A high-affinity aptamer with base-appended base-modified DNA bound to isolated authentic SARS-CoV-2 strains wild-type and B.1.617.2 (delta variant).

Simple, highly sensitive detection technologies for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are crucial for the effective implementation of public health policies. We used the systematic evolution of ligands by exponential enrichment with a modified DNA library, including a base-appended base (uracil with a guanine base at its fifth position), to create an aptamer with a high affinity for the receptor-binding domain (RBD) of the SARS-CoV-2 spike glycoprotein. The aptamer had a dissociation constant of 1.2 and < 1 nM for the RBD and spike trimer, respectively. Furthermore, enzyme-linked aptamer assays confirmed that the aptamer binds to isolated authentic SARS-CoV-2 wild-type and B.1.617.2 (delta variant). The binding signal was larger that of commercially available anti-SARS-CoV-2 RBD antibody. Thus, this aptamer as a sensing element will enable the highly sensitive detection of SARS-CoV-2.

Construction of a Risk Prediction Model of Extended Release Oxycodone Tablet-Induced Nausea and Clarification of Predictive Factors.

Nausea is a typical adverse event associated with opioids. In this study, we performed logistic regression analysis with the aim of clarifying the risk factors for nausea induced by extended-release oxycodone (ER-OXY). Furthermore, we constructed a decision tree (DT) model, a typical data mining method, to estimate the risk of oxycodone-induced nausea by combining multiple factors. A retrospective study was conducted on patients who newly received ER-OXY for cancer pain during hospitalization at Hokkaido University Hospital in Japan from April 2015 to March 2018. In logistic regression and DT analyses, the dependent variable was the presence or absence of nausea. Independent variables were the potential risk factors. First, univariate analyses were performed to screen potential factors associated with oxycodone-induced nausea. Then, multivariate and DT analyses were performed using factors with p-values <0.1 in the univariate analysis. Of 267 cases included in this study, nausea was observed in 30.3% (81/267). In multivariate logistic regression analysis, only female sex was extracted as an independent factor affecting nausea (odds ratio, 1.98). In the DT analysis, we additionally revealed that an age <50 years was a risk factor for nausea in female patients. Thus, our DT model indicated that the risk of ER-OXY-induced nausea was highest in the subgroup comprising females <50 years of age (66.7%) and lowest in male patients (25.1%). The DT model suggested that the factor of young women may be an increased risk of ER-OXY-induced nausea.

FSBC: fast string-based clustering for HT-SELEX data.

BACKGROUND: The combination of systematic evolution of ligands by exponential enrichment (SELEX) and deep sequencing is termed high-throughput (HT)-SELEX, which enables searching aptamer candidates from a massive amount of oligonucleotide sequences. A clustering method is an important procedure to identify sequence groups including aptamer candidates for evaluation with experimental analysis. In general, aptamer includes a specific target binding region, which is necessary for binding to the target molecules. The length of the target binding region varies depending on the target molecules and/or binding styles. Currently available clustering methods for HT-SELEX only estimate clusters based on the similarity of full-length sequences or limited length of motifs as target binding regions. Hence, a clustering method considering the target binding region with different lengths is required. Moreover, to handle such huge data and to save sequencing cost, a clustering method with fast calculation from a single round of HT-SELEX data, not multiple rounds, is also preferred. RESULTS: We developed fast string-based clustering (FSBC) for HT-SELEX data. FSBC was designed to estimate clusters by searching various lengths of over-represented strings as target binding regions. FSBC was also designed for fast calculation with search space reduction from a single round, typically the final round, of HT-SELEX data considering imbalanced nucleobases of the aptamer selection process. The calculation time and clustering accuracy of FSBC were compared with those of four conventional clustering methods, FASTAptamer, AptaCluster, APTANI, and AptaTRACE, using HT-SELEX data (>15 million oligonucleotide sequences). FSBC, AptaCluster, and AptaTRACE could complete the clustering for all sequence data, and FSBC and AptaTRACE performed higher clustering accuracy. FSBC showed the highest clustering accuracy and had the second fastest calculation speed among all methods compared. CONCLUSION: FSBC is applicable to a large HT-SELEX dataset, which can facilitate the accurate identification of groups including aptamer candidates. AVAILABILITY OF DATA AND MATERIALS: FSBC is available at http://www.aoki.ecei.tohoku.ac.jp/fsbc/.

