Utility of Screening Chest Radiographs in Patients with Asymptomatic or Minimally Symptomatic COVID-19 in Singapore.

Background Singapore saw an escalation of coronavirus disease 2019 (COVID-19) cases from fewer than 4000 in April 2020 to more than 40 000 in June 2020, with most of these cases attributed to spread within shared facilities housing foreign workers. Appropriate triage and escalation of clinical care are crucial for this patient group managed in community care facilities (CCFs). Purpose To evaluate the imaging guideline recommendations for COVID-19 from the Fleischner Society and to analyze the clinical utility of screening chest radiography for asymptomatic or minimally symptomatic patients with COVID-19. Materials and Methods In this retrospective study, patients with reverse-transcription polymerase chain reaction-confirmed COVID-19 who were admitted to a designated CCF for continuation of their treatment during May 3-31, 2020, were identified. Upon admission, patients aged 36 years and older without any baseline chest images underwent chest radiography. All chest radiographs and clinical outcomes of patients, including those who were subsequently transferred to acute hospitals for escalation of care, were reviewed. Key proportions of patients with findings of pulmonary infection and those requiring further inpatient treatment were calculated, and 95% binomial proportion CIs were obtained using the Clopper-Pearson method. Results The study included 5621 patients. All patients were men (100%; 5621 of 5621), and the mean patient age was 37 years ± 8 (range, 17-60 years). A total of 1964 chest radiographs were obtained, of which normal images accounted for 98.0% (1925 of 1964 radiographs) and findings of pulmonary infection represented 2.0% (39 of 1964 radiographs). Only 0.2% of patients (four of 1964) with findings of pulmonary infection at chest radiography (all of whom were symptomatic) required supplemental oxygenation and inpatient treatment. None of the asymptomatic patients with findings of pulmonary infection required supplemental oxygenation, and they received only symptomatic treatment. Conclusion In accordance with Fleischner Society recommendations, screening chest radiography is not indicated in patients with coronavirus disease 2019 who are aged 17-60 years with mild or no symptoms unless there is risk of clinical deterioration. © RSNA, 2021 See also the editorial by Schaefer-Prokop and Prokop in this issue.

Parallel Reaction Monitoring Mass Spectrometry Method for Detection of Both Casein and Whey Milk Allergens from a Baked Food Matrix.

Milk allergy is among the most common food allergies present in early childhood, which in some cases may persist into adulthood as well. Proteins belonging to both casein and whey fractions of milk can trigger an allergic response in susceptible individuals. Milk is present as an ingredient in many foods, and it can also be present as casein- or whey-enriched milk-derived ingredients. As whey proteins are more susceptible to thermal processing than caseins, conventional methods often posed a challenge in accurate detection of whey allergens, particularly from a processed complex food matrix. In this study, a targeted mass spectrometry method has been developed to detect the presence of both casein and whey allergens from thermally processed foods. A pool of 19 candidate peptides representing four casein proteins and two whey proteins was identified using a discovery-driven target selection approach from various milk-derived ingredients. These target peptides were evaluated by parallel reaction monitoring of baked cookie samples containing known amounts of nonfat dry milk (NFDM). The presence of milk could be detected from baked cookies incurred with NFDM at levels as low as 1 ppm using seven peptides representing α-, ß-, and κ-casein proteins and three peptides representing a whey protein, ß-lactoglobulin, by this consensus PRM method.

Detection of Six Commercially Processed Soy Ingredients in an Incurred Food Matrix Using Parallel Reaction Monitoring.

Soybeans are one of the major allergenic foods in many countries. Soybeans are commonly processed into different types of soy ingredients to achieve the desired properties. The processing, however, may affect the protein profiles and protein structure, thus affecting the detection of soy proteins. Mass spectrometry (MS) is a potential alternative to the traditional immunoassays for the detection of soy-derived ingredients in foods. This study aims to develop a liquid chromatography-tandem MS method that uniformly detects different types of soy-derived ingredients. Target peptides applicable to the detection of six commercial soy ingredients were identified based on the results of MS label-free quantification and a set of selection criteria. The results indicated that soy ingredient processing can result in different protein profiles. A total of six soy ingredients were then individually incurred into cookie matrices at different levels. Sample preparation methods were optimized, and a distinct improvement in peptide performance was observed after optimization. Cookies and dough incurred with different soy ingredients at 100 ppm total soy protein showed a similar level of peptide recovery (90% mean signal relative to unroasted soy flour), demonstrating the ability of the MS method to detect processed soy ingredients in a uniform manner.

