bluuebunny/crossref_metadata_2025 huggingface dataset:
Genetic diagnosis of fetal microcephaly at a single tertiary center in China
Authors: Wang You , Fu Fang , Lei Tingying , Zhen Li , Deng Qiong , Zhou Hang , Ma Chunling , Cheng Ken , Huang Ruibin , Li Ru , Yu Qiuxia , Li Lushan , Han Jin , Yang Xin , Li Dongzhi , Liao Can
Published: May 2023

Background: Microcephaly is common in patients with neuropsychiatric problems, and it is usually closely related to genetic causes. However, studies on chromosomal abnormalities and single-gene disorders associated with fetal microcephaly are limited.

Objective: We investigated the cytogenetic and monogenic risks of fetal microcephaly and evaluated their pregnancy outcomes.

Methods: We performed a clinical evaluation, high-resolution chromosomal microarray analysis (CMA), and trio exome sequencing (ES) on 224 fetuses with prenatal microcephaly and closely followed the pregnancy outcome and prognosis.

Results: Among 224 cases of prenatal fetal microcephaly, the diagnosis rate was 3.74% (7/187) for CMA and 19.14% (31/162) for trio-ES. Exome sequencing identified 31 pathogenic or likely pathogenic (P/LP) single nucleotide variants (SNVs) in 25 genes associated with fetal structural abnormalities in 37 microcephaly fetuses; 19 (61.29%) of which occurred de novo. Variants of unknown significance (VUS) was found in 33/162 (20.3%) fetuses. The gene variant involved included the single gene MPCH 2 and MPCH 11, which is associated with human microcephaly, and HDAC8, TUBGCP6, NIPBL, FANCI, PDHA1, UBE3A, CASK, TUBB2A, PEX1, PPFIBP1, KNL1, SLC26A4, SKIV2L, COL1A2, EBP, ANKRD11, MYO18B, OSGEP, ZEB2, TRIO, CLCN5, CASK, and LAGE3. The live birth rate of fetal microcephaly in the syndromic microcephaly group was significantly higher than that in the primary microcephaly group [62.9% (117/186) vs 31.56% (12/38), p = 0.000].

Conclusion: We conducted a prenatal study by conducting CMA and ES for the genetic analysis of fetal microcephaly cases. CMA and ES had a high diagnostic rate for the genetic causes of fetal microcephaly cases. In this study, we also identified 14 novel variants, which expanded the disease spectrum of microcephaly-related genes.
Genomic characterization of the NAC transcription factors, directed at understanding their functions involved in endocarp lignification of iron walnut (Juglans sigillata Dode)
Authors: Yu Anmin , Zou Hanyu , Li Ping , Yao Xiaowei , Zhou Zekun , Gu Xu , Sun Rui , Liu Aizhong
Published: May 2023

The NAC (NAM, ATAF1/2, and CUC2) transcription factors (TF), one of the largest plant-specific gene families, play important roles in the regulation of plant growth and development, stress response and disease resistance. In particular, several NAC TFs have been identified as master regulators of secondary cell wall (SCW) biosynthesis. Iron walnut (Juglans sigillata Dode), an economically important nut and oilseed tree, has been widely planted in the southwest China. The thick and high lignified shell derived endocarp tissues, however, brings troubles in processing processes of products in industry. It is indispensable to dissect the molecular mechanism of thick endocarp formation for further genetic improvement of iron walnut. In the present study, based on genome reference of iron walnut, 117 NAC genes, in total, were identified and characterized in silico, which involves only computational analysis to provide insight into gene function and regulation. We found that the amino acids encoded by these NAC genes varied from 103 to 1,264 in length, and conserved motif numbers ranged from 2 to 10. The JsiNAC genes were unevenly distributed across the genome of 16 chromosomes, and 96 of these genes were identified as segmental duplication genes. Furthermore, 117 JsiNAC genes were divided into 14 subfamilies (A-N) according to the phylogenetic tree based on NAC family members of Arabidopsis thaliana and common walnut (Juglans regia). Furthermore, tissue-specific expression pattern analysis demonstrated that a majority of NAC genes were constitutively expressed in five different tissues (bud, root, fruit, endocarp, and stem xylem), while a total of 19 genes were specifically expressed in endocarp, and most of them also showed high and specific expression levels in the middle and late stages during iron walnut endocarp development. Our result provided a new insight into the gene structure and function of JsiNACs in iron walnut, and identified key candidate JsiNAC genes involved in endocarp development, probably providing mechanistic insight into shell thickness formation across nut species.
Identification of the lncRNA–miRNA‒mRNA regulatory network for middle cerebral artery occlusion-induced ischemic stroke
Authors: Shi Guixin , He Dong , Xiao Hua , Liu Yu’e , Liu Chuanyong , Cao Fang
Published: May 2023

Stroke known as a neurological disease has significant rates of disability and mortality. Middle cerebral artery occlusion (MCAO) models in rodents is crucial in stroke research to mimic human stroke. Building the mRNA and non-conding RNA network is essential for preventing MCAO-induced ischemic stroke occurrence. Herein, genome-wide mRNA, miRNA, and lncRNA expression profiles among the MCAO group at 3 h, 6 h, and 12 h after surgery and controls using high-throughput RNA sequencing. We detected differentially expressed mRNAs (DE-mRNAs), miRNAs (DE-miRNAs), and lncRNAs (DE-lncRNAs) between the MCAO and control groups. In addition, biological functional analyses were conducted, including GO/KEGG enrichment analysis, and protein-protein interaction analysis (PPI). GO analysis indicated that the DE-mRNAs were mainly enriched in several important biological processes as lipopolysaccharide, inflammatory response, and response to biotic stimulus. The PPI network analysis revealed that the 12 DE-mRNA target proteins showed more than 30° with other proteins, and the top three proteins with the highest node degree were Alb, IL-6, and TNF. In the DE-mRNAs, we found the mRNA of Gp6 and Elane interacting with two miRNAs (novel_miR_879 and novel_miR_528) and two lncRNAs (MSTRG.348134.3 and MSTRG.258402.19). As a result of this study, a new perspective can be gained into the molecular pathophysiology leading to the formation of MCAO. The mRNA-miRNA‒lncRNA regulatory networks play an important role in MCAO-induced ischemic stroke pathogenesis and could be applied to the treatment and prevention of ischemic stroke in the future.
NK cell defects: implication in acute myeloid leukemia
Authors: D’Silva Selma Z. , Singh Meenakshi , Pinto Andrea S.
Published: May 2023

