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1.
J Invertebr Pathol ; 204: 108118, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38679369

ABSTRACT

Portunid crabs are distributed worldwide and highly valued in aquaculture. Viral infections are the main limiting factor for the survival of these animals and, consequently, for the success of commercial-scale cultivation. However, there is still a lack of knowledge about the viruses that infect cultured portunid crabs worldwide. Herein, the genome sequence and phylogeny of Callinectes sapidus reovirus 2 (CsRV2) are described, and the discovery of a new bunyavirus in Callinectes danae cultured in southern Brazil is reported. The CsRV2 genome sequence consists of 12 dsRNA segments (20,909 nt) encode 13 proteins. The predicted RNA-dependent RNA polymerase (RdRp) shows a high level of similarity with that of Eriocheir sinensis reovirus 905, suggesting that CsRV2 belongs to the genus Cardoreovirus. The CsRV2 particles are icosahedral, measuring approximately 65 nm in diameter, and exhibit typical non-turreted reovirus morphology. High throughput sequencing data revealed the presence of an additional putative virus genome similar to bunyavirus, called Callinectes danae Portunibunyavirus 1 (CdPBV1). The CdPBV1 genome is tripartite, consisting of 6,654 nt, 3,120 nt and 1,656 nt single-stranded RNA segments that each encode a single protein. Each segment has a high identity with European shore crab virus 1, suggesting that CdPBV1 is a new representative of the family Cruliviridae. The putative spherical particles of CdPBV1 measure ∼120 nm in diameter and present a typical bunyavirus morphology. The results of the histopathological analysis suggest that these new viruses can affect the health and, consequently, the survival of C. danae in captivity. Therefore, the findings reported here should be used to improve prophylactic and pathogen control practices and contribute to the development and optimization of the production of soft-shell crabs on a commercial scale in Brazil.


Subject(s)
Brachyura , Genome, Viral , Phylogeny , Reoviridae , Animals , Brachyura/virology , Reoviridae/genetics , Reoviridae/classification , Orthobunyavirus/genetics , Aquaculture
2.
J Agric Food Chem ; 72(18): 10469-10476, 2024 May 08.
Article in English | MEDLINE | ID: mdl-38659344

ABSTRACT

Lacto-N-difucohexaose II (LNDFH II) is a typical fucosylated human milk oligosaccharide and can be enzymatically produced from lacto-N-tetraose (LNT) by a specific α1,3/4-fucosyltransferase from Helicobacter pylori DMS 6709, referred to as FucT14. Previously, we constructed an engineered Escherichia coli BL21(DE3) with a single plasmid for highly efficient biosynthesis of LNT. In this study, two additional plasmids harboring the de novo GDP-L-fucose pathway module and FucT14, respectively, were further introduced to construct the strain for successful biosynthesis of LNDFH II. FucT14 was actively expressed, and the engineered strain produced LNDFH II as the major product, lacto-N-fucopentaose (LNFP) V as the minor product, and a trace amount of LNFP II and 3-fucosyllactose as very minor products. Additional expression of the α1,3-fucosyltransferase FutM1 from a Bacteroidaceae bacterium from the gut metagenome could obviously enhance the LNDFH II biosynthesis. After optimization of induction conditions, the maximum titer reached 3.011 g/L by shake-flask cultivation. During the fed-batch cultivation, LNDFH II was highly efficiently produced with the highest titer of 18.062 g/L and the productivity yield of 0.301 g/L·h.


Subject(s)
Bacterial Proteins , Escherichia coli , Fucosyltransferases , Guanosine Diphosphate Fucose , Metabolic Engineering , Humans , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Biosynthetic Pathways , Escherichia coli/genetics , Escherichia coli/metabolism , Fucosyltransferases/genetics , Fucosyltransferases/metabolism , Guanosine Diphosphate Fucose/metabolism , Helicobacter pylori/genetics , Helicobacter pylori/metabolism , Helicobacter pylori/enzymology , Oligosaccharides/metabolism , Oligosaccharides/biosynthesis
3.
J Agric Food Chem ; 72(13): 7179-7186, 2024 Apr 03.
Article in English | MEDLINE | ID: mdl-38520358

