Detection of calcitonin gene-related peptide based on increased antigen-driven interaction with antibody variable regions.

Introduction: Calcitonin gene-related peptide (CGRP) is involved in trigeminal neuralgia and migraine, and measuring the CGRP concentration in the serum is crucial for the early prediction of these conditions. Current methods for CGRP detection are primarily radioimmunoassay, which needs radioactive substances and enzyme-linked immunosorbent assays (ELISAs) which need long detection time and some have a narrow detection range. Methods: The genes of anti-CGRP antibody variable regions were cloned into pDong1 vector to obtain pDong1/Fab-CGRP, with which phage-Fab was prepared, and the concentration of CGRP was detected by competitive ELISA. The pDong1/Fab-CGRP was modified to obtain pDong1/OS-CGRP, with which the co-expression solution containing phage-displayed heavy chain variable fragments (phage-VH) and light chain was obtained. CGRP was detected by OS-ELISA based on phage-VH, antibody light chain, and anti-light chain antibody. The VL gene was cloned into the pMAL vector to obtain pMAL-VL (CGRP), with which maltose binding protein fused with VL (MBP-VL) was prepared. CGRP was detected by OS-ELISA employing MBP-VL and phage-VH. Results: OS-ELISAs that measure the CGRP concentration by quantifying the interaction between variable regions were investigated. OS-ELISA using phage-VH and secreted light chains in the same culture system exhibited a limit of detection (LOD) of 0.05 nM, offering higher sensitivity than competitive assay with an LOD of 0.75 nM, whereas using phage-VH and separately prepared MBP-VL exhibited an LOD of 0.15 nM and a broader detection range of 0.15-500 nM than competitive ELISA, whose detection range was 0.75-10 nM. Discussion: The combination of the two OS assays achieved high sensitivity and a broad detection range for CGRP, which may have significance in clinical applications.

A New Strategy for the Design of Triple-Phase Eutectic High-Entropy Alloys Based on Infinite Solid Solution and Pseudo-Ternary Method.

Eutectic high-entropy alloys (EHEAs) have combined both high-entropy alloys and eutectic alloy contributions, with excellent castability and high-temperature application potential. Yet, multielement/triple-phase eutectic high-entropy alloy (TEHEA) designs remain puzzling. This work proposed a new strategy based on an infinite solid solution and pseudo-ternary model to reveal the puzzle of TEHEAs. The designed triple-phase eutectic high-entropy alloys (TEHEAs) with more than seven elements were identified as face-centered cubic (FCC), ordered body-centered cubic (B2), and Laves phase structures. In this work, the alloy C showcases outstanding comprehensive mechanical properties, offering a novel avenue for the design of high-performance EHEAs.

CircBCAR3 sponges miR-27a-3p and mediates ferroptosis in human B-prolymphocytic leukaemia cells via SLC7A11.

B-lymphocytic leukaemia is one of the most commonly diagnosed blood malignancies, and our knowledge of B-prolymphocytic leukaemia remained barely comprehensive. CircRNAs and miRNAs were identified of important regulatory roles in tumours. This study focused on the possibly existing interaction of circBCAR3 and miR-27a-3p, and downstream molecules thereafter in B-prolymphocytic leukaemia cells. CircBCAR3 and miR-27a-3p expression was evaluated in JVM-2 cell line and normal lymphocytes. Dual-luciferase luminescence assay was conducted for validation of circBCAR3 and miR-27a-3p interaction, as well as western blot and flow cytometry for evaluation and validation of their association with SLC7A11, reactive-oxygen species and Fe2+ regarding ferroptosis. CircBCAR3 were upregulated in JVM-2 cells and were reversely correlated with the expression of miR-27a-3p. CircBCAR3 targeted at miR-27a-3p and was consequently associated with SLC7A11 expression positively, inhibiting ferroptosis and peroxidative damage in JVM-2 cells. This study identified a circBCAR3-miR-27a-3p-SLC7A11 axis regulating ferroptosis and peroxidation of B-prolymphocytic leukaemia cells which might be a key mechanism facilitating the survival of tumour cells. However, further validation based on more diverse cell lines and animal models might be required.

The haplotype-resolved T2T reference genome highlights structural variation underlying agronomic traits of melon.