Acute Myeloid Leukemia (AML) is a complex disease with rapid progression and poor/unsatisfactory outcomes. In the past few years, the focus has been on developing newer therapies for AML; however, relapse remains a significant problem. Natural Killer cells have strong anti-tumor potential against AML. This NK-mediated cytotoxicity is often restricted by cellular defects caused by disease-associated mechanisms, which can lead to disease progression. A stark feature of AML is the low/no expression of the cognate HLA ligands for the activating KIR receptors, due to which these tumor cells evade NK-mediated lysis. Recently, different Natural Killer cell therapies have been implicated in treating AML, such as the adoptive NK cell transfer, Chimeric antigen receptor-modified NK (CAR-NK) cell therapy, antibodies, cytokine, and drug treatment. However, the data available is scarce, and the outcomes vary between different transplant settings and different types of leukemia. Moreover, remission achieved by some of these therapies is only for a short time. In this mini-review, we will discuss the role of NK cell defects in AML progression, particularly the expression of different cell surface markers, the available NK cell therapies, and the results from various preclinical and clinical trials.
A rapid and efficient platform for antiviral crRNA screening using CRISPR-Cas13a-based nucleic acid detection
Authors: Yang Lan , Zhang Youcui , Yi Wenyanbo , Dong Xue , Niu Mengwei , Song Yingjie , Han Yao , Li Hao , Sun Yansong
Published: May 2023

Introduction

Rapid and high-throughput screening of antiviral clustered regularly interspaced short palindromic repeat (CRISPR) RNAs (crRNAs) is urgently required for the CRISPR-Cas13a antiviral system. Based on the same principle, we established an efficient screening platform for antiviral crRNA through CRISPR-Cas13a nucleic acid detection.

Method

In this study, crRNAs targeting PA, PB1, NP, and PB2 of the influenza A virus (H1N1) were screened using CRISPR-Cas13a nucleic acid detection, and their antiviral effects were confirmed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The RNA secondary structures were predicted by bioinformatics methods.

Results

The results showed that crRNAs screened by CRISPR-Cas13a nucleic acid detection could effectively inhibit viral RNA in mammalian cells. Besides, we found that this platform for antiviral crRNA screening was more accurate than RNA secondary structure prediction. In addition, we validated the feasibility of the platform by screening crRNAs targeting NS of the influenza A virus (H1N1).

Discussion

This study provides a new approach for screening antiviral crRNAs and contributes to the rapid advancement of the CRISPR-Cas13a antiviral system.
Exploiting endocytosis for transfection of mRNA for cytoplasmatic delivery using cationic gold nanoparticles
Authors: Gustà Muriel F. , Edel Michael J. , Salazar Vivian A. , Alvarez-Palomo Belén , Juan Manel , Broggini Massimo , Damia Giovanna , Bigini Paolo , Corbelli Alessandro , Fiordaliso Fabio , Barbul Alexander , Korenstein Rafi , Bastús Neus G. , Puntes Víctor
Published: May 2023

Introduction

Gene therapy holds promise to cure various diseases at the fundamental level. For that, efficient carriers are needed for successful gene delivery. Synthetic ‘non-viral’ vectors, as cationic polymers, are quickly gaining popularity as efficient vectors for transmitting genes. However, they suffer from high toxicity associated with the permeation and poration of the cell membrane. This toxic aspect can be eliminated by nanoconjugation. Still, results suggest that optimising the oligonucleotide complexation, ultimately determined by the size and charge of the nanovector, is not the only barrier to efficient gene delivery.

Methods

We herein develop a comprehensive nanovector catalogue comprising different sizes of Au NPs functionalized with two different cationic molecules and further loaded with mRNA for its delivery inside the cell.

Results and Discussion

Tested nanovectors showed safe and sustained transfection efficiencies over 7 days, where 50 nm Au NPs displayed the highest transfection rates. Remarkably, protein expression was increased when nanovector transfection was performed combined with chloroquine. Cytotoxicity and risk assessment demonstrated that nanovectors are safe, ascribed to lesser cellular damage due to their internalization and delivery via endocytosis. Obtained results may pave the way to design advanced and efficient gene therapies for safely transferring oligonucleotides.
Single-cell landscape dissecting the transcription and heterogeneity of innate lymphoid cells in ischemic heart
Authors: Jiang Shijiu , Zheng Yuqi , Lv Bingjie , He Shaolin , Yang Wenling , Wang Boyuan , Zhou Jin , Liu Shangwei , Li Dazhu , Lin Jibin
Published: May 2023

Background

Until now, few articles have revealed the potential roles of innate lymphoid cells (ILCs) in cardiovascular diseases. However, the infiltration of ILC subsets in ischemic myocardium, the roles of ILC subsets in myocardial infarction (MI) and myocardial ischemia-reperfusion injury (MIRI) and the related cellular and molecular mechanisms have not been described with a sufficient level of detail.

