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Background/Objectives: Static reference intervals (RIs) fail to capture the dynamic changes in bioanalytes. This study aimed to develop gender-based continuous age- and season-related RIs for neuron-specific enolase (NSE) using real-world data and to compare them with partitioned RIs. Methods: The NSE results from 4097 individuals were included after rigorous screening. Partitioned RIs were determined using the Hoffmann method. Generalized additive models for location, scale and shape (GAMLSS) were selected to develop continuous RIs. Results: The partitioned RIs are as follows: <16.4 µg/L for males aged ≥19 years; <14.47 µg/L for females aged 19–49 years; and <17.25 µg/L for females aged ≥50 years. For continuous RIs, NSE levels in males remain stable with age, while in females, NSE levels evidently increase around the age of 50. Although less impactful than age, seasonal changes still affect NSE levels. Dynamic changes and continuous RIs for NSE are visualized in this study. Conclusions: We developed gender-based continuous age- and season-integrated RIs for NSE in North China, highlighting the variation in NSE levels in females with age and season. Compared to static RIs, continuous RIs are more responsive to NSE, potentially enhancing the precision and individualization of health assessments. [ABSTRACT FROM AUTHOR]

With the aim of improving the uncertainties associated with the correct diagnosis of seronegative rheumatoid arthritis (RA) and identifying those at risk of developing interstitial lung disease (ILD), we have designed new peptide antigens bearing three post-translational modifications (PTMs) (citrulline, homocitrulline and acetyl-lysine) related to RA that could complement existing tests based on anti-citrullinated peptide/protein antibodies (ACPAs). Several chimeric peptides were synthesized and comparatively tested as antigens in ELISAs with two cohorts of sera: 178 RAs and 110 healthy blood donors. The results indicated that although chimeric peptides containing all three PTMs and vimentin and enolase domains do not significantly outperform existing ACPA tests in terms of sensitivity and specificity, they show potential to complement current assays, especially when detecting antibodies in some seronegative patients. Furthermore, the presence of these autoantibodies significantly identified patients with RA and ILD. We can conclude that the identification of specific autoantibody profiles using synthetic antigens containing peptide domains derived from proteins present in the human joint could help in the early detection of the risk of ILD in patients with RA and be useful for adapting follow-up strategies and guiding decisions during treatment. [ABSTRACT FROM AUTHOR]

Organic carbon fixed in chloroplasts through the Calvin–Benson–Bassham Cycle can be diverted toward different metabolic fates, including cytoplasmic and mitochondrial respiration, gluconeogenesis, and synthesis of diverse plastid metabolites via the pyruvate hub. In plants, pyruvate is principally produced via cytoplasmic glycolysis, although a plastid-targeted lower glycolytic pathway is known to exist in non-photosynthetic tissue. Here, we characterized a lower plastid glycolysis–gluconeogenesis pathway enabling the direct interconversion of glyceraldehyde-3-phosphate and phospho-enol-pyruvate in diatoms, ecologically important marine algae distantly related to plants. We show that two reversible enzymes required to complete diatom plastid glycolysis–gluconeogenesis, Enolase and bis-phosphoglycerate mutase (PGAM), originated through duplications of mitochondria-targeted respiratory isoforms. Through CRISPR-Cas9 mutagenesis, integrative 'omic analyses, and measured kinetics of expressed enzymes in the diatom Phaeodactylum tricornutum, we present evidence that this pathway diverts plastid glyceraldehyde-3-phosphate into the pyruvate hub, and may also function in the gluconeogenic direction. Considering experimental data, we show that this pathway has different roles dependent in particular on day length and environmental temperature, and show that the cpEnolase and cpPGAM genes are expressed at elevated levels in high-latitude oceans where diatoms are abundant. Our data provide evolutionary, meta-genomic, and functional insights into a poorly understood yet evolutionarily recurrent plastid metabolic pathway. Environmental and experimental methods reveal the importance of plastid-targeted Enolase and bis-phosphoglycerate mutase enzymes to diatoms, marine algae that contribute to global primary production. [ABSTRACT FROM AUTHOR]

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Heat stress severely affects the yield and quality of maize. Melatonin (N-acetyl-5-methoxy-tryptamin, MT) plays an important role in various types of stress resistance in plants, including heat tolerance. Enolase (ENO, 2-phospho-D-glycerate hydrolyase) contributes to plant growth, development, and stress response. As of now, the molecular mechanisms by which MT and ENO1 affect heat tolerance are unknown. In our research, we have revealed that heat stress (H) and heat stress + MT (MH) treatment upregulate ZmENO1 expression levels by 15 and 20 times, respectively. ZmENO1 overexpression and mutant maize lines were created by transgenic and genome editing. These results illustrate that heat stress has a significant impact on the growth of maize at the seedling stage. However, ZmENO1-OE lines showed a lower degree of susceptibility to heat stress, whereas the mutant exhibited the most severe effects. Under heat stress, exogenous application of MT improves heat resistance in maize. The ZmENO1-OE lines exhibited the best growth and highest survival rate, while the zmeno1 mutants showed the least desirable results. Following treatment with H and MH, the level of MT in ZmENO1-OE lines exhibited the greatest increase and reached the maximum value, whereas the level of MT in the zmeno1 mutant was the lowest. Heat stress decreased the maize's relative water content and fresh weight, although ZmENO1-OE lines had the highest and zmeno1 mutants had the lowest. Heat stress led to an increase in the levels of MDA, hydrogen peroxide, and superoxide in all plants. Additionally, the ionic permeability and osmotic potential of the plants were significantly increased. However, the levels of MT were decreased in all plants, with the greatest decrease observed in the ZmENO1-OE lines. Interestingly, the zmeno1 mutant plants had the highest expression levels of MT. Heat stress-induced upregulation of ZmSOD, ZmPOD, ZmAPX, ZmCAT, ZmP5CS, and ZmProDH in all plants. However, the ZmENO1-OE lines exhibited the greatest increase in expression levels, while the zmeno1 mutants showed the lowest increase following MT spraying. The patterns of SOD, POD, APX, and CAT enzyme activity, as well as proline and soluble protein content, aligned with the variations in the expression levels of these genes. Our findings indicate that MT can upregulate the expression of the ZmENO1 gene. Upregulating the ZmENO1 gene resulted in elevated expression levels of ZmSOD, ZmPOD, ZmAPX, ZmCAT, ZmP5CS, and ZmProDH. This led to increased activity of antioxidant enzymes and higher levels of osmoregulatory substances. Consequently, it mitigated the cell membrane damage caused by heat stress and ultimately improved the heat resistance of maize. The results of this study provide genetic resources for molecular design breeding and lay a solid foundation for further exploring the molecular mechanism of MT regulation of heat stress tolerance in maize. [ABSTRACT FROM AUTHOR]

Background and Objectives: Most patients who are successfully resuscitated from cardiac arrest remain comatose, and only half regain consciousness 72 h after the arrest. Neuroprognostication methods can be complex and even inconclusive. As mitochondrial components have been identified as markers of post-cardiac-arrest injury and associated with survival, we aimed to investigate cytochrome c and mtDNA in comatose patients after cardiac arrest to compare neurological outcomes and to evaluate the markers' neuroprognostic value. Materials and Methods: This prospective observational study included 86 comatose post-cardiac-arrest patients and 10 healthy controls. Cytochrome c and mtDNA were determined at admission. Neuron-specific enolase (NSE) was measured after 72 h. Additional neuroprognostication methods were performed when patients remained unconscious. Cerebral performance category (CPC) was determined. Results: Cytochrome c was elevated in patients compared to healthy controls (2.029 [0.85–4.97] ng/mL vs. 0 [0.0–0.16], p < 0.001) but not mtDNA (95,228 [52,566–194,060] vs. 41,466 [28,199–104,708] copies/μL, p = 0.074). Compared to patients with CPC 1–2, patients with CPC 3–5 had higher cytochrome c (1.735 [0.717–3.40] vs. 4.109 [1.149–8.457] ng/mL, p = 0.011), with no differences in mtDNA (87,855 [47,598–172,464] vs. 126,452 [69,447–260,334] copies/μL, p = 0.208). Patients with CPC 1–2 and CPC 3–5 differed in all neuroprognostication methods. In patients with good vs. poor neurological outcome, ROC AUC was 0.664 (p = 0.011) for cytochrome c, 0.582 (p = 0.208) for mtDNA, and 0.860 (p < 0.001) for NSE. The correlation between NSE and cytochrome c was moderate, with a coefficient of 0.576 (p < 0.001). Conclusions: Cytochrome c was higher in comatose patients after cardiac arrest compared to healthy controls and higher in post-cardiac-arrest patients with poor neurological outcomes. Although cytochrome c correlated with NSE, its neuroprognostic value was poor. We found no differences in mtDNA. [ABSTRACT FROM AUTHOR]

