Novel electrochemical platform based on C3N4-graphene composite for the detection of neuron-specific enolase as a biomarker for lung cancer.

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.

Identification of biomarkers for the early detection of non-small cell lung cancer: a systematic review and meta-analysis.

Lung cancer (LC) causes few symptoms in the earliest stages, leading to one of the highest mortality rates among cancers. Low-dose computerised tomography (LDCT) is used to screen high-risk individuals, reducing the mortality rate by 20%. However, LDCT results in a high number of false positives and is associated with unnecessary follow-up and cost. Biomarkers with high sensitivities and specificities could assist in the early detection of LC, especially in patients with high-risk features. Carcinoembryonic antigen (CEA), cytokeratin 19 fragments and cancer antigen 125 have been found to be highly expressed during the later stages of LC but have low sensitivity in the earliest stages. We determined the best biomarkers for the early diagnosis of LC, using a systematic review of eight databases. We identified 98 articles that focussed on the identification and assessment of diagnostic biomarkers and achieved a pooled area under curve of 0.85 (95% CI 0.82-0.088), indicating that the diagnostic performance of these biomarkers when combined was excellent. Of the studies, 30 focussed on single/antigen panels, 22 on autoantibodies, 31 on miRNA and RNA panels, and 15 suggested the use of circulating DNA combined with CEA or neuron-specific enolase (NSE) for early LC detection. Verification of blood biomarkers with high sensitivities (Ciz1, exoGCC2, ITGA2B), high specificities (CYFR21-1, antiHE4, OPNV) or both (HSP90α, CEA) along with miR-15b and miR-27b/miR-21 from sputum may improve early LC detection. Further assessment is needed using appropriate sample sizes, control groups that include patients with non-malignant conditions, and standardised cut-off levels for each biomarker.

A novel construction strategy of photoelectrochemical immunosensor for detecting neuron-specific enolase: Sensing mode integrating photoanode and photocathode.

In the realm of photoelectrochemical (PEC) analysis, it is customary to employ a lone photoanode or photocathode system for analyte detection. Nonetheless, such a single detection scheme inherently carries some deficiencies. While photoanode-based PEC immunoassay methods do exhibit conspicuous photocurrent responses and heightened sensitivity, they do suffer from inadequate resistance to interference when detecting in real sample detection. Photocathode-based analysis methods are capable of effectively surmounting the limitations of photoanode-based analysis methods, but their stability is poor. Based on the above reasons, this paper reports a novel immunosensing system combining ITO/WO3/Bi2S3 photoanode and ITO/CuInS2 photocathode. The system that incorporates both photoanode and photocathode exhibits a steady and discernible photocurrent, possesses robust resistance to external interference, and has effectively accomplished the quantification of NSE in the linear range from 5 pg/mL to 30 ng/mL. Remarkably, the detection limit has been determined to be 1.59 pg/mL. Besides the notable advantages of satisfactory stability, exceptional specificity, and outstanding reproducibility, the sensing system also introduces an innovative approach to fabricate PEC immunosensors.

Biological Transformation of AgI on MOF-on-MOF-Derived Heterostructures: Toward Polarity-Switchable Photoelectrochemical Biosensors for Neuron-Specific Enolase.

The sensitive detection of neuron-specific enolase (NSE) as a biomarker for lung cancer at an early stage is critical but has long been a challenge. The emergence of polarity-switchable photoelectrochemical (PEC) bioanalysis has opened up new avenues for developing highly sensitive NSE sensors. In this study, we present such a biosensor depending on the bioinduced AgI transition on MOF-on-MOF-derived semiconductor heterojunctions. Specifically, treatment of ZnO@In2O3@AgI by bioproduced H2S can in situ generate the ZnO@In2O3@In2S3@Ag2S heterojunction, with the photocurrent switching from the cathodic to anodic one due to the changes in the carrier transfer pathway. Linking an NSE-targeted sandwich immunorecognition with labeled alkaline phosphatase (ALP) catalyzed generation of H2S, such a phenomenon was correlated to NSE concentration with good performance in terms of selectivity and sensitivity and a low detection limit of 0.58 pg/mL. This study offered a new perspective on the use of MOF-on-MOF-derived heterostructures for advanced polarity-switchable PEC bioanalysis.

Physicochemical and immunological characterization of Amb a 12, a novel ragweed (Ambrosia artemisiifolia) pollen allergen.