Method

In the current study, 8-week-old male C57BL/6J mice were divided into three groups: MI, MIRI and sham group. Single-cell sequencing technology was used to perform dimensionality reduction clustering of ILC to analyze the ILC subset landscape at a single-cell resolution, and finally flow cytometry was used to confirm the existence of the new ILC subsets in different disease groups.

Results

Five ILC subsets were found, including ILC1, ILC2a, ILC2b, ILCdc and ILCt. It is worth noting that ILCdc, ILC2b and ILCt were identified as new ILC subclusters in the heart. The cellular landscapes of ILCs were revealed and signal pathways were predicted. Furthermore, pseudotime trajectory analysis exhibited different ILC statuses and traced related gene expression in normal and ischemic conditions. In addition, we established a ligand–receptor–transcription factor–target gene regulatory network to disclose cell communications among ILC clusters. Moreover, we further revealed the transcriptional features of the ILCdc and ILC2a subsets. Finally, the existence of ILCdc was confirmed by flow cytometry.

Conclusion

Collectively, by characterizing the spectrums of ILC subclusters, our results provide a new blueprint for understanding ILC subclusters’ roles in myocardial ischemia diseases and further potential treatment targets.
TME-targeted approaches of brain metastases and its clinical therapeutic evidence
Authors: Khan Ibrar Muhammad , Khan Safir Ullah , Sala Hari Siva Sai , Khan Munir Ullah , Ud Din Muhammad Azhar , Khan Samiullah , Hassan Syed Shams ul , Khan Nazir Muhammad , Liu Yong
Published: May 2023

The tumor microenvironment (TME), which includes both cellular and non-cellular elements, is now recognized as one of the major regulators of the development of primary tumors, the metastasis of which occurs to specific organs, and the response to therapy. Development of immunotherapy and targeted therapies have increased knowledge of cancer-related inflammation Since the blood-brain barrier (BBB) and blood-cerebrospinal fluid barrier (BCB) limit immune cells from entering from the periphery, it has long been considered an immunological refuge. Thus, tumor cells that make their way “to the brain were believed to be protected from the body’s normal mechanisms of monitoring and eliminating them. In this process, the microenvironment and tumor cells at different stages interact and depend on each other to form the basis of the evolution of tumor brain metastases. This paper focuses on the pathogenesis, microenvironmental changes, and new treatment methods of different types of brain metastases. Through the systematic review and summary from macro to micro, the occurrence and development rules and key driving factors of the disease are revealed, and the clinical precision medicine of brain metastases is comprehensively promoted. Recent research has shed light on the potential of TME-targeted and potential treatments for treating Brain metastases, and we’ll use that knowledge to discuss the advantages and disadvantages of these approaches.
Innate and adaptive immunity to SARS-CoV-2 and predisposing factors
Authors: Shen Jiaying , Fan Junyan , Zhao Yue , Jiang Doming , Niu Zheyun , Zhang Zihan , Cao Guangwen
Published: May 2023

The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus (SARS-CoV-2), has affected all countries worldwide. Although some symptoms are relatively mild, others are still associated with severe and even fatal clinical outcomes. Innate and adaptive immunity are important for the control of SARS-CoV-2 infections, whereas a comprehensive characterization of the innate and adaptive immune response to COVID-19 is still lacking and the mechanisms underlying immune pathogenesis and host predisposing factors are still a matter of scientific debate. Here, the specific functions and kinetics of innate and adaptive immunity involved in SARS-CoV-2 recognition and resultant pathogenesis are discussed, as well as their immune memory for vaccinations, viral-mediated immune evasion, and the current and future immunotherapeutic agents. We also highlight host factors that contribute to infection, which may deepen the understanding of viral pathogenesis and help identify targeted therapies that attenuate severe disease and infection.
Type 1 interferon signature in peripheral blood mononuclear cells and monocytes of idiopathic inflammatory myopathy patients with different myositis-specific autoantibodies
Authors: Li Mengdi , Zhang Yusheng , Zhang Wenzhe , Sun Jinlei , Liu Rui , Pan Zhou , Zhang Panpan , Liu Shengyun
Published: May 2023

Background

Myositis-specific autoantibodies (MSAs) are clinically used to diagnose and define idiopathic inflammatory myopathy (IIM) subsets. However, the underlying pathogenic mechanisms of patients with different MSAs remain unclear.

Methods

A total of 158 Chinese patients with IIM and 167 gender- and age-matched healthy controls (HCs) were enrolled. Transcriptome sequencing (RNA-Seq) was performed with peripheral blood mononuclear cells (PBMCs), followed by the identification of differentially expressed genes (DEGs) and analysis of gene set enrichment analysis, immune cell infiltration, and WGCNA. Monocyte subsets and related cytokines/chemokines were quantified. The expressions of interferon (IFN)-related genes were validated using qRT-PCR and Western blot in both PBMCs and monocytes. We also performed correlation analysis and ROC analysis to explore the potential clinical significance of the IFN-related genes.

Results

There were 1,364 genes altered in patients with IIM, including 952 upregulated and 412 downregulated genes. The type I interferon (IFN-I) pathway was remarkably activated in patients with IIM. Compared with patients with other MSAs, IFN-I signatures were significantly activated in patients with anti-melanoma differentiation-associated gene 5 (MDA5) antibodies. In total, 1,288 hub genes associated with IIM onset were identified using WGCNA, including 29 key DEGs associated with IFN signaling. The patients had more CD14brightCD16- classical, CD14brightCD16+ intermediate, and fewer CD14dimCD16+ non-classical monocyte subsets. Plasma cytokines like IL-6 and TNF and chemokines including CCL3 and MCPs increased. The validation of IFN-I-related gene expressions was consistent with the findings from RNA-Seq. The IFN-related genes were correlated with laboratory parameters and helpful for IIM diagnosis.