ABSTRACT

Lacto-N-neotetraose (LNnT), a prominent neutral human milk oligosaccharide (HMO), serves as a pivotal structural element in complex HMO biosynthesis. Given its promising health effects for infants, the biosynthesis of LNnT is garnering greater interest. Using a previously engineered strain as a chassis, a highly effective LNnT producer was constructed. First, LNnT synthesis in Escherichia coli MG1655 was achieved by introducing ß1,3-N-acetylglucosaminyltransferase LgtA and ß1,4-galactosyltransferase CpsIaJ, coupled with the optimization of enzyme expression levels using various promoters. Subsequently, ugd underwent disruption, and the galE gene was enhanced by replacing its promoter with PJ23119 or Ptac. Then, a lux-type quorum sensing (QS) system was applied to achieve varied metabolic regulation. Additionally, systematic optimization of the QS promoters was conducted to further improve the LNnT titer in the shake flask. Finally, the extracellular titer of LNnT was 20.33 g/L, accompanied by a productivity of 0.41 g/L/h.


Subject(s)
Escherichia coli , Quorum Sensing , Humans , Escherichia coli/genetics , Escherichia coli/metabolism , Oligosaccharides/metabolism , Milk, Human/chemistry
4.
Holist Nurs Pract ; 2024 Mar 19.
Article in English | MEDLINE | ID: mdl-38451845

ABSTRACT

To investigate the status of social jet lag (SJL) through sociodemographic factors among clinical nurses and examine the correlation with burnout. There has been relatively little research on the possible factors resulting in SJL among nurses in China and its role in burnout. A multicenter cross-sectional study recruited 596 nurses from 7 Chinese hospitals. Online questionnaires were delivered to assess sociodemographics, shift work, SJL, chronotypes, and the burnout of nurses. Nurses had severe levels of SJL. The number of children, forms of employment, specialty area, length of professional service, and chronotypes were the main predictors of SJL. Moreover, SJL affected burnout (emotional exhaustion and deindividuation), and reducing the nurses' SJL could relieve their burnout. Additional evidence-based interventions indicate that reducing the SJL is essential as the nurses are suffering severe job burnout.

5.
J Agric Food Chem ; 72(8): 4317-4324, 2024 Feb 28.
Article in English | MEDLINE | ID: mdl-38364314

ABSTRACT

Lacto-N-fucopentaose I (LNFP I) has recently been approved as generally recognized as safe, demonstrating its great commercial potential in the food industry. Microbial synthesis through metabolic engineering strategies is an effective approach for large-scale production of LNFP I. Biosynthesis of LNFP I requires consideration of two key points: high titer with low byproduct 2'-fucosyllactose (2'-FL) generation and high purity with low lacto-N-triose II (LNTri II) and lacto-N-tetraose (LNT) residues. Herein, α1,2-fucosyltransferase from Thermoanaerobacterium sp. RBIITD was screened from 16 selected LNFP I-producing glycosyltransferase candidates, showing the highest in vivo LNFP I productivity. Chromosomal integration of wbgO enhanced the LNFP I production by improving the precursor conversion from LNTri II to LNT. The best engineered strain produced 4.42 and 35.1 g/L LNFP I in shake-flask and fed-batch cultivation, respectively. The residual LNTri II and LNT were eliminated by further cultivation with a recombinant strain coexpressing Bifidobacterium bifidum ß-N-acetylhexosaminidase and lacto-N-biosidase. A strategy for LNFP I biosynthesis with high yield and purity was finally realized, providing support for its practical application in large-scale production.


Subject(s)
Glycosyltransferases , Milk, Human , Trisaccharides , Glycosyltransferases/genetics , Glycosyltransferases/metabolism , Milk, Human/chemistry , Oligosaccharides/chemistry
6.
J Alzheimers Dis ; 97(3): 1275-1288, 2024.
Article in English | MEDLINE | ID: mdl-38277290