BACKGROUND: Ragweed is an invasive plant in Europe, causing hay fever and asthma in allergic patients. Climate change is predicted to increase expansion and allergenicity. Elevated NO2 induced upregulation of a new allergen in ragweed pollen, an enolase, Amb a 12. OBJECTIVE: of this study was producing ragweed enolase as a recombinant protein and characterizing its physicochemical and immunological features. METHODS: Amb a 12 was designed for E. coli and insect cell expression. Physicochemical features were determined by mass spectrometry, circular dichroism measurements and enzymatic activity assay. Immunological characteristics were determined in ELISA, in a mediator release assay and by investigation of association with clinical symptoms. Common allergen sources were screened for similar proteins. RESULTS: Ragweed enolase was produced as a 48 kDa protein forming oligomers in both expression systems, showing differences in secondary structure content and enzymatic activity depending on expression system. IgE frequency and allergenicity were low regardless of expression system. Enolase-specific serum bound to similar sized molecules in mugwort, timothy grass and birch pollen, as well as food allergen sources, while highest IgE inhibition was achieved with peach pulp extract. CONCLUSIONS: Amb a 12 had high sequence similarity and comparable IgE frequency to enolase allergens from different sources. 50 kDa proteins were found in other pollen and food allergen sources, suggesting that enolases might be pan-allergens in pollen and plant foods.

Blood-Pressure Targets in Comatose Survivors of Cardiac Arrest.

BACKGROUND: Evidence to support the choice of blood-pressure targets for the treatment of comatose survivors of out-of-hospital cardiac arrest who are receiving intensive care is limited. METHODS: In a double-blind, randomized trial with a 2-by-2 factorial design, we evaluated a mean arterial blood-pressure target of 63 mm Hg as compared with 77 mm Hg in comatose adults who had been resuscitated after an out-of-hospital cardiac arrest of presumed cardiac cause; patients were also assigned to one of two oxygen targets (reported separately). The primary outcome was a composite of death from any cause or hospital discharge with a Cerebral Performance Category (CPC) of 3 or 4 within 90 days (range, 0 to 5, with higher categories indicating more severe disability; a category of 3 or 4 indicates severe disability or coma). Secondary outcomes included neuron-specific enolase levels at 48 hours, death from any cause, scores on the Montreal Cognitive Assessment (range, 0 to 30, with higher scores indicating better cognitive ability) and the modified Rankin scale (range, 0 to 6, with higher scores indicating greater disability) at 3 months, and the CPC at 3 months. RESULTS: A total of 789 patients were included in the analysis (393 in the high-target group and 396 in the low-target group). A primary-outcome event occurred in 133 patients (34%) in the high-target group and in 127 patients (32%) in the low-target group (hazard ratio, 1.08; 95% confidence interval [CI], 0.84 to 1.37; P = 0.56). At 90 days, 122 patients (31%) in the high-target group and 114 patients (29%) in the low-target group had died (hazard ratio, 1.13; 95% CI, 0.88 to 1.46). The median CPC was 1 (interquartile range, 1 to 5) in both the high-target group and the low-target group; the corresponding median modified Rankin scale scores were 1 (interquartile range, 0 to 6) and 1 (interquartile range, 0 to 6), and the corresponding median Montreal Cognitive Assessment scores were 27 (interquartile range, 24 to 29) and 26 (interquartile range, 24 to 29). The median neuron-specific enolase level at 48 hours was also similar in the two groups. The percentages of patients with adverse events did not differ significantly between the groups. CONCLUSIONS: Targeting a mean arterial blood pressure of 77 mm Hg or 63 mm Hg in patients who had been resuscitated from cardiac arrest did not result in significantly different percentages of patients dying or having severe disability or coma. (Funded by the Novo Nordisk Foundation; BOX ClinicalTrials.gov number, NCT03141099.).

Oxygen Targets in Comatose Survivors of Cardiac Arrest.