Conclusion

Gene expressions were remarkably altered in the PBMCs of IIM patients. Anti-MDA5+ IIM patients had a more pronounced activated IFN signature than others. Monocytes exhibited a proinflammatory feature and contributed to the IFN signature of IIM patients.
Uncovering new insights into the role of the ubiquitin ligase Smurf1 on the regulation of innate immune signaling and resistance to infection
Authors: Souza-Costa Luiz Pedro , Andrade-Chaves Josiane Teixeira , Andrade Juvana Moreira , Costa Vivian Vasconcelos , Franco Luis Henrique
Published: May 2023

Innate immunity is the body’s first line of defense against infections. Innate immune cells express pattern recognition receptors in distinct cellular compartments that are responsible to detect either pathogens-associated molecules or cellular components derived from damaged cells, to trigger intracellular signaling pathways that lead to the activation of inflammatory responses. Inflammation is essential to coordinate immune cell recruitment, pathogen elimination and to keep normal tissue homeostasis. However, uncontrolled, misplaced or aberrant inflammatory responses could lead to tissue damage and drive chronic inflammatory diseases and autoimmunity. In this context, molecular mechanisms that tightly regulate the expression of molecules required for the signaling of innate immune receptors are crucial to prevent pathological immune responses. In this review, we discuss the ubiquitination process and its importance in the regulation of innate immune signaling and inflammation. Then, we summarize the roles of Smurf1, a protein that works on ubiquitination, on the regulation of innate immune signaling and antimicrobial mechanisms, emphasizing its substrates and highlighting its potential as a therapeutic target for infectious and inflammatory conditions.
Immunity and reproduction protective effects of Chitosan Oligosaccharides in Cyclophosphamide/Busulfan-induced premature ovarian failure model mice
Authors: Li Xiaoyan , Ye Haifeng , Su Tie , Hu Chuan , Huang Yaoqi , Fu Xinxin , Zhong Zhisheng , Du Xuelian , Zheng Yuehui
Published: May 2023

Introduction

Premature ovarian failure (POF) is a major cause of infertility among women of reproductive age. Unfortunately, there is no effective treatment available currently. Researchers have shown that immune disorders play a significant role in the development of POF. Moreover, growing evidence suggest that Chitosan Oligosaccharides (COS), which act as critical immunomodulators, may have a key role in preventing and treating a range of immune related reproductive diseases.

Methods

KM mice (6-8 weeks) received a single intraperitoneal injection of cyclophosphamide (CY, 120mg/kg) and busulfan (BUS, 30mg/kg) to establish POF model. After completing the COS pre-treatment or post-treatment procedures, peritoneal resident macrophages (PRMs) were collected for neutral erythrophagocytosis assay to detect phagocytic activity. The thymus, spleen and ovary tissues were collected and weighed to calculate the organ indexes. Hematoxylin-eosin (HE) staining was performed to observe the histopathologic structure of those organs. The serum levels of estrogen (E2) and progesterone (P) were measured via the enzyme-linked immunosorbent assay (ELISA). The expression levels of immune factors including interleukin 2 (IL-2), interleukin 4 (IL-4), and tumor necrosis factor α (TNF-α), as well as germ cell markers Mouse Vasa Homologue (MVH) and Fragilis in ovarian tissue, were analyzed by Western blotting and qRT-PCR. In addition, ovarian cell senescence via p53/p21/p16 signaling was also detected.

Results

The phagocytic function of PRMs and the structural integrity of thymus and spleen were preserved by COS treatment. The levels of certain immune factors in the ovaries of CY/BUS- induced POF mice were found to be altered, manifested as IL-2 and TNF-α experiencing a significant decline, and IL-4 presenting a notable increase. Both pre-treatment and post-treatment with COS were shown to be protective effects against the damage to ovarian structure caused by CY/BUS. Senescence-associated β-galactosidase (SA-β-Gal) staining results showed that COS prevents CY/BUS-induced ovarian cell senescence. Additionally, COS regulated estrogen and progesterone levels, enhanced follicular development, and blocked ovarian cellular p53/p21/p16 signaling which participating in cell senescence.

Conclusion

COS is a potent preventative and therapeutic medicine for premature ovarian failure by enhancing both the ovarian local and systemic immune response as well as inhibiting germ cell senescence.
Unveiling a Virulence-Regulating Mechanism in Aeromonas hydrophila: a Quantitative Exoproteomic Analysis of an AraC-Like Protein
Authors: Zhang Lishan , Sun Lina , Srinivasan Ramanathan , Lin Meizhen , Gong Lanqing , Lin Xiangmin
Published: May 2023

