2-day Protocol Research Articles

Effects of two carbonic anhydrase inhibitors on exercise performance in acute hypoxia.

Acute mountain sickness occurs due to rapid altitude ascents and/or insufficient acclimatization. Acetazolamide (AZ) is commonly prescribed for AMS prophylaxis but inhibits exercise performance. Methazolamide (MZ), an analogous drug, has similar prophylactic benefits but does not impair isolated muscle mass exercise performance in normoxia. We sought to compare whole-body exercise performance in acute hypoxia (FIO2 = 0.15) between AZ, MZ and placebo (PLA). Fifteen healthy participants completed 5 testing visits: day 1 maximal exercise test, day 2 a familiarization, and days 3-5 were the experimental visits. Each experimental visit involved a 5-km hypoxic cycling time trial performed after a 2-day dosing protocol of either AZ (250 mg t.i.d.), MZ (100 mg b.i.d.) or PLA (t.i.d.); the order was randomized and double-blinded. Prior to exercise, capillary blood samples were taken, and maximal voluntary contractions of quadriceps were performed. AZ and MZ resulted in a partially compensated metabolic acidosis at rest compared to PLA (capillary H+ 47±3, 43±2, 39±2 nmol for AZ, MZ and PLA respectively, p<0.01). Time to complete 5-km with PLA (562±32 seconds, p<0.01) was significantly faster than AZ and MZ (577±38 vs. 581±37s respectively), with no differences between AZ and MZ (p=0.96). There were no differences in average ventilation (124±27, 127±24, 127±19 l/min) and oxyhemoglobin saturation (87±2, 88±2, 88±3%) between AZ, MZ and PLA respectively (p>0.05). Overall, both AZ and MZ impair whole-body exercise performance in acute normobaric hypoxia.

Feeding protocol with live feed and inert diet to determine the ideal age/size for weaning mullet Mugil liza (Valenciennes, 1836).

The Lebranche mullet Mugil liza is a marine fish of great importance for artisanal and industrial fishing, as well as aquaculture. The use of live feeds during the larviculture phase of marine fish is a significant component of production costs. The present study evaluated the effects of the feeding transition on different larval stages of M. liza, from the combined supply of live feed (Brachionus rotundiformis + Artemia franciscana) and live + inert feed (Artemia franciscana + inert diet) until the weaning phase to only the inert diet. A total of 3240 M. liza larvae (weight 0.0133 ± 0.0062 g and length 0.793 ± 0.160 cm) were distributed among the 12 experimental units (n = 270), resulting in four groups with three replicates each. Treatment groups consisted of feed transition with A. franciscana (enriched metanauplii) to commercial inert feed starting weaning at four different larval ages: 28, 31, 34, and 37 days post hatching. Zootechnical performance indexes and intestinal histomorphometry were evaluated. Mortality, condition factor, and length variation coefficient did not show significant differences between treatments. Final weight, final length, weight gain, and length gain were significantly greater in larvae that started weaning at 31, 34, and 37 days post hatching. Weight coefficient of variation was significantly higher in larvae that started weaning at 28 days (67.51 ± 11.70) compared to 37 days (34.40 ± 7.30). In intestinal histology, villi height (180.3 ± 4.4) was significantly higher in larvae that started weaning at 37 days post hatching. Considering the evidence found in the present study, it is recommended to start weaning M. liza on the 31st day post-hatching, using a 2-day co-feeding protocol (31st and 32nd days). From the 33rd day after hatching, M. liza larvae can receive only commercial feed.

Exploring the impact of recent COVID-19 infection on perfusion and functional parameters derived from gated myocardial perfusion imaging in patients undergoing evaluation for coronary artery disease.