BACKGROUND: The appropriate oxygenation target for mechanical ventilation in comatose survivors of out-of-hospital cardiac arrest is unknown. METHODS: In this randomized trial with a 2-by-2 factorial design, we randomly assigned comatose adults with out-of-hospital cardiac arrest in a 1:1 ratio to either a restrictive oxygen target of a partial pressure of arterial oxygen (Pao2) of 9 to 10 kPa (68 to 75 mm Hg) or a liberal oxygen target of a Pao2 of 13 to 14 kPa (98 to 105 mm Hg); patients were also assigned to one of two blood-pressure targets (reported separately). The primary outcome was a composite of death from any cause or hospital discharge with severe disability or coma (Cerebral Performance Category [CPC] of 3 or 4; categories range from 1 to 5, with higher values indicating more severe disability), whichever occurred first within 90 days after randomization. Secondary outcomes were neuron-specific enolase levels at 48 hours, death from any cause, the score on the Montreal Cognitive Assessment (ranging from 0 to 30, with higher scores indicating better cognitive ability), the score on the modified Rankin scale (ranging from 0 to 6, with higher scores indicating greater disability), and the CPC at 90 days. RESULTS: A total of 789 patients underwent randomization. A primary-outcome event occurred in 126 of 394 patients (32.0%) in the restrictive-target group and in 134 of 395 patients (33.9%) in the liberal-target group (hazard ratio, 0.95; 95% confidence interval, 0.75 to 1.21; P = 0.69). At 90 days, death had occurred in 113 patients (28.7%) in the restrictive-target group and in 123 (31.1%) in the liberal-target group. On the CPC, the median category was 1 in the two groups; on the modified Rankin scale, the median score was 2 in the restrictive-target group and 1 in the liberal-target group; and on the Montreal Cognitive Assessment, the median score was 27 in the two groups. At 48 hours, the median neuron-specific enolase level was 17 µg per liter in the restrictive-target group and 18 µg per liter in the liberal-target group. The incidence of adverse events was similar in the two groups. CONCLUSIONS: Targeting of a restrictive or liberal oxygenation strategy in comatose patients after resuscitation for cardiac arrest resulted in a similar incidence of death or severe disability or coma. (Funded by the Novo Nordisk Foundation; BOX ClinicalTrials.gov number, NCT03141099.).

Comparative Content of Neuron-Specific Enolase in Human Blood Serum and Seminal Plasma.

We studied the content of neuron-specific enolase (NSE) in 69 paired samples of blood serum and seminal plasma from men with azoospermia (n=11) and oligoastenozoospermia (n=10) and from men with fertile ejaculate (n=48). NSE concentration was determined by ELISA (Vector-Best kit). The median concentration and the interquartile range of the NSE content in seminal plasma were 65.7 (47.9; 83.4) ng/ml and 24.33 times (р<0.000001) exceeded those for blood serum 2.7 (1.45; 4.0) ng/ml. There were no differences in the content of NSE between the groups for both seminal plasma and blood serum. The content of NSE in seminal plasma did not correlate with the content of NSE in blood serum, and also did not depend on the content of spermatozoa. A weak negative correlation (r=-0.341; p=0.0057) was found between the age of the examinees and the level of NSE in seminal plasma, but not in blood serum.

Correlation between lung cancer markers and air pollutants in western China population.

The relationship between serum lung cancer markers and the air pollution remains unclear. To further reveal the correlation between air pollutants and lung cancer, a retrospective analysis of 446,032 asymptomatic healthy people and symptomatic healthy people from the Health Management Center of the First Affiliated Hospital of Chongqing Medical University from 2014 to 2019 was performed. The distribution characteristics of serum lung cancer markers, cancer embryo antigens (CEA), cytokeratin 19 fragment (CYFRA211), squamous cell carcinoma antigen (SCC), and nerve-specific enolase (NSE) was analyzed in these population. Two independent sample man-Whitney U test was used to analyze the correlation of lung cancer markers and age, and a Chi-square test was used to analyze the relationship between lung cancer markers and gender. The daily change trend was profiled for six main air quality indicators PM10, PM2.5, SO2, NO2, CO, O3 during the same period. The correlation between lung markers and air pollutants was investigated by Spearman and multiple linear regression. The results showed that CYFRA211 had the highest excess rate in the screening population. There were differences in the number of cases with concentrated expression of lung cancer markers in the different age groups. Among them, the people with NSE exceeding the standard were the youngest, and most of them were 40-55 years old. Besides SCC, the expression levels of other markers increased with age, and the expression levels of the four markers in males were significantly higher than those in females. Although the levels of PM10 and PM2.5 exceeded the WHO standard (World Health Organization. 2011), they were not correlated with lung cancer markers. Multiple comparisons showed that the air pollutants SO2 and CYFRA211, as well as NO2 and NSE were closely related, but there was no significant linear relationship between CEA, SCC, and air pollutants. In conclusion, among the four lung cancer markers, CYFRA211 had the highest abnormal excess rate in total screening population, and the expression levels of these markers varied by gender and age, with males showing significantly higher expression levels than females, and they increased significantly with age except for SCC. The differential expression of these lung cancer markers may provide more strategies for lung cancer screening in the corresponding population. Lung cancer markers, CYFRA211 and NSE, can be used as sensitive biomarkers for exposure to certain air pollutants and provide references for the prevention and management of air pollution.