Objective: This study aimed to investigate the serum levels of neuron-specific enolase (NSE) in sepsis-associated encephalopathy (SAE) and perform a meta-analysis to assess the diagnostic and prognostic potential of serum NSE in SAE patients. Methods: We searched English and Chinese databases for studies related to SAE that reported serum NSE levels until November 2023. We extracted information from these studies including the first author and year of publication, the number of samples, the gender and age of patients, the collection time of blood samples in patients, the assay method of serum NSE, the study methods, and the levels of serum NSE with units of ng/mL. The quality assessment of diagnostic accuracy studies 2 (QUADAS-2) tool was used to evaluate the study quality. A meta-analysis was performed using Review Manager version 5.3, employing either a random effects model or a fixed effects model. Results: A total of 17 studies were included in the final meta-analysis, including 682 SAE patients and 946 NE patients. The meta-analysis demonstrated significantly higher serum NSE levels in SAE patients compared to NE patients (Z = 5.97, P < 0.001, MD = 7.79, 95%CI 5.23–10.34), irrespective of the method used for serum NSE detection (Z = 6.15, P < 0.001, mean difference [MD] = 7.75, 95%CI 5.28–10.22) and the study methods (Z = 5.97, P < 0.001, MD = 7.79, 95%CI 5.23–10.34). Furthermore, sepsis patients with a favorable outcome showed significantly lower levels of serum NSE compared to those with an unfavorable outcome (death or adverse neurological outcomes) (Z = 5.44, P < 0.001, MD = − 5.34, 95%CI − 7.26–3.42). Conclusion: The Serum level of NSE in SAE patients was significantly higher than that in septic patients without encephalopathy. The higher the serum NSE level in SAE patients, the higher their mortality rate and incidence of adverse neurological outcomes. [ABSTRACT FROM AUTHOR]

Numerous studies have demonstrated that neuron-specific enolase (NSE) serves as a distinctive indicator of neuronal injury, with its concentration in blood reflecting the extent and magnitude of nervous system damage, and the expression of serum NSE is correlated with cognitive dysfunction. The assessment of NSE holds significant importance in diagnosing cognitive dysfunction, assessing disease severity, predicting prognosis, and guiding treatment. In this review, the research progress of NSE in cognitive dysfunction was reviewed, and the value of serumNSE level in predicting disease severity and prognosis of patients with cognitive dysfunction was discussed. [ABSTRACT FROM AUTHOR]

Background: Any illness that disrupts the central nervous system in the first few days of life can induce encephalopathy among neonates. This ailment can have a variety of causes. Of all the CSF neurobiochemical markers in babies with HIE, NSE has received the greatest amount of research. Up to four percent of all soluble brain proteins are NSE, a dimeric glycolytic enzyme. Neurons and neuroendocrine cells contain it primarily in their cytoplasm. When neurons suffer damage to their axons, NSE is found in high concentrations in their cytoplasm. Thus, it is sensitive for axonal injury and neuronal cell death and has a fairly high specificity for detecting axonal damage. Traditional terminology for neonatal encephalopathy (NE) has focused on hypoxic-ischemic encephalopathy, which occurs when an intrapartum incident causes perinatal hypoxia-ischemia. Objective: The purpose of this study was to review the diagnostic value of the Neuron-Specific Enolase biomarker in the evaluation of early detection and its role in prognosis of neonatal encephalopathy. Methods: In our search for information on NSE and its level in neonates with encephalopathy, we used Google Scholar, Science Direct, PubMed, and other internet databases. Additionally, the writers combed through relevant literature for references, however they only included researches that were either very recent or covering the years from 2010 to 2023. Conclusion: Serum NSE is considered to be a reliable marker for the diagnosis of different stages of NE. [ABSTRACT FROM AUTHOR]

Background: Neuron-specific enolase is a dimeric isoform of the glycolytic enzyme that is mostly present in neurons is important as a diagnostic and a prognostic value in pediatric encephalopathy which is common and is potentially life threatening, previous studies were done worldwide to detect its diagnostic and prognostic value in acute encephalopathy among adult and pediatric age groups. The current study aimed at evaluation of serum neuron specific enolase for early diagnosis of pediatric acute encephalopathy. Methods: This case control study was carried out in pediatric intensive care unit and the inpatient pediatric department, faculty of medicine, Zagazig University and was conducted on 24 patients suffering from acute encephalopathy and 24 patients presented with seizures without encephalopathy. serum neuron-specific enolase was measured. Results: Serum neuron-specific enolase level had sensitivity 95%, specificity 88.5%, PPV 94%, NPV 90% and accuracy 93% in prediction of encephalopathy. Conclusions: There was statistically significant increase in serum NSE levels in encephalopathy group more than seizures group. [ABSTRACT FROM AUTHOR]

SUMMARY: The protein‐repairing enzyme (PRE) PROTEIN L‐ISOASPARTYL METHYLTRANSFERASE (PIMT) influences seed vigor by repairing isoaspartyl‐mediated protein damage in seeds. However, PIMTs function in other seed traits, and the mechanisms by which PIMT affects such seed traits are still poorly understood. Herein, through molecular, biochemical, and genetic studies using overexpression and RNAi lines in Oryza sativa and Arabidopsis thaliana, we demonstrate that PIMT not only affects seed vigor but also affects seed size and weight by modulating enolase (ENO) activity. We have identified ENO2, a glycolytic enzyme, as a PIMT interacting protein through Y2H cDNA library screening, and this interaction was further validated by BiFC and co‐immunoprecipitation assay. We show that mutation or suppression of ENO2 expression results in reduced seed vigor, seed size, and weight. We also proved that ENO2 undergoes isoAsp modification that affects its activity in both in vivo and in vitro conditions. Further, using MS/MS analyses, amino acid residues that undergo isoAsp modification in ENO2 were identified. We also demonstrate that PIMT repairs such isoAsp modification in ENO2 protein, protecting its vital cellular functions during seed maturation and storage, and plays a vital role in regulating seed size, weight, and seed vigor. Taken together, our study identified ENO2 as a novel substrate of PIMT, and both ENO2 and PIMT in turn implicate in agronomically important seed traits. [ABSTRACT FROM AUTHOR]

The storage of serum tumor markers (STM) at −18 °C for one year has been a legal requirement in France since 1999, but has been abolished in 2022. This raises the question of the relevance of maintaining these biobanks in terms of conditions of storage. These should only be implemented after validation; in order to maintain the integrity of the biological sample and must be controlled over time according to the laboratoryʼs procedures. The aim of the study was to assess the suitability of storing 10 STMs by evaluating their stability after one year of storage at −18 °C. A new immuno-enzymatic assay (A+1) was conducted on samples that had been stored at −18 °C for one year after an initial assay (A) of one of the following STMs: carcino-embryonic antigen (CEA), alpha-fetoprotein (AFP), carbohydrate antigen 125 (CA125), carbohydrate antigen 15-3 (CA15-3), carbohydrate antigen 19-9 (CA19-9), total (TPSA), and free (FPSA) prostate-specific antigen, calcitonin (CT), thyroglobulin (TG), and neuro-specific enolase (NSE). The results were confronted to four different permissible error sources. In total, 1148 A+1 assays were performed. A strong correlation between A+1 and A values was found for all analytes, but with a statistically significant reduction in the mean A+1 concentration compared to the mean A concentration in 7/10 STMs. The bias induced by conservation seems to be technically unsustainable if we rely on the repositories closest to the current analytical performances. These results support the discontinuation of mandatory STM biobank storage at −18 °C, which requires considerable technical time and organizational effort. [ABSTRACT FROM AUTHOR]

Neuron-specific enolase (NSE) derived from neurons and peripheral neuroendocrine cells is a biomarker for neuroendocrine tumors and for prognostication in comatose cardiac arrest survivors. However, as platelets and erythrocytes contain NSE, hemolysis causes falsely elevated NSE. We used native serum and hemolysate derived from the same patients to make serial dilutions, and subsequently measured NSE (mNSE) and hemolytic index (HI) in each dilution. An algorithm suitable for the laboratory information system was developed based on the mNSE, HI and the estimated gradient of hemolytic interference from 30 patients. We estimated the associated uncertainty of the corrected NSE (cNSE) results based on the observed range of the gradient and derived an equation for cNSE for samples with limited hemolysis (i.e. 5 < HI ≤ 30): cNSE = mNSE − HI × (0.34 ± 0.23) µg/L. By semi-quantitatively grading the contribution from limited hemolysis, a texted result noting the hemolysis-associated degree of uncertainty can accompany the cNSE result. The major challenge of hemolysis when using serum NSE as a biomarker can be managed using an automated algorithm for correction of NSE results based on degree of hemolysis. However, laboratorians and clinicians should be aware of the limitations associated with in vivo hemolysis. [ABSTRACT FROM AUTHOR]

Research on prognostic factors for good outcomes in out-of-hospital cardiac arrest (OHCA) survivors is lacking. We assessed whether normal levels of normal neuron-specific enolase (NSE) value would be useful for predicting good neurological outcomes in comatose OHCA survivors treated with targeted temperature management (TTM). This registry-based observational study with consecutive adult (≥18 years) OHCA survivors with TTM who underwent NSE measurement 48 hours after cardiac arrest was conducted from October 2015 to November 2022. Normal NSE values defined as the upper limit of the normal range by the manufacturer (NSE <16.3 μg/L) and guideline-suggested (NSE < 60 μg/L) were examined for good neurologic outcomes, defined as Cerebral Performance Categories ≤2, at 6 months post-survival. Among 226 OHCA survivors with TTM, 200 patients who underwent NSE measurement were enrolled. The manufacturer-suggested normal NSE values (<16.3 μg/L) had a specificity of 99.17% for good neurological outcomes with a very low sensitivity of 12.66%. NSE <60 μg/L predicted good outcomes with a sensitivity of 87.34% and specificity of 72.73%. However, excluding 14 poor-outcome patients who died from multi-organ dysfunction excluding hypoxic brain injury, the sensitivity and specificity of normal NSE values were 12.66% and 99.07% of NSE < 16.3 μg/L, and 87.34% and 82.24% of NSE < 60 μg/L. The manufacturer-suggested normal NSE had high specificity with low sensitivity, but the guideline-suggested normal NSE value had a comparatively low specificity for good outcome prediction in OHCA survivors. Our data demonstrate normal NSE levels can be useful as a tool for multimodal appropriation of good outcome prediction. [ABSTRACT FROM AUTHOR]

