Normal Sequence Research Articles

Whole-genome resequencing identifies candidate genes and SNPs in genomic regions associated with shell color selection in the Pacific oyster, Crassostrea gigas

The various shell colors of bivalves not only result in a better visual perception but also show great commercial value as a highly potential breeding trait. Four shell color strains of Pacific oyster (Crassostrea gigas) characterized by black (SB), white (SW), gold (SG), and orange (SO) were bred purposefully for more than ten generations. In this study, whole-genome resequencing was performed on 20 oysters from each of the four strains, and a total of 14.67 million single nucleotide polymorphisms (SNPs) were obtained after stringent filtering, with an average sequencing depth of >10×. The population structure analysis provided support for the subdivision of all samples into four major genetic clusters corresponding to shell color phenotypes, indicating a high level of genomic differentiation driven by artificial selection. Then, a composite measure of fixation index (FST) and cross-population composite likelihood ratio (XP-CLR) allowed us to identify 336 to 414 candidate divergent regions (CDRs) in pairwise comparisons of different shell colors, with a total of 480 to 614 corresponding genes distributed. Of particular interest were tyrosinase (TYR) and perilipin2 (PLIN2) occurring with extremely high frequency, which participates in shell color determination by regulating melanin production and carotenoid metabolism, respectively. In addition, two genes, namely porphobilinogen deaminase (PBGD) and hephaestin (HEPH), may have a potential function in the accumulation of specific porphyrin in orange shells, while the kynurenine 3-monooxygenase (KMO) gene, known to be involved in ommochrome biosynthesis, may contribute to the darker coloration in black shells. Further, a total of 304 SNPs were extracted from the exons of candidate genes mentioned above, of which 25 loci had highly significant differences among populations (p-value ≤0.01). After Sanger sequencing in another population, 14 SNPs were successfully validated as specific loci closely linked to different shell colors of black, white, gold, and orange, respectively. All in all, the results of this study shed light on the molecular basis of pigmentation and facilitate the selective breeding of C. gigas with desired phenotypic color.

Long-term outcomes of infants with severe BPD

Preterm birth disrupts the normal sequence of lung development. Additionally, interventions that support gas exchange, including positive pressure ventilation and supplemental oxygen can further exacerbate lung injury, increasing the risk of developing bronchopulmonary dysplasia (BPD) in infants born preterm. Approximately 50,000 preterm infants each year in the United States develop BPD. Heterogeneous lung pathology involving the upper and lower respiratory tract can contribute to the BPD phenotype and can be age-dependent. These phenotypes include alveolar, upper airway, large airways, small airways, and vascular. Each of these phenotypes may improve, resolve, or persist at different ages, throughout childhood. The development of BPD endotypes can be influenced by gestational age and length and type of respiratory support. Although, long-term pulmonary outcomes of infants with severe BPD are variable, the presence of small airway disease is a common phenotype in school age and adolescent children. In this review we examine the more common respiratory endotypes found in infants and children with severe BPD and discuss the long-term prognosis for cardiovascular, neurological, and gastrointestinal morbidities in this patient population.

Discovery of broadly-neutralizing antibodies against brown recluse spider and Gadim scorpion sphingomyelinases using consensus toxins as antigens.

Broadly-neutralizing monoclonal antibodies are becoming increasingly important tools for treating infectious diseases and animal envenomings. However, designing and developing broadly-neutralizing antibodies can be cumbersome using traditional low-throughput iterative protein engineering methods. Here, we present a new high-throughput approach for the standardized discovery of broadly-neutralizing monoclonal antibodies relying on phage display technology and consensus antigens representing average sequences of related proteins. We showcase the utility of this approach by applying it to toxic sphingomyelinases from the venoms of species from very distant orders of the animal kingdom, the recluse spider and Gadim scorpion. First, we designed a consensus sphingomyelinase and performed three rounds of phage display selection, followed by DELFIA-based screening and ranking, and benchmarked this to a similar campaign involving cross-panning against recombinant versions of the native toxins. Second, we identified two scFvs that not only bind the consensus toxins, but which can also neutralize sphingomyelinase activity of native whole venom in vitro. Finally, we conclude that the phage display campaign involving the use of the consensus toxin was more successful in yielding cross-neutralizing scFvs than the phage display campaign involving cross-panning.