Fluorescence Polarization-Based Rapid Detection System for Salivary Biomarkers Using Modified DNA Aptamers Containing Base-Appended Bases.

The field of care testing toward the analysis of blood and saliva lacks nowadays simple test techniques for biomarkers. In this study, we have developed a novel nucleobase analog, Ugu, which is a uracil derivative bearing a guanine base at the 5-position. Moreover, we attempted the development of aptamers that can bind to secretory immunoglobulin A (SIgA), which has been examined as a stress marker in human saliva. It was observed that the acquired aptamer binds strongly and selectively to the SIgA dimer (Kd = 13.6 nM) without binding to the IgG and IgA monomers of human serum. Reduction of the aptamer length (41 mer) successfully improved 4-fold the binding affinity (Kd = 3.7 nM), compared to the original, longer aptamer (78 mer). Furthermore, the development of a simple detection system for human saliva samples by fluorescence polarization was investigated, using the reported human salivary α-amylase (sAA) and the SIgA-binding aptamer. Comparison of the present method with conventional enzyme-linked immunosorbent assay techniques highlighted a significant Pearson's correlation of 0.94 and 0.83 when targeting sAA and SIgA, respectively. It is thus strongly suggested that a new simple test of stress markers in human saliva can be quantified quickly without bound/free (B/F) separation.

An Aptamer-Based Biosensor for Direct, Label-Free Detection of Melamine in Raw Milk.

Melamine, a nitrogen-rich compound, has been used as a food and milk additive to falsely increase the protein content. However, melamine is toxic, and high melamine levels in food or in milk can cause kidney and urinary problems, or even death. Hence, the detection of melamine in food and milk is desirable, for which numerous detection methods have been developed. Several methods have successfully detected melamine in raw milk; however, they require a sample preparation before the analyses. This study aimed to develop an aptamer-DNAzyme conjugated biosensor for label-free detection of melamine, in raw milk, without any sample preparation. An aptamer-DNAzyme conjugated biosensor was developed via screening using microarray analysis to identify the candidate aptamers followed by an optimization, to reduce the background noise and improve the aptamer properties, thereby, enhancing the signal-to-noise (S/N) ratio of the screened biosensor. The developed biosensor was evaluated via colorimetric detection and tested with raw milk without any sample preparation, using N-methylmesoporphyrin IX for fluorescence detection. The biosensor displayed significantly higher signal intensity at 2 mM melamine (S/N ratio, 20.2), which was sufficient to detect melamine at high concentrations, in raw milk.

Association of growth differentiation factor 11/8, putative anti-ageing factor, with cardiovascular outcomes and overall mortality in humans: analysis of the Heart and Soul and HUNT3 cohorts.

AIMS: Growth differentiation factor 11 and/or its homologue growth differentiation factor 8 (GDF11/8) reverses age-related cardiac hypertrophy and vascular ageing in mice. We investigated whether GDF11/8 associates with cardiovascular outcomes, left ventricular hypertrophy (LVH), or age in humans. METHODS AND RESULTS: We measured plasma GDF11/8 levels in 928 participants with stable ischaemic heart disease in the Heart and Soul study. We adjudicated heart failure hospitalization, stroke, myocardial infarction, death, and their composite endpoint. Left ventricular hypertrophy was evaluated by echocardiography. We used multivariable Cox proportional hazards models to compare rates of cardiovascular events and death across GDF11/8 quartiles and logistic regression models to evaluate the association between GDF11/8 and LVH. Four hundred and fifty participants (48.5%) experienced a cardiovascular event or death during 8.9 years of follow-up. The adjusted risk of the composite endpoint was lower in the highest compared with the lowest GDF11/8 quartile [hazard ratio (HR), 0.45; 95% confidence interval (CI), 0.33-0.60; P < 0.001]. We replicated this relationship of GDF11/8 to adverse events in 971 participants in the HUNT3 cohort (adjusted HR, 0.34; 95% CI, 0.23-0.51; P < 0.001). Left ventricular hypertrophy was present in 368 participants (39.7%) at baseline. Participants in the highest quartile of GDF11/8 were less likely to have LVH than those in the lowest quartile (adjusted OR, 0.55; 95% CI, 0.35-0.86; P = 0.009). GDF11/8 levels were lower in older individuals (P < 0.001). CONCLUSION: In patients with stable ischaemic heart disease, higher GDF11/8 levels are associated with lower risk of cardiovascular events and death. Our findings suggest that GDF11/8 has similar cardioprotective properties in humans to those demonstrated in mice.