Bacterial AraC is a transcription factor family that initiates transcription by recruiting RNA polymerase to the promoter and directly regulating various bacterial phenotypes. It also directly regulates various bacterial phenotypes. However, how this transcription factor regulates bacterial virulence and affects host immunity is still largely unknown. In this study, deleting the orf02889 (AraC-like transcription factor) gene in virulent Aeromonas hydrophila LP-2 affected several important phenotypes, such as increasing biofilm formation and siderophore production abilities. Moreover, Δorf02889 also significantly decreased the virulence of A. hydrophila and has promising attenuated vaccine potential. To better understand the effects of orf02889 on biological functions, a data independent acquisition (DIA)-based quantitative proteomics method was performed to compare the differentially expressed proteins between Δorf02889 and the wild-type strain in extracellular fractions. The following bioinformatics analysis suggested that ORF02889 may regulate various metabolic pathways, such as quorum sensing and ATP binding cassette (ABC) transporter metabolism. Moreover, 10 selected genes from the top 10 decreasing abundances in proteomics data were deleted, and their virulence to zebrafish was evaluated, respectively. The results showed that ΔcorC, Δorf00906, and Δorf04042 significantly reduced bacterial virulence. Finally, the following chromatin immunoprecipitation and polymerase chain reaction (ChIP-PCR) assay validated that the promoter of corC was directly regulated by ORF02889. Overall, these results provide insight into the biological function of ORF02889 and demonstrate its inherent regulatory mechanism for the virulence of A. hydrophila.
Invasion potential of hornets (Hymenoptera: Vespidae: Vespa spp.)
Authors: Otis Gard W. , Taylor Benjamin A. , Mattila Heather R.
Published: May 2023

Hornets are large, predatory wasps that have the potential to alter biotic communities and harm honey bee colonies once established in non-native locations. Mated, diapausing females (gynes) can easily be transported to new habitats, where their behavioral flexibility allows them to found colonies using local food and nest materials. Of the 22 species in the genusVespa, five species are now naturalized far from their endemic populations and another four have been detected either in nature or during inspections at borders of other countries. By far the most likely pathway of long-distance dispersal is the transport of gynes in transoceanic shipments of goods. Thereafter, natural dispersal of gynes in spring and accidental local transport by humans cause shorter-range expansions and contribute to the invasion process. Propagule pressure of hornets is unquantified, although it is likely low but unrelenting. The success of introduced populations is limited by low propagule size and the consequences of genetic founder effects, including the extinction vortex linked to single-locus, complementary sex determination of most hymenopterans. Invasion success is enhanced by climatic similarity between source locality and introduction site, as well as genetic diversity conferred by polyandry in some species. These and other factors that may have influenced the successful establishment of invasive populations ofV. velutina,V. tropica,V. bicolor,V. orientalis, andV. crabroare discussed. The highly publicized detections ofV. mandariniain North America and research into its status provide a real-time example of an unfolding hornet invasion.
Differential methylation between sex in adult green sea turtle skin biopsies
Authors: Mayne Benjamin , Mustin Walter , Baboolal Vandanaa , Casella Francesca , Ballorain Katia , Barret Mathieu , Vanderklift Mathew A. , Tucker Anton D. , Berry Oliver
Published: May 2023

In marine turtles, the sex of hatchlings is determined by their egg incubation temperature. Global warming may increase the extinction risk by skewing hatchling sex ratios. Assessment of this risk at the population level requires the identification of sex in hatchlings and juveniles. However, available methods are typically lethal, highly invasive, or difficult to conduct at a large scale. Changes in DNA methylation, an epigenetic modification, have been characterized as part of sex differentiation pathways in some species with environmentally determined sex, but so far not in marine turtles. Neither have epigenetic biomarkers for sex been developed into rapid assays suited to research on wildlife. In this study, we aimed to develop a rapid, minimally invasive, and inexpensive method to identify the sex of marine turtles. We used reduced representation bisulfite sequencing DNA methylation data from adult green sea turtle (Chelonia mydas) skin biopsies to identify 16 genomic regions exhibiting differential methylation between males and females (adjusted p-value < 0.01). We designed methylation sensitive qPCR assays for these regions and tested their capacity to identify the sex of turtles ranging in age between 3-34 years. The qPCR assay identified the correct sex in turtles > 17 years. However, the sex of younger turtles could not be accurately identified. This suggests the sex differences distinguishable by the assay were adult specific, reflecting the training data on which the sex-specific regions were identified, and likely linked to late-stage ontogenetic changes associated with sexual maturity. Epigenetic biomarkers are a promising tool for wildlife research because they can be minimally invasive and high throughput. Future research into sex-specific differentially methylated regions in hatchlings and juveniles should be based on genome-wide DNA methylation data from a wider age range, which includes hatchlings.
GADD34 activates p53 and may have utility as a marker of atherosclerosis
Authors: Tomiyoshi Go , Nakamura Rika , Shinmen Natsuko , Yoshida Yoichi , Mine Seiichiro , Machida Toshio , Iwase Katsuro , Iwadate Yasuo , Hiwasa Takaki , Kuroda Hideyuki
Published: May 2023

We previously identified growth arrest and DNA-damage-inducible gene 34 (GADD34) as a marker of ischemic stroke. In the present study, serum levels of anti-GADD34 antibodies were found to be significantly higher in patients with acute ischemic stroke or chronic kidney disease compared to healthy donors. We then examined the biological function of GADD34 by transfection into U2OS human osteosarcoma and U87 human glioblastoma cells. Knockdown of GADD34 by siRNA resulted in enhanced cell proliferation, which was reversed by co-knockdown of MDM2. Luciferase reporter assays revealed that the transactivation ability of p53 enhanced by genotoxic anticancer drugs such as camptothecin and etoposide was further potentiated by enforced expression of GADD34 but attenuated by co-transfection with p53 shRNA expression plasmids. Western blotting demonstrated increased p53 protein levels after treatment with camptothecin, which was also potentiated by GADD34 but suppressed by GADD34 siRNA, ATM siRNA, and ATM inhibitor wortmannin. GADD34 levels also increased in response to treatment with camptothecin or adriamycin, and this increase was attenuated by MDM2 siRNA. Immunoprecipitation with anti-GADD34 antibody followed by Western blotting with anti-MDM2 antibodies indicated ubiquitination of GADD34 is mediated by MDM2. Accordingly, GADD34 may function as a ubiquitination decoy to reduce p53 ubiquitination and increase p53 protein levels. Increased neuronal cell death due to activation of p53 by GADD34 may account for the elevated serum levels of anti-GADD34 antibodies observed in patients with acute ischemic stroke.
Prolonged mechanical ventilation after lung transplantation: risks factors and consequences on recipient outcome
Authors: Atchade Enora , Boughaba Aimane , Dinh Alexy Tran , Jean-Baptiste Sylvain , Tanaka Sébastien , Copelovici Léa , Lortat-Jacob Brice , Roussel Arnaud , Castier Yves , Messika Jonathan , Mal Hervé , Tymowski Christian de , Montravers Philippe
Published: May 2023

Background

Risk factors and the incidence of prolonged mechanical ventilation (PMV) after lung transplantation (LT) have been poorly described. The study assessed predictive factors of PMV after LT.