Feasibility of whole-exome sequencing in fine-needle aspiration specimens of papillary thyroid microcarcinoma for the identification of novel gene mutations.

Genetic profiling is important for assisting the management of papillary thyroid microcarcinoma (PTMC). Although whole-exome sequencing (WES) of surgically resected PTMC tissue has been performed and revealed potential prognostic biomarkers, its application in PTMC fine-needle aspiration (FNA) specimens has not been explored. This study aimed to evaluate the feasibility of WES using FNA specimens of PTMC. Five PTMC patients were enrolled with clinical characteristics gathered. Fine aspiration cytology needle (23 gauges) was used to collect FNA biopsy with ultrasound guidance. WES analysis of FNA specimens from five PTMC patients and matched blood samples was performed. The WES of FNA samples yielded an average sequencing depth of 281× and average coverage of 99.5%. We identified 534 somatic single-nucleotide variants and 13 indels in total, and per sample, we found a mean of 24 exonic mutations, which affected a total of 120 genes. In the PTMC FNA samples, the most frequently mutated genes were BRAF and ANKRD18B, and the four driver genes were BRAF, AFF3, SRCAP, and EGFR. We also identified several germline cancer predisposing gene mutations. The results suggest that WES of FNA specimens is feasible for PTMC and can identify novel genetic mutations.

A SLAF-based high-density genetic map construction and genetic architecture of thermotolerant traits in maize (Zea mays L.).

The leaf scorching trait at flowering is a crucial thermosensitive phenotype in maize under high temperature stress (HS), yet the genetic basis of this trait remains poorly understood. In this study, we genotyped a 254 RIL-F2:8 population, derived from the leaf scorch-free parental inbred line Abe2 and the leaf scorching maternal inbred line B73, using the specific-locus amplified fragment sequencing (SLAF-seq) method. A total of 10,112 polymorphic SLAF markers were developed, and a high-density genetic map with a total length of 1,475.88 cM was constructed. The average sequencing depth of the parents was 55.23X, and that of the progeny was 12.53X. Then, we identified a total of 16 QTLs associated with thermotolerant traits at flowering, of which four QTLs of leaf scorching damage (LS) were distributed on chromosomes 1 (qLS1), 2 (qLS2.1, qLS2.2) and 3 (qLS3), which could explain 19.73% of phenotypic variation. Combining one qLS1 locus with QTL-seq results led to the identification of 6 candidate genes. Expression experiments and sequence variation indicated that Zm00001d033328, encoding N-acetyl-gamma-glutamyl-phosphate reductase, was the most likely candidate gene controlling thermotolerant traits at flowering. In summary, the high-density genetic map and genetic basis of thermotolerant traits lay a critical foundation for mapping other complex traits and identifying the genes associated with thermotolerant traits in maize.

Divergent HLA variations and heterogeneous expression but recurrent HLA loss-of- heterozygosity and common HLA-B and TAP transcriptional silencing across advanced pediatric solid cancers.