Development and Validation of a Protein-Based Risk Score for Cardiovascular Outcomes Among Patients With Stable Coronary Heart Disease.

IMPORTANCE: Precise stratification of cardiovascular risk in patients with coronary heart disease (CHD) is needed to inform treatment decisions. OBJECTIVE: To derive and validate a score to predict risk of cardiovascular outcomes among patients with CHD, using large-scale analysis of circulating proteins. DESIGN, SETTING, AND PARTICIPANTS: Prospective cohort study of participants with stable CHD. For the derivation cohort (Heart and Soul study), outpatients from San Francisco were enrolled from 2000 through 2002 and followed up through November 2011 (≤11.1 years). For the validation cohort (HUNT3, a Norwegian population-based study), participants were enrolled from 2006 through 2008 and followed up through April 2012 (5.6 years). EXPOSURES: Using modified aptamers, 1130 proteins were measured in plasma samples. MAIN OUTCOMES AND MEASURES: A 9-protein risk score was derived and validated for 4-year probability of myocardial infarction, stroke, heart failure, and all-cause death. Tests, including the C statistic, were used to assess performance of the 9-protein risk score, which was compared with the Framingham secondary event model, refit to the cohorts in this study. Within-person change in the 9-protein risk score was evaluated in the Heart and Soul study from paired samples collected 4.8 years apart. RESULTS: From the derivation cohort, 938 samples were analyzed, participants' median age at enrollment was 67.0 years, and 82% were men. From the validation cohort, 971 samples were analyzed, participants' median age at enrollment was 70.2 years, and 72% were men. In the derivation cohort, C statistics were 0.66 for refit Framingham, 0.74 for 9-protein, and 0.75 for refit Framingham plus 9-protein models. In the validation cohort, C statistics were 0.64 for refit Framingham, 0.70 for 9-protein, and 0.71 for refit Framingham plus 9-protein models. Adding the 9-protein risk score to the refit Framingham model increased the C statistic by 0.09 (95% CI, 0.06-0.12) in the derivation cohort, and in the validation cohort, the C statistic was increased by 0.05 (95% CI, 0.02-0.09). Compared with the refit Framingham model, the integrated discrimination index for the 9-protein model was 0.12 (95% CI, 0.08-0.16) in the derivation cohort and 0.08 (95% CI, 0.05-0.10) in the validation cohort. In analysis of paired samples among 139 participants with cardiovascular events after the second sample, absolute within-person annualized risk increased more for the 9-protein model (median, 1.86% [95% CI, 1.15%-2.54%]) than for the refit Framingham model (median, 1.00% [95% CI, 0.87%-1.19%]) (P = .002), while among 375 participants without cardiovascular events, both scores changed less and similarly (P = .30). CONCLUSIONS AND RELEVANCE: Among patients with stable CHD, a risk score based on 9 proteins performed better than the refit Framingham secondary event risk score in predicting cardiovascular events, but still provided only modest discriminative accuracy. Further research is needed to assess whether the score is more accurate in a lower-risk population.

Angiotensin II type 1A receptor signaling facilitates tumor metastasis formation through P-selectin-mediated interaction of tumor cells with platelets and endothelial cells.

Angiotensin II is involved in tumor growth; however, the precise mechanism is not known. Platelets also contribute to tumor growth, and angiotensin II type 1 receptor (AT1) is expressed on the platelet surface. We hypothesized that interaction of platelets with tumor cells through AT1 receptor signaling promotes tumor metastasis. B16F1 melanoma cells were intravenously injected into Agtr1a knockout mice (AT1a(-/-)) and wild-type littermates (WT); the AT1a(-/-) mice exhibited a reduction in lung colonies. Angiotensin II induced expression of P-selectin on platelets in WT but not in AT1a(-/-) mice. A selective P-selectin neutralizing antibody decreased lung colony numbers in WT but not in AT1a(-/-) mice. Levels of vascular endothelial growth factor (VEGF) and stromal cell-derived factor 1 (SDF-1) receptor in platelets at metastatic locus were lower in AT1a(-/-) mice. Treatment of neutralizing antibodies against VEGF and CXCR4 decreased lung colony numbers in WT but not in AT1a(-/-) mice. In AT1a(-/-) mice, and both mobilization of progenitor cells expressing CXCR4(+)VEGFR1(+) cells from bone marrow and their recruitment to lung tissues were suppressed. These results suggest that AT1A signaling plays a critical role in tumor metastasis through P-selectin-mediated interactions of platelets with tumor and endothelial cells and through the AT1A signaling-dependent production of VEGF and SDF-1, which may be involved in mobilization of CXCR4(+)VEGFR1(+) cells.