ABSTRACT

BACKGROUND: Alzheimer's disease (AD), a major dementia cause, lacks effective treatment. MRI-based hippocampal volume measurement using artificial intelligence offers new insights into early diagnosis and intervention in AD progression. OBJECTIVE: This study, involving 483 AD patients, 756 patients with mild cognitive impairment (MCI), and 968 normal controls (NC), investigated the predictive capability of MRI-based hippocampus volume measurements for AD risk using artificial intelligence and evidence-based medicine. METHODS: Utilizing data from ADNI and OASIS-brains databases, three convolutional neural networks (InceptionResNetv2, Densenet169, and SEResNet50) were employed for automated AD classification based on structural MRI imaging. A multitask deep learning model and a densely connected 3D convolutional network were utilized. Additionally, a systematic meta-analysis explored the value of MRI-based hippocampal volume measurement in predicting AD occurrence and progression, drawing on 23 eligible articles from PubMed and Embase databases. RESULTS: InceptionResNetv2 outperformed other networks, achieving 99.75% accuracy and 100% AUC for AD-NC classification and 99.16% accuracy and 100% AUC for MCI-NC classification. Notably, at a 512×512 size, InceptionResNetv2 demonstrated a classification accuracy of 94.29% and an AUC of 98% for AD-NC and 97.31% accuracy and 98% AUC for MCI-NC. CONCLUSIONS: The study concludes that MRI-based hippocampal volume changes effectively predict AD onset and progression, facilitating early intervention and prevention.


Subject(s)
Alzheimer Disease , Cognitive Dysfunction , Deep Learning , Humans , Artificial Intelligence , Alzheimer Disease/diagnosis , Evidence-Based Medicine , Magnetic Resonance Imaging/methods , Hippocampus/diagnostic imaging , Cognitive Dysfunction/diagnostic imaging
7.
Cell Stem Cell ; 30(12): 1624-1639.e8, 2023 12 07.
Article in English | MEDLINE | ID: mdl-37989316

ABSTRACT

Reactivating silenced γ-globin expression through the disruption of repressive regulatory domains offers a therapeutic strategy for treating ß-hemoglobinopathies. Here, we used transformer base editor (tBE), a recently developed cytosine base editor with no detectable off-target mutations, to disrupt transcription-factor-binding motifs in hematopoietic stem cells. By performing functional screening of six motifs with tBE, we found that directly disrupting the BCL11A-binding motif in HBG1/2 promoters triggered the highest γ-globin expression. Via a side-by-side comparison with other clinical and preclinical strategies using Cas9 nuclease or conventional BEs (ABE8e and hA3A-BE3), we found that tBE-mediated disruption of the BCL11A-binding motif at the HBG1/2 promoters triggered the highest fetal hemoglobin in healthy and ß-thalassemia patient hematopoietic stem/progenitor cells while exhibiting no detectable DNA or RNA off-target mutations. Durable therapeutic editing by tBE persisted in repopulating hematopoietic stem cells, demonstrating that tBE-mediated editing in HBG1/2 promoters is a safe and effective strategy for treating ß-hemoglobinopathies.


Subject(s)
Gene Editing , Hemoglobinopathies , Humans , Fetal Hemoglobin/genetics , Fetal Hemoglobin/metabolism , gamma-Globins/genetics , gamma-Globins/metabolism , CRISPR-Cas Systems , Mutation/genetics , Hemoglobinopathies/genetics , Hemoglobinopathies/metabolism , Hematopoietic Stem Cells/metabolism , Transcription Factors/metabolism
8.
Chin J Cancer Res ; 35(4): 354-364, 2023 Aug 30.
Article in English | MEDLINE | ID: mdl-37691897

ABSTRACT

Objective: As laparoscopic surgery is widely applied for primarily treated gastric cancer (GC)/gastroesophageal junction cancer (GEJC) and gains many advantages, the feasibility of laparoscopic total gastrectomy (LTG) for GC/GEJC patients who have received preoperative therapy (PT) has come to the fore. This study aims to analyze the safety and feasibility of LTG after PT for GC/GEJC patients. Methods: We retrospectively analyzed the data of 511 patients with GC/GEJC undergoing LTG, of which 405 received LTG (LTG group) and 106 received PT+LTG (PT-LTG group) at Nanfang Hospital between June 2018 and September 2022. The surgical outcomes were compared between the two groups. Results: The surgical duration was significantly longer in the PT-LTG group (P<0.001), while the incidence of intraoperative complications (P=1.000), postoperative complications (LTG group vs. PT-LTG group: 26.2% vs. 23.6%, P=0.587), the classification of complication severity (P=0.271), and postoperative recovery was similar between two groups. Notably, the incidence of anastomotic complications of esophagojejunostomy was also comparable between the two groups (LTG group vs. PT-LTG group: 5.9% vs. 5.7%, P=0.918). The univariate and multivariate analysis confirmed that positive proximal margin [positive vs. negative: odds ratio (OR)=14.094, 95% confidence interval (95% CI): 2.639-75.260, P=0.002], rather than PT, has an impact on anastomotic complications after LTG (OR=0.945, 95% CI: 0.371-2.408, P=0.905). Conclusions: PT did not increase the surgical risk of LTG for GC/GEJC. Therefore, considering the positive effect of PT on long-term survival, the broader application of PT and LTG for GC/GEJC is supported by our findings.