Melon (Cucumis melo L.) is an important vegetable crop that has an extensive history of cultivation. However, the genome of wild and semi-wild melon types that can be used for the analysis of agronomic traits is not yet available. Here we report a chromosome-level T2T genome assembly for 821 (C. melo ssp. agrestis var. acidulus), a semi-wild melon with two haplotypes of ~373 Mb and ~364 Mb, respectively. Comparative genome analysis discovered a significant number of structural variants (SVs) between melo (C. melo ssp. melo) and agrestis (C. melo ssp. agrestis) genomes, including a copy number variation located in the ToLCNDV resistance locus on chromosome 11. Genome-wide association studies detected a significant signal associated with climacteric ripening and identified one candidate gene CM_ac12g14720.1 (CmABA2), encoding a cytoplasmic short chain dehydrogenase/reductase, which controls the biosynthesis of abscisic acid. This study provides valuable genetic resources for future research on melon breeding.

Genome-Wide Identification and Characterization Analysis of WUSCHEL-Related Homeobox Family in Melon (Cucumis melo L.).

WUSCHEL-related homeobox (WOX) proteins are very important in controlling plant development and stress responses. However, the WOX family members and their role in response to abiotic stresses are largely unknown in melon (Cucumis melo L.). In this study, 11 WOX (CmWOX) transcript factors with conserved WUS and homeobox motif were identified and characterized, and subdivided into modern clade, ancient clade and intermediate clade based on bioinformatic and phylogenetic analysis. Evolutionary analysis revealed that the CmWOX family showed protein variations in Arabidopsis, tomato, cucumber, melon and rice. Alignment of protein sequences uncovered that all CmWOXs had the typical homeodomain, which consisted of conserved amino acids. Cis-element analysis showed that CmWOX genes may response to abiotic stress. RNA-seq and qRT-PCR results further revealed that the expression of partially CmWOX genes are associated with cold and drought. CmWOX13a and CmWOX13b were constitutively expressed under abiotic stresses, CmWOX4 may play a role in abiotic processes during plant development. Taken together, this study offers new perspectives on the CmWOX family's interaction and provides the framework for research on the molecular functions of CmWOX genes.

Development and validation of a nomogram to predict suicidal behavior in female patients with mood disorder.

Introduction: This study aims to explore the risk factors associated with suicidal behavior and establish predictive models in female patients with mood disorders, specifically using a nomogram of the least absolute shrinkage and selection operator (LASSO) regression. Methods: A cross-sectional survey was conducted among 396 female individuals diagnosed with mood disorders (F30-F39) according to the International Classification of Diseases and Related Health Problems 10th Revision (ICD-10). The study utilized the Chi-Squared Test, t-test, and the Wilcoxon Rank-Sum Test to assess differences in demographic information and clinical characteristics between the two groups. Logistic LASSO Regression Analyses were utilized to identify the risk factors associated with suicidal behavior. A nomogram was constructed to develop a prediction model. The accuracy of the prediction model was evaluated using a Receiver Operating Characteristic (ROC) curve. Result: The LASSO regression analysis showed that psychotic symptoms at first-episode (ß = 0.27), social dysfunction (ß = 1.82), and somatic disease (ß = 1.03) increased the risk of suicidal behavior. Conversely, BMI (ß = -0.03), age of onset (ß = -0.02), polarity at onset (ß = -1.21), and number of hospitalizations (ß = -0.18) decreased the risk of suicidal behavior. The area under ROC curve (AUC) of the nomogram predicting SB was 0.778 (95%CI: 0.730-0.827, p < 0.001). Conclusion: The nomogram based on demographic and clinical characteristics can predict suicidal behavior risk in Chinese female patients with mood disorders.

Root-secreted bitter triterpene modulates the rhizosphere microbiota to improve plant fitness.