Candida albicans is the predominant cause of systemic candidiasis, although other non albicans Candida species are progressively becoming more widespread nowadays. Candida auris has emerged as a deadly multidrug-resistant fungal pathogen, posing a significant threat to global public health. In the absence of effective antifungal therapies, the development of a vaccine against C. auris infections is imperative. Enolase, a key glycolytic enzyme, has emerged as a promising vaccine candidate due to its immunogenic properties and essential role in fungal virulence. Herein, full-length Enolase gene sequences from C. albicans and C. auris were cloned into suitable expression vector and transformed into Escherichia coli expression hosts. Recombinant Enolase proteins were successfully expressed and purified using affinity chromatography under native conditions, followed by SDS-PAGE characterization and Western blot analysis. CD spectroscopy verified the existence of expressed proteins in soluble native conformation. Preliminary in silico studies verified the immunogenicity of recombinant Enolase proteins isolated from both C. albicans and C. auris. Furthermore, bioinformatics analysis revealed conserved B-cell and T-cell epitopes across C. albicans and C. auris Enolase proteins, suggesting potential cross-reactivity and broad-spectrumvaccine efficacy. Our findings are anticipated to play a role in advancing therapeutic as well as diagnostic strategies against systemic candidiasis. [ABSTRACT FROM AUTHOR]

A study published in Systematic Reviews by Zhi et al. explores the use of neuron-specific enolase (NSE) as a diagnostic tool for sepsis-associated encephalopathy (SAE). The study found that SAE patients had significantly higher levels of serum NSE compared to septic patients without encephalopathy. Higher levels of serum NSE were associated with increased mortality rates and adverse neurological outcomes. While the study provides valuable insights, further research is needed to fully understand the relationship between NSE and SAE. [Extracted from the article]

Background: Neurospecific Enolase (NSE), a multifunctional protein, is present in various tissues of the body and plays an important role in many disease processes, such as infection, inflammation, tumours, injury, and immunity. In recent years, the application of NSE in respiratory diseases has become increasingly widespread and a research hotspot. Objective: This study aims to explore the relationship between NSE and childhood pneumonia, providing assistance for the diagnosis and assessment of pneumonia. Methods: Using prospective research and case-control methods, We selected 129 children with pneumonia hospitalised in Weifang People's Hospital from September 2020 to April 2022 as the case group. Among them were 67 cases of Mycoplasma pneumoniae pneumonia (MP+), 62 cases of non-Mycoplasma pneumoniae pneumonia (MP -), and 21 cases of severe pneumonia. At the same time, 136 children who underwent outpatient health examinations were selected as the control group. The levels of NSE, ESR, CRP in cases group and NSE in control group were measured separately. Result: The NSE levels in the MP + group were 17.86 (14.29–22.54) ng/mL, while those in the MP- group were 17.89 (14.10–21.66) ng/mL, both of which were higher than the control group's NSE levels of 13.26(12.18,14.44) ng/mL (H = 46.92, P = 0.000). There was no statistically significant difference in NSE levels between the MP + and MP - groups (P > 0.05). The NSE level in the severe pneumonia group was 27.38 (13.95–34.06) ng/mL, higher than that in the mild pneumonia group, which was 17.68 (14.27–21.04) ng/mL, (P = 0.024). The AUC values for diagnosing pneumonia are NSE0.714, CRP0.539, and ESR0.535, with NSE having the highest diagnostic value. Conclusion: Serum NSE can serve as an inflammatory indicator for paediatric pneumonia, which has important clinical guidance significance for the diagnosis, condition evaluation, and prognosis of paediatric pneumonia. [ABSTRACT FROM AUTHOR]

Hybrid breeding is an effective approach to generate better varieties and prevent variety degradation. The present study investigated the metabolic changes underlying growth superiority in the novel Gymnocypris hybrid (GH), Gymnocypris przewalskii ♀ (GP) × G. eckloni ♂ (GE). The ranking of survival rate was GH > GE > GP, whereas the ranking of growth rate was GE > GH > GP. A proteomic analysis of G. hybrid and its parents was conducted to elucidate the metabolic changes underlying growth superiority. Identified pathways were primarily associated with amino acid, carbohydrate, energy, lipid, and nucleotide metabolism. These metabolic pathways, which are closely associated with growth, are controlled through regulation of the expression of numerous proteins, including adenosylhomocysteinase, hydroxypyruvate reductase, glutamate-cysteine ligase, L-lactate dehydrogenase, creatine kinase, GDP-L-fucose synthase, pyruvate kinase, fructose-bisphosphate aldolase, carbonic anhydrase, phosphopyruvate hydratase, phosphoglycerate kinase, S-(hydroxymethyl) glutathione dehydrogenase, and AMP deaminase. Real-time PCR assays showed that the level of mRNA expression of differentially expressed genes was positively correlated with growth. Proteins that were differentially expressed in GH exhibited fewer differences from GP and more differences from GE. These data are the first to reveal the molecular mechanism whereby growth is regulated in G. hybrid and its parents at the protein level. The study thus provides important information for genetic breeding and improvement of G. hybrid for aquaculture production. [ABSTRACT FROM AUTHOR]

[Objective]The objective of this study is to identify the genes(TaENO2s)encoding the enolase family member ENO2 in the important crop plant of wheat(Triticum aestivum L.)and to determine the alternative translation feature and protein enolase activity of TaENO2s.[Method]Wheat TaENO2s were identified by bioinformatics. One of the identified TaENO2s was selected as a representative for protoplast-based exogenous expression and characterization of alternative translation products. This selected TaENO2 was also subjected to prokaryotic expression for purification of recombinant protein. The enolase activity of the purified recombinant protein was determined with in vitro assays.[Result]The hexaploid wheat genome had three homeologous TaENO2s which sequences encoded a conserved enolase catalytic center and had a putative alternative translation start site at the internal ATG codon encoding the residue M94(ATGM94) of the enolase sequence. When exogenously expressed in protoplasts, the representative TaENO2 encoded two proteins, one full-length form of cytoplasmic and nuclear enolase protein and one N-terminal truncated form of nuclear transcriptional repressor TaMBP-1, while the variant of the same gene containing mutated ATGM94 only encoded the full-length form of enolase protein. Soluble recombinant GST-TaENO2 protein expressed in Escherichia coli was successfully purified and verified to have an in vitro enzymatic activity to catalyze the conversion from 2-phosphoglycerate (2-PGA)to phosphoenolpyruvate(PEP)[Conclusion]The wheat genome has three TaENO2 genes encoding the enolase protein ENO2. Under the condition of exogenous expression, TaENO2 encodes two protein products via mRNA alternative translation, one enolase protein translated from the first start codon and one TaMBP-1 protein translated from the internal start codon(2-PGA)to phosphoenolpyruvate(PEP).[Conclusion[The wheat genome has three TaENO2 genes encoding the enolase protein ENO2. Under the condition of exogenous expression, TaENO2 encodes two protein products via mRNA alternative translation, one enolase protein translated from the first start codon and one TaMBP-1 protein translated from the internal start codon ATGM94. The prokaryotically expressed recombinant protein GST-TaENO2 possesses in vitro enolase activity.. The prokaryotically expressed recombinant protein GST-TaENO2 possesses in vitro enolase activity. [ABSTRACT FROM AUTHOR]

Since the discovery of the neuron-specific protein by Moore and McGregor in 1965, tens of thousands of studies have investigated the basic and applied significance of neuron-specific enolase (NSE). This promising biomarker, according to many researchers, has not found widespread use in clinical practice, particularly in acute cerebrovascular accidents. Moreover, the several studies refuting the usefulness of serum NSE measurement in critically ill patients leads us to consider the reasons for such contradictory conclusions. In this article, we have analyzed the main directions in the study of NSE and expressed our perspective on the reasons for the contradictory results and the difficulties in implementing the results of these studies in clinical practice. In our opinion, the method of the enzyme-linked immunosorbent assay (ELISA) used in the majority of the studies is inappropriate for the evaluation of NSE as a marker of central nervous system damage, because it does not allow for the differentiation of heterodimers of enolases and the assessment of the enzymatic activity of this group of enzymatic proteins. Therefore, the methodological approach for the evaluation of NSE (γγ-enolase) as a biomarker needs to be elaborated and improved. Furthermore, the specificity of the applied research methods and the appropriateness of the continued use of the term "neuron-specific enolase" must be addressed. [ABSTRACT FROM AUTHOR]