The human leukocyte antigen (HLA) system is a major factor controlling cancer immunosurveillance and response to immunotherapy, yet its status in pediatric cancers remains fragmentary. We determined high-confidence HLA genotypes in 576 children, adolescents and young adults with recurrent/refractory solid tumors from the MOSCATO-01 and MAPPYACTS trials, using normal and tumor whole exome and RNA sequencing data and benchmarked algorithms. There was no evidence for narrowed HLA allelic diversity but discordant homozygosity and allele frequencies across tumor types and subtypes, such as in embryonal and alveolar rhabdomyosarcoma, neuroblastoma MYCN and 11q subtypes, and high-grade glioma, and several alleles may represent protective or susceptibility factors to specific pediatric solid cancers. There was a paucity of somatic mutations in HLA and antigen processing and presentation (APP) genes in most tumors, except in cases with mismatch repair deficiency or genetic instability. The prevalence of loss-of-heterozygosity (LOH) ranged from 5.9 to 7.7% in HLA class I and 8.0 to 16.7% in HLA class II genes, but was widely increased in osteosarcoma and glioblastoma (~15-25%), and for DRB1-DQA1-DQB1 in Ewing sarcoma (~23-28%) and low-grade glioma (~33-50%). HLA class I and HLA-DR antigen expression was assessed in 194 tumors and 44 patient-derived xenografts (PDXs) by immunochemistry, and class I and APP transcript levels quantified in PDXs by RT-qPCR. We confirmed that HLA class I antigen expression is heterogeneous in advanced pediatric solid tumors, with class I loss commonly associated with the transcriptional downregulation of HLA-B and transporter associated with antigen processing (TAP) genes, whereas class II antigen expression is scarce on tumor cells and occurs on immune infiltrating cells. Patients with tumors expressing sufficient HLA class I and TAP levels such as some glioma, osteosarcoma, Ewing sarcoma and non-rhabdomyosarcoma soft-tissue sarcoma cases may more likely benefit from T cell-based approaches, whereas strategies to upregulate HLA expression, to expand the immunopeptidome, and to target TAP-independent epitopes or possibly LOH might provide novel therapeutic opportunities in others. The consequences of HLA class II expression by immune cells remain to be established. Immunogenetic profiling should be implemented in routine to inform immunotherapy trials for precision medicine of pediatric cancers.

Cross-modality integration framework with prediction, perception and discrimination for video anomaly detection

Video anomaly detection is an important task for public security in the multimedia field. It aims to distinguish events that deviate from normal patterns. As important semantic representation, the textual information can effectively perceive different contents for anomaly detection. However, most existing methods primarily rely on visual modality, with limited incorporation of textual modality in anomaly detection. In this paper, a cross-modality integration framework (CIForAD) is proposed for anomaly detection, which combines both textual and visual modalities for prediction, perception and discrimination. Firstly, a feature fusion prediction (FUP) module is designed to predict the target regions by fusing the visual features and textual features for prompting, which can amplify the discriminative distance. Then an image-text semantic perception (ISP) module is developed to judge semantic consistency by associating the fine-grained visual features with textual features, where a strategy of local training and global inference is introduced to perceive local details and global semantic correlation. Finally, a self-supervised time attention discrimination (TAD) module is built to explore the inter-frame relation and further distinguish abnormal sequences from normal sequences. Extensive experiments on the three challenging benchmarks indicate that our CIForAD obtains state-of-the-art anomaly detection performance.

Decisive role of mRNA sequencing to solve a suspicion of Fabry-disease in a patient with hypertrophic cardiomyopathy and normal GLA DNA sequencing

We report the case of a 44-year-old-male presenting with hypertrophic cardiomyopathy and a biochemical analysis suggestive of a Fabry-disease but with a negative coding-regions sequencing of GLA gene (Next Generation Sequencing (NGS), Sanger sequencing and GLA large rearrangement assay).

Multi-Omics Analyses Unravel Genetic Relationship of Chinese Coffee Germplasm Resources

The genetic relationships between Coffea arabica resources were analyzed via specific length amplified fragment sequencing (SLAF-seq) and transcriptome sequencing to provide the theoretical basis for breeding new varieties. Twenty C. arabica accessions were used to analyze genetic diversity on the basis of SNPs identified in SLAFs and the transcriptome data. For the SLAF-seq analysis of 20 C. arabica accessions, two Coffea canephora accessions, one Coffea liberica accession, and one Coffea racemosa accession, the number of reads ranged from 2,665,424 to 7,210,310, with a GC content of 38.49%–40.91% and a Q30 value of 94.99%–96.36%. A total of 3,347,069 SLAF tags were obtained, with an average sequencing depth of 13.90×. Moreover, the 1,048,575 SNPs identified in the polymorphic SLAFs were filtered, then the remaining 198,955 SNPs were used to construct a phylogenetic tree, perform a principal component analysis, and characterize the population structure. For the transcriptome analysis, 128.50 Gb clean reads were generated for the 20 C. arabica accessions, with a GC content of 44.36%–51.09% and a Q30 value of 94.55%–95.40%. Furthermore, 25,872 genes’ expression levels were used for the correlation analysis. The phylogenetic relationships as well as the results of the principal component analysis, population structure analysis, and correlation analysis clearly distinguished C. arabica Typica-type accessions from the C. arabica Bourbon-type accessions. Notably, several C. arabica local selections with unknown genetic backgrounds were classified according to all four clustering results.