The Value of the Biomarkers Neuron-Specific Enolase and S100 Calcium-Binding Protein for Prediction of Mortality in Children Resuscitated After Cardiac Arrest.

The aim of the present study was to assess the ability of the biomarkers neuron-specific enolase (NSE) and S100 calcium-binding protein b (S100b) to predict 30 day mortality in children resuscitated from cardiac arrest (CA). It was a prospective observational study at a single tertiary heart centre. Consecutive children were admitted after resuscitated in-hospital and out-of-hospital CA. Levels of NSE and S100b were analyzed from 12 to 24 hours, from 24 to 48 hours, and from 48 to 72 hours after admission. The primary endpoint was 30-day mortality. Differences in biomarker levels between survivors and non-survivors were analyzed with the Mann-Whitney U test. Receiver operating characteristics (ROC) curves were applied to assess the predictive ability of the biomarkers and the areas under the ROC curves (AUC) were presented. A total of 32 resuscitated CA patients were included, and 12 (38%) patients died within 30 days after resuscitation. We observed significantly higher levels of NSE and S100b in non-survivors compared to survivors at all timepoints from 12 to 72 hours after CA. NSE achieved AUCs from 0.91-0.98 for prediction of 30 day mortality, whereas S100b achieved AUCs from 0.93-0.94. An NSE cut-off of 61 µg/L sampled between 12-24 hours from admission achieved a sensitivity of 80% and a specificity of 100% for prediction of 30 day mortality. In children resuscitated from CA, the biomarkers NSE and S100b appear to be solid predictors of mortality after 30 days.

Proteomics of exhaled breath condensate in lung cancer and controls using data-independent acquisition (DIA): a pilot study.

Lung cancer, the leading cause of cancer mortality worldwide has a poor prognosis. To develop a non-invasive method for early lung cancer detection, exhaled breath condensate (EBC) was explored in this study. EBC samples were collected from lung cancer patients (n= 10) and healthy controls (n= 10), and a proteomic study was performed to identify potential biomarkers. Data-dependent acquisition was used to build the spectral library, and a data-independent acquisition (DIA) approach was applied for quantification of EBC proteomics. A total of 1151 proteins were identified, and several proteins were significantly upregulated in the lung cancer group compared to the control group. The Gene Ontology analysis revealed that most of the proteins were located within several organelles in the cells and were involved in binding and catalytic activity, and the Kyoto Encyclopedia Genes and Genomes results revealed that the proteins were mainly related to organismal systems and human disease. And S100A11, ANXA1, ENO1, and FABP5 might play a vital role in the EBC proteome. In summary, we demonstrated that the DIA-based quantification method was efficient in performing proteomic analysis in individual EBC samples, and some of the proteins might be novel biomarkers for lung cancer.

Neuroinflammatory Biomarkers Associated With Mild Traumatic Brain Injury History in Special Operations Forces Combat Soldiers.

BACKGROUND: Special Operations Forces (SOF) combat soldiers are frequently exposed to blast and blunt neurotrauma, most often classified as mild traumatic brain injury (mTBI). Repetitive mTBI may increase the risk of developing long-term neurological sequelae. Identifying changes in neuroinflammatory biomarkers before chronic conditions emerge could serve as preliminary evidence of developing neuropathology. OBJECTIVE: To determine the effects of mTBI history, lifetime mTBI incidence, and recency on blood biomarker concentrations of axonal protein neurofilament light (NfL), glycolytic enzyme neuron-specific enolase (NSE), astrocyte-expressed S100 calcium-binding protein B (S100B), and neurotrophic cytokine interleukin-6 (IL-6) in healthy, active duty SOF combat soldiers. METHODS: Self-reported mTBI history/recency and fasted blood samples were collected in this cross-sectional study of 104 asymptomatic SOF combat soldiers. Biomarker concentrations were quantified using commercial enzyme-linked immunosorbent assays. Mann-Whitney U and Kruskal-Wallis tests were used to compare groups. Post hoc tests with appropriate corrections were conducted as warranted. RESULTS: Soldiers with mTBI history had higher NSE concentrations than those without (z = -2.60, P = .01). We also observed significant main effects of lifetime mTBI incidence on NSE (χ(3) = 9.52, P = .02) and S100B (χ(3) = 8.21, P = .04) concentrations and a significant main effect of mTBI recency on NfL concentration (χ(2) = 6.02, P = .049). CONCLUSION: The SOF combat soldiers with mTBI history had increased NSE. Longitudinal studies in this population are needed due to between-subject heterogeneity in biomarker concentrations. The NfL concentrations in our SOF combat soldiers-regardless of mTBI history or recency-were similar to values previously reported in civilian acute TBI patients.