This study seeks to evaluate how recent COVID-19 infection affects myocardial perfusion and functional parameters derived from gated myocardial perfusion imaging in patients undergoing evaluation for coronary artery disease. The goal is to enhance our understanding of COVID-19's influence on the cardiovascular system. Conducted at Farshchian Heart Hospital from 2022 to 2023, this case-control study enrolled patients suspected of coronary artery disease, stratified into two groups: those with confirmed COVID-19 infection within the past 6months (study group) and those without prior COVID-19 infection (control group). Employing a 2-day protocol, stress testing and gated SPECT MPI were performed. Statistical analysis included descriptive statistics, Chi-square test, Student's t test, and Mann-Whitney U test. Among the 86 patients included, 43 were in each group. Significantly higher summed stress core and summed difference score values were observed in the study group compared to the control group (p < 0.05). In addition, the study group exhibited significantly altered global left ventricular ejection fraction, end-diastolic volume, and end-systolic volume (p < 0.05). Non-perfusion findings, including transient ischemic dilation and transient right ventricular visualization, were more prevalent in the study group. Recent COVID-19 infection is associated with impaired myocardial perfusion and altered functional parameters as detected by MPI. These findings underscore the intricate interplay between COVID-19 and cardiovascular health, emphasizing the importance of comprehensive evaluation and management strategies to address cardiac complications in affected individuals. Further research is warranted to elucidate the underlying mechanisms and optimize patient care in the context of COVID-19-associated cardiovascular manifestations.

Systemic immunological responses are dependent on sex and ovarian hormone presence following acute inhaled woodsmoke exposure

BackgroundRural regions of the western United States have experienced a noticeable surge in both the frequency and severity of acute wildfire events, which brings significant challenges to both public safety and environmental conservation efforts, with impacts felt globally. Identifying factors contributing to immune dysfunction, including endocrinological phenotypes, is essential to understanding how hormones may influence toxicological susceptibility.MethodsThis exploratory study utilized male and female C57BL/6 mice as in vivo models to investigate distinct responses to acute woodsmoke (WS) exposure with a focus on sex-based differences. In a second set of investigations, two groups were established within the female mouse cohort. In one group, mice experienced ovariectomy (OVX) to simulate an ovarian hormone-deficient state similar to surgical menopause, while the other group received Sham surgery as controls, to investigate the mechanistic role of ovarian hormone presence in driving immune dysregulation following acute WS exposure. Each experimental cohort followed a consecutive 2-day protocol with daily 4-h exposure intervals under two conditions: control HEPA-filtered air (FA) and acute WS to simulate an acute wildfire episode.ResultsMetals analysis of WS particulate matter (PM) revealed significantly increased levels of 63Cu, 182W, 208Pb, and 238U, compared to filtered air (FA) controls, providing insights into the specific metal components most impacted by the changing dynamics of wildfire occurrences in the region. Male and female mice exhibited diverse patterns in lung mRNA cytokine expression following WS exposure, with males showing downregulation and females displaying upregulation, notably for IL-1β, TNF-α, CXCL-1, CCL-5, TGF-β, and IL-6. After acute WS exposure, there were notable differences in the responses of macrophages, neutrophils, and bronchoalveolar lavage (BAL) cytokines IL-10, IL-6, IL-1β, and TNF-α. Significant diverse alterations were observed in BAL cytokines, specifically IL-1β, IL-10, IL-6, and TNF-α, as well as in the populations of immune cells, such as macrophages and polymorphonuclear leukocytes, in both Sham and OVX mice, following acute WS exposure. These findings elucidated the profound influence of hormonal changes on inflammatory outcomes, delineating substantial sex-related differences in immune activation and revealing altered immune responses in OVX mice due to ovarian hormone deficiency. In addition, the flow cytometry analysis highlighted the complex interaction between OVX surgery, acute WS exposure, and their collective impact on immune cell populations within the hematopoietic bone marrow niche.ConclusionsIn summary, both male and female mice, alongside females subjected to OVX and those who had sham surgery, exhibit significant variations in the expression of proinflammatory cytokines, chemokines, lung mRNA gene expression, and related functional networks linked to signaling pathways. These differences potentially act as mediators of sex-specific and hormonal influences in the systemic inflammatory response to acute WS exposure during a wildfire event. Understanding the regulatory roles of genes expressed differentially under environmental stressors holds considerable implications, aiding in identifying sex-specific therapeutic targets for addressing acute lung inflammation and injury.

Safety and feasibility of intravenous sotalol loading for the prevention of ventricular arrhythmias.