An insight into tissue culture-induced variation origin shared between anther culture-derived triticale regenerants

BackgroundThe development of the plant in vitro techniques has brought about the variation identified in regenerants known as somaclonal or tissue culture-induced variation (TCIV). S-adenosyl-L-methionine (SAM), glutathione (GSH), low methylated pectins (LMP), and Cu(II) ions may be implicated in green plant regeneration efficiency (GPRE) and TCIV, according to studies in barley (Hordeum vulgare L.) and partially in triticale (× Triticosecale spp. Wittmack ex A. Camus 1927). Using structural equation models (SEM), these metabolites have been connected to the metabolic pathways (Krebs and Yang cycles, glycolysis, transsulfuration), but not for triticale. Using metabolomic and (epi)genetic data, the study sought to develop a triticale regeneration efficiency statistical model. The culture’s induction medium was supplemented with various quantities of Cu(II) and Ag(I) ions for regeneration. The period of plant regeneration has also changed. The donor plant, anther-derived regenerants, and metAFLP were utilized to analyze TCIV concerning DNA in symmetric (CG, CHG) and asymmetric (CHH) sequence contexts. Attenuated Total Reflectance–Fourier Transfer Infrared (ATR-FTIR) spectroscopy was used to gather the metabolomic information on LMP, SAM, and GSH. To frame the data, a structural equation model was employed.ResultsAccording to metAFLP analysis, the average sequence change in the CHH context was 8.65%, and 0.58% was de novo methylation. Absorbances of FTIR spectra in regions specific for LMP, SAM, and GSH were used as variables values introduced to the SEM model. The average number of green regenerants per 100 plated anthers was 2.55.ConclusionsThe amounts of pectin demethylation, SAM, de novo methylation, and GSH are connected in the model to explain GPRE. By altering the concentration of Cu(II) ions in the medium, which influences the amount of pectin, triticale’s GPRE can be increased.

BAYESIAN STUDY OF NORMAL SEQUENCE APPLYING A PRIOR MIXTURE TO THE LOCATION PARAMETER

This article discusses Bayesian Analysis of Normal Sequence using Mixture of Priors. Using the Bayesian methodology, one can generate Bayes estimates by assuming a newly created Mixture combination of priors for location and correct prior for scale parameters. In order to get Bayes estimates for our study, we have assumed that one innovative form of prior, such as a double exponential prior combined with the usual type prior, viz., Normal prior for the mean parameter and correct prior, viz., Inverted gamma prior for the location parameter. Examples of the recently developed methodology in numerical studies are shown by the mean square error of the Bayes estimates of both parameters computed with different known and unknown nature of the parameters and results given with full discussion.

Whole-genome resequencing of Russian sturgeon (Acipenser gueldenstaedtii) reveals selection signatures associated with caviar color