Ticks, blood-sucking ectoparasites, spread diseases to humans and animals. Haemaphysalis longicornis is a significant vector for tick-borne diseases in medical and veterinary contexts. Identifying protective antigens in H. longicornis for an anti-tick vaccine is a key tick control strategy. Enolase, a multifunctional protein, significantly converts D-2-phosphoglycerate and phosphoenolpyruvate in glycolysis and gluconeogenesis in cell cytoplasm. This study cloned a complete open reading frame (ORF) of enolase from the H. longicornis tick and characterized its transcriptional and silencing effect. We amplified the full-length cDNA of the enolase gene using rapid amplification of cDNA ends. The complete cDNA, with an ORF of 1,297 nucleotides, encoded a 432-amino acid polypeptide. Enolase of the Jeju strain H. longicornis exhibited the highest sequence similarity with H. flava (98%), followed by Dermacentor silvarum (82%). The enolase motifs identified included N-terminal and C-terminal regions, magnesium binding sites, and several phosphorylation sites. Reverse transcription-polymerase chain reaction (RT-PCR) analysis indicated that enolase mRNA transcripts were expressed across all developmental stages of ticks and organs such as salivary gland and midgut. RT-PCR showed higher transcript levels in syn-ganglia, suggesting that synganglion nerves influence enolase's role in tick salivary glands. We injected enolase double-stranded RNA into adult unfed female ticks, after which they were subsequently fed with normal unfed males until they spontaneously dropped off. RNA interference significantly (P<0.05) reduced feeding and reproduction, along with abnormalities in eggs (no embryos) and hatching. These findings suggest enolase is a promising target for future tick control strategies. [ABSTRACT FROM AUTHOR]

Importance: Haemaphysalis longicornis is an obligate blood-sucking ectoparasite that has gained attention due its role of transmitting medically and veterinary significant pathogens and it is the most common tick species in Republic of Korea. The preferred strategy for controlling ticks is a multi-antigenic vaccination. Testing the efficiency of a combination antigen is a promising method for creating a tick vaccine. Objective: The aim of the current research was to analyze the role of subolesin and enolase in feeding and reproduction of H. longicornis by gene silencing. Methods: In this study, we used RNA interference to silence salivary enolase and subolesin in H. longicornis. Unfed female ticks injected with double-stranded RNA targeting subolesin and enolase were attached and fed normally on the rabbit's ear. Real-time polymerase chain reaction was used to confirm the extent of knockdown. Results: Ticks in the subolesin or enolase dsRNA groups showed knockdown rates of 80% and 60% respectively. Ticks in the combination dsRNA (subolesin and enolase) group showed an 80% knockdown. Knockdown of subolesin and enolase resulted in significant depletion in feeding, blood engorgement weight, attachment rate, and egg laying. Silencing of both resulted in a significant (p < 0.05) reduction in tick engorgement, egg laying, egg hatching (15%), and reproduction. Conclusions and Relevance: Our results suggest that subolesin and enolase are an exciting target for future tick control strategies. [ABSTRACT FROM AUTHOR]

Background: We compared the performance of 21 different assays performed by the Wantai Wan200+ (Wantai BioPharm, Beijing, China) with respect to other methods in use at the University Hospital of Padova (AOPD), Italy. Methods: The plasma (P) or serum (S) of 5027 leftover samples, collected from May to Sept 2023, was either analyzed or frozen at −20 °C. Beckman DXI800 (DXI), Roche Cobas 8000 e801 (RC), Snibe Maglumi 4000 plus (SM), DiaSorin Liaison XL (DL) and Binding Site Optilite (BS) equipment were used at the AOPD. P-procalcitonin (PCT), DXI; P-Troponin I (TnI), DXI; S-CA125, DXI; S-free PSA (f-PSA), DXI; S-total PSA (t-PSA), DXI; S-IL6, SM; P-Troponin T (TnT), RC; P-NT-proBNP, RC; P-Neuron-Specific Enolase (NSE), RC; S-CA15-3, DL; S-CA19-9, DL; S-AFP, DL; and S-CEA, DL were tested in fresh samples. P-Myoglobin (Myo), DXI; P-Cyfra21-1, RC; S-β2 microglobulin (B2MIC), BS; S-HE4, SM; S-PGI, SM; S-PGII, SM; S-CA72-4, SM; and S-CA50, SM were analyzed in frozen and thawed samples. Bland–Altman (BA), Passing–Bablok (PB) and Cohen's Kappa (CKa) metrics were used as statistics. Results: An excellent comparability profile was found for 11 analytes. For example, the t-PSA CKa was 0.94 (95%CI: 0.90 to 0.98), and the PB slope and intercept were 1.02 (95%CI: 0.99 to 1.03) and 0.02 (95%CI: 0.01 to 0.03), respectively; the BA bias was 2.25 (95%CI: −0.43 to 4.93). Ten tested measurands demonstrated a suboptimal comparability profile. Biological variation in EFLM (EuBIVAS) performance specifications was evaluated to assess the clinical relevance of measured biases. Conclusions: Evaluation of the Wantai Wan200+'s performance suggests that between-method differences did not exceed the calculated bias. Metrological traceability may influence the comparisons obtained for some measurands. [ABSTRACT FROM AUTHOR]

Background: Sepsis-associated encephalopathy (SAE) is one of the most ubiquitous complications of sepsis and is characterized by cognitive impairment, poor prognosis, and a lack of uniform clinical diagnostic criteria. Therefore, this study investigated the early diagnostic and prognostic value of serum neuronspecific enolase (NSE) in SAE. Methods: This systematic review and meta-analysis systematically searched for clinical trials with serum NSE information in patients with sepsis in the PubMed, Web of Science, Embase, and Cochrane databases from their inception to April 10, 2023. Included studies were assessed for quality and risk of bias using The Quality Assessment of Diagnostic Accuracy-2 tool. The meta-analysis of the included studies was performed using Stata 17.0 and Review Manager version 5.4. Findings: Eleven studies were included in this meta-analysis involving 1259 serum samples from 947 patients with sepsis. Our results showed that the serum NSE levels of patients with SAE were higher than those of the non-encephalopathy sepsis group (mean deviation, MD,12.39[95% CI 8.27-16.50, Z = 5.9, p < 0.00001]), and the serum NSE levels of patients with sepsis who died were higher than those of survivors (MD,4.17[95% CI 2.66-5.68, Z = 5.41, p < 0.00001]). Conclusion: Elevated serum NSE levels in patients with sepsis are associated with the early diagnosis of SAE and mortality; therefore, serum NSE probably is a valid biomarker for the early diagnosis and prognosis of patients with SAE. [ABSTRACT FROM AUTHOR]

Neonatal brain injury (NBI) is a critical condition for preterm neonates with potential long-term adverse neurodevelopmental outcomes. This prospective longitudinal case–control study aimed at investigating the levels and prognostic value of serum neuron-specific enolase (NSE) during the first 3 days of life in preterm neonates (<34 weeks) that later developed brain injury in the form of either periventricular leukomalacia (PVL) or intraventricular hemorrhage (IVH) during their hospitalization. Participants were recruited from one neonatal intensive care unit, and on the basis of birth weight and gestational age, we matched each case (n = 29) with a neonate who had a normal head ultrasound scan (n = 29). We report that serum NSE levels during the first three days of life do not differ significantly between control and preterm neonates with NBI. Nevertheless, subgroup analysis revealed that neonates with IVH had significantly higher concentrations of serum NSE in comparison to controls and neonates with PVL on the third day of life (p = 0.014 and p = 0.033, respectively). The same pattern on the levels of NSE on the third day of life was also observed between (a) neonates with IVH and all other neonates (PVL and control; p = 0.003), (b) neonates with II–IV degree IVH and all other neonates (p = 0.003), and (c) between control and the five (n = 5) neonates that died from the case group (p = 0.023). We conclude that NSE could be an effective and useful biomarker on the third day of life for the identification of preterm neonates at high risk of developing severe forms of IVH. [ABSTRACT FROM AUTHOR]

Introduction COVID-19 is a relevant issue for scientists around the world. Considering the development of numerous complications due to COVID-19, the search for diagnostic approaches for the timely detection of probable lesions of the nervous system is urgent. Objectives was to determine the diagnostic and prognostic value of the NSE marker in children with COVID-19 regarding the complicated course and symptoms of nervous system damage. Materials and Methods We conducted a cohort, observational, retrospective study involving 88 children aged 1 month to 18 years with laboratory-confirmed COVID-19 by PCR. Blood serum was collected for the study to assess the level of the neurobiomarker NSE by enzyme immunoassay. We divided patients into two cohorts according to the course of the disease - the main one, which included 42 patients with a complicated course of COVID-19, and a control group - 46 patients with an uncomplicated course of the disease. The study protocol was approved by the Local Ethics Committee of the hospital. Results: In the patients of the control group, NSE was observed at the level of 12.1±1.2 µg/l, while in the children of the main one, the indicator reached 16.9±1.5 µg/l (p=0.087). Increase in the level of NSE above 15 µg/L associated with a significant increase in the risk of the appearance of clinical symptoms of damage to the nervous system and a complicated course in children with COVID-19 (p<0.05). We noted a tendency towards an increase in NSE with increased ESR indicators >10mm/h (p<0.1), a decrease in PTI <85% (p=0,03) and an increase in D-dimer >2.5mg/l (p<0.1). Conclusions We found the diagnostic and prognostic value of the NSE marker in children with COVID-19 regarding the complicated course and symptoms of nervous system damage. [ABSTRACT FROM AUTHOR]