Underground microbial ecosystems have profound impacts on plant health1-5. Recently, essential roles have been shown for plant specialized metabolites in shaping the rhizosphere microbiome6-9. However, the potential mechanisms underlying the root-to-soil delivery of these metabolites remain to be elucidated10. Cucurbitacins, the characteristic bitter triterpenoids in cucurbit plants (such as melon and watermelon), are synthesized by operon-like gene clusters11. Here we report two Multidrug and Toxic Compound Extrusion (MATE) proteins involved in the transport of their respective cucurbitacins, a process co-regulated with cucurbitacin biosynthesis. We further show that the transport of cucurbitacin B from the roots of melon into the soil modulates the rhizosphere microbiome by selectively enriching for two bacterial genera, Enterobacter and Bacillus, and we demonstrate that this, in turn, leads to robust resistance against the soil-borne wilt fungal pathogen, Fusarium oxysporum. Our study offers insights into how transporters for specialized metabolites manipulate the rhizosphere microbiota and thereby affect crop fitness.

First clinical isolation report of Shewanella xiamenensis from Chinese giant salamander.

Shewanella xiamenensis, a newly virulent zoonotic pathogen belonging to the genus Shewanella is the causative organism of emerging intra-abdominal infection, acute skin ulceration, rotten limbs and ascites in humans and animals. The global spread of S. xiamenensis entails severe economic impact. However, it was rarely reported as a cause of infection and no reports were found that S. xiamenensis isolated from clinical samples. The isolate was identified as a S . xiamenensis strain by 16S rDNA amplification and DNA sequencing identification method. Even if co-infection by other bacteria could not be ruled out, this is the first report of acute disease caused by S . xiamenensis in the Chinese giant salamander in China. By using the Kirby-Bauer disc diffusion method, the sensitivity of the isolate to clinical antibiotics was evaluated. Antibiotic susceptibility test indicated that the isolate was resistant to 32 antibacterial drugs such as kanamycin, florfenicol and ceftriaxone suggesting that the isolate was a multi-drug resistant strain.

Application of metagenomic sequencing toward rapid and sensitive diagnosis of goose avastrovirus infection in China.

The gosling gout, a newly emerged disease, has widely broken out in China since 2017. Typical signs for the disease include diarrhea, anorexia, depression, dehydration, emaciation and paralysis. At autopsy, uratosis was the main pathological change which could be found at kidney, pericardium, air sac, muscle and leg joint. In this study, gosling gout was firstly diagnosed by metagenomic analysis. Samples of kidney, Fabricius bursa, spleen and jejunum were collected and submitted to next-generation DNA sequencing. Our results demonstrated that goose avastrovirus was highly related with this disease. We confirmed the sequencing results by reverse transcription polymerase chain reaction method and artificial infection experiment and got consistent results. In summary, metagenomic sequencing method combined with traditional molecular identification was applied toward diagnosis of a novel gosling gout disease in China and revealed that goose avastrovirus was highly related with this disease. It has been proved to be a powerful tool for rapid and sensitive diagnosis of animal diseases, especially for some exceptional pathogens. In addition, host range, variation, molecular pathogenesis and potential zoonotic infection of this novel goose astrovirus need to be further studied.

Rapid conversion of IgG to biosensor using an antibody-binding protein-based probe.

We successfully developed a fluorescent probe that can quickly convert full-length antibodies to Quenchbodies, which represent a type of fluorescent immunosensor with high binding affinity and specificity depending on the reaction of antigens and antibodies. An anti-testosterone IgG was successfully converted to an immunosensor that detects testosterone with a limit of detection (LOD) of 0.76 nM and concentration for 50% of maximal effect (EC50) of 61.5 nM. Another IgG-based immunosensor detected ractopamine with an LOD of 15.5 pM and EC50 of 48.6 nM.

QTLs and candidate genes analyses for fruit size under domestication and differentiation in melon (Cucumis melo L.) based on high resolution maps.

BACKGROUND: Melon is a very important horticultural crop produced worldwide with high phenotypic diversity. Fruit size is among the most important domestication and differentiation traits in melon. The molecular mechanisms of fruit size in melon are largely unknown. RESULTS: Two high-density genetic maps were constructed by whole-genome resequencing with two F2 segregating populations (WAP and MAP) derived from two crosses (cultivated agrestis × wild agrestis and cultivated melo × cultivated agrestis). We obtained 1,871,671 and 1,976,589 high quality SNPs that show differences between parents in WAP and MAP. A total of 5138 and 5839 recombination events generated 954 bins in WAP and 1027 bins in MAP with the average size of 321.3 Kb and 301.4 Kb respectively. All bins were mapped onto 12 linkage groups in WAP and MAP. The total lengths of two linkage maps were 904.4 cM (WAP) and 874.5 cM (MAP), covering 86.6% and 87.4% of the melon genome. Two loci for fruit size were identified on chromosome 11 in WAP and chromosome 5 in MAP, respectively. An auxin response factor and a YABBY transcription factor were inferred to be the candidate genes for both loci. CONCLUSION: The high-resolution genetic maps and QTLs analyses for fruit size described here will provide a better understanding the genetic basis of domestication and differentiation, and provide a valuable tool for map-based cloning and molecular marker assisted breeding.