Molecular and biochemical characterization of enolase from Dermanyssus gallinae.

Enolase, a multifunctional glycolytic enzyme, is known to act as a plasminogen receptor in many species, involved in the pivotal processes such as motility, adhesion, invasion, growth, and differentiation of the parasites. Knowledge on the function of enolase from Dermanyssus gallinae is very limited. Here we report on the molecular cloning, enzymatic activity, tissue distribution and plasminogen binding activity of enolase from D. gallinae (DgENO). The full-length of cDNA was 1305 bp, specifying a peptide of 434 amino acids. Bioinformatics analysis showed that DgENO was highly conserved compared with a range of organisms, indicating the potentially similar functions in D. gallinae. A recombinant DgENO (rDgENO) protein was produced and characterized, it catalyzed the dehydration of 2-phospho-D-glycerate to phosphoenolpyruvate, the optimal pH was 7.5. Polyclonal antibodies were generated in mice and western blotting indicated that antiserum specifically recognized the native enolase in the somatic extracts from D. gallinae. Immunohistochemical staining of mite sections revealed that the distribution of DgENO was ubiquitous with high level in salivary gland, mite digestive tissues and fat bodies in D. gallinae. Expression level of DgENO was observed mostly in engorged adult mites. Moreover, ELISA binding assay showed that rDgENO could bind plasminogen, and lysine analog ε-aminocaproic acid significantly inhibited this binding activity, indicating that D. gallinae enolase is a receptor of plasminogen. The present study provided foundation for understanding of the biological functions of DgENO and its application in development of vaccines against D. gallinae.

[Relationship between body temperature, neuron-specific enolase, and clinical course in patients after out-of-hospital cardiac arrest]. / Zusammenhang zwischen Körpertemperatur, neurononspezifischer Enolase und klinischem Verlauf bei Patienten nach prähospitaler Reanimation.

BACKGROUND: According to ILCOR (International Liaison Committee on Resuscitation) recommendations (released in 2003), use of therapeutic hypothermia is recommended for unconscious adult patients who have survived a cardiac arrest regardless of the initial monitored cardiac rhythm. Thereby, the treatment goal is to achieve and maintain a body temperature of 32-34 °C for a period of 12-24 h. According to the October 2015 recommendations of the European Resuscitation Council (ERC), targeted temperature management (TTM) remains part of treatment, but, as an option, it is advised that the targeted body temperature be 36 °C rather than 32-34 °C. PATIENT POPULATION AND METHODS: For a non-randomized retrospective observational study, a total of 149 patients were treated with cardiopulmonary resuscitation (CPR) between May 1999 and September 2009. For the first 4 days after CPR, data associated with demography, resuscitation, therapy (temperature course, neuron-specific enolase [NSE]) and clinical-neurological development (Glasgow Outcome Scale [GOS]) were collected. In the study, patients receiving mild hyperthermia were compared with those who did not receive hypothermia. RESULTS: Of the 149 patients included, 90 were treated with mild hypothermia (as decided by the attending physician), while 59 received no hypothermia therapy. Assessment reveals that mild hypothermia positively influences clinical-neurological progression, but not survival. On day three and four, patients with an unfavorable neurological progression exhibited significantly increased serum levels of NSE (day 4: 108.7 ± 137.3 ng/ml versus 25.5 ± 15.4 ng/ml). Patients receiving hypothermia showed lower average NSE levels compared with persons not receiving hypothermia. Furthermore, during the first 4 days, their NSE values tended to increase slower (NSE value at day 4: 55.9 ± 64.9 ng/ml versus 129.9 ± 174.9 ng/ml). The best cut-off-value for an unfavorable neurological result was 74.2 ng/ml at day four (specificity 100%, sensitivity 48.6%). For the group of patients who received hypothermia, the best cut-off-value was 74.2 ng/ml at day four (specificity 100%, sensitivity 40.9%), and, for the comparison group, best cut-off-value was 25.5 ng/ml at day three (specificity 100%, sensitivity 88.2%). CONCLUSION: After out-of-hospital resuscitation, there is a trend for improved clinical-neurological progression with mild hypothermia but it does not influence the prognostic significance of serum NSE. After assessment of available data, it is not possible to recommend uniform cut-off values for patients who received mild therapeutic hypothermia and for those who did not receive hypothermia treatment.