Utility of salivary cortisol profile as a predictive biomarker in nurses' turnover risk: a preliminary study.

BACKGROUND: Predicting nurse turnover risk is crucial due to the global nursing shortage; however, existing predictors, such as fatigue and burnout, lack objectivity. Salivary cortisol is a non-invasive marker of stress and fatigue, but its utility in predicting nurse turnover risk is unknown. We examined whether salivary cortisol profiles across three different day shifts in a month are predictors of the extent of nurses' reluctance to stay in their current jobs. METHODS: This preliminary longitudinal study followed forty female nurses who engaged in shift work at a university hospital for 3 months. Data at enrollment were collected including demographics, working conditions, chronic fatigue (the Japanese version of the Occupational Fatigue/Exhaustion Recovery Scale), and burnout (Japanese Burnout scale). Salivary cortisol was measured before the three different day shifts (after awakening) during the first month, and the means of these measurements were used as the cortisol profile. The extent of reluctance to stay was assessed using the numerical rating scale at 3 months. RESULTS: Among the forty female nurses (mean [SD] age, 28.3 [5.1]), all completed follow-up and were included in the analysis. The cortisol profile was associated with the extent of reluctance to stay (P = 0.017), and this association was significant despite adjustments for chronic fatigue and burnout (P = 0.005). A multiple regression model with chronic fatigue, burnout, and job tenure explained 41.5% of the variation in reluctance to stay. When the cortisol profile was added to this model, the association of the cortisol profile was significant (P = 0.006) with an R2 of 0.529 (ΔR2 = 0.114). CONCLUSIONS: This preliminary study conducted in an actual clinical setting indicated the potential of the salivary cortisol profile across three different day shifts in a month to predict nurses' reluctance to stay in their current jobs. The combination of subjective indicators and the cortisol profile would be useful in predicting nurses' turnover risk.

Quantitative Analysis and Manipulation of Alkali Metal Cations at the Cathode Surface in Membrane Electrode Assembly Electrolyzers for CO2 Reduction Reactions.

The stable operation of the CO2 reduction reaction (CO2RR) in membrane electrode assembly (MEA) electrolyzers is known to be hindered by the accumulation of bicarbonate salt, which are derived from alkali metal cations in anolytes, on the cathode side. In this study, we conducted a quantitative evaluation of the correlation between the CO2RR activity and the transported alkali metal cations in MEA electrolyzers. As a result, although the presence of transported alkali metal cations on the cathode surface significantly contributes to the generation of C2+ compounds, the rate of K+ ion transport did not match the selectivity of C2+, suggesting that a continuous supply of high amount of K+ to the cathode surface is not required for C2+ formation. Based on these findings, we achieved a faradaic efficiency (FE) and a partial current density for C2+ of 77 % and 230 mA cm-2, respectively, even after switching the anode solution from 0.1 M KHCO3 to a dilute K+ solution (<7 mM). These values were almost identical to those when 0.1 M KHCO3 was continuously supplied. Based on this insight, we successfully improved the durability of the system against salt precipitation by intermittently supplying concentrated KHCO3, compared with the continuous supply.

High-throughput quantitative screening of peroxidase-mimicking DNAzymes on a microarray by using electrochemical detection.