Satisfiability Attack-Resistant Camouflaged Two-Dimensional Heterostructure Devices.

Reverse engineering (RE) is one of the major security threats to the semiconductor industry due to the involvement of untrustworthy parties in an increasingly globalized chip manufacturing supply chain. RE efforts have already been successful in extracting device level functionalities from an integrated circuit (IC) with very limited resources. Camouflaging is an obfuscation method that can thwart such RE. Existing work on IC camouflaging primarily involves transformable interconnects and/or covert gates where variation in doping and dummy contacts hide the circuit structure or build cells that look alike but have different functionalities. Emerging solutions, such as polymorphic gates based on a giant spin Hall effect and Si nanowire field effect transistors (FETs), are also promising but add significant area overhead and are successfully decamouflaged by the satisfiability solver (SAT)-based RE techniques. Here, we harness the properties of two-dimensional (2D) transition-metal dichalcogenides (TMDs) including MoS2, MoSe2, MoTe2, WS2, and WSe2 and their optically transparent transition-metal oxides (TMOs) to demonstrate area efficient camouflaging solutions that are resilient to SAT attack and automatic test pattern generation attacks. We show that resistors with resistance values differing by 5 orders of magnitude, diodes with variable turn-on voltages and reverse saturation currents, and FETs with adjustable conduction type, threshold voltages, and switching characteristics can be optically camouflaged to look exactly similar by engineering TMO/TMD heterostructures, allowing hardware obfuscation of both digital and analog circuits. Since this 2D heterostructure devices family is intrinsically camouflaged, NAND/NOR/AND/OR gates in the circuit can be obfuscated with significantly less area overhead, allowing 100% logic obfuscation compared to only 5% for complementary metal oxide semiconductor (CMOS)-based camouflaging. Finally, we demonstrate that the largest benchmarking circuit from ISCAS'85, comprised of more than 4000 logic gates when obfuscated with the CMOS-based technique, is successfully decamouflaged by SAT attack in <40 min; whereas, it renders to be invulnerable even in more than 10 h when camouflaged with 2D heterostructure devices, thereby corroborating our hypothesis of high resilience against RE. Our approach of connecting material properties to innovative devices to secure circuits can be considered as a one of a kind demonstration, highlighting the benefits of cross-layer optimization.

Genetic dissection of climacteric fruit ripening in a melon population segregating for ripening behavior.

Melon is as an alternative model to understand fruit ripening due to the coexistence of climacteric and non-climacteric varieties within the same species, allowing the study of the processes that regulate this complex trait with genetic approaches. We phenotyped a population of recombinant inbred lines (RILs), obtained by crossing a climacteric (Védrantais, cantalupensis type) and a non-climcteric variety (Piel de Sapo T111, inodorus type), for traits related to climacteric maturation and ethylene production. Individuals in the RIL population exhibited various combinations of phenotypes that differed in the amount of ethylene produced, the early onset of ethylene production, and other phenotypes associated with ripening. We characterized a major QTL on chromosome 8, ETHQV8.1, which is sufficient to activate climacteric ripening, and other minor QTLs that may modulate the climacteric response. The ETHQV8.1 allele was validated by using two reciprocal introgression line populations generated by crossing Védrantais and Piel de Sapo and analyzing the ETHQV8.1 region in each of the genetic backgrounds. A Genome-wide association study (GWAS) using 211 accessions of the ssp. melo further identified two regions on chromosome 8 associated with the production of aromas, one of these regions overlapping with the 154.1 kb interval containing ETHQV8.1. The ETHQV8.1 region contains several candidate genes that may be related to fruit ripening. This work sheds light into the regulation mechanisms of a complex trait such as fruit ripening.