Methods

This observational, retrospective, monocentric study included all patients who received LT in Bichat Claude Bernard Hospital between January 2016 and December 2020. PMV was defined as a duration of MV > 14 days. Independent risk factors for PMV were studied using multivariate analysis. One-year survival depending on PMV was studied using Kaplan Meier and log-rank tests. A p value <0.05 was defined as significant.

Results

224 LT recipients were analysed. 64 (28%) of them received PMV for a median duration of 34 [26–52] days versus 2 [1–3] days without PMV. Independent risk factors for PMV were higher body mass index (BMI) (p = 0.031), diabetes mellitus of the recipient (p = 0.039), ECMO support during surgery (p = 0.029) and intraoperative transfusion >5 red blood cell units (p < 0.001). Increased mortality rates were observed at one-year in recipients who received PMV (44% versus 15%, p < 0.001).

Conclusion

PMV was associated with increased morbidity and mortality one-year after LT. Preoperative risk factors (BMI and diabetes mellitus) must be considered when selecting and conditioning the recipients.
Mental health and sleep quality among patients with asthma and COPD
Authors: Aldabayan Yousef Saad
Published: May 2023

This study aims to compare the mental health of patients with asthma and COPD in terms of anxiety, depression, and sleep quality and to examine the factors that predict sleep disturbance, anxiety, and depressive symptoms.

Methods

This quantitative cross-sectional study employed convenience sampling to enroll 200 patients with asthma and 190 patients with COPD. Data were gathered using a standardized self-administered questionnaire that contained sections on patients’ characteristics, the Sleep Quality, Anxiety, and Depression.

Results

The prevalence of poor sleep quality was 17.5 and 32.6% among asthmatic and COPD patients, respectively. The incidence of anxiety and depression was 38 and 49.5% among the patients with asthma, respectively. Their prevalence in patients with COPD was 48.9 and 34.7%, respectively. The multivariate regression analysis showed that marital status (married), BMI, education level (pre-university level), presence of comorbid illness, and depression were significant predictors of PSQI in asthmatic patients. Moreover, age, gender (male), marital status (married), education level (pre-university level), depression, and anxiety were significant predictors of PSQI among COPD participants. According to this study, COPD, and asthma pose serious health risks, including reduced sleep quality, anxiety, and depression.
Baohe pill decoction treats diarrhea induced by high-fat and high-protein diet by regulating lactase-producing bacteria in intestinal mucosa
Authors: Zhou Kang , Yi Xin , Tan Zhoujin , Peng Maijiao , Xiao Nenqun
Published: May 2023

Introduction

This study aimed to investigate the effects of Baohe pill decoction (BPD) on microbial, lactase activity, and lactase-producing bacteria in the intestinal mucosa of mice with diarrhea induced by high-fat and high-protein diet (HFHPD).

Methods

Thirty male Kunming (KM) mice were randomly divided into normal (NM), model (MD), and BPD groups. Diarrhea models were manufactured using HFHPD combined with a gavage of vegetable oil. At the end of modeling, the BPD group was given BPD (6.63 g·kg−1d−1) intervention twice daily for 3 d. The NM and MD groups were given equal amounts of sterile water. Subsequently, the intestinal mucosa of the mice was collected, one portion was used for microbial and lactase activity measurement, and the other portion was used for its lactase-producing bacterial characteristics by high-throughput sequencing technology.

Results

Our results showed that microbial and lactase activity of intestinal mucosa decreased significantly following diarrhea in mice (Pmicrobial < 0.05, Plactase < 0.001). After BPD intervention, microbial and lactase activity increased significantly (P < 0.01). The number of operational taxonomic units (OTUs), richness, and diversity index of lactase-producing bacteria increased in the BPD group compared to the MD group (P > 0.05), and the community structure were significant differences (P < 0.01). Compared to other groups, Saccharopolyspora, Rhizobium, Cedecea, and Escherichia were enriched in the BPD group. Notably, the relative abundance of the dominant lactase-producing genus Bifidobacterium decreased after BPD intervention.