Some guanine-rich DNA sequences, which are called DNAzymes, can adopt G-quadruplex structures and exhibit peroxidase activity by binding with hemin. Although known DNAzymes show less activity than horseradish peroxidase, they have the potential to be widely used for the detection of target molecules in enzyme-linked immunosorbent assays if sequences that exhibit higher activity can be identified. However, techniques for achieving this have not yet been described. Therefore, we compared the DNAzyme activities of more than 1000 novelistically designed sequences with that of the original DNAzyme by using an electrochemical detection system on a 12K DNA microarray platform. To the best of our knowledge, this is the first description of an array-based assessment of peroxidase activity of G-quadruplex-hemin complexes. By using this novel assay system, more than 200 different mutants were found that had significantly higher activities than the original DNAzyme sequence. This microarray-based DNAzyme evaluation system is useful for identifying highly active new DNAzymes that might have potential as tools for developing DNA-based biosensors with aptamers.

Selective and high-rate CO2 electroreduction by metal-doped covalent triazine frameworks: a computational and experimental hybrid approach.

The electrochemical CO2 reduction reaction (CO2RR) has attracted intensive attention as a technology to achieve a carbon-neutral society. The use of gas diffusion electrodes (GDEs) enables the realization of high-rate CO2RRs, which is one of the critical requirements for social implementation. Although both a high reaction rate and good selectivity are simultaneously required for electrocatalysts on GDEs, no systematic study of the relationship among active metal centers in electrocatalysts, reaction rate, and selectivity under high-rate CO2RR conditions has been reported. In the present study, we employed various metal-doped covalent triazine frameworks (M-CTFs) as platforms for CO2 reduction reaction (CO2RR) electrocatalysts on GDEs and systematically investigated them to deduce sophisticated design principles using a combined computational and experimental approach. The Ni-CTF showed both high selectivity (faradaic efficiency (FE) > 98% at -0.5 to -0.9 V vs. reversible hydrogen electrode) and a high reaction rate (current density < -200 mA cm-2) for CO production. By contrast, the Sn-CTF exhibited selective formic acid production, and the FE and partial current density reached 85% and 150 mA cm-2, respectively. These results for the CO2RR activity and selectivity at high current density with respect to metal centers correspond well with predictions based on first-principles calculations. This work is the first demonstration of a clear relationship between the computational adsorption energy of intermediates depending on metal species and the experimental high-rate gaseous CO2RR.

Geographic Distribution and Genetic Structures of the Tideland Snails Pirenella nipponica and P.asiatica in Taiwan and Japan.

The tideland snails Pirenellanipponica and Pirenellaasiatica are distributed north of the central Ryukyu Islands and in South Ryukyu, respectively, in Japan. To reveal their distribution and genetic characteristics in Taiwan, we sampled Pirenella snails along the western coast of Taiwan Island and analyzed the nucleotide sequences of their mitochondrial DNAs. Pirenella nipponica and P. asiatica inhabit the northern and southern parts of the western coast of Taiwan, respectively, and coexist only in the central part. Taiwanese and Japanese populations of P. asiatica showed significant genetic differentiation. The former showed higher genetic diversity and a larger effective population size than the latter. However, the Taiwanese population of P. nipponica was not genetically deviated from the local Japanese population on Kyushu Island. Both the Taiwan and Kyushu populations of P. nipponica showed significant genetic differences from local populations in other regions of Japan, namely, Honshu Island (the Japanese mainland) and Central Ryukyu. They also showed higher genetic diversity and a larger effective population size than the others. The Taiwanese populations of both species might be part of a large panmictic population with individuals from the Asian continent and Kyushu Island.

Preferred catalysis distinctly determined by metals doped with nitrogen in three-dimensionally ordered porous carbon materials.

Three-dimensionally ordered nanoporous structures were generated in carbon materials doped with metals and nitrogen as catalytically active sites for electrochemical reactions. Free-base and metal phthalocyanines with a strategically designed molecular structure were used as carbon sources to obtain an ordered porous structure via homogeneous self-assembly with Fe3O4 nanoparticles as the pore template and the prevention of melting away during carbonization. The doping of Fe and nitrogen was achieved by a reaction between the free-base phthalocyanine and Fe3O4 through carbonization at 550 °C, while Co and Ni were doped using the corresponding metal phthalocyanines. The preference of these three types of ordered porous carbon materials for catalytic reactions was distinctly determined by the doped metals. Fe-N-doped carbon showed the highest activity for O2 reduction. Additional heat treatment at 800 °C enhanced this activity. CO2 reduction and H2 evolution were preferred by the Ni- and Co-N-doped carbon materials, respectively. A change in the template particle size was capable of controlling the pore size to enhance mass transfer and improve performance. The technique presented in this study enabled systematic metal doping and pore size control in the ordered porous structures of carbonaceous catalysts.