Russian sturgeon, Acipenser gueldenstaedtii, a primitive freshwater fish, holds high economic value as the source of caviar. Remarkably, the economic significance of gold caviar surpasses that of its black counterpart. However, the genetic basis underlying the color variations in caviar remains unexplored. In this study, we conducted whole-genome resequencing of 62 individuals from gold caviar (G) and black caviar (B) populations, achieving an average sequencing depth of approximately 12.21×, and each individual produced approximately 32.47 million high-quality single nucleotide polymorphisms (SNPs). By combining FST and XPEHH methods, we identified 1138 candidate selection regions spanning 26.7 Mb, encompassing 771 genes. Furthermore, significantly enriched GO terms and KEGG pathways were related to pigmentation (e.g., Protein digestion and absorption and Retinol metabolism, etc.) and immunity (e.g., Purine metabolism, Phagosome and Vibrio cholerae infection). Based on the findings, 12 genes (ACTA1, ADCY3, ADCY5, ATP6V1C1, ATP6V1H, CHST11, COL6A3, GART, KCNJ13, RDH10, KCNN4 and XDH) were identified as candidate genes that may affect caviar color. Furthermore, four genes (ATP6V1H, GART, RDH10 and KCNN4) detected simultaneously by FST and XPEHH methods were validated using KASP. This study lays the groundwork for unraveling the molecular mechanisms underlying caviar color and provides insights for the selective breeding of sturgeons with the desired gold caviar trait.

HLA Typing and Mutation Calling from Normal and Tumor Whole Genome Sequencing Data with ALPHLARD-NT.

HLA somatic mutations can alter the expression and function of HLA molecules, which in turn affect the ability of the immune system to recognize and respond to cancer cells. Therefore, it is crucial to accurately identify HLA somatic mutations to enhance our understanding of the interaction between cancer and the immune system and improve cancer treatment strategies. ALPHLARD-NT is a reliable tool that can accurately identify HLA somatic mutations as well as HLA genotypes from whole genome sequencing data of paired normal and tumor samples. Here, we provide a comprehensive guide on how to use ALPHLARD-NT and interpret the results.

ProteoMutaMetrics: machine learning approaches for solute carrier family 6 mutation pathogenicity prediction.

The solute carrier transporter family 6 (SLC6) is of key interest for their critical role in the transport of small amino acids or amino acid-like molecules. Their dysfunction is strongly associated with human diseases such as including schizophrenia, depression, and Parkinson's disease. Linking single point mutations to disease may support insights into the structure-function relationship of these transporters. This work aimed to develop a computational model for predicting the potential pathogenic effect of single point mutations in the SLC6 family. Missense mutation data was retrieved from UniProt, LitVar, and ClinVar, covering multiple protein-coding transcripts. As encoding approach, amino acid descriptors were used to calculate the average sequence properties for both original and mutated sequences. In addition to the full-sequence calculation, the sequences were cut into twelve domains. The domains are defined according to the transmembrane domains of the SLC6 transporters to analyse the regions' contributions to the pathogenicity prediction. Subsequently, several classification models, namely Support Vector Machine (SVM), Logistic Regression (LR), Random Forest (RF), and Extreme Gradient Boosting (XGBoost) with the hyperparameters optimized through grid search were built. For estimation of model performance, repeated stratified k-fold cross-validation was used. The accuracy values of the generated models are in the range of 0.72 to 0.80. Analysis of feature importance indicates that mutations in distinct regions of SLC6 transporters are associated with an increased risk for pathogenicity. When applying the model on an independent validation set, the performance in accuracy dropped to averagely 0.6 with high precision but low sensitivity scores.

The evolution of the Mesozoic lithosphere of northwestern Neotethys: a petrogenetic and geodynamic perspective

A complex and chaotic assemblage of the north Croatian inselbergs, considered a geological conundrum, is marked by omnipresent ophiolite mélange. This is a vestige of the unbroken formation of the lithosphere in the northwestern segment of Neotethys during the Mesozoic, spanning from the late Anisian to the late Tithonian. In this contribution, we present detailed mineralogical, petrological, geochemical and isotopic data on the mélange collected from 1991 to 2023. These data include the entire normal ophiolite sequence, from mantle tectonites and cumulate ultramafic rocks through cumulate and isotropic gabbro and sheeted dyke complex to massive and pillow lavas with interbedded radiolarian cherts. We found that the continuous development of oceanic lithosphere during the Mesozoic is recorded by seven geochemical groups of ophiolitic rocks, each representing a distinct basalt–gabbro suite. Based on their geochemical and isotopic characteristics, a sequence of petrogenetic processes and tectonomagmatic events has been reconstructed. Proposed geodynamic models that shed light on the Mesozoic evolution of the northwestern segment of Neotethys are consistent with the current geodynamic understanding of the broader Mediterranean region. The similarities in the tectonomagmatic and geodynamic history of NW Neotethyan ophiolites and ophiolites from the Dinaridic–Albanide–Hellenide belts suggest that they evolved together, probably within a single branch of the Neotethys Ocean during Triassic to Jurassic time. Thematic collection: This article is part of the Ophiolites, melanges and blueschists collection available at: https://www.lyellcollection.org/topic/collections/ophiolites-melanges-and-blueschists

ConLBS: An Attack Investigation Approach Using Contrastive Learning with Behavior Sequence.