Intracellular Binding Site for a Positive Allosteric Modulator of the Dopamine D1 Receptor.

The binding site for DETQ [2-(2,6-dichlorophenyl)-1-((1S,3R)-3-(hydroxymethyl)-5-(2-hydroxypropan-2-yl)-1-methyl-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one], a positive allosteric modulator (PAM) of the dopamine D1 receptor, was identified and compared with the binding site for CID 2886111 [N-(6-tert-butyl-3-carbamoyl-4,5,6,7-tetrahydro-1-benzothiophen-2-yl)pyridine-4-carboxamide], a reference D1 PAM. From D1/D5 chimeras, the site responsible for potentiation by DETQ of the increase in cAMP in response to dopamine was narrowed down to the N-terminal intracellular quadrant of the receptor; arginine-130 in intracellular loop 2 (IC2) was then identified as a critical amino acid based on a human/rat species difference. Confirming the importance of IC2, a ß2-adrenergic receptor construct in which the IC2 region was replaced with its D1 counterpart gained the ability to respond to DETQ. A homology model was built from the agonist-state ß2-receptor structure, and DETQ was found to dock to a cleft created by IC2 and adjacent portions of transmembrane helices 3 and 4 (TM3 and TM4). When residues modeled as pointing into the cleft were mutated to alanine, large reductions in the potency of DETQ were found for Val119 and Trp123 (flanking the conserved DRY sequence in TM3), Arg130 (located in IC2), and Leu143 (TM4). The D1/D5 difference was found to reside in Ala139; changing this residue to methionine as in the D5 receptor reduced the potency of DETQ by approximately 1000-fold. None of these mutations affected the activity of CID 2886111, indicating that it binds to a different allosteric site. When combined, DETQ and CID 2886111 elicited a supra-additive response in the absence of dopamine, implying that both PAMs can bind to the D1 receptor simultaneously.

A small molecule activator of Nav 1.1 channels increases fast-spiking interneuron excitability and GABAergic transmission in vitro and has anti-convulsive effects in vivo.

Nav 1.1 (SCN1A) channels primarily located in gamma-aminobutyric acid (GABA)ergic fast-spiking interneurons are pivotal for action potential generation and propagation in these neurons. Inappropriate function of fast-spiking interneurons, leading to disinhibition of pyramidal cells and network desynchronization, correlates with decreased cognitive capability. Further, reduced functionality of Nav 1.1 channels is linked to various diseases in the central nervous system. There is, at present, however no subtype selective pharmacological activators of Nav 1.1 channels available for studying pharmacological modulation of interneuron function. In the current study, we identified a small molecule Nav 1.1 activator, 3-amino-5-(4-methoxyphenyl)thiophene-2-carboxamide, named AA43279, and provided an in vitro to in vivo characterization of the compound. In HEK-293 cells expressing human Nav 1.1 channels, AA43279 increased the Nav 1.1-mediated current in a concentration-dependent manner mainly by impairing the fast inactivation kinetics of the channels. In rat hippocampal brain slices, AA43279 increased the firing activity of parvalbumin-expressing, fast-spiking GABAergic interneurons and increased the spontaneous inhibitory post-synaptic currents (sIPSCs) recorded from pyramidal neurons. When tested in vivo, AA43279 had anti-convulsive properties in the maximal electroshock seizure threshold test. AA43279 was tested for off-target effects on 72 different proteins, including Nav 1.2, Nav 1.4, Nav 1.5, Nav 1.6 and Nav 1.7 and exhibited reasonable selectivity. Taken together, AA43279 might constitute a valuable tool compound for revealing biological functions of Nav 1.1 channels.

An Allosteric Potentiator of the Dopamine D1 Receptor Increases Locomotor Activity in Human D1 Knock-In Mice without Causing Stereotypy or Tachyphylaxis.

