overexpression contributed to the hypoxia-induced H9c2 cell injury by sponging miR-9-5p to upregulate BNIP3 expression, showing a novel molecular network of MI pathomechanism.