9.
J Agric Food Chem ; 71(33): 12390-12402, 2023 Aug 23.
Article in English | MEDLINE | ID: mdl-37552889

ABSTRACT

Human milk oligosaccharides (HMOs) have received increasing attention because of their special effects on infant health and commercial value as the new generation of core components in infant formula. Currently, large-scale production of HMOs is generally based on microbial synthesis using metabolically engineered cell factories. Introduction of the specific glycosyltransferases is essential for the construction of HMO-producing engineered strains in which the HMO-producing glycosyltransferases are generally sugar nucleotide-dependent. Four types of glycosyltransferases have been used for typical glycosylation reactions to synthesize HMOs. Soluble expression, substrate specificity, and regioselectivity are common concerns of these glycosyltransferases in practical applications. Screening of specific glycosyltransferases is an important research topic to solve these problems. Molecular modification has also been performed to enhance the catalytic activity of various HMO-producing glycosyltransferases and to improve the substrate specificity and regioselectivity. In this article, various sugar nucleotide-dependent glycosyltransferases for HMO synthesis were overviewed, common concerns of these glycosyltransferases were described, and the future perspectives of glycosyltransferase-related studies were provided.


Subject(s)
Glycosyltransferases , Milk, Human , Infant , Humans , Milk, Human/chemistry , Glycosyltransferases/genetics , Glycosyltransferases/chemistry , Oligosaccharides/chemistry , Glycosylation , Sugars/metabolism
10.
J Agric Food Chem ; 71(33): 12511-12518, 2023 Aug 23.
Article in English | MEDLINE | ID: mdl-37555443

ABSTRACT

Lacto-N-tetraose (LNT) is an important neutral human milk oligosaccharide (HMO) and acts as a significant core structure for complex HMO biosynthesis. We previously achieved high-yield LNT biosynthesis (57.5 g/L) using fed-batch fermentation; however, residual lacto-N-triose II (LNTri II) was also found (21.58 g/L). Here, we re-engineered an efficient LNT-producing Escherichia coli with low LNTri II accumulation using genetically stable LNTri II-producing strains with a genomic insertion of lgtA (encoding ß1,3-N-acetylglucosaminyltransferase). Comparable and low titers of LNT (3.73-4.61 g/L) and LNTri II (0.33-0.63 g/L), respectively, were obtained by introducing ß1,3-galactosyltransferase. To reduce residual LNTri II, the E. coli transporter gene setA was disrupted, obviously reducing the accumulation of LNTri II and LNT. Next, the gene encoding ß-N-acetylhexosaminidase (BbhI) was introduced into LNT-producing strains or E. coli BL21(DE3) for single- or mixed-strain cultivation, respectively. Finally, LNT was obtained (30.13 g/L) in a cocultivation system of mixed engineered strains without undesired LNTri II.


Subject(s)
Escherichia coli , Oligosaccharides , Humans , Escherichia coli/genetics , Oligosaccharides/chemistry , Trisaccharides/analysis , Milk, Human/chemistry
12.
Synth Syst Biotechnol ; 8(3): 509-519, 2023 Sep.
Article in English | MEDLINE | ID: mdl-37502821