Attack investigation is an important research field in forensics analysis. Many existing supervised attack investigation methods rely on well-labeled data for effective training. While the unsupervised approach based on BERT can mitigate the issues, the high degree of similarity between certain real-world attacks and normal behaviors makes it challenging to accurately identify disguised attacks. This paper proposes ConLBS, an attack investigation approach that combines the contrastive learning framework and multi-layer transformer network to realize the classification of behavior sequences. Specifically, ConLBS constructs behavior sequences describing behavior patterns from audit logs, and a novel lemmatization strategy is proposed to map the semantics to the attack pattern layer. Four different augmentation strategies are explored to enhance the differentiation between attack and normal behavior sequences. Moreover, ConLBS can perform unsupervised representation learning on unlabeled sequences, and can be trained either supervised or unsupervised depending on the availability of labeled data. The performance of ConLBS is evaluated in two public datasets. The results show that ConLBS can effectively identify attack behavior sequences in the cases of unlabeled data or less labeled data to realize attack investigation, and can achieve superior effectiveness compared to existing methods and models.

A randomized controlled trial with extended long-term follow-up: Quality of cervical spine motion after anterior cervical discectomy (ACD) or anterior cervical discectomy with arthroplasty (ACDA)

IntroductionIn previous research, a consistent sequence of segmental contributions during dynamic extension X-rays of the cervical spine was observed in 80–90% of healthy participants. Research questionTo investigate whether this previously defined ‘normal’ sequence of segmental contributions was present in patients who underwent anterior cervical discectomy with arthroplasty (ACDA) or anterior cervical discectomy (ACD). Materials & methodsA randomized controlled trial with extended follow-up was conducted. Patients with single level cervical degenerative radiculopathy with a surgical indication were included and randomized. Dynamic X-ray recordings were made before surgery, one-year post-operative, and at long term follow-up. ResultsA total of 27 patients were included, three in an ACDA pilot group and 24 were randomized to receive ACDA (N = 12) or ACD (N = 12). A total of 20 patients were available for follow-up. Preoperatively, 16.7% of patients in the ACDA group and 58.3% of patients in the ACD group showed a normal sequence. One-year post-operative, 66.7% showed a normal sequence in the ACDA group versus 30.0% in the ACD group (p = 0.036). After an average of 11-years follow-up, a normal sequence was observed in 9.1% of patients in the ACDA group and in none of the patients in the ACD group (p = 0.588). Discussion & conclusionThese findings suggest that while ACDA can restore and preserve a normal sequence of segmental contributions in the short term, this effect is not maintained in the long term. Throughout the process of ageing, not only the quantity, but also the quality of motion changes.

Genome-wide selective signatures mining the candidate genes for egg laying in goose

BackgroundImproving the egg production of goose is a crucial goal of breeding, because genetics is the key factor affecting egg production. Thus, we sequenced the genomes of 55 Chinese indigenous geese from six breeds, which were divided into the high egg-laying group (ZE, HY, and SC) and low egg-laying group (ZD, LH, and ST). Based on the results of the inter-population selection signal analysis, we mined the selected genome regions in the high egg-laying germplasm population to identify the key candidate genes affecting the egg-laying traits.ResultsAccording to the whole-genome sequencing data, the average sequencing depth reached 11.75X. The genetic relationships among those six goose breeds coincided with the breed’s geographical location. The six selective signal detection results revealed that the most selected regions were located on Chr2 and Chr12. In total, 12,051 single-nucleotide polymorphism (SNP) sites were selected in all six methods. Using the enrichment results of candidate genes, we detected some pathways involved in cell differentiation, proliferation, and female gonadal development that may cause differences in egg production. Examples of these pathways were the PI3K-Akt signaling pathway (IGF2, COMP, and FGFR4), animal organ morphogenesis (IGF2 and CDX4), and female gonad development (TGFB2).ConclusionOn analyzing the genetic background of six local goose breeds by using re-sequencing data, we found that the kinship was consistent with their geographic location. 107 egg-laying trait-associated candidate genes were mined through six selection signal analysis. Our study provides a critical reference for analyzing the molecular mechanism underlying differences in reproductive traits and molecular breeding of geese.