Allosteric potentiators amplify the sensitivity of physiologic control circuits, a mode of action that could provide therapeutic advantages. This hypothesis was tested with the dopamine D1 receptor potentiator DETQ [2-(2,6-dichlorophenyl)-1-((1S,3R)-3-(hydroxymethyl)-5-(2-hydroxypropan-2-yl)-1-methyl-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one]. In human embryonic kidney 293 (HEK293) cells expressing the human D1 receptor, DETQ induced a 21-fold leftward shift in the cAMP response to dopamine, with a Kb of 26 nM. The maximum response to DETQ alone was ∼12% of the maximum response to dopamine, suggesting weak allosteric agonist activity. DETQ was ∼30-fold less potent at rat and mouse D1 receptors and was inactive at the human D5 receptor. To enable studies in rodents, an hD1 knock-in mouse was generated. DETQ (3-20 mg/kg orally) caused a robust (∼10-fold) increase in locomotor activity (LMA) in habituated hD1 mice but was inactive in wild-type mice. The LMA response to DETQ was blocked by the D1 antagonist SCH39166 and was dependent on endogenous dopamine. LMA reached a plateau at higher doses (30-240 mg/kg) even though free brain levels of DETQ continued to increase over the entire dose range. In contrast, the D1 agonists SKF 82958, A-77636, and dihydrexidine showed bell-shaped dose-response curves with a profound reduction in LMA at higher doses; video-tracking confirmed that the reduction in LMA caused by SKF 82958 was due to competing stereotyped behaviors. When dosed daily for 4 days, DETQ continued to elicit an increase in LMA, whereas the D1 agonist A-77636 showed complete tachyphylaxis by day 2. These results confirm that allosteric potentiators may have advantages compared with direct-acting agonists.

Ontogeny and phylogeny of language.

How did language evolve? A popular approach points to the similarities between the ontogeny and phylogeny of language. Young children's language and nonhuman primates' signing both appear formulaic with limited syntactic combinations, thereby suggesting a degree of continuity in their cognitive abilities. To evaluate the validity of this approach, as well as to develop a quantitative benchmark to assess children's language development, I propose a formal analysis that characterizes the statistical profile of grammatical rules. I show that very young children's language is consistent with a productive grammar rather than memorization of specific word combinations from caregivers' speech. Furthermore, I provide a statistically rigorous demonstration that the sign use of Nim Chimpsky, the chimpanzee who was taught American Sign Language, does not show the expected productivity of a rule-based grammar. Implications for theories of language acquisition and evolution are discussed.

Bilateral Paravertebral Blockade (T7-10) Versus Incisional Local Anesthetic Administration for Pediatric Laparoscopic Cholecystectomy: A Prospective, Randomized Clinical Study.

BACKGROUND: Single-injection paravertebral nerve blocks (PVBs) provide effective postoperative analgesia after adult laparoscopic cholecystectomy (LC). We sought to compare PVBs with local anesthetic injections at laparoscopic port sites in a pediatric population. METHODS: Eighty-three patients (8-17 years old) scheduled for LC were randomized prospectively to 2 treatment groups: the PVB group received ropivacaine 0.5% injected in the paravertebral space and normal saline injections at laparoscopic instrument sites, and the port infiltration group received normal saline in the paravertebral space and ropivacaine 0.5% at instrument sites. Postoperative analgesia was provided with hydromorphone via patient-controlled analgesia for up to 12 hours, followed by oxycodone and hydromorphone. The total amount of analgesic, serial visual analog scale scores for pain and subject pain control satisfaction, type and characteristics of pain, and complications were recorded for 24 hours. RESULTS: The intraoperative fentanyl requirement (ng/kg/min) was lower in the PVB group than in the port infiltration group (12.81 vs 16.57, P = 0.007). Total postoperative analgesic consumption and mean visual analog scale scores were not different between the groups. Baseline pain recorded before surgery correlated with self-reported postoperative pain scores only in the port infiltration group. The rate of complications was low and similar between groups. There was no difference in incidence of patient-reported incisional, visceral, or gas pain. Shoulder pain, however, was 49% less (95% confidence interval, 0.269-0.893) in the port infiltration group. CONCLUSIONS: PVBs did not reduce postoperative pain associated with pediatric LC but decreased intraoperative fentanyl requirements.

Artemin growth factor increases nicotinic cholinergic receptor subunit expression and activity in nociceptive sensory neurons.