Genetic mapping and candidate gene analysis for melon resistance to Phytophthora capsici.

Phytophthora blight is one of the most serious diseases affecting melon (Cucumis melo) production. Due to the lack of highly resistant germplasms, the progress on disease-resistant research is slow. To understand the genetics of melon resistance to Phytophthora capsici, an F2 population containing 498 individuals was developed by crossing susceptible line E31 to highly resistant line ZQK9. Genetic analysis indicated that the resistance in ZQK9 was controlled by a dominant gene, tentatively named MePhyto. Through bulked-segregant analysis (BSA-Seq) and chromosome walking techniques, the MePhyto gene was mapped to a 52.44 kb interval on chromosome 12. In this region, there were eight genes and their expression patterns were validated by qRT-PCR. Among them, one wall-associated receptor kinase (WAK) gene MELO3C002430 was significantly induced in ZQK9 after P. capsici inoculation, but not in E31. Based on the non-synonymous mutation site in MELO3C002430, a cleaved amplified polymorphic sequence (CAPS) marker, CAPS2430, was developed and this maker was co-segregated with MePhyto in both F2 population and a collection of 36 melon accessions. Thus MELO3C002430 was considered as the candidate gene and CAPS2430 was a promising marker for marker-assisted selection (MAS) in breeding. These results lay a foundation for revealing the resistance mechanism of melon to P. capsici.

Development of a fluorescent probe for the detection of hPD-L1.

Interaction of human programmed death factor-1 (hPD-1) of T cells and one of its ligands hPD-L1 which is expressed on cancer cells suppresses effector T cell functions. Studies showed that the hPD-1/hPD-L1 pathway is associated with killing mechanisms of tumor cells evading the immune system. Immunotherapy based on the checkpoint inhibitor on hPD-1 has been an important approach to treat cancer; however, not all cancer cells over-express hPD-L1. Detection of hPD-L1 over-expression in cancer cells may be a key factor for deciding on whether immunotherapy should be conducted. In the present study, we produced recombinant hPD-1 using Escherichia coli, and created a fluorescent probe termed quenched hPD-1 (QPD-1) for the detection of hPD-L1. We found that hPD-1 can quench fluorescence of carboxytetramethylrhodamine labeled on its N-terminal and QPD-1 is a convenient tool to rapidly detect hPD-L1 with a limit of detection of 10 nM and detectable range of 10 nM-1000 nM. QPD-1 may also function as a probe to screen for hPD-L1 over-expressing tumor cells and promote appropriate medical procedure through tumor immunotherapy.

Transcriptome analysis clarified genes involved in resistance to Phytophthora capsici in melon.

Phytophthora blight caused by Phytophthora capsici is a devastating disease for melon plant. However, the underlying resistance mechanisms are still poorly understood. In this study, the transcriptome differences between the resistant ZQK9 and susceptible E31 at 0, 3, and 5 days post-inoculation (dpi) were identified by RNA-seq. A total of 1,195 and 6,595 differentially expressed genes (DEGs) were identified in ZQK9 and E31, respectively. P. capsici infection triggered massive transcript changes in the inoculated tissues. Genes related to plant defense responses were activated, which was reflected by a lot of up-regulated DEGs involved in pathogenesis-related (PR) genes, hormones biosynthesis and signal transduction, secondary metabolites biosynthesis and cell wall modification in resistant ZQK9. The dataset generated in this study may provide a basis for identifying candidate resistant genes in melon against P. capsici and lay a foundation for further research on the molecular mechanisms.

Draft genome sequencing and annotation of a low-virulence Morganella morganii strain CQ-M7, a multidrug-resistant isolate from the giant salamander in China.