Thromboxane A2 receptor signaling facilitates tumor colonization through P-selectin-mediated interaction of tumor cells with platelets and endothelial cells.

Thromboxane A(2) (TXA(2) ) is a prostanoid formed by thromboxane synthase using the cyclooxygenase product, prostaglandin H(2), as the substrate. TXA(2) was shown to enhance tumor metastasis, but the underlying mechanism remains unclear. B16F1 melanoma cells were intravenously injected into TXA(2) receptor (TP) knockout mice (TP(-/-) ) and wild-type littermates (WT). TP(-/-) showed a reduction in B16F1 lung colonization and mortality rate, which were associated with a decreased number of platelets. Platelet activation as assessed by P-selectin expression was suppressed in TP(-/-) . A selective P-selectin neutralizing antibody decreased the lung colonization in WT mice, but not in TP(-/-) . The expression of P-selectin glycoprotein ligand-1 in B16F1 and HUVEC were enhanced by treatment with U46619, a thromboxane analog. The plasma levels of vascular endothelial growth factor (VEGF) and stromal-derived factor (SDF)-1 were lower in TP(-/-) . In TP(-/-) , the mobilization of progenitor cells expressing CXCR4(+) VEGFR1(+) from bone marrow and the recruitment of those cells to lung tissues were suppressed. These results suggest that TP signaling plays a critical role in tumor colonization through P-selectin-mediated interactions between platelets-tumor cells and tumor cells-endothelial cells through the TP signaling-dependent production of VEGF and SDF-1, which might be involved in the mobilization of VEGFR1(+) CXCR4(+) cells. Blockade of TP signaling might be useful in the treatment of tumor metastasis.

[Efficacy Survey of Naldemedine in the Poor-performance Status Group].

Naldemedine (Nal) is widely used as a therapeutic drug against opioid-induced constipation. However, patients in phase III trials are limited to those with good performance status (PS). Cancer patients may have inferior PS owing to progression of symptoms and adverse events from chemotherapy. Therefore, it is important to survey the efficacy of Nal in patients with poor PS. This study aimed to evaluate Nal efficacy in patients with poor PS. We retrospectively investigated patients from July 2017 to June 2019 and compared Nal efficacy between patients with good and poor PS. The efficacy of Nal was evaluated using changes in the number of spontaneous bowel movements 7 days before and after the introduction of Nal with reference to previous reports. Multivariate analysis was performed to reveal whether poor PS affects Nal efficacy. In total, 141 patients at the Hokkaido University Hospital were analyzed. The effective rate of Nal from day 1 to day 7 of administration was 71.7% and 71.4% in the patients with good and poor PS, respectively, that from day 1 to day 2 of administration was 61.1% and 57.1%, respectively, and that from day 3 to day 7 of administration was 60.2% and 71.4%, respectively, suggesting an absence of significant differences. Furthermore, results of multivariate analysis showed that "best supportive care" and "body weight (55 kg and above)" reduced Nal efficacy. In conclusion, Nal showed similar effectiveness in patients with poor PS as that in those with good PS.

The mitochondrial genome of the threatened tideland snail Pirenella pupiformis (Mollusca: Caenogastropoda: Potamididae) determined by shotgun sequencing.

The nearly complete mitochondrial genome of the threatened tideland snail Pirenella pupiformis (Mollusca: Cerithioidea: Potamididae) was determined by shotgun next-generation sequencing. The mitogenome is comprised of 13 protein-coding genes (PCGs), two ribosomal RNA (12S and 16S) genes, and 22 transfer RNA genes (tRNAs). This gene order is consistent with the previously published mitochondrial genomes of other species belonging to the family Potamididae. The family Potamididae including P. pupiformis was recovered as a monophyletic group in the superfamily Cerithioidea.

Salivary Biomarker Profiles and Chronic Fatigue among Nurses Working Rotation Shifts: An Exploratory Pilot Study.