BACKGROUND: Artemin (Artn), a member of the glial cell line-derived growth factor (GDNF) family, supports the development and function of a subpopulation of peptidergic, TRPV1-positive sensory neurons. Artn (enovin, neublastin) is elevated in inflamed tissue and its injection in skin causes transient thermal hyperalgesia. A genome wide expression analysis of trigeminal ganglia of mice that overexpress Artn in the skin (ART-OE mice) showed elevation in nicotinic acetylcholine receptor (nAChR) subunits, suggesting these ion channels contribute to Artn-induced sensitivity. Here we have used gene expression, immunolabeling, patch clamp electrophysiology and behavioral testing assays to investigate the link between Artn, nicotinic subunit expression and thermal hypersensitivity. RESULTS: Reverse transcriptase-PCR validation showed increased levels of mRNAs encoding the nAChR subunits α3 (13.3-fold), ß3 (4-fold) and ß4 (7.7-fold) in trigeminal ganglia and α3 (4-fold) and ß4 (2.8-fold) in dorsal root ganglia (DRG) of ART-OE mice. Sensory ganglia of ART-OE mice had increased immunoreactivity for nAChRα3 and exhibited increased overlap in labeling with GFRα3-positive neurons. Patch clamp analysis of back-labeled cutaneous afferents showed that while the majority of nicotine-evoked currents in DRG neurons had biophysical and pharmacological properties of α7-subunit containing nAChRs, the Artn-induced increase in α3 and ß4 subunits resulted in functional channels. Behavioral analysis of ART-OE and wildtype mice showed that Artn-induced thermal hyperalgesia can be blocked by mecamylamine or hexamethonium. Complete Freund's adjuvant (CFA) inflammation of paw skin, which causes an increase in Artn in the skin, also increased the level of nAChR mRNAs in DRG. Finally, the increase in nAChRs transcription was not dependent on the Artn-induced increase in TRPV1 or TRPA1 in ART-OE mice since nAChRs were elevated in ganglia of TRPV1/TRPA1 double knockout mice. CONCLUSIONS: These findings suggest that Artn regulates the expression and composition of nAChRs in GFRα3 nociceptors and that these changes contribute to the thermal hypersensitivity that develops in response to Artn injection and perhaps to inflammation.

Genetic variants at 13q12.12 are associated with high myopia in the Han Chinese population.

High myopia, which is extremely prevalent in the Chinese population, is one of the leading causes of blindness in the world. Genetic factors play a critical role in the development of the condition. To identify the genetic variants associated with high myopia in the Han Chinese, we conducted a genome-wide association study (GWAS) of 493,947 SNPs in 1088 individuals (419 cases and 669 controls) from a Han Chinese cohort and followed up on signals that were associated with p < 1.0 × 10(-4) in three independent cohorts (combined, 2803 cases and 5642 controls). We identified a significant association between high myopia and a variant at 13q12.12 (rs9318086, combined p = 1.91 × 10(-16), heterozygous odds ratio = 1.32, and homozygous odds ratio = 1.64). Furthermore, five additional SNPs (rs9510902, rs3794338, rs1886970, rs7325450, and rs7331047) in the same linkage disequilibrium (LD) block with rs9318086 also proved to be significantly associated with high myopia in the Han Chinese population; p values ranged from 5.46 × 10(-11) to 6.16 × 10(-16). This associated locus contains three genes-MIPEP, C1QTNF9B-AS1, and C1QTNF9B. MIPEP and C1QTNF9B were found to be expressed in the retina and retinal pigment epithelium (RPE) and are more likely than C1QTNF9B-AS1 to be associated with high myopia given the evidence of retinal signaling that controls eye growth. Our results suggest that the variants at 13q12.12 are associated with high myopia.

The effect of cisatracurium and rocuronium on lung function in anesthetized children.