To explore the clinical significance of dynamic monitoring of cerebrospinal fluid (CSF) and serum Lactic acid（Lac), neuron-specific enolase (NSE), and the blood–brain barrier (BBB) index in evaluating the condition and prognosis after a severe traumatic brain injury (TBI). A total of 52 severe TBI patients admitted to the Department of Neurosurgery within 24 hours after injury were dynamically monitored. CSF and serum samples were collected on the 1st, 3rd, and 7th day after a severe TBI to monitor the changes in Lac, NSE, and the BBB index. Intracranial pressure (ICP), Glasgow coma scale (GCS), and 6-month Glasgow outcome scale-extended (GOS-E) were tested. According to the results of GOS-E, the patients were divided into two groups (i.e. the poor prognosis group and good prognosis group). Statistical analysis was conducted to investigate the clinical significance of dynamic monitoring of CSF and serum Lac, NSE, and BBB index after a severe TBI. After a severe TBI, the levels of Lac, NSE, and BBB in CSF and serum were significantly higher than those in the normal range. Lac, NSE, and the BBB index did not correlate with ICP (except serum Lac) but had correlations with GCS and post-injury 6 months post-injury (except serum Lac). Moreover, the correlations between Lac, NSE, and BBB index were statistically significant (p < 0.05): CSF Lac and CSF NSE; CSF Lac and serum NSE; Lac and BBB index of CSF; Lac and BBB index of CSF; NSE and CSE of serum; CSF NSE and BBB index; and serum NSE and BBB index. Additionally, serum NSE is correlated with NSE in CSF (p < 0.05). After a severe TBI, dynamic monitoring of CSF and serum Lac, NSE, and BBB index has the potential to assess the condition, predict the prognosis, and have clinical significance. [ABSTRACT FROM AUTHOR]

This study evaluates the protective effect of Hesperidin (HES), flavanoglycone, during pre-eclampsia developed pregnancy. The pregnant rats were randomly divided into five groups as Control, L-NAME (25 mg/kg), HES (100 mg/kg), L-NAME-HES and L-NAME P-HES. After the 13th day of pregnancy, the chemicals were administrated until the 20th day. 20 days after the offspring's birth, they were separated from their mothers and all rats were sacrificed after 6 weeks' time. Serum samples were analyzed for levels of Glutathione (GSH), lipid peroxidation (LPO), and neuron-specific enolase (NSE) activity. Urine samples were analyzed for the proteinuria. While Proteinuria and LPO level increasing, GSH levels decreased in the L-NAME group when compared to the Control. As we compared L-NAME group with the L-NAMEHES and L-NAME P-HES groups, the GSH levels increased significantly while LPO levels in the HES, L-NAME-HES, L-NAME P-HES groups were decreasing. The NSE activity significantly increased in the L-NAME group compared to the Control. In contrast to the L-NAME group, its activity decreased in HES, L-NAME-HES and L-NAME P-HES groups significantly. Briefly, it can be said that HES has a significant effect on both the LPO levels and the NSE activity in the case of pre-eclampsia. [ABSTRACT FROM AUTHOR]

Lung cancer remains the leading cause of cancer mortality worldwide. Small cell lung cancer (SCLC) accounts for 10–15% of cases and has an overall 5-years survival rate of only 15%. Neuron-specific enolase (NSE) has been identified as a useful biomarker for early SCLC diagnosis and therapeutic monitoring. This work reports an electrochemical immunosensing platform based on a graphene-graphitic carbon nitride (g-C3N4) nanocomposite for ultrasensitive NSE detection. The g-C3N4 nanosheets and graphene nanosheets were synthesized via liquid exfoliation and integrated through self-assembly to form the nanocomposite. This nanocomposite was used to modify screen-printed carbon electrodes followed by covalent immobilization of anti-NSE antibodies. The unique properties of the graphene-g-C3N4 composite facilitated efficient antibody loading while also enhancing electron transfer efficiency and electrochemical response. Systematic optimization of experimental parameters was performed. The immunosensor exhibited a wide linear detection range of 10 pg/mL to 100 ng/mL and low limit of detection of 3 pg/mL for NSE along with excellent selectivity against interferences. Real serum matrix analysis validated the applicability of the developed platform for sensitive and accurate NSE quantifica-tion at clinically relevant levels. This novel graphene-g-C3N4 nanocomposite based electro-chemical immunoassay demonstrates great promise for early diagnosis of SCLC. [ABSTRACT FROM AUTHOR]

Objective: To investigate the efficacy of high-frequency repetitive transcranial magnetic stimulation (HF-rTMS) combined with galantamine in patients with cognitive impairment after stroke and its effect on serum homocysteine (Hcy) and neuron-specific enolase (NSE) levels. Methods: A total of 90 patients with cognitive impairment after the first ischemic stroke were enrolled. They were randomly divided into rTMS+ cognitive rehabilitation group, Galantamine + cognitive rehabilitation group, and rTMS+ Galantamine + cognitive rehabilitation group. All groups received routine medical treatment and limb rehabilitation treatment. The rTMS stimulation site was the left dorsolateral prefrontal cortex (left DLPFC), the magnetic stimulation frequency was 5 Hz, the magnetic stimulation intensity was 80% of the motor threshold level, and 3,000 pulses were given every day. The Mini-Mental State Examination (MMSE), Montreal Cognitive Assessment (MoCA), Fugl-Meyer scale, and modified Barthel index, as well as rehabilitation scale and serum NSE and Hcy were evaluated before and after treatment (after 4 weeks). Results: After 4 weeks of treatment, the scores of MMSE, MoCa scale, Fugl-Meyer scale, and modified Barthel index in the three groups were significantly higher than those before treatment (all p < 0.05), while the serum NSE and Hcy levels of the three groups were decreased. rTMS+ Galantamine + cognitive rehabilitation group had higher scale scores, and the difference between the three groups was statistically significant compared with the other two groups (all p < 0.05). Conclusion: Cognitive rehabilitation combined with HF-rTMS and galantamine could improve the cognitive function of patients to the greatest extent, promote the recovery of physical activity, improve the self-care ability of daily life, and effectively reduce the serum HCY and NSE levels in patients with cognitive impairment after stroke. No randomized controlled trials of similar combination treatments have been reported. The better therapeutic effect may be related to the fact that galantamine combined with repetitive transcranial magnetism can activate the brain cholinergic system more extensively, promote brain neural remodeling through long-term potentiation and inhibit local neuroinflammatory responses in brain injury. [ABSTRACT FROM AUTHOR]

In the current study a simple and highly specific label-free impedimetric neuron specific enolase (NSE) immunosensor based on a copolymer matrix-coated disposable electrode was designed and tested. The copolymer matrix was prepared using a very conductive EDOT monomer and semi-conductive thiophene-bearing epoxy groups (ThEp), and the combination of the two monomers enhanced the conductivity and protein loading capacity of the electrode surface. The P(ThEp-co-EDOT) copolymer matrix was prepared via a drop-casting process and anti-NSE recognition biomolecules were immobilized directly on the epoxy groups of the copolymer. After the coupling of NSE molecules on the P(ThEp-co-EDOT) copolymer matrix-coated electrode surface, the charge transfer resistance (Rct) of the biosensor changed dramatically. These changes in Rct were proportional to the NSE molecule amounts captured by anti-NSE molecules. Under optimized experimental conditions, the increment in the Rct value was proportional to the NSE concentration over a range of 0.01 to 25 pg mL−1 with a detection limit (LOD) of 2.98 × 10−3 pg mL−1. This copolymer-coated electrode provided a lower LOD than the other biosensors. In addition, the suggested electrochemical immuno-platform showed good selectivity, superior reproducibility, long-term stability, and high recovery of NSE in real serum (95.64–102.20%) and saliva (95.28–105.35%) samples. These results showed that the present system had great potential for electrochemical biosensing applications. [ABSTRACT FROM AUTHOR]

Streptococcus suis is an emerging zoonotic pathogen that can cause invasive disease commonly associated with meningitis in pigs and humans. To cause meningitis, S. suis must cross the blood-brain barrier (BBB) comprising blood vessels that vascularize the central nervous system (CNS). The BBB is highly selective due to interactions with other cell types in the brain and the composition of the extracellular matrix (ECM). Purified streptococcal surface enolase, an essential enzyme participating in glycolysis, can bind human plasminogen (Plg) and plasmin (Pln). Plg has been proposed to increase bacterial traversal across the BBB via conversion to Pln, a protease which cleaves host proteins in the ECM and monocyte chemoattractant protein 1 (MCP1) to disrupt tight junctions. The essentiality of enolase has made it challenging to unequivocally demonstrate its role in binding Plg/Pln on the bacterial surface and confirm its predicted role in facilitating translocation of the BBB. Here, we report on the CRISPR/Cas9 engineering of S. suis enolase mutants eno261, eno252/253/255, eno252/261, and eno434/435 possessing amino acid substitutions at in silico predicted binding sites for Plg. As expected, amino acid substitutions in the predicted Plg binding sites reduced Plg and Pln binding to S. suis but did not affect bacterial growth in vitro compared to the wild-type strain. The binding of Plg to wild-type S. suis enhanced translocation across the human cerebral microvascular endothelial cell line hCMEC/D3 but not for the eno mutant strains tested. To our knowledge, this is the first study where predicted Plg-binding sites of enolase have been mutated to show altered Plg and Pln binding to the surface of S. suis and attenuation of translocation across an endothelial cell monolayer in vitro. [ABSTRACT FROM AUTHOR]