ABSTRACT

N-Acetylneuraminic acid (Neu5Ac), the most common type of Sia, generally acts as the terminal sugar in cell surface glycans, glycoconjugates, oligosaccharides, lipo-oligosaccharides, and polysaccharides, thus exerting numerous physiological functions. The extensive applications of Neu5Ac in the food, cosmetic, and pharmaceutical industries make large-scale production of this chemical desirable. Biosynthesis which is associated with important application potential and environmental friendliness has become an indispensable approach for large-scale synthesis of Neu5Ac. In this review, the physiological roles of Neu5Ac was first summarized in detail. Second, the safety evaluation, regulatory status, and applications of Neu5Ac were discussed. Third, enzyme-catalyzed preparation, whole-cell biocatalysis, and microbial de novo synthesis of Neu5Ac were comprehensively reviewed. In addition, we discussed the main challenges of Neu5Ac de novo biosynthesis, such as screening and engineering of key enzymes, identifying exporters of intermediates and Neu5Ac, and balancing cell growth and biosynthesis. The corresponding strategies and systematic strategies were proposed to overcome these challenges and facilitate Neu5Ac industrial-scale production.

13.
J Agric Food Chem ; 71(28): 10701-10709, 2023 Jul 19.
Article in English | MEDLINE | ID: mdl-37409796

ABSTRACT

N-Acetylneuraminic acid (NeuAc) is the predominant sialic acid found in human cells and a human-identical milk monosaccharide. Due to its numerous health benefits, it has great commercial potential in the pharmaceutical, cosmetic, and food industries. Microbial synthesis via metabolic engineering strategies is an important approach to its large-scale production. In this study, a NeuAc synthetic pathway was constructed in Escherichia coli BL21(DE3) by deleting the competitive pathway genes and introducing two genes encoding UDP-N-acetylglucosamine (GlcNAc) 2-epimerase (NeuC) and NeuAc synthase (NeuB). UDP-GlcNAc pathway genes, glmS, glmM, and glmU, were overexpressed to strengthen precursor supply for enhancement of NeuAc synthesis. The microbial source of neuC and neuB was optimized, and their expression was fine-tuned. In addition, glycerol as the carbon source showed a much better effect on NeuAc synthesis than glucose. The final engineered strain produced 7.02 g/L NeuAc by shake-flask cultivation. The titer was enhanced to 46.92 g/L by fed-batch cultivation, with the productivity of 0.82 g/L/h and 1.05 g/g DCW.


Subject(s)
Acetylglucosamine , N-Acetylneuraminic Acid , Humans , Acetylglucosamine/metabolism , Biosynthetic Pathways , Escherichia coli/genetics , Escherichia coli/metabolism , Metabolic Engineering , Uridine Diphosphate/metabolism
14.
Viruses ; 15(2)2023 02 18.
Article in English | MEDLINE | ID: mdl-36851777

ABSTRACT

The movement of viruses in aquatic systems is rarely studied over large geographic scales. Oceanic currents, host migration, latitude-based variation in climate, and resulting changes in host life history are all potential drivers of virus connectivity, adaptation, and genetic structure. To expand our understanding of the genetic diversity of Callinectes sapidus reovirus 1 (CsRV1) across a broad spatial and host life history range of its blue crab host (Callinectes sapidus), we obtained 22 complete and 96 partial genomic sequences for CsRV1 strains from the US Atlantic coast, Gulf of Mexico, Caribbean Sea, and the Atlantic coast of South America. Phylogenetic analyses of CsRV1 genomes revealed that virus genotypes were divided into four major genogroups consistent with their host geographic origins. However, some CsRV1 sequences from the US mid-Atlantic shared high genetic similarity with the Gulf of Mexico genotypes, suggesting potential human-mediated movement of CsRV1 between the US mid-Atlantic and Gulf coasts. This study advances our understanding of how climate, coastal geography, host life history, and human activity drive patterns of genetic structure and diversity of viruses in marine animals and contributes to the capacity to infer broadscale host population connectivity in marine ecosystems from virus population genetic data.