BACKGROUND: Neuromuscular blocking drugs have been implicated in intraoperative bronchoconstrictive episodes. We examined the effects of clinically relevant doses of cisatracurium and rocuronium on the lung mechanics of pediatric subjects. We hypothesized that cisatracurium and rocuronium would have bronchoconstrictive effects. METHODS: We studied ASA physical status I and II pediatric subjects having elective dental or urological procedures, requiring general anesthesia with endotracheal intubations with either cisatracurium or rocuronium. Pulmonary function tests were performed before and after neuromuscular blocking drug dosing and again after albuterol administration. Using forced deflation and passive deflation techniques, forced vital capacity (FVC) and maximum expiratory flow rate at 10% (MEF10) of FVC were obtained. Fractional changes from the baseline were used to compare subjects. Changes in MEF10 of >30% were considered clinically significant. A Shapiro-Wilk test, paired t test, and Wilcoxon rank sum test were used to analyze the data. RESULTS: Twenty-five subjects (median age = 5.25 years; range = 9 months-9.9 years) were studied; 12 subjects received cisatracurium and 13 subjects received rocuronium. Data are shown as mean proportional change ± SD or, in the case of not normally distributed, median proportional change (first, third quartile) with P values. In the cisatracurium group, there were no differences between baseline and postneuromuscular blocker administration in the fractional change from the baselines of FVC (1.00 ± 0.04, P = 0.5), but there was a significant decrease in MEF10 (0.80 ± 0.18, P = 0.002). In the rocuronium group, there were small yet significant decreases of FVC (0.99 [first quartile 0.97, third quartile 1], P = 0.02) and significant decreases in MEF10 (0.78 ± 0.26, P = 0.008). After administration of albuterol in the cisatracurium group, FVC increased slightly but significantly from baseline values (1.02 ± 0.02, P = 0.005). MEF10 increased significantly beyond baseline values (1.24 ± 0.43, P =0.04). In the rocuronium group, there were also significant differences between baseline and postalbuterol administration from the baseline value of FVC (1.02 ± 0.02, P = 0.004) and MEF10 (1.23 ± 0.29, P = 0.01). CONCLUSIONS: At clinically relevant doses, both cisatracurium and rocuronium caused changes in lung function, indicating constriction of smaller airways. In general, these changes were mild and not clinically detectable. However, in the rocuronium group, 3 of 13 patients showed more noticeable decreases in MEF10 (≤50%), demonstrating the potential for significant broncho-bronchiolar constriction in susceptible patients.

Effects of DPTQ, a novel positive allosteric modulator of the dopamine D1 receptor, on spontaneous eye blink rate and spatial working memory in the nonhuman primate.

RATIONALE: Dopamine (DA) signaling through the D1 receptor has been shown to be integral to multiple aspects of cognition, including the core process of working memory. The discovery of positive allosteric modulators (PAMs) of the D1 receptor has enabled treatment modalities that may have alternative benefits to orthosteric D1 agonists arising from a synergism of action with functional D1 receptor signaling. OBJECTIVES: To investigate this potential, we have studied the effects of the novel D1 PAM DPTQ on a spatial delayed response working memory task in the rhesus monkey. Initial studies indicated that DPTQ binds to primate D1R with high affinity and selectivity and elevates spontaneous eye blink rate in rhesus monkeys in a dose-dependent manner consistent with plasma ligand exposures and central D1activation. RESULTS: Based on those results, DPTQ was tested at 2.5 mg/kg IM in the working memory task. No acute effect was observed 1 h after dosing, but performance was impaired 48 h later. Remarkably, this deficit was immediately followed by a significant enhancement in cognition over the next 3 days. In a second experiment in which DPTQ was administered on days 1 and 5, the early impairment was smaller and did not reach statistical significance, but statistically significant enhancement of performance was observed over the following week. Lower doses of 0.1 and 1.0 mg/kg were also capable of producing this protracted enhancement without inducing any transient impairment. CONCLUSIONS: DPTQ exemplifies a class of D1PAMs that may be capable of providing long-term improvements in working memory.

Design and synthesis of procollagen C-proteinase inhibitors.

Non-peptidic inhibitors of procollagen C-proteinase (PCP) were designed from substrate leads. Compounds were optimized for potency and selectivity, with N-substituted aryl sulfonamide hydroxamates having the best combination of these properties. Compounds 89 and 60 have IC(50) values of 10 and 80 nM, respectively, against PCP; excellent selectivity over MMP's 1, 2, and 9; and activity in cell-based collagen deposition assays.

Simultaneous thermoosmotic and thermoelectric responses in nanoconfined electrolyte solutions: Effects of nanopore structures and membrane properties.