The use of intravenous (IV) sotalol loading following recent U.S. Food and Drug Administration (FDA) approval of a 1-day loading protocol has reduced the obligatory 3-day hospital stay for sotalol initiation when given orally. Several studies have recently demonstrated the safety and feasibility of IV loading for patients with atrial arrhythmias. However, there is a paucity of data on the feasibility and safety of IV sotalol loading for patients with ventricular arrhythmias. This study aims to assess the safety, feasibility, and length of stay (LOS) outcomes of IV sotalol loading for the prevention of ventricular arrhythmias. A retrospective analysis was performed of all patients undergoing IV sotalol loading and oral sotalol initiation for ventricular arrhythmias, or IV sotalol loading for atrial arrhythmias between August 2021 and December 2023 at Northwestern University. Baseline characteristics, success of sotalol initiation/loading, changes in heart rate (HR) and QT/QTc, safety, and LOS were compared between patients undergoing sotalol loading/initiation for ventricular arrhythmias (IV vs. PO) and between patients undergoing IV sotalol loading for ventricular arrhythmias vs. for atrial arrhythmias. A total of 28 patients underwent sotalol loading/initiation for ventricular arrhythmias (N = 15 IV and N = 13 PO) and 41 patients underwent IV sotalol loading for atrial arrhythmias. Baseline characteristics of congestive heart failure history and left ventricular ejection fraction were worse in the ventricular arrhythmias group. There was no significant difference in the successful completion of IV sotalol loading for ventricular arrhythmias compared to oral sotalol initiation for ventricular arrhythmias or IV sotalol loading for atrial arrhythmias (86.7% vs. 92.3% vs. 90.2%, p = 0.88). There was a significant increase in ΔQTc following IV sotalol infusion for ventricular arrhythmias compared to following PO sotalol initiation for ventricular arrhythmias (46.4 ± 29.2ms vs. 8.9 ± 32.6ms, p = 0.004) and following IV sotalol infusion for atrial arrhythmias (46.4 ± 29.2ms vs. 24.0 ± 25.1ms, p = 0.018). ΔHR following IV sotalol infusion for ventricular arrhythmias was similar to ΔHR following PO sotalol initiation for ventricular arrhythmias and ΔHR following IV sotalol infusion for atrial arrhythmias (- 7.5 ± 8.7bpm vs. - 8.5 ± 13.9bpm vs. - 8.3 ± 13.2bpm, p = 0.87). There were no significant differences in discontinuation for QTc prolongation (6.7% vs. 1.7% vs. 2.4%, p = 0.64) and bradycardia (13.3% vs. 7.7% vs. 9.8%, p = 0.88) between IV sotalol loading for ventricular arrhythmias, PO sotalol initiation for ventricular arrhythmias, and IV sotalol loading for atrial arrhythmias. There were no instances of hypotension, life-threatening ventricular arrhythmias, heart failure, or death. Length of stay was significantly shorter for IV sotalol loading compared to PO sotalol initiation for ventricular arrhythmias (1.1 ± 0.36days vs. 4.2 ± 1.0days, p < 0.0001). IV sotalol loading appears feasible and safe for use in ventricular arrhythmias and results in a decreased length of stay. Despite increased comorbidities and greater increase in QTc interval following IV sotalol infusion in the ventricular arrhythmias group, there were no significant differences in successful completion of loading or adverse outcomes when compared to PO sotalol initiation for ventricular arrhythmias and IV loading for atrial arrhythmias.

Clinical-Hematological Changes and Predictors of Severity in Acute Food Protein–Induced Enterocolitis Syndrome Reactions at Oral Food Challenge: A Multicenter Observational Study

BackgroundOral food challenge (OFC) is the gold standard for diagnosis of acute Food Protein-Induced Enterocolitis Syndrome (FPIES). No diagnostic/prognostic biomarkers are available, and OFC assessment criteria are not validated. Objective. To assess clinical-haematological changes and predictors of severity of FPIES reactions at OFC. MethodsObservational multicentre prospective study. Children aged 0-18 years diagnosed with acute FPIES were recruited at follow-up OFC in 12 tertiary centres in Spain and Italy. OFC Outcomes (as positive/negative/inconclusive and mild/moderate/severe) were assessed based on published ‘2017 FPIES Consensus’ criteria. Clinical characteristics were recorded, and full blood count was done at baseline, reaction onset and 4 hours later. Regression analysis was performed to assess predictors of severe reactions at OFC. Results81 children had positive OFC (mild in 11% (9/81), moderate in 61% (49/81), severe in 28% (23/81)). Increase in neutrophils and reduction in eosinophils, basophils and lymphocytes was observed (P-value<0.05). OFC was inconclusive in 19 cases despite objective signs or neutrophilia. Regression analysis showed a 2-day OFC protocol where only 25% of an age-appropriate portion is given on day 1 (not gender, age, culprit food, cumulative dose and previous reaction severity) was associated with reduced odds of severe reaction compared to giving multiple doses in a single day. ConclusionDistinct haematological changes may help support FPIES diagnosis. Current OFC assessment criteria may not capture the broad spectrum of acute FPIES presentations. This 2-day protocol may associate a reduced risk of severe reactions. Future work should aim to develop safer OFC and non-OFC diagnostics for FPIES.