Background/Aim. Sepsis-associated encephalopathy (SAE) is a severe complication of sepsis, characterized by brain dysfunction and associated with a poor prognosis. SAE has a complex pathogenesis, and its severity is in close association with the levels of various serum factors. The aim of the study was to investigate the correlation of tumor necrosis factor (TNF)-a, monocyte chemoattractant protein (MCP)-1, and neuron-specific enolase (NSE) levels with the severity of SAE and to analyze the prognostic values of the three parameters. Methods. This prospective study enrolled 126 patients treated for SAE from June 2020 to June 2022. The levels of TNF-α, MCP-1, and NSE were measured, and the severity of SAE was evaluated using the Sequential Organ Failure Assessment (SOFA) score. Based on the SOFA score, the patients were assigned to two groups: a group with a bad prognosis and a group with a good prognosis. The correlations of TNF-α, MCP-1, and NSE levels with the severity of SAE were analyzed, and their prognostic values were evaluated during a 28-day follow-up. Results. The mean levels of TNF-α, MCP-1, and NSE and the SOFA score of the 126 patients with SAE were 6.52 ± 1.48 pg/mL, 62.53 ± 18.49 pg/mL, 8.61 ± 2.17 ng/mL, and 10.24 ± 2.86 points, respectively. Pearson's analysis demonstrated significant correlations between TNF-α, MCP-1, and NSE levels and the SOFA score of patients with SAE (r > 0, p < 0.05). Of the 126 patients, 61 (48.4%) had a poor prognosis, while 65 (51.6%) had a good prognosis. Increased serum TNF-α, MCP-1, and NSE levels were risk factors for the poor prognosis of patients with SAE [odds ratio (OR) > 1, p < 0.05]. The areas under the receiver operating characteristic (ROC) curves of serum TNF-α, MCP-1, and NSE levels were all > 0.7, suggesting high predictive values of these parameters. Conclusion. Serum TNF-α, MCP-1, and NSE levels are closely correlated with the severity of SAE and may work as valuable predictors of treatment outcome. [ABSTRACT FROM AUTHOR]

Background: The high number of accidents and traumatic brain injuries, especially in the productive age group, causes a lot of morbidity and mortality. A fast and accurate examination method is needed for the diagnosis and treatment of traumatic brain injury. Nerve damage biomarkers such as Neuron Specific Enolase, S100B, Glial Fibrillary Acidic Protein, and Myelin Basic Protein, have been used globally both for research and daily use to determine the severity of traumatic brain injury. Methods: Searches and journal searches were carried out from Science Direct, Scopus, Springer Link, and PubMed, with the keywords "Neuron Specific Enolase", "S100B", "Glial Fibrillary Acidic Protein", "Myelin Basic Protein", and "Traumatic Brain Injury ". Screening was carried out using PRISMA 2021 to look for studies that met the criteria and were of sufficient study quality according to the Newcastle-Ottawa Scale. Results: Twenty-three studies were collected and further grouped based on outcomes, both prognostic and survival outcomes. Neuron Specific Enolase, S100B, and Glial Fibrillary Acidic Protein values were higher in poor outcomes (all p values < 0.001) and poor survival (all p values < 0.001) in traumatic brain injury. Myelin Basic Protein was not significant in poor outcome (p = 0.35), but was higher in poor survival (p < 0.001) in traumatic brain injury. Conclusion: Neuron Specific Enolase, S100B, and Glial Fibrillary Acidic Protein, can be used as markers for prognostic and survival value in traumatic brain injury. Myelin Basic Protein can be used as a marker for survival value in traumatic brain injury. [ABSTRACT FROM AUTHOR]

Abstract Background Ribosomal protein SA (RPSA) of human brain microvascular endothelial cells (HBMECs) can transfer from the cytosol to the cell surface and act as a receptor for some pathogens, including Streptococcus suis serotype 2 (SS2), a zoonotic pathogen causing meningitis in pigs and humans. We previously reported that SS2 virulence factor enolase (ENO) binds to RPSA on the cell surface of HBMECs and induces apoptosis. However, the mechanism that activates RPSA translocation to the cell surface and induces ENO-mediated HBMEC apoptosis is unclear. Results Here, we show that RPSA localization and condensation on the host cell surface depend on its internally disordered region (IDR). ENO binds to the IDR of RPSA and promotes its interaction with RPSA and vimentin (VIM), which is significantly suppressed after 1,6-Hexanediol (1,6-Hex, a widely used tool to disrupt phase separation) treatment, indicating that ENO incorporation and thus the concentration of RPSA/VIM complexes via co-condensation. Furthermore, increasing intracellular calcium ions (Ca2+) in response to SS2 infection further facilitates the liquid-like condensation of RPSA and aggravates ENO-induced HBMEC cell apoptosis. Conclusions Together, our study provides a previously underappreciated molecular mechanism illuminating that ENO-induced RPSA condensation activates the migration of RPSA to the bacterial cell surface and stimulates SS2-infected HBMEC death and, potentially, disease progression. This study offers a fresh avenue for investigation into the mechanism by which other harmful bacteria infect hosts via cell surfaces’ RPSA.

Heat stress severely affects the yield and quality of maize. Melatonin (N-acetyl-5-methoxy-tryptamin, MT) plays an important role in various types of stress resistance in plants, including heat tolerance. Enolase (ENO, 2-phospho-D-glycerate hydrolyase) contributes to plant growth, development, and stress response. As of now, the molecular mechanisms by which MT and ENO1 affect heat tolerance are unknown. In our research, we have revealed that heat stress (H) and heat stress + MT (MH) treatment upregulate ZmENO1 expression levels by 15 and 20 times, respectively. ZmENO1 overexpression and mutant maize lines were created by transgenic and genome editing. These results illustrate that heat stress has a significant impact on the growth of maize at the seedling stage. However, ZmENO1-OE lines showed a lower degree of susceptibility to heat stress, whereas the mutant exhibited the most severe effects. Under heat stress, exogenous application of MT improves heat resistance in maize. The ZmENO1-OE lines exhibited the best growth and highest survival rate, while the zmeno1 mutants showed the least desirable results. Following treatment with H and MH, the level of MT in ZmENO1-OE lines exhibited the greatest increase and reached the maximum value, whereas the level of MT in the zmeno1 mutant was the lowest. Heat stress decreased the maize’s relative water content and fresh weight, although ZmENO1-OE lines had the highest and zmeno1 mutants had the lowest. Heat stress led to an increase in the levels of MDA, hydrogen peroxide, and superoxide in all plants. Additionally, the ionic permeability and osmotic potential of the plants were significantly increased. However, the levels of MT were decreased in all plants, with the greatest decrease observed in the ZmENO1-OE lines. Interestingly, the zmeno1 mutant plants had the highest expression levels of MT. Heat stress-induced upregulation of ZmSOD, ZmPOD, ZmAPX, ZmCAT, ZmP5CS, and ZmProDH in all plants. However, the ZmENO1-OE lines exhibited the greatest increase in expression levels, while the zmeno1 mutants showed the lowest increase following MT spraying. The patterns of SOD, POD, APX, and CAT enzyme activity, as well as proline and soluble protein content, aligned with the variations in the expression levels of these genes. Our findings indicate that MT can upregulate the expression of the ZmENO1 gene. Upregulating the ZmENO1 gene resulted in elevated expression levels of ZmSOD, ZmPOD, ZmAPX, ZmCAT, ZmP5CS, and ZmProDH. This led to increased activity of antioxidant enzymes and higher levels of osmoregulatory substances. Consequently, it mitigated the cell membrane damage caused by heat stress and ultimately improved the heat resistance of maize. The results of this study provide genetic resources for molecular design breeding and lay a solid foundation for further exploring the molecular mechanism of MT regulation of heat stress tolerance in maize.