Although nurses' fatigue affects their well-being and patient safety, no effective objective measurements exist. We explored the profiles of salivary biomarkers associated with nurses' chronic fatigue across several shifts. This longitudinal study involved 45 shiftwork nurses and collected their saliva samples before two night and two day shifts for a month. Chronic fatigue was measured using the Cumulative Fatigue Symptom Index before the first night shift. Biomarker profiles were analyzed using hierarchical cluster analysis, and chronic fatigue levels were compared between the profiles. Cortisol profiles were classified into high- and low-level groups across two day shifts; the low-level group presented significantly higher irritability and unwillingness to work. Secretory immunoglobulin A (s-IgA) profiles across the four shifts were classified into high- and low-level groups; the high-level group had significantly higher depressive feelings, decreased vitality, irritability, and unwillingness to work. Cortisol (two day shifts) and s-IgA (four shifts) profiles were combined, and (i) cortisol low-level and s-IgA high-level and (ii) cortisol high-level and s-IgA low-level groups were identified. The former group had significantly higher chronic fatigue sign and irritability than the latter group. The profiles of salivary cortisol and s-IgA across several shifts were associated with nurses' chronic fatigue.

A proteomic surrogate for cardiovascular outcomes that is sensitive to multiple mechanisms of change in risk.

A reliable, individualized, and dynamic surrogate of cardiovascular risk, synoptic for key biologic mechanisms, could shorten the path for drug development, enhance drug cost-effectiveness and improve patient outcomes. We used highly multiplexed proteomics to address these objectives, measuring about 5000 proteins in each of 32,130 archived plasma samples from 22,849 participants in nine clinical studies. We used machine learning to derive a 27-protein model predicting 4-year likelihood of myocardial infarction, stroke, heart failure, or death. The 27 proteins encompassed 10 biologic systems, and 12 were associated with relevant causal genetic traits. We independently validated results in 11,609 participants. Compared to a clinical model, the ratio of observed events in quintile 5 to quintile 1 was 6.7 for proteins versus 2.9 for the clinical model, AUCs (95% CI) were 0.73 (0.72 to 0.74) versus 0.64 (0.62 to 0.65), c-statistics were 0.71 (0.69 to 0.72) versus 0.62 (0.60 to 0.63), and the net reclassification index was +0.43. Adding the clinical model to the proteins only improved discrimination metrics by 0.01 to 0.02. Event rates in four predefined protein risk categories were 5.6, 11.2, 20.0, and 43.4% within 4 years; median time to event was 1.71 years. Protein predictions were directionally concordant with changed outcomes. Adverse risks were predicted for aging, approaching an event, anthracycline chemotherapy, diabetes, smoking, rheumatoid arthritis, cancer history, cardiovascular disease, high systolic blood pressure, and lipids. Reduced risks were predicted for weight loss and exenatide. The 27-protein model has potential as a "universal" surrogate end point for cardiovascular risk.

Calcitonin gene-related peptide facilitates revascularization during hindlimb ischemia in mice.

It is known that the neural system plays a fundamental role in neovascularization. A neuropeptide, calcitonin gene-related peptide (CGRP), is widely distributed in the central and peripheral neuronal systems. However, it remains to be elucidated the role of CGRP in angiogenesis during ischemia. The present study examined whether endogenous CGRP released from neuronal systems facilitates revascularization in response to ischemia using CGRP knockout mice (CGRP-/-). CGRP-/- or their wild-type littermates (CGRP+/+) were subjected to unilateral hindlimb ischemia. CGRP-/- exhibited impaired blood flow recovery from ischemia and decreased capillary density expressed in terms of the number of CD-31-positive cells in the ischemic tissues compared with CGRP+/+. In vivo microscopic studies showed that the functional capillary density in CGRP-/- was reduced. Hindlimb ischemia increased the expression of pro-CGRP mRNA and of CGRP protein in the lumbar dorsal root ganglia. Lack of CGRP decreased mRNA expression of growth factors, including CD31, vascular endothelial growth factor-A, basic fibroblast growth factor, and transforming growth factor-ß, in the ischemic limb tissue. The application of CGRP enhanced the mRNA expression of CD31 and VEGF-A in human umbilical vein endothelial cells (HUVECs) and fibroblasts. Subcutaneous infusion of CGRP8-37, a CGRP antagonist, using miniosmotic pumps delayed angiogenesis and reduced the expression of proangiogenic growth factors during hindlimb ischemia. These results indicate that endogenous CGRP facilitates angiogenesis in response to ischemia. Targeting CGRP may provide a promising approach for controlling angiogenesis related to pathophysiological conditions.