Relationship between transient ischemic dilatation and changes in heart rate during gated SPECT acquisition in a low-risk population without perfusion defects.

Transient ischemic dilatation (TID) in myocardial perfusion single photon emission computed tomography (SPECT) is considered a marker of poor prognosis. However, it has been suggested that some cases are due to apparent volumetric changes secondary to differences in heart rate (HR) at the time of acquisition. We assessed the correlation between transient dilatation and HR in low risk patients with no perfusion defects. We retrospectively analyzed patients sent for 99mTc-MIBI SPECT using a 2-day protocol. We recorded the median HR during acquisition and the HR difference (HRD) between the rest and post-stress. We obtained the medium ventricular volume, end-diastolic volume (EDV), and end-systolic volume (ESV). We included patients in which TID using medium ventricular volume (TIDMV) was ≥1.2. TID was also calculated for the EDV and ESV (TIDEDV, TIDESV). We excluded patients with known coronary artery disease, perfusion defects, various ECG disorders, positive stress test, or ESV < 10 ml. From a total of 2006 patients, 63 (50 exercise, 13 dipyridamole) met the criteria for analysis (age 63.8 ± 9.7, 44 men). TIDMV was 1.29 ± 0.09 and HRD 9.8 beats per minute (BPM) (range -10 to 41). There was positive correlation between HRD and TIDMV ( r = 0.51, P < 0.001) and TIDEDV ( r = 0.5, P < 0.001), but not TIDESV ( r = 0.23, P = 0.07). Correlation was stronger when HRD was ≥10 BPM ( r = 0.67, P < 0.001). TID without perfusion defects should be interpreted with caution in the presence of HRD ≥ 10 BPM during post-stress acquisition.

Effect of exercise intensity and apnea on splenic contraction and hemoglobin increase in well-trained cross-country skiers

The human spleen acts as a reservoir for red blood cells, which is mobilized into the systemic circulation during various conditions such as hypoxia and physical exertion. Cross-country (XC) skiers, renowned for their exceptional aerobic capacity, are regularly exposed to high-intensity exercise and local oxygen deficits. We investigated a putative dose-dependent relationship between splenic contraction and concomitant hemoglobin concentration ([Hb]) elevation across four exercise intensities in well-trained XC skiers. Fourteen male XC skiers voluntarily participated in a 2-day protocol, encompassing a serial apnea test and a V˙\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$\\dot{V}$$\\end{document}O2max test (day 1), followed by three submaximal exercise intensities on a roller skiing treadmill corresponding to 55, 70, and 85% of V˙\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$\\dot{V}$$\\end{document}O2max (day 2). Spleen volume was measured via ultrasonic imaging, and venous blood samples were used to determine [Hb] levels. Baseline spleen volume was similar (266(35) mL) for all conditions (NS). Notably, all conditions induced significant splenic contractions and transient [Hb] elevations. The V˙\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$\\dot{V}$$\\end{document}O2max test exhibited the most pronounced splenic contraction (35.8%, p < 0.001) and a [Hb] increase of 8.1%, while the 85% exercise intensity led to 27.1% contraction and the greatest [Hb] increase (8.3%, < 0.001) compared to baseline. The apnea test induced relatively smaller responses (splenic contraction: 20.4%, [Hb] = 3.3%, p < 0.001), akin to the response observed at the 70% exercise intensity (splenic contraction = 23%, [Hb] = 6.4%, p < 0,001) and 55% (splenic contraction = 20.0%, [Hb] = 4.8%, p < 0.001). This study shows a discernible dose-dependent relationship between splenic contraction and [Hb] increase with levels of exercise, effectively distinguishing between submaximal and maximal exercise intensity.

One- versus 2-day aspirin desensitization in aspirin exacerbated respiratory disease: Aquality improvement project.