HYPOTHESIS: Nanofluidic systems provide an emerging and efficient platform for thermoelectric conversion and fluid pumping with low-grade heat energy. As a basis of their performance enhancement, the effects of the structures and properties of the nanofluidic systems on the thermoelectric response (TER) and the thermoosmotic response (TOR) are yet to be explored. METHODS: The simultaneous TER and TOR of electrolyte solutions in nanofluidic membrane pores on which an axial temperature gradient is exerted are investigated numerically and semi-analytically. A semi-analytical model is developed with the consideration of finite membrane thermal conductivity and the reservoir/entrance effect. FINDINGS: The increase in the access resistance due to the nanopore-reservoir interfaces accounts for the decrease of short circuit current at the low concentration regime. The decrease in the thermal conductivity ratio can enhance the TER and TOR. The maximum power density occurring at the nanopore radius twice the Debye length ranges from several to dozens of mW K-2 m-2 and is an order of magnitude higher than typical thermo-supercapacitors. The surface charge polarity can heavily affect the sign and magnitude of the short-circuit current, the Seebeck coefficient and the open-circuit thermoosmotic coefficient, but has less effect on the short-circuit thermoosmotic coefficient. Furthermore, the membrane thickness makes different impacts on TER and TOR for zero and finite membrane thermal conductivity.

Sexual function during long-term duloxetine treatment in patients with recurrent major depressive disorder.

INTRODUCTION: Sexual dysfunction (SD) is frequently associated with major depressive disorder (MDD) in the untreated state and may be worsened by antidepressant treatment. AIM: We evaluated SD in duloxetine-treated patients during an MDD recurrence prevention study. METHOD: Patients (N = 514) received open-label duloxetine 60-120 mg/day for up to 34 weeks. Responders (N = 288) were randomly assigned to duloxetine or placebo during a further 52-week double-blind maintenance phase. MAIN OUTCOME MEASURES: The Arizona Sexual Experience Scale (ASEX) was used to assess sexual functioning. RESULTS: At study entry, 73.4% of patients met ASEX criteria for SD. After open-label duloxetine treatment, the probability of continued SD was 77.9% for nonresponders and 53.2% for responders. In patients without SD at study entry, the probability of emergent SD was 49.6% (nonresponders) and 33.2% (responders). In the double-blind maintenance phase, there was no significant difference (P = 0.105) in the probability of emergent SD between placebo-treated (49.2%) and duloxetine-treated (27.9%) patients without SD at baseline, with no significant treatment-by-gender interaction. In patients with a recurrence of MDD, the probability of emergent SD was similar between placebo- (71.3%) and duloxetine-treated (82.7%) patients. However, in patients with no recurrence of MDD, the probability of emergent SD in placebo patients (40.0%) was numerically higher than in duloxetine patients (12.9%). Spontaneous reports of adverse events related to sexual function were infrequent and no patients discontinued due to these events. CONCLUSIONS: In patients with MDD, the probability of continued or emergent SD after up to 34 weeks of open-label duloxetine treatment was associated with the response status of the patients. In patients who responded to duloxetine treatment, after up to a further 52 weeks of double-blind treatment either with duloxetine or placebo, the probability of continued or emergent SD appeared to be more related to MDD itself than the treatments that the patients received.

Ultrasound-guided bilateral paravertebral continuous nerve blocks for a mildly coagulopathic patient undergoing exploratory laparotomy for bowel resection.

Regional anesthesia techniques commonly utilized in post-operative pain management are often considered contraindicated in coagulopathic patients. We report on successful postoperative pain control utilizing peripheral nerve blockade after exploratory laparotomy with small bowel resection in a mildly coagulopathic patient. In our case, complicated by abnormal PT, PTT and INR, a thromboelastogram (TEG) was performed before the procedure and found to be normal. An ultrasound-guided bilateral paravertebral blockade with continuous paravertebral catheters was then performed in this pediatric patient without complications. The patient expressed satisfaction with his pain control. More studies are needed to evaluate the validity of TEG in the prediction of bleeding risk and the safety of this regional technique in a mildly coagulopathic patients.

Targeted mass spectrometry quantification of total soy protein residues from commercially processed ingredients for food allergen management.