[The possibility of using neuron-specific enolase as a biomarker in the acute period of stroke]. / Vozmozhnost' ispol'zovaniia neiron-spetsificheskoi enolazy kak biomarkera v ostrom periode insul'ta.

The article presents a review of the literature on neuron-specific enolase (NSE) as a biomarker of stroke. It is shown that NSE does not allow differentiation of the ischemic and hemorrhagic process in stroke, but is suitable for determining the extent of brain tissue destruction both in the first hours of stroke and in the dynamics. The HSE analysis can be useful for monitoring the course of the disease, control of the dynamics of the pathological process, including when the size of the lesion increases, for evaluating the effectiveness of therapy and as a prognostic biomarker.

NSE & S100B protein blood level assessment during a long-distance trail race.

The acute and chronic consequences of long-distance running on brain function have received little attention. The impact of such a hard-physical burden associated with sleep privation during such events such has never been explored in terms of neuropsychological function and brain damage. METHODS: Blood samples were collected from 4 athletes before, during and at the end of one of two races: Grand Raid de la Réunion 2017 (GRR: 165 km, elevation gain: 9529 m, 2 runners) and Trail de la Bourbon 2017 (TB: 111 km, elevation gain: 6433 m, 2 runners). Serum S100B and NSE levels were measured for each runner before, during and after the race. RESULTS: Serum S100B levels (normal range: < 0.15 µg/L) increased early during the race and remained high up to the end of the race in all 4 runners (range: 0.17-0.59 µg/L). NSE level (normal range: < 15 µg/L) increased in 3 of the 4 runners (range: 16.8-39.2 µg/L). CONCLUSIONS: This preliminary study shows the potential interest of S100B and NSE serum assessment during long-distance races. Further studies are needed to confirm these results and to investigate the origins and significance of this increase in brain injury markers.

Optimized combination of multiple biomarkers to improve diagnostic accuracy in male fertility.

Artificial insemination is the general method of breeding for genetic improvement in offspring. However, almost half of the insemination cases fail to achieve full-term pregnancy, due to male infertility or subfertility. To maximize the success of insemination, accurate semen quality testing is required prior to insemination. Even though basic semen analyses have been used to provide preliminary information, it cannot fully identify the superior or inferior fertility bulls. Therefore, more powerful and easy to use methods for the prediction of male fertility are required, such as proteomic or microarray chips. During past decades, omics approaches have been developed and suggested the numerous fertility-related potential biomarkers. Our previous study identified the fertility related protein markers, enolase1 (ENO1), ATP synthase, H+ transporting, mitochondrial F1 complex, beta subunit (ATP5B), voltage-dependent anion channel 2 (VDAC2), phospholipid hydroperoxide glutathione peroxide (GPx4), and ubiquinol-cytochrome-c reductase complex core protein 2 (UQCRC2) in bovine spermatozoa. In the present study, we perform a marker combination assay using the western blot data of ENO1, ATP5B, VDAC2, GPx4, and UQCRC2 from 20 individual bull semen samples. And then, we identified the predictive ability of these markers for normal (non-return rate (NRR) ≥ 70%) and normal fertility (NRR<70%) in bulls. ENO1, a single protein marker, achieved an area under the curve (AUC) of 0.86 and 90% discriminatory power between normal and below-normal fertility bulls, with 90% sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV). Although no meaningful changes existed in overall accuracy (70-85%) to discriminate the normal and below-normal fertility between ENO1 single marker and combined marker panels, multiple marker combination methods using ENO1, VDAC2, GPx4, and UQCRC2 provided absolute sensitivity and NPV, with higher specificity (70%) and PPV (77%). ENO1 can be used as a fertility marker candidate, but there were limitations for providing absolute information about normal and below-normal fertility. Although the combined use of fertility-related markers cannot provide absolute accuracy, it can help in indicating below-normal fertility in bulls. These results may contribute to the maintenance cost in the animal industry, via selection of bulls with inferior fertility.