Current aspirin desensitization protocols for aspirin-exacerbated respiratory disease (AERD) require from 1 to 3 days to complete. Our aim was to assess the implementation of a 1-day versus 2-day aspirin desensitization protocol in patients with aspirin-exacerbated respiratory disease. We used a preintervention-postintervention quality improvement design to compare the completion rates, reaction rates, and estimated costs of a 2-day versus 1-day aspirin desensitization. The cost for each desensitization was estimated on the basis of 2017-2020 US Medicare standards. We included the predesensitization variables for FEV1 value, urinary leukotriene E4 level, absolute eosinophil count (AEC), and total IgE level for each group. A total of 15 patients underwent a 2-day aspirin desensitization in the 4-year (2017-2020) preintervention period and were compared with 8 patients who underwent a 1-day aspirin desensitization in the 1-year (2021) postintervention period. The desensitization completion rate (93% vs 100% [P= 1]) and the mean number of reactions requiring intervention during the desensitization protocols (0.26 vs 0.8 [P= .14]) were similar between groups. The average time frame between last polypectomy and desensitization was longer in the 2-day group (1946 vs 39.2 days [P= .03]). The mean values for FEV1 level, urinary leukotriene E4 level, absolute eosinophil count, and total IgE level were 76% vs 83% (P= .6), 1084 vs 385 pg/mg (P= .2), 686 vs 306 cells/μL (P= .74), and 735 vs 278 kU/L (P= .5), respectively. The estimated direct cost reduction was $762 per aspirin desensitization for using 1-day vs 2-day aspirin desensitization. Compared with a 2-day protocol, the implementation of a 1-day aspirin desensitization was characterized by similar completion and reaction rates as well as lower costs.

Safety of 2-day rush induction protocol in sublingual immunotherapy with Pru p 3

Aim: Allergy to lipid transfer proteins (LTPs) clinically manifests from oral allergy syndrome (OAS) to anaphylaxis. The risk of systemic symptoms and cross-reactivity make it an important target for allergen immunotherapy. Sublingual immunotherapy (SLIT) with Pru p 3 is effective and safe, but the induction phase (IP) of standard protocol (SP) is time consuming. Rush protocols (RPs) are described without serious adverse effects. The aim was to compare the safety of RP with SP and assess the existence of predictive factors for adverse reactions (ARs). Methods: Retrospective study of patients with LTP syndrome followed at the Food Allergy Unit undergoing SLIT with Pru p 3 between 2012 and 2021. SP has an IP of 4 days and an RP of 2 days. The safety of the IP was assessed by recording the AR. Results: Fifty-one patients: 41 (73.2% women) in SP group (SPG) and 10 (80% women) in RP group (RPG). Anaphylaxis as a presentation of LTP syndrome was overlapping in both groups (SPG 34.1%, RPG 33.3%). There were 5 (12.2%) ARs in SPG: 3 (60%) OAS, 1 (20%) oropharyngeal tightness, and 1 (20%) uvula edema; and 5 (50%) ARs in RPG: 4 (80%) OAS and 1 (20%) palmar pruritus and cough. All patients completed IP. Mean Pru p 3 specific immunoglobulin E (sIgE) value (kUA/L) of patients with ARs in IP: 6.7 kUA/L in SPG and 5.7 kUA/L in RPG. No group showed significant differences (P &gt; 0.05) between Pru p 3 sIgE value, presence of atopy or greater severity in LTP syndrome presentation, and greater probability of AR/more severe ARs in IP. Conclusions: ARs in IP were similar in both groups. No association was found between Pru p 3 value, atopy and higher probability of ARs in IP. RP appears to be a safe and less expensive option.

Machine learning-based diagnosis and risk classification of coronary artery disease using myocardial perfusion imaging SPECT: A radiomics study