OBJECTIVES: A multidrug-resistant Morganella morganii strain (CQ-M7), isolated from the kidney of a diseased Chinese giant salamander in China, was examined with whole genome sequencing to better understand drug tolerance and its pathogenicity. METHODS: The draft genome of the investigated strain was assembled using HGA assembler and annotated using Rapid Annotations Subsystems Technology (RAST) server. The contigs were annotated by the appropriate bioinformatics tools available on the National Center for Biotechnology Information (NCBI) website. Antibiotic resistance genes were detected by PCR. Pathogenicity of the isolate was performed on 30 healthy Chinese giant salamanders with different infection dosages. RESULTS: The CQ-M7 strain showed resistance to multiple antimicrobials, especially to aminoglycoside and ß-lactam antibiotics. Seventeen drug-resistance genes were detected, which were related to ß-lactams, aminoglycosides, fluoroquinolones, tetracyclines, peptide antibiotic, and fosfomycin resistance. Sequence analysis showed the assembled genome size to be 4 966 326bp with 51.16% of GC content, containing 4587 protein-coding genes, 71 pseudogenes, five rRNAs, 80 tRNAs, and five noncoding RNAs. The genome sequence was deposited in GenBank under accession number RQIJ00000000. Artificial infection results indicated that the CQ-M7 strain was a low-virulence strain for the Chinese giant salamander. CONCLUSION: It is believed that this is the first draft genome of Chinese giant salamander original Morganella morganii strain harbouring multiple antibiotic resistance genes in China. The reported genome sequence could provide insights into antibiotic resistance mechanisms and control strategies of Morganella morganii.

QTL mapping of resistance to Fusarium oxysporum f. sp. niveum race 2 and Papaya ringspot virus in Citrullus amarus.

KEY MESSAGE: A Citrullus amarus mapping population segregating for resistance to Fusarium oxysporum f. sp. niveum race 2 and Papaya ringspot virus was used to identify novel QTL, important for the improvement in watermelon disease resistance. Multiple disease screens of the USDA Citrullus spp. germplasm collection have highlighted the value of Citrullus amarus (citron melon or wild watermelon) as a resource for enhancing modern watermelon cultivars (Citrullus lanatus) with resistance to a broad range of fungal, bacterial and viral diseases of watermelon. We have generated a genetic population of C. amarus segregating for resistance to two important watermelon diseases: Fusarium wilt (caused by the fungus Fusarium oxysporum f. sp. niveum; Fon race 2) and Papaya ringspot virus-watermelon strain (PRSV-W). QTL mapping of Fon race 2 resistance identified seven significant QTLs, with the major QTL representing a novel genetic source of resistance and an opportunity for gene pyramiding. A single QTL was associated with resistance to PRSV-W, which adhered to expectations of a prior study indicating a single-gene recessive inheritance in watermelon. The resistance loci identified here provide valuable genetic resources for introgression into cultivated watermelon for the improvement in disease resistance.

A comprehensive genome variation map of melon identifies multiple domestication events and loci influencing agronomic traits.

Melon is an economically important fruit crop that has been cultivated for thousands of years; however, the genetic basis and history of its domestication still remain largely unknown. Here we report a comprehensive map of the genomic variation in melon derived from the resequencing of 1,175 accessions, which represent the global diversity of the species. Our results suggest that three independent domestication events occurred in melon, two in India and one in Africa. We detected two independent sets of domestication sweeps, resulting in diverse characteristics of the two subspecies melo and agrestis during melon breeding. Genome-wide association studies for 16 agronomic traits identified 208 loci significantly associated with fruit mass, quality and morphological characters. This study sheds light on the domestication history of melon and provides a valuable resource for genomics-assisted breeding of this important crop.

Pneumonia infection by Morganella morganii in a male alpaca.

After sudden death with a history of about two weeks ruminal tympany, a 3-year-old, male alpaca from Huantaiqi Zoo, Chongqing, China was presented to the Animal Diseases Rapid Diagnosis Center, Southwest University, Chongqing, China for diagnosis of the death causes. At necropsy, the primary pathological lesions were found in the lung. A pronounced hemorrhage with topical congestion and lobular pneumonia was identified. Sero-fibrinogenous pleural effusion was also detected in the thoracic cavity. After necropsy, the lung sample was processed for histological examination, while lung, hydropericardium, and heart-blood samples were processed for bacteriological examination. From the lung tissue, abundant fluid exudate was found in the pulmonary alveoli. Meanwhile, a mild to moderate hemorrhage and inflammatory cells infiltrations were also observed in the lung sections. Pure isolates on the 5.00% defibrinated sheep blood agar were submitted for identification by morphological and molecular methods. Sequencing results indicated that the Gram-negative sporadic bacilli were all belonged to Morganella morganii. To the best of our knowledge, this is the first record of M. morganii induced pneumonia in an alpaca.