Background: Ribosomal protein SA (RPSA) of human brain microvascular endothelial cells (HBMECs) can transfer from the cytosol to the cell surface and act as a receptor for some pathogens, including Streptococcus suis serotype 2 (SS2), a zoonotic pathogen causing meningitis in pigs and humans. We previously reported that SS2 virulence factor enolase (ENO) binds to RPSA on the cell surface of HBMECs and induces apoptosis. However, the mechanism that activates RPSA translocation to the cell surface and induces ENO-mediated HBMEC apoptosis is unclear. Results: Here, we show that RPSA localization and condensation on the host cell surface depend on its internally disordered region (IDR). ENO binds to the IDR of RPSA and promotes its interaction with RPSA and vimentin (VIM), which is significantly suppressed after 1,6-Hexanediol (1,6-Hex, a widely used tool to disrupt phase separation) treatment, indicating that ENO incorporation and thus the concentration of RPSA/VIM complexes via co-condensation. Furthermore, increasing intracellular calcium ions (Ca2+) in response to SS2 infection further facilitates the liquid-like condensation of RPSA and aggravates ENO-induced HBMEC cell apoptosis. Conclusions: Together, our study provides a previously underappreciated molecular mechanism illuminating that ENO-induced RPSA condensation activates the migration of RPSA to the bacterial cell surface and stimulates SS2-infected HBMEC death and, potentially, disease progression. This study offers a fresh avenue for investigation into the mechanism by which other harmful bacteria infect hosts via cell surfaces' RPSA. [ABSTRACT FROM AUTHOR]

A photoelectrochemical (PEC) immunosensor was designed based on MgIn2S4-decorated inorganic halide perovskite CsPbBr3 combined with the signal polarity conversion strategy for neuron-specific enolase (NSE) detection. CsPbBr3 was applied as the basic photoactive material owing to its excellent optical and electronic properties, which provide a good PEC performance for sensor construction. In order to improve the stability of this perovskite, the three-dimensional flower-like MgIn2S4 with a desirable direct band gap was applied to enhance the PEC response. Also, the excellent structure of MgIn2S4 provides large surface-active sites for CsPbBr3 loaded. For enhancing the detection sensitivity of PEC immunosensor, p-type CuInS2 was used as a signal probe which fixed on detection antibody (Ab2). When the target NSE was present, the photogenerated electrons produced by CuInS2 were transferred to the test solution, and the polarity of PEC signal changes. Based on the above photosensitive materials and signal conversion strategy, the proposed PEC immunosensor showed favorable detection performance, and the linear detection range is 0.0001 ~ 100 ng/mL with a 38 fg/mL of detection limit. The proposed strategy improved the adhibition of CsPbBr3 in the analytical chemistry field as well as provided a reference method for other protein detections. [ABSTRACT FROM AUTHOR]

A photoelectrochemical (PEC) immunosensor was designed based on MgIn2S4-decorated inorganic halide perovskite CsPbBr3 combined with the signal polarity conversion strategy for neuron-specific enolase (NSE) detection. CsPbBr3 was applied as the basic photoactive material owing to its excellent optical and electronic properties, which provide a good PEC performance for sensor construction. In order to improve the stability of this perovskite, the three-dimensional flower-like MgIn2S4 with a desirable direct band gap was applied to enhance the PEC response. Also, the excellent structure of MgIn2S4 provides large surface-active sites for CsPbBr3 loaded. For enhancing the detection sensitivity of PEC immunosensor, p-type CuInS2 was used as a signal probe which fixed on detection antibody (Ab2). When the target NSE was present, the photogenerated electrons produced by CuInS2 were transferred to the test solution, and the polarity of PEC signal changes. Based on the above photosensitive materials and signal conversion strategy, the proposed PEC immunosensor showed favorable detection performance, and the linear detection range is 0.0001 ~ 100 ng/mL with a 38 fg/mL of detection limit. The proposed strategy improved the adhibition of CsPbBr3 in the analytical chemistry field as well as provided a reference method for other protein detections. [ABSTRACT FROM AUTHOR]

Mycoplasma synoviae (MS) is a primary avian pathogen prevalent worldwide that causes airsacculitis and synovitis in birds. Vaccination is recommended as the most cost-effective strategy in the control of MS infection. Novel alternative vaccines are needed for eradicating and controlling MS infection in flocks. DnaK, enolase, elongation factor Tu (EF-Tu), MSPB, NADH oxidase and LP78 are the major immunogenic antigens of MS and are promising targets for subunit vaccine candidates. In the present study, genes encoding DnaK, enolase, EF-Tu, MSPB, LP78, and NADH oxidase were cloned and expressed in Escherichia coli. Enzyme-linked immunosorbent assay showed that the six recombinant proteins were recognized by convalescent sera, indicating that they were expressed during infection. Two injections of the six subunit vaccines induced a robust antibody response and increased the concentrations of IFN-γ and IL-4, especially rEnolase and rEF-Tu. The proliferation of peripheral blood lymphocytes was enhanced in all of the immunized groups. Chickens immunized with rEnolase, rEF-Tu, rLP78, and rMSPB conferred significant protection against MS infection, as indicated by significantly lower DNA copies in the trachea, lower scores of air sac lesions, and lesser tracheal mucosal thickness than that in the challenge control. Especially, rEnolase provided the best protective efficacy, followed by rEFTu, rMSPB, and rLP78. Our finds demonstrate that the subunit vaccines and bacterin can only reduce the lesions caused by MS infection, but not prevent colonization of the organism. Our findings may contribute to the development of novel vaccine agents against MS infection. [ABSTRACT FROM AUTHOR]

Background: Early diagnosis of malignant pleural effusion (MPE) is of great significance. Current prediction models are not simple enough to be widely used in heavy clinical work. Objectives: We aimed to develop a simple and efficient clinical prediction scoring system to distinguish MPE from benign pleural effusion (BPE). Design: This retrospective study involved patients with MPE or BPE who were admitted in West China Hospital from December 2010 to September 2016. Methods: Patients were divided into training, testing, and validation set. Prediction model was developed from training set and modified to a scoring system. The diagnostic efficacy and clinical benefits of the scoring system were estimated in all three sets. Results: Finally, 598 cases of MPE and 1094 cases of BPE were included. Serum neuron-specific enolase, serum cytokeratin 19 fragment (CYFRA21-1), pleural carcinoembryonic antigen (CEA), and ratio of pleural CEA to serum CEA were selected to establish the prediction models in training set, which were modified to the scoring system with scores of 6, 8, 10, and 9 points, respectively. Patients with scores >12 points have high MPE risk while ⩽12 points have low MPE risk. The scoring system has a high predictive value and good clinical benefits to differentiate MPE from BPE or lung-specific MPE from BPE. Conclusion: This study developed a simple clinical prediction scoring system and was proven to have good clinical benefits, and it may help clinicians to separate MPE from BPE. [ABSTRACT FROM AUTHOR]

Here, we employ coelution experiments and far‐western blotting to identify stable interactions between the main components of the B. subtilis degradosome and the small proteins SR1P and SR7P. Our data indicate that B. subtilis has a degradosome comprising at least RNases Y and PnpA, enolase, phosphofructokinase, glycerol‐3‐phosphate dehydrogenase GapA, and helicase CshA that can be co‐purified without cross‐linking. All interactions were corroborated by far‐western blotting with proteins purified from E. coli. Previously, we discovered that stress‐induced SR7P binds enolase to enhance its interaction with and activity of enolase‐bound RNase Y (RnY), while SR1P transcribed under gluconeogenic conditions interacts with GapA to stimulate its interaction with and the activity of RnjA (RnjA). We show that SR1P can directly bind RnjA, RnY, and PnpA independently of GapA, whereas SR7P only interacts with enolase. Northern blotting suggests that the degradation of individual RNAs in B. subtilis under gluconeogenic or stress conditions depends on either RnjA or RnY alone or on RnjA‐SR1P, RnY‐SR1P, or RnY‐Eno. In vitro degradation assays with RnY or RnjA substrates corroborate the in vivo role of SR1P. Currently, it is unknown which substrate property is decisive for the utilization of one of the complexes. [ABSTRACT FROM AUTHOR]

Background and Objectives: Survival with favorable neurologic outcomes after out-of-hospital cardiac arrest (OHCA) remains elusive. Post-cardiac arrest syndrome (PCAS) involves myocardial and neurological injury, ischemia-reperfusion response, and underlying pathology. Neurologic injury is a crucial determinant of survival and functional outcomes, with damage caused by free radicals among the responsible mechanisms. This study explores the feasibility of adding intravenous vitamin C to the treatment of OHCA survivors, aiming to mitigate PCAS. Vitamin C, a nutrient with antioxidative and free radical-scavenging properties, is often depleted in critically ill patients. Materials and Methods: This randomized, double-blinded trial was conducted at a tertiary-level university hospital with adult OHCA survivors. Participants received either standard care or the addition of 1.5 g of intravenous vitamin C every 12 h for eight consecutive doses. Neurologic injury was assessed using neuron-specific enolase (NSE) levels, with additional clinical and laboratory outcomes, such as enhanced neuroprognostication factors, inflammatory markers, and cardiac parameters. Results: NSE levels were non-significantly higher in patients who received vitamin C compared to the placebo group (55.05 µg/L [95% confidence interval (CI) 26.7–124.0] vs. 39.4 µg/L [95% CI 22.6–61.9], p > 0.05). Similarly, a non-significantly greater proportion of patients in the vitamin C group developed myoclonus in the first 72 h. We also observed a non-significantly shorter duration of mechanical ventilation, fewer arrhythmias, and reduced length of stay in the intensive care unit in the group of patients who received vitamin C (p = 0.031). However, caution is warranted in interpretation of our results due to the small number of participants. Conclusions: Our findings suggest that intravenous vitamin C should not be used outside of clinical trials for OHCA survivors. Due to the small sample size and conflicting results, further research is needed to determine the potential role of vitamin C in post-cardiac arrest care. [ABSTRACT FROM AUTHOR]