This study aimed to investigate the diagnostic performance of machine learning-based radiomics analysis to diagnose coronary artery disease status and risk from rest/stress Myocardial Perfusion Imaging (MPI) single-photon emission computed tomography (SPECT). A total of 395 patients suspicious of coronary artery disease who underwent 2-day stress-rest protocol MPI SPECT were enrolled in this study. The left ventricle myocardium, excluding the cardiac cavity, was manually delineated on rest and stress images to define a volume of interest. Added to clinical features (age, sex, family history, diabetes status, smoking, and ejection fraction), a total of 118 radiomics features, were extracted from rest and stress MPI SPECT images to establish different feature sets, including Rest-, Stress-, Delta-, and Combined-radiomics (all together) feature sets. The data were randomly divided into 80% and 20% subsets for training and testing, respectively. The performance of classifiers built from combinations of three feature selections, and nine machine learning algorithms was evaluated for two different diagnostic tasks, including 1) normal/abnormal (no CAD vs. CAD) classification, and 2) low-risk/high-risk CAD classification. Different metrics, including the area under the ROC curve (AUC), accuracy (ACC), sensitivity (SEN), and specificity (SPE), were reported for models’ evaluation. Overall, models built on the Stress feature set (compared to other feature sets), and models to diagnose the second task (compared to task 1 models) revealed better performance. The Stress-mRMR-KNN (feature set-feature selection-classifier) reached the highest performance for task 1 with AUC, ACC, SEN, and SPE equal to 0.61, 0.63, 0.64, and 0.6, respectively. The Stress-Boruta-GB model achieved the highest performance for task 2 with AUC, ACC, SEN, and SPE of 0.79, 0.76, 0.75, and 0.76, respectively. Diabetes status from the clinical feature family, and dependence count non-uniformity normalized, from the NGLDM family, which is representative of non-uniformity in the region of interest were the most frequently selected features from stress feature set for CAD risk classification. This study revealed promising results for CAD risk classification using machine learning models built on MPI SPECT radiomics. The proposed models are helpful to alleviate the labor-intensive MPI SPECT interpretation process regarding CAD status and can potentially expedite the diagnostic process.

Eye-Tracking–Based Measurement of Social Visual Engagement Compared With Expert Clinical Diagnosis of Autism

In the US, children with signs of autism often experience more than 1 year of delay before diagnosis and often experience longer delays if they are from racially, ethnically, or economically disadvantaged backgrounds. Most diagnoses are also received without use of standardized diagnostic instruments. To aid in early autism diagnosis, eye-tracking measurement of social visual engagement has shown potential as a performance-based biomarker. To evaluate the performance of eye-tracking measurement of social visual engagement (index test) relative to expert clinical diagnosis in young children referred to specialty autism clinics. In this study of 16- to 30-month-old children enrolled at 6 US specialty centers from April 2018 through May 2019, staff blind to clinical diagnoses used automated devices to measure eye-tracking-based social visual engagement. Expert clinical diagnoses were made using best practice standardized protocols by specialists blind to index test results. This study was completed in a 1-day protocol for each participant. Primary outcome measures were test sensitivity and specificity relative to expert clinical diagnosis. Secondary outcome measures were test correlations with expert clinical assessments of social disability, verbal ability, and nonverbal cognitive ability. Eye-tracking measurement of social visual engagement was successful in 475 (95.2%) of the 499 enrolled children (mean [SD] age, 24.1 [4.4] months; 38 [8.0%] were Asian; 37 [7.8%], Black; 352 [74.1%], White; 44 [9.3%], other; and 68 [14.3%], Hispanic). By expert clinical diagnosis, 221 children (46.5%) had autism and 254 (53.5%) did not. In all children, measurement of social visual engagement had sensitivity of 71.0% (95% CI, 64.7% to 76.6%) and specificity of 80.7% (95% CI, 75.4% to 85.1%). In the subgroup of 335 children whose autism diagnosis was certain, sensitivity was 78.0% (95% CI, 70.7% to 83.9%) and specificity was 85.4% (95% CI, 79.5% to 89.8%). Eye-tracking test results correlated with expert clinical assessments of individual levels of social disability (r = -0.75 [95% CI, -0.79 to -0.71]), verbal ability (r = 0.65 [95% CI, 0.59 to 0.70]), and nonverbal cognitive ability (r = 0.65 [95% CI, 0.59 to 0.70]). In 16- to 30-month-old children referred to specialty clinics, eye-tracking-based measurement of social visual engagement was predictive of autism diagnoses by clinical experts. Further evaluation of this test's role in early diagnosis and assessment of autism in routine specialty clinic practice is warranted. ClinicalTrials.gov Identifier: NCT03469986.

Fluorescent In Situ Hybridization for miRNA Combined with Staining of Proteins.