Soybean is one of the most commonly allergenic foods in the U.S. However, the variety of commercial soy ingredients used in the food industry makes soybean a challenging allergen to detect and quantify. The processing methods used to produce soy-derived ingredients result in protein modifications that often substantially impact detection and quantification with commonly used antibody-based methods. This study aimed to develop a mass spectrometry (MS)-based method capable of quantifying commercially processed soy ingredients in food matrices. A quantification strategy using external standards with internal calibrants was developed and evaluated, resulting in the ultimate use of a matrix-independent standard curve of non-roasted soy flour with milk proteins as carrier proteins. The method performance was evaluated by quantifying six soy-derived ingredients in incurred food matrices using three quantification strategies. Out of the twelve ingredient-matrix combinations with 10 ppm incurred total soy protein, eight had maximum recoveries between 60 and 120% using the full standard curve strategy. Other quantification strategies may be useful for internal quality control and interlaboratory calibrations. Compared with three commercial ELISA kits, the MS method showed a substantial advantage in quantifying the highly processed soy proteins in food matrices. SIGNIFICANCE: The ability to quantify undeclared soy protein in food products regardless of the soy ingredient source is essential for food allergen management, risk assessment, and regulatory enforcement. The MS-based method described here is able to reliably quantify six different soy-derived ingredients incurred in a model processed food. When compared with existing commercial ELISA methods, the MS method is much less affected by matrices and ingredient types, indicating its wider applicability to a range of soy-derived ingredients and processed products.

Integrating additional factors into the TNM staging for cutaneous melanoma by machine learning.

BACKGROUND: Integrating additional factors into the TNM staging system is needed for more accurate risk classification and survival prediction for patients with cutaneous melanoma. In the present study, we introduce machine learning as a novel tool that incorporates additional prognostic factors to improve the current TNM staging system. METHODS AND FINDINGS: Cancer-specific survival data for cutaneous melanoma with at least a 5 years follow-up were extracted from the Surveillance, Epidemiology, and End Results Program of the National Cancer Institute and split into the training set (40,781 cases) and validation set (5,390 cases). Five factors were studied: the primary tumor (T), regional lymph nodes (N), distant metastasis (M), age (A), and sex (S). The Ensemble Algorithm for Clustering Cancer Data (EACCD) was applied to the training set to generate prognostic groups. Utilizing only T, N, and M, a basic prognostic system was built where patients were stratified into 10 prognostic groups with well-separated survival curves, similar to 10 AJCC stages. These 10 groups had a significantly higher accuracy in survival prediction than 10 stages (C-index = 0.7682 vs 0.7643; increase in C-index = 0.0039, 95% CI = (0.0032, 0.0047); p-value = 7.2×10-23). Nevertheless, a positive association remained between the EACCD grouping and the AJCC staging (Spearman's rank correlation coefficient = 0.8316; p-value = 4.5×10-13). With additional information from A and S, a more advanced prognostic system was established using the training data that stratified patients into 10 groups and further improved the prediction accuracy (C-index = 0.7865 vs 0.7643; increase in C-index = 0.0222, 95% CI = (0.0191, 0.0254); p-value = 8.8×10-43). Both internal validation using the training set and temporal validation using the validation set showed good stratification and a high predictive accuracy of the prognostic systems. CONCLUSIONS: The EACCD allows additional factors to be integrated into the TNM to create a prognostic system that improves patient stratification and survival prediction for cutaneous melanoma. This integration separates favorable from unfavorable clinical outcomes for patients and improves both cohort selection for clinical trials and treatment management.

Current perspectives of the roles of the central norepinephrine system in anxiety and depression.

Norepinephrine (NE) is a major monoamine neurotransmitter that has widespread effects across multiple brain areas to regulate arousal and stress responses. The underlying function of the NE cortical system is to balance vigilance/scanning behavior with focused attention on novel environmental stimuli and the state of arousal. The central NE system is involved intrinsically with the stress response system, and dysregulation within the NE system has been implicated in the pathogenesis of anxiety and depressive disorders. Central NE activity paradoxically has either anxiogenic or anxiolytic effects, depending on whether the time course of the stress is acute or chronic, whether the stress is predictable or unpredictable, and which underlying brain regions are affected. Under conditions of chronic stress, NE system activity dysregulation of the hypothalamic-pituitary-adrenal system may turn a homeostatic stress response into a pathological stress response. Data suggest that the NE interplay with the serotonin system may exert neurobiological normalization of the pathophysiological state of anxious depression. Accordingly, pharmacological interventions targeting the NE system can result in anxiolytic, rather than anxiogenic, outcomes when used to treat patients with anxiety and depression.