Effect of spinal orthosis on clinical outcomes of patients after oblique lumbar interbody fusion: a randomized controlled trial study protocol

BackgroundOblique lumbar interbody fusion (OLIF) is an internationally popular minimally invasive technology for the treatment of various lumbar diseases. Since its introduction to China in 2014, OLIF technology has clearly shown its superiority in reconstructing intervertebral stability, restoring intervertebral space height, achieving indirect decompression, and restoring normal lumbar sequence. However, some patients still suffer from persistent symptoms after OLIF, including low back pain and soreness, which indirectly affect the overall surgical efficacy and patient satisfaction. Therefore, some clinicians recommend that patients routinely use spinal orthoses after OLIF to reduce the stress on the lower back muscles and ligaments, thereby relieving or avoiding postoperative residual symptoms or new symptoms. Accordingly, spinal orthosis use after OLIF has emerged as an essential option. However, the role of spinal orthoses in OLIF and their specific impact on postoperative patient clinical outcomes have remained unclear, and there is a lack of strong clinical evidence to indirectly or directly support the role of spinal orthoses in OLIF and demonstrate their impact on patient clinical outcomes. This study aims to investigate the role of spinal orthoses in OLIF by grouping patients based on the use or nonuse of spinal orthosis after OLIF, thus providing a better basis for the majority of patients and physicians.Methods/designWe plan to conduct a 1-year randomized controlled trial involving 60 subjects. The subjects will be randomized into two groups: group A (those wearing spinal orthoses after surgery) and group B (those not wearing spinal orthoses after surgery). The clinical outcomes of these patients will be evaluated using the Oswestry disability index, visual analog scale, and Brantigan, Steffee, Fraser 1 day before surgery and 2 weeks and 1, 6, and 12 months after surgery.DiscussionThis randomized controlled trial aims to provide a reference for further comprehensive trial design. The findings of this study will provide a better and more scientific basis for the choice of postoperative rehabilitation and treatment for patients undergoing such a procedure.Trial registrationThis study has been registered in the Chinese Clinical Trial Registry (Registration No.: ChiCTR2200059000). Registration date: April 22, 2022.Registration website: http://www.chictr.org.cn/showproj.aspx?proj=166310

Molecular Structure Characterization of Micro/Nanoplastics by 193 nm Ultraviolet Photodissociation Mass Spectrometry.

The degradation of macroplastics results in micro/nanoplastics (MNPs) in the natural environment, inducing high health risks worldwide. It remains challenging to characterize the accurate molecular structures of MNPs. Herein, we integrate 193 nm ultraviolet photodissociation (UVPD) with mass spectrometry to interrogate the molecular structures of poly(ethylene glycol) terephthalate and polyamide (PA) MNPs. The backbones of the MNP polymer can be efficiently dissociated by UVPD, producing rich types of fragment ions. Compared to high-energy collision dissociation (HCD), the structural informative fragment ions and corresponding sequence coverages obtained by UVPD were all improved 2.3 times on average, resulting in almost complete sequence coverage and precise structural interrogation of MNPs. We successfully determine the backbone connectivity differences of MNP analogues PA6, PA66, and PA610 by improving the average sequence coverage from 26.8% by HCD to 89.4% by UVPD. Our results highlight the potential of UVPD in characterizing and discriminating backbone connectivity and chain end structures of different types of MNPs.