The prediction of outcomes following cardiac arrest continues to provide significant difficulties. A preferred strategy involves adopting a multimodal approach, which encompasses the careful evaluation of the biomarker neuron-specific enolase (NSE). This systematic review and meta-analysis aimed to gather and summarize new and existing evidence on the prediction effect of neuron-specific enolase for survival to hospital discharge among adult patients with cardiac arrest. We searched PubMed Central, Scopus, EMBASE databases, and the Cochrane Library without language restrictions from their inceptions until 30 October 2023 and checked the reference lists of the included studies. Pooled results were reported as standardized mean differences (SMDs) and were presented with corresponding 95% confidence intervals (CIs). The primary outcome was survival to hospital discharge (SHD). Eighty-six articles with 10,845 participants were included. NSE showed a notable degree of specificity in its ability to predict mortality as well as neurological status among individuals who experienced cardiac arrest (p < 0.05). This study demonstrates the ability to predict fatality rates and neurological outcomes, both during the time of admission and at various time intervals after cardiac arrest. The use of NSE in a multimodal neuroprognostication algorithm has promise in improving the accuracy of prognoses for persons who have undergone cardiac arrest. [ABSTRACT FROM AUTHOR]

Neuron-specific enolase (NSE), which is a highly specific marker for neurons, could be a predictor for prognosis in patients with symptomatic intracranial hemorrhage (sICH) with acute ischemic stroke who are receiving endovascular treatment (EVT). This study aimed to investigate the relationship between NSE and sICH in patients with acute anterior circulation stroke undergoing EVT. A total of 215 consecutive patients with acute stroke treated with EVT were included. Patients with stroke and acute anterior circulation occlusion, receiving EVT treated at our hospital, were enrolled between January 2017 and August 2021. NSE level was measured on arrival at the neurology intensive care unit after EVT. The patients were divided into 2 groups according to whether sICH was present. Univariate and multivariate analyses were performed. NSE level was also incorporated into the TAG score (modified Thrombolysis in Cerebral Infarction score, Alberta Stroke Program Early CT Score, and glucose level), which was developed as a scoring system to predict sICH, and the prediction capability was compared with the TAG score alone. Causal inference was performed using the package DoWhy in Python to evaluate the causal relationship between NSE and sICH. The area under the curve (AUC) value of NSE showed moderate accuracy, with an AUC value of 0.729 (95% confidence interval, 0.655–0.795; P < 0.001). The NSE cutoff value was set at 23.88 ng/mL. When the NSE level ≥23.88 ng/mL, the sensitivity was 58.33% and the specificity was 78.72% (P < 0.001). The AUC for the TAG + NSE score was 0.801 compared with an AUC of 0.632 for the TAG score (Z = 2.034; P = 0.042). A causal inference model using the DoWhy library shows a proportional relationship between NSE and the diagnosis of sICH. This study is the first to show that increased NSE level is an independent predictor of sICH in patients with acute anterior circulation stroke who are undergoing endovascular treatment. [ABSTRACT FROM AUTHOR]

The early stages of diabetic peripheral neuropathy (DPN) are symptomless. A reliable dependable and sensitive biomarker is needed for the purpose of early identification of diabetic peripheral neuropathy. The main objective of the study was to evaluate the accuracy of serum neuron-specific enolase (NSE) as a biomarker for early identification of diabetic peripheral neuropathy. Patient samples were collected from the National Diabetes Center, Mustansiriyah University; a case control study was done from April 2022 to November 2022, in Baghdad, Iraq. One hundred sixty individuals between 30 to 60 years-old were included. Participants were divided into three groups: group one included 40 type 2 diabetic patients with peripheral neuropathy, group two consisted of 40 type 2 diabetic patients without peripheral neuropathy and group three included 80 apparently in good health as the control. Toronto Clinical Neuropathy Scoring System (TCSS) was used for clinical evaluation of peripheral neuropathy. Glycated hemoglobin (HbA1c) was measured by the CLOVER A1c system. In addition, serum NSE levels were measured by Enzyme Linked Immunosorbent Assay (ELISA) technique. Age, sex, and other standard variables were used as a basis for comparisons between groups. Statistically, diabetic patients with peripheral neuropathy demonstrated higher level of NSE (28.42±6.93 ng/ml) than did either diabetic patients without peripheral neuropathy (21.07±2.0 ng/ml) or controls (12.54±2.34 ng/ml) with a high degree of significance (p <0.001). In the context of Discrimination between DPN patients and diabetic patients without neuropathy, the area under curve for neuron-specific enolase was 0.812, 95% confidence interval [CI] = 0.716-0.909, p <0.001. Cut-off value of serum neuron-specific enolase was 22.53 ng/ml, sensitivity and specificity were 70% and 77%, respectively. In the context of discrimination between DPN and controls, the area under curve for neuron-specific enolase was 1.00, 95% confidence interval was 1.0-1.0, p <0.001. At a cut-off value of serum neuron-specific enolase = 18.3 ng/ml, both the sensitivity and specificity were 100%. Neuron-specific enolase could potentially be used as a biomarker to detect early diabetic peripheral neuropathy and prevent it from developing to an advanced state. [ABSTRACT FROM AUTHOR]

Objective To explore the clinical significance of monitoring neuron specific enolase (NSE), carbohydrate antigen 125 (CA125), and carbohydrate antigen 199 (CA199) in patients with pneumoconiosis. Methods A retrospective analysis was conducted among 222 patients with pneumoconiosis admitted to this hospital from January 2018 to December 2022. The patients were divided into three groups according to the stage of pneumoconiosis (stage I, II, and III). The serum levels and abnormal rates of NSE, CA125, and CA199 among these groups were compared. Results Among the 222 patients with pneumoconiosis, 51 (22.97%) were in stage I, 52 (23.42%) were in stage II, and 119 (53.61%) were in stage III. There were statistically significant differences in serum NSE and CA125 levels among pneumoconiosis patients at different stages (P < 0.001), and serum NSE and CA125 levels in patients at stage III were higher than those in stage I and stage II (P < 0.001). There was no statistically significant difference in serum CA199 level among pneumoconiosis patients at different stages (P = 0.163). The abnormal rates of serum NSE, CA125, and CA199 levels among 222 patients were 45.05%, 27.93%, and 2.70%, respectively. There were statistically significant differences in serum NSE and CA125 abnormal rates among pneumoconiosis patients at different stages (both P < 0.001), and both showed an increasing trend with the increase of stage. Conclusions Monitoring serum NSE and CA125 in patients with pneumoconiosis can indicate the progression of their condition and provide references for the treatment plan. [ABSTRACT FROM AUTHOR]

Objective: To explore the application value of magnetic resonance imaging(MRI) diffusion tensor imaging(DTI)parameters combined with serum neuron specific enolase(NSE) and lipoprotein-associated phospholipase A2(Lp-PLA2) in the diagnosis and risk assessment of poor prognosis of patients with cerebral infarction. Methods: 106 patients with cerebral infarction who were admitted to China Japan Friendship Hospital of Jilin University from March 2021 to September 2022 were selected as cerebral infarction group, another 62 healthy volunteers who were underwent physical examinations during the same period were selected as control group,the DTI parameters, serum NSE and Lp-PLA2 levels were compared between two groups. 90d after discharge from cerebral infarction group, the assessment of prognosis by the modified Rankin scale(m RS), they were divided into good prognosis group and poor prognosis group, and the above indicators levels were compared between two groups. The value of DTI parameters combined with serum NSE and Lp-PLA2 for diagnosed cerebral infarction and predicted the prognosis of patients with cerebral infarction was analyzed by the receiver operating characteristic(ROC) curve. Results: The apparent diffusion coefficient (ADC) value and fractional anisotropy index(FA) value of cerebral infarction group were lower than those of control group (P＜0.05), and the serum NSE and Lp-PLA2 levels were higher than those of control group(P＜0.05). The FA value and ADC value of poor prognosis group were lower than those of good prognosis group(P＜0.05), and the serum NSE and Lp-PLA2 levels were higher than those of good prognosis group(P＜0.05). The area under the curve(AUC) for diagnosed cerebral infarction and predicted poor prognosis of patients with cerebral infarction by combined with FA value,ADC value, NSE and Lp-PLA2 were 0.852 and 0.874, respectively, and which were higher than individual diagnosed and predicted of each factor. Conclusion: The DTI parameters FA value and ADC value of cerebral infarction decrease, while serum NSE and Lp-PLA2levels increase, combine with DTI parameters and serum NSE and Lp-PLA2 detection has high value for the diagnosis and prognosis predict of cerebral infarction [ABSTRACT FROM AUTHOR]

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Candida albicans is the predominant cause of systemic candidiasis, although other non albicans Candida species are progressively becoming more widespread nowadays. Candida auris has emerged as a deadly multidrug-resistant fungal pathogen, posing a significant threat to global public health. In the absence of effective antifungal therapies, the development of a vaccine against C. auris infections is imperative. Enolase, a key glycolytic enzyme, has emerged as a promising vaccine candidate due to its immunogenic properties and essential role in fungal virulence. Herein, full-length Enolase gene sequences from C. albicans and C. auris were cloned into suitable expression vector and transformed into Escherichia coli expression hosts. Recombinant Enolase proteins were successfully expressed and purified using affinity chromatography under native conditions, followed by SDS-PAGE characterization and Western blot analysis. CD spectroscopy verified the existence of expressed proteins in soluble native conformation. Preliminary in silico studies verified the immunogenicity of recombinant Enolase proteins isolated from both C. albicans and C. auris. Furthermore, bioinformatics analysis revealed conserved B-cell and T-cell epitopes across C. albicans and C. auris Enolase proteins, suggesting potential cross-reactivity and broad-spectrum vaccine efficacy. Our findings are anticipated to play a role in advancing therapeutic as well as diagnostic strategies against systemic candidiasis.