Effect of infectious bursal disease virus infection on energy metabolism in embryonic chicken livers.

1. The purpose of this study was to investigate ATP levels and the activities of important enzymes involved in glycolysis and TCA cycle in livers of embryonated chicken eggs infected by infectious bursal disease virus (IBDV).2. Embryonated chicken eggs (9 days) were randomly divided into two groups (50 eggs per group). The first group was inoculated with a very virulent IBDV (vvIBDV) isolate into the chorioallantoic membrane. The second group was maintained as uninfected control eggs and inoculated with physiological saline. Embryo survival was assessed daily, and six embryos were sacrificed at 24, 48, 72, 96, and 120 hpi for examining livers. Viral loads in the livers were evaluated by qRT-PCR. A comparative analysis of markers associated with the regulation of energy metabolism across several functional classes (ATP, pyruvic and lactic acids, mitochondrial protein, NAD+/NADH ratios, and enolase, lactic acid dehydrogenase and the respiratory chain complex I activities) were examined in the context of IBDV infection.3. The results indicated that increases in the enzymatic activities associated with glycolytic metabolism in turn affected the synthesis and cytoplasmic concentrations of ATP at early timepoints in infected chicken embryos. Subsequently, energy metabolism was inhibited through the pathological perturbations of metabolic enzymes and mitochondrial damage, as inferred from reduced ATP generation.4. These results suggested impaired bioenergetics, which may lead to liver dysfunction consequent to IBDV infection, contributing to the disease pathogenesis.

Neuron-specific enolase (NSE) improves clinical risk scores for prediction of neurological outcome and death in cardiac arrest patients: Results from a prospective trial.

AIM: Neuron-specific enolase (NSE) increases in response to brain injury and is recommended for outcome prediction in cardiac arrest patients. Our aim was to investigate whether NSE measured at different days after a cardiac arrest and its kinetics would improve the prognostic ability of two cardiac arrest specific risk scores. METHODS: Within this prospective observational study, we included consecutive adult patients after cardiac arrest. We calculated the Out-of-hospital cardiac arrest (OHCA) score and the Cardiac Arrest Hospital Prognosis (CAHP) score upon ICU admission and measured serum NSE upon admission and days 1, 2, 3, 5 and 7. We calculated logistic regression models to study associations of scores and NSE levels with neurological outcome defined by Cerebral Performance Category (CPC) scale and in-hospital death. RESULTS: From 336 included patients, 180 (54%) survived until hospital discharge, of which 150 (45%) had a good neurological outcome. NSE at day 3 showed the highest prognostic accuracy (discrimination) for neurological outcome (area under the curve (AUC) 0.89) and in-hospital mortality (AUC 0.88). These results were robust in reclassification statistics and across different subgroups. NSE kinetics with admission levels serving as a baseline did not further improve prognostication. NSE on day 3 significantly improved discrimination of both clinical risk scores (CAHP from AUC 0.81 to 0.91; OHCA from AUC 0.79 to 0.89). CONCLUSION: NSE measured at day 3 significantly improves clinical risk scores for outcome prediction in cardiac arrest patients and may thus add to clinical decision making about escalation or withdrawal of therapy in this vulnerable patient population.

Recent advances in biosensor for detection of lung cancer biomarkers.

Lung cancer is primary cancer threatening human life worldwide with the highest mortality rate. The early detection of lung cancer plays a critical role in the early diagnosis and subsequent treatment. However, the conventional methodologies limit the applications due to the low sensitivity, being expensive, and invasive procedure. Tumor markers as biochemical parameters can reflect cancer occurrence and progression, which show sensitivity, convenience, and low cost in developing biosensors, and act as good candidates for fabricating biosensors of detecting lung cancer. This review describes various biosensors (2013-2019) for detection of lung cancer biomarkers. Firstly, the various reported tumor markers of lung cancer are briefly described. Then, the advancements of designing biosensors for sensitive, stable, and selective identification of lung cancer biomarkers are systematically provided, with a specific focus on the main clinical biomarkers such as neuron-specific enolase (NSE), cytokeratin 19 fragment (CYFRA 21-1). Finally, the recent challenges and further opportunities for developing effective biosensors for early diagnosis of lung cancer are discussed.