Discussion

The mechanism of BPD in relieving diarrhea induced by HFHPD is closely related to the promotion of lactase activity in the intestinal mucosa, which may be achieved by regulating the structure of lactase-producing bacteria.
What happens in the brain when we die? Deciphering the neurophysiology of the final moments in life
Authors: Shlobin Nathan A. , Aru Jaan , Vicente Raul , Zemmar Ajmal
Published: May 2023

When do we die and what happens in the brain when we die? The mystery around these questions has engaged mankind for centuries. Despite the challenges to obtain recordings of the dying brain, recent studies have contributed to better understand the processes occurring during the last moments of life. In this review, we summarize the literature on neurophysiological changes around the time of death. Perhaps the only subjective description of death stems from survivors of near-death experiences (NDEs). Hallmarks of NDEs include memory recall, out-of-body experiences, dreaming, and meditative states. We survey the evidence investigating neurophysiological changes of these experiences in healthy subjects and attempt to incorporate this knowledge into the existing literature investigating the dying brain to provide valuations for the neurophysiological footprint and timeline of death. We aim to identify reasons explaining the variations of data between studies investigating this field and provide suggestions to standardize research and reduce data variability.
Increased expression of cell adhesion molecules in myofasciitis
Authors: Ma Xue , Gao Huajie , Xu Li , Bi Zhuajin , Ji Suqiong , Bu Bitao
Published: May 2023

Background

Myofasciitis is a heterogeneous group of diseases pathologically characterized by inflammatory cell infiltration into the fascia. Endothelial activation plays a critical role in the pathogenesis of the inflammatory response. However, the expression of cellular adhesion molecules (CAMs) in myofasciitis has not been investigated.

Methods

Data on clinical features, thigh magnetic resonance imaging, and muscle pathology were collected from five patients with myofasciitis. Immunohistochemical (IHC) staining and Western blot (WB) of the muscle biopsies from patients and healthy controls were performed.

Results

Increased levels of serum pro-inflammatory cytokines, including IL-6, TNF-α, and IL-2R, were detected in four patients. IHC staining and WB indicated significantly increased expression of cell adhesion molecules in blood vessels or inflammatory cells within the perimysium in muscle and fascia tissues of patients with myofasciitis compared to controls.

Conclusion

The up-regulation of CAMs in myofasciitis indicates endothelial activation, which may be potential therapy targets for the treatment of myofasciitis.
Validation of a portable monitor compared with polysomnography for screening of obstructive sleep apnea in polio survivors
Authors: Ding Qidi , Liu Jianghua , Wu Jinxian , Du Juan , Li Xiao , Wang Meng , Sun Yunliang , Yu Yan , Wang Jingyu , Sun Ting , Zhang Chi , Lv Changjun , Strohl Kingman P. , Han Fang , Dong Xiaosong
Published: May 2023

Subjective

Sleep-disordered breathing (SDB) is highly prevalent in polio survivors. Obstructive sleep apnea (OSA) is the most frequent type. Full polysomnography (PSG) is recommended for OSA diagnosis in patients with comorbidities by current practice guidelines, but it is not always accessible. The purpose of this study was to evaluate whether type 3 portable monitor (PM) or type 4 PM might be a viable alternative to PSG for the diagnosis of OSA in postpolio subjects.

Methods

A total of 48 community-living polio survivors (39 men and 9 women) with an average age of 54.4 ± 5.3 years referred for the evaluation of OSA and who volunteered to participate were recruited. First, they completed the Epworth Sleepiness Scale (ESS) questionnaire and underwent pulmonary function testing and blood gas tests the day before PSG night. Then, they underwent an overnight in-laboratory PSG with a type 3 PM and type 4 PM recording simultaneously.

Results

The AHI from PSG, respiratory event index (REI) from type 3 PM, and ODI3 from type 4 PM was 30.27 ± 22.51/h vs. 25.18 ± 19.11/h vs. 18.28 ± 15.13/h, respectively (P < 0.001). For AHI ≥ 5/h, the sensitivity and specificity of REI were 95.45 and 50%, respectively. For AHI ≥ 15/h, the sensitivity and specificity of REI were 87.88% and 93.33%, respectively. The Bland–Altman analysis of REI on PM vs. AHI on PSG showed a mean difference of −5.09 (95% confidence interval [CI]: −7.10, −3.08; P < 0.001) with limits of agreement ranging from −18.67 to 8.49 events/h. ROC curve analysis for patients with REI ≥ 15/h showed an area under the curve (AUC) of 0.97. For AHI ≥ 5/h, the sensitivity and specificity of ODI3 from type 4 PM were 86.36 and 75%, respectively. For patients with AHI ≥ 15/h, the sensitivity was 66.67%, and the specificity was 100%.

Conclusion

Type 3 PM and Type 4 PM could be alternative ways to screen OSA for polio survivors, especially for moderate to severe OSA.
Stroke recurrence is associated with unfavorable intracranial venous outflow in patients with symptomatic intracranial atherosclerotic large vessel severe stenosis or occlusion
Authors: Gao Jiali , Zhang Liang , Lin Jiaxin , Yang Jiajie , Yao Mingzheng , Cheng Zhongyuan , Cai Xiangran , Huang Li’an
Published: May 2023

Objective

The purpose of this study was to investigate the predictive value of intracranial venous outflow for recurrent cerebral ischemic events (RCIE) in patients with symptomatic intracranial atherosclerotic large-vessel severe stenosis or occlusion (sICAS-S/O).

Methods

This retrospective study included sICAS-S/O patients with anterior circulation who underwent dynamic computed tomography angiography (dCTA) and computed tomography perfusion (CTP). Arterial collaterals were evaluated using the pial arterial filling score for dCTA data, tissue-level collaterals (TLC) were assessed using the high-perfusion intensity ratio (HIR, Tmax >10 s/Tmax >6 s), and cortical veins were evaluated using the multi-phase venous score (MVS) for the vein of Labbé (VOL), sphenoparietal sinus (SPS), and superficial cerebral middle vein (SCMV). The relationships between multi-phase venous outflow (mVO), TLC, and 1-year RCIE were analyzed.