Subject(s)
Brachyura , Orthoreovirus, Mammalian , Reoviridae , Animals , Humans , Ecosystem , Phylogeny , Genetic Structures , Genetic Variation
15.
Ann Surg Oncol ; 30(5): 2729-2738, 2023 May.
Article in English | MEDLINE | ID: mdl-36658250

ABSTRACT

INTRODUCTION: Overlap guiding tube (OGT)-assisted overlap oesophagojejunostomy (EJS), which was first designed and reported by our team, has shown feasibility. However, its safety and efficiency have not yet been compared with the conventional overlap approach. METHODS: We retrospectively analysed the data of 155 gastric/gastroesophageal junction (G/GEJ) cancer patients who underwent laparoscopic total gastrectomy by conventional (conventional group, n = 83) or OGT-assisted (OGT group, n = 72) overlap methods at Nanfang Hospital. The anastomotic efficiency and surgical outcomes were compared between the two groups. RESULTS: The success rate of inserting an anvil fork into the oesophageal lumen at the first attempt in the OGT group was much higher than in the conventional group (86.7% vs. 97.2%, P = 0.019). Consistently, the duration of EJS (P < 0.001) in the OGT group was significantly shorter than that in the conventional group. Operatively, there was one case in which oesophageal pseudocanals developed; another case was converted to thoracoscopic surgery in the conventional group, but there were no such cases in the OGT group. In terms of postoperative recovery, the OGT group was superior to the conventional group. The incidence of postoperative complications (28.9% vs. 20.8%, P = 0.247) and the classification of complication severity (P = 0.450) were milder in the OGT group, although the difference was not statistically significant. Notably, the conventional group had four cases (4.8%) of oesophagojejunal anastomotic leakage (EJAL) and one case (1.2%) of anastomotic stenosis. In the OGT group, two patients (2.8%) developed EJAL, but none developed anastomotic stenosis or anastomotic bleeding. Neither group had any cases of unplanned secondary surgery or perioperative deaths. CONCLUSIONS: The OGT-assisted method reduced the surgical difficulty of overlap EJS with good safety. This study provides new perspectives for optimizing EJS.


Subject(s)
Laparoscopy , Stomach Neoplasms , Humans , Retrospective Studies , Constriction, Pathologic , Laparoscopy/methods , Anastomosis, Surgical/adverse effects , Gastrectomy/methods , Stomach Neoplasms/pathology , Postoperative Complications/etiology , Esophagogastric Junction/surgery , Esophagogastric Junction/pathology , Treatment Outcome
16.
Med Phys ; 49(10): 6505-6516, 2022 Oct.
Article in English | MEDLINE | ID: mdl-35758644

ABSTRACT

BACKGROUND: Endometrial carcinoma (EC) is one of the most common gynecological malignancies with an increasing incidence, and an accurate preoperative diagnosis of deep myometrial invasion (DMI) is crucial for personalized treatment. OBJECTIVE: To determine the predictive value of a magnetic resonance imaging (MRI)-based radiomics nomogram for the presence of DMI in the International Federation of Gynecology and Obstetrics (FIGO) stage I EC. METHODS: We retrospectively collected 163 patients with pathologically confirmed stage I EC from two centers and divided all samples into a training group (Center 1) and a validation group (Center 2). Clinical and routine imaging indicators were analyzed by logistical regression to construct a conventional diagnostic model (M1). Radiomics features extracted from the axial T2-weighted and axial contrast-enhanced T1-weighted (CE-T1W) images were treated with the intraclass correlation coefficient, Mann-Whitney U test, least absolute shrinkage and selection operator, and logistic regression analysis with Akaike information criterion to build a combined radiomics signature (M2). A nomogram (M3) was constructed by M1 and M2. Calibration and decision curves were drawn to evaluate the nomogram in the training and validation cohorts. The diagnostic performance of each indicator and model was evaluated by the area under the receiver operating characteristic curve (AUC). RESULT: The four most significant radiomics features were finally selected from the CE-T1W MRI. For the diagnosis of DMI, the AUCT /AUCV of M1 was 0.798/0.738, the AUCT /AUCV of M2 was 0.880/0.852, and the AUCT /AUCV of M3 was 0.936/0.871 in the training and validation groups, respectively. The calibration curves showed that M3 was in good agreement with the ideal values. The decision curve analysis suggested potential clinical application values of the nomogram. CONCLUSION: A nomogram based on MRI radiomics and clinical imaging indicators can improve the diagnosis of DMI in patients with FIGO stage I EC.


Subject(s)
Endometrial Neoplasms , Nomograms , Endometrial Neoplasms/diagnostic imaging , Endometrial Neoplasms/surgery , Female , Humans , Magnetic Resonance Imaging/methods , Reproducibility of Results , Retrospective Studies
17.
Front Microbiol ; 13: 855750, 2022.
Article in English | MEDLINE | ID: mdl-35369474

ABSTRACT

The advancement of high throughput sequencing has greatly facilitated the exploration of viruses that infect marine hosts. For example, a number of putative virus genomes belonging to the Totiviridae family have been described in crustacean hosts. However, there has been no characterization of the most newly discovered putative viruses beyond description of their genomes. In this study, two novel double-stranded RNA (dsRNA) virus genomes were discovered in the Atlantic blue crab (Callinectes sapidus) and further investigated. Sequencing of both virus genomes revealed that they each encode RNA dependent RNA polymerase proteins (RdRps) with similarities to toti-like viruses. The viruses were tentatively named Callinectes sapidus toti-like virus 1 (CsTLV1) and Callinectes sapidus toti-like virus 2 (CsTLV2). Both genomes have typical elements required for -1 ribosomal frameshifting, which may induce the expression of an encoded ORF1-ORF2 (gag-pol) fusion protein. Phylogenetic analyses of CsTLV1 and CsTLV2 RdRp amino acid sequences suggested that they are members of two new genera in the family Totiviridae. The CsTLV1 and CsTLV2 genomes were detected in muscle, gill, and hepatopancreas of blue crabs by real-time reverse transcription quantitative PCR (RT-qPCR). The presence of ~40 nm totivirus-like viral particles in all three tissues was verified by transmission electron microscopy, and pathology associated with CsTLV1 and CsTLV2 infections were observed by histology. PCR assays showed the prevalence and geographic range of these viruses, to be restricted to the northeast United States sites sampled. The two virus genomes co-occurred in almost all cases, with the CsTLV2 genome being found on its own in 8.5% cases, and the CsTLV1 genome not yet found on its own. To our knowledge, this is the first report of toti-like viruses in C. sapidus. The information reported here provides the knowledge and tools to investigate transmission and potential pathogenicity of these viruses.

18.
Front Immunol ; 13: 783695, 2022.
Article in English | MEDLINE | ID: mdl-35401534

ABSTRACT

Background: Antibodies against programmed death 1 (PD-1) and its ligand, programmed death-ligand 1 (PD-L1) have recently shown promising results in gastric cancer (GC). However, clinicians still lack predictive biomarkers for the efficacy of anti-PD-1 therapy; thus, we investigated the expression of PD-L1 in GC and further assessed its clinical relevance with other clinicopathological features. Methods: We retrospectively collected clinical data on 968 consecutive GC cases from Nanfang Hospital between November 2018 and August 2021. Discrepancy in the combined positive score (CPS) of PD-L1 protein expression between gastric mucosa biopsy and postoperative pathology were investigated. Correlations between CPS and clinicopathological parameters were determined using chi-squared test, multiple logistic aggression analysis, and linear regression analysis. Results: Among the 968 consecutive GC patients, 199 who did not receive preoperative chemotherapy or immunotherapy were tested for CPS both in gastric mucosa biopsy and postoperative pathology, and the results showed that the CPS of gastric mucosa biopsy was significantly lower than that of postoperative pathology [mean ± SD: 5.5 ± 9.4 vs. 13.3 ± 17.4; M(IQR): 2(5) vs. 5(12), p<0.001)]. 62.3% of patients (579/930) had CPS≥ 1, 49.2% of patients (458/930) had CPS≥5, and 33.3% of patients (310/930) had CPS≥10. Mismatch repair deficiency (dMMR) status was seen in 6.1% of patients (56 of 919). Positive Epstein-Barr virus (EBV) status was detected in 4.4% of patients (38 of 854). The patients with CPS≥1/CPS≥5/CPS≥10 were significantly independently correlated with age, Lauren classification, Ki-67 index, and EBV status. According to linear regression analysis, PD-L1 expression was correlated with age (p<0.001), Ki-67 index (p<0.001), EBV (p<0.001), and Lauren classification (p=0.002). Conclusions: Our results confirmed that PD-L1 expression has Intratumoral heterogeneity in GC. Furthermore, the variables of age, Ki-67 index, and Lauren classification, which are common and accessible in most hospitals, are worth exploring as potential biomarkers for anti-PD-1 therapy in GC.