Results

Ninety-nine patients were included, 37 of whom had unfavorable mVO (mVO−) and 62 of whom had favorable mVO (mVO+). Compared with the mVO+ patients, mVO- patients had a higher admission National Institutes of Health Stroke Scale (NIHSS) score (median, 4 [interquartile range (IQR), 0–9] vs. 1 [IQR, 0–4]; p = 0.048), larger ischemic volume (median, 74.3 [IQR, 10.1–177.9] vs. 20.9 [IQR, 5–86.4] mL; p = 0.042), and worse tissue perfusion (median, 0.04 [IQR, 0–0.17] vs. 0 [IQR, 0–0.03]; p = 0.007). Multivariate regression analysis showed that mVO− was an independent predictor of 1-year RCIE.

Conclusion

For patients with sICAS-S/O of the anterior circulation, unfavorable intracranial venous outflow is a potential imaging indicator for predicting higher 1-year RCIE risk.
Qualitative and quantitative meta-analysis of acupuncture effects on the motor function of Parkinson's disease patients
Authors: Lei Suying , Fan Jingqi , Liu Xin , Xv Xiaoyan , Zhang Jiayan , Zhou Zipu , Zhuang Lixing
Published: May 2023

Objective

To explore the association between acupuncture sessions and its effects on the motor function of Parkinson's Disease (PD).

Methods

Eight databases and two clinical trials registries were searched from inception to August 2022. Randomized controlled trials (RCTs) that compared acupuncture with sham acupuncture, or antiparkinsonian drugs, were included. After qualitative meta-analysis, a non-linear meta regression approach with restricted cubic spline was used to investigate the dose-response relationship between acupuncture sessions and their efficacy on the Unified Parkinson's Disease Rating Scale Part III (UPDRS-III) score. Subgroup meta-analysis was performed of the included studies according to the weekly acupuncture frequency. And finally, the included studies containing the determination of intermediate efficacy were compared.

Results

Of the 268 citations screened, 16 studies (462 patients of PD) were included. The qualitative meta-analysis showed that the acupuncture group had better effect on UPDRS-III scores than the control group. And the quantitative meta-analysis suggested that acupuncture dose was correlated with the reduction of UPDRS-III score in PD patients with motor symptoms. In subgroup analysis, on the one hand, when the frequency of acupuncture was no more than 3 times a week, with the increase of acupuncture session, the changes of UPDRS-III score decreased and then increased (P = 0.000). On the other hand, when acupuncture for more than 3 times a week and the dose of acupuncture treatment was <60 times, the changes of UPDRS-III score increased with the increase of acupuncture dose, but the score stopped to decrease if the dose continued to increase (P = 0.020). The comparative analysis of two quantitative RCTs found that the score improvement was more significant at the higher weekly acupuncture frequency.

Interpretation

This study found that when treating PD patients with motor symptoms, acupuncture treatment may need to reach a certain dose to obtain better therapeutic effect and excessive acupuncture stimulation may cause the body to develop a certain tolerance. However, the above results still need to be verified by more high-quality clinical studies. The protocol was registered on PROSPERO International Prospective Register of Systematic Reviews (CRD42022351428).
A systematic review and meta-analysis of voxel-based morphometric studies of fibromyalgia
Authors: Xin Ming , Qu Yuzhu , Peng Xingfu , Zhu Deliang , Cheng Shirui
Published: May 2023

Objective

Although neuroimaging investigations have revealed significant changes in brain structure in fibromyalgia (FM) patients, these findings are inconsistent. The current study conducted a systematic review and meta-analysis of voxel-based morphometric studies in order to comprehend those alterations in brain structure in FM patients.

Methods

Voxel-based morphometric (VBM) studies published up to January 17, 2023 were searched in the Web of Science, PubMed, EMBASE, Cochrane Library (CENTRAL), China National Knowledge Infrastructure (CNKI), Chongqing VIP, Wanfang Database. Two independent researchers carried out study screening, quality assessment, clinical data and neuroimaging data extraction. The whole-brain voxel-based gray matter (GM) data of FM patients were collected from eligible studies, and meta-analyzed using anisotropic effect size-signed differential mapping (AES-SDM).

Results

Twelve researches were included in this study, including 289 FM patients (mean age: 47.36 years) and 272 HS (mean age: 47.34 years). According to the meta-analysis, FM patients had increased GM in the right postcentral gyrus and left angular gyrus, and decreased GM in the right cingulate gyrus, right paracingulate gyrus, left cerebellum, and left gyrus rectus.

Conclusion

Our study suggests that fibromyalgia patients have altered gray matter in several brain regions that are involved in affective, cognitive functions, and in motor adaptations to pain processing.
Recessive aminoacyl-tRNA synthetase disorders: lessons learned from in vivo disease models
Authors: Kalotay Elizabeth , Klugmann Matthias , Housley Gary D. , Fröhlich Dominik
Published: May 2023

Protein synthesis is a fundamental process that underpins almost every aspect of cellular functioning. Intriguingly, despite their common function, recessive mutations in aminoacyl-tRNA synthetases (ARSs), the family of enzymes that pair tRNA molecules with amino acids prior to translation on the ribosome, cause a diverse range of multi-system disorders that affect specific groups of tissues. Neurological development is impaired in most ARS-associated disorders. In addition to central nervous system defects, diseases caused by recessive mutations in cytosolic ARSs commonly affect the liver and lungs. Patients with biallelic mutations in mitochondrial ARSs often present with encephalopathies, with variable involvement of peripheral systems. Many of these disorders cause severe disability, and as understanding of their pathogenesis is currently limited, there are no effective treatments available. To address this, accurate in vivo models for most of the recessive ARS diseases are urgently needed. Here, we discuss approaches that have been taken to model recessive ARS diseases in vivo, highlighting some of the challenges that have arisen in this process, as well as key results obtained from these models. Further development and refinement of animal models is essential to facilitate a better understanding of the pathophysiology underlying recessive ARS diseases, and ultimately to enable development and testing of effective therapies.