Subject(s)
Epstein-Barr Virus Infections , Stomach Neoplasms , B7-H1 Antigen/metabolism , Biomarkers, Tumor/metabolism , Epstein-Barr Virus Infections/complications , Herpesvirus 4, Human/genetics , Herpesvirus 4, Human/metabolism , Humans , Ki-67 Antigen , Programmed Cell Death 1 Receptor , Retrospective Studies , Stomach Neoplasms/pathology
19.
J Invertebr Pathol ; 190: 107739, 2022 05.
Article in English | MEDLINE | ID: mdl-35248555

ABSTRACT

Crabs can be infected by a variety of pathogenic micro-organisms but the most damaging are viruses. Naturally-occurring Callinectes sapidus reovirus 1 (CsRV1) is thought to contribute to mortality of Callinectes sapidus in soft crab culture in the USA. In Brazil, soft crabs are frequently produced using Callinectes danae, which suffers a similar rate of mortality in culture as C. sapidus. This study investigated whether CsRV1 could be detected in healthy or dead Callinectes danae from Paraná, Brazil and kept in captivity, we also evaluated the relationship between viral infection, and biochemical and behavioral parameters. C. danae from Paranaguá Bay were kept in a recirculation system for 14 days and subjected to weekly biochemical analyses and a reflex action mortality predictors (RAMP) test. RT-qPCR assays for CsRV1 were negative for all samples. However, electrophoretic analysis of extracted RNA from some crabs showed a pattern of 12 dsRNA bands that indicated intense infection by a reovirus with a genome organization different from CsRV1. The banding pattern was indistinguishable from a putative novel reovirus detected in C. sapidus in Rio Grande do Sul, Brazil, provisionally called CsRV2. The prevalence of dsRNA of CsRV2 showed no significant difference between crabs that died and survived. Interestingly, the presence of CsRV2 dsRNA was correlated with a significant reduction in glycogen concentration in hepatopancreas and a decrease in reflex action. The results obtained in this study are an early glimpse of the occurrence of reoviruses in C. danae and their potential effects in soft-shell crab systems in Brazil.


Subject(s)
Brachyura , Reoviridae , Animals , Brazil/epidemiology , Hepatopancreas , Prevalence , RNA, Double-Stranded
20.
Front Microbiol ; 13: 1082565, 2022.
Article in English | MEDLINE | ID: mdl-36687653

ABSTRACT

Myasthenia gravis (MG) is an autoimmune disease caused by autoantibodies that is dependent on T-cell immunity and complement participation and mainly involves neuromuscular junctions. In this study, 30 patients with myasthenia gravis were selected and divided into pretreatment (Case group) and posttreatment (Treatment group) and 30 healthy volunteers (CON group) were included. Among them, the treatment group was treated with Modified Buzhong Yiqi Decoction (MBZYQD), and the levels of antibodies such as AChR, Musk and Titin in blood and intestinal microbiota were compared before treatment (Case group), after treatment (Treatment group) and in healthy volunteers (CON group). The results showed that after treatment with MBZYQD, the antibody levels of AChR, MuSK, and Titin and the inflammatory factor level of IL-6, IL-1ß, and IL-22 in MG patients decreased significantly and nearly returned to a healthy level. In addition, after treatment with MBZYQD, the diversity, structure and function of intestinal microorganisms in MG patients also recovered to a healthy level. At the phylum level, the relative abundance of Proteobacteria in the Case group increased significantly, accompanied by a significant decrease in the relative abundance of Bacteroides compared with that in the CON group, the relative abundance of Proteobacteria and Bacteroides in the Treatment group was similar to that in the CON group. At the genus level, the relative abundance of Shigella in the Case group was significantly increased, accompanied by a significant decrease in the relative abundance of Prevotella, and the relative abundance of Shigella and Prevotella in Treatment group was similar to that in the CON group. Moreover, the fluorobenzoate degradation pathway (KO00364) was significantly increased in the Case group, while this pathway was significantly decreased in the Treatment group. In conclusion, MBZYQD can improve the immune function of the host by regulating the diversity, structure and function of the intestinal microbiota to treat myasthenia gravis.

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