The last decades have illustrated the importance of microRNAs (miRNAs) in various biological and pathological processes. The combined visualization of miRNAs using fluorescent in situ hybridization (FISH) and proteins using immunofluorescence (IF) can reveal their spatiotemporal distribution in relation to the cell and tissue morphology and can provide interesting insights into miRNA-protein interactions. However, standardized protocols for co-localization of miRNAs and proteins are currently lacking, and substantial technical obstacles still need to be addressed. In particular, the incompatibility of protein IF protocols with steps required for miRNA FISH, such as proteolytic pretreatments and ethylcarbodiimide post-fixation, as well as hurdles related to low signal intensity of low-copy miRNAs, remains challenging. Our technique may considerably enhance miRNA-based research, as current detection techniques lack the ability to elucidate cellular and subcellular localization. Here, we describe an optimized 2-day protocol for combined detection of low-abundant miRNAs and proteins in cryosections of cardiac tissue, without the need for protease-dependent pretreatment or post-fixation treatment. We successfully demonstrate endothelial-specific localization of low-abundant miR-181c-5p in cardiac tissue. © 2023 Wiley Periodicals LLC. Basic Protocol: Fluorescent in situ hybridization for miRNA combined with staining of proteins.

SPECT and PET myocardial perfusion imaging in Austria, Germany, and Switzerland results of the first joint survey of 2021

PurposeThis paper presents the results of the first joint survey on the use of SPECT and PET myocardial perfusion imaging (MPI) and cardiac amyloidosis imaging in Austria, Germany, and Switzerland of the year 2021.MethodsA questionnaire was sent in 2022 to centres practicing nuclear medicine.ResultsData from 14 Austrian (10,710 SPECT), 218 German (133,047 SPECT), and 16 Swiss centres (11,601 MPI (6,879 SPECT, 4722 PET)) were analysed. In Austria and Germany, the PET MPI numbers were close to zero and not considered. Official MPS numbers from 2015 to 2021 from Austria and Germany revealed a decline in Austria by about 40% in the pandemic years 2020 to 2021, but an increase in Germany by 9%. Ambulatory care cardiologists represented the major referral group (56–71%). Mostly, stress tests were performed pharmacologically (58–92%). Contrary to Germany, a 1-day protocol was predominant (58–97%) in Austria and Switzerland. The leading camera systems were SPECT-CT in Austria and Switzerland (57–79%) and multi-head systems in Germany (58%). Switzerland had the highest proportion of SPECT MPI with attenuation correction (84%), followed by Austria (43%), and Germany (33%). Electrocardiogram-gated SPECT MPI showed an overall high penetration of 87–99%. Scoring was most frequently applied in Germany (72%), followed by Austria (64%), and Switzerland (60%). Related to the population, the number of cardiac amyloidosis imaging was highest in Austria, followed by Switzerland and Germany.ConclusionsThis first joint survey of 2021 shows considerable differences among the countries. The Swiss situation is outstanding due to the wide use of PET MPI. In terms of camera equipment, Switzerland is also leading, followed by Austria and Germany. Despite the differences in procedural issues, the results reveal an overall high standard of MPI imaging.

Transient ischaemic dilation ratio thresholds in patients with zero coronary calcium score undergoing exercise or dipyridamole stress SPECT myocardial perfusion imaging using a cadmium-zinc-telluride camera.

Transient ischaemic dilation (TID) is a marker of underlying extensive coronary artery disease (CAD) during myocardial perfusion imaging (MPI). The cut-off for a normal TID ratio (TIDr) value is often derived from a cohort of individuals with no apparent CAD. Varying criteria have been used to define the absence of CAD. We aim to derive TIDr cut-offs using patients with normal MPI and coronary artery calcium (CAC) score of zero, and compare the TIDr obtained from different software packages. We studied 232 patients with zero CAC and normal MPI undergoing exercise or dipyridamole stress using either a 1- or 2-day protocol. All patients were scanned in the supine position with a cadmium-zinc-telluride camera. TIDr was automatically generated using quantitative perfusion SPECT (QPS) software initially, and subsequently using Myometrix for comparison. The TIDr cut-offs calculated using the mean + 2 standard deviation were 1.29 and 1.24 for the 1- and 2-day protocol groups, respectively. In patients undergoing a 2-day protocol, dipyridamole stress resulted in significantly higher mean TIDr when compared to exercise stress (1.07 ± 0.13 vs. 1.01 ± 0.12, P = 0.035). Myometrix-derived TIDr were also significantly lower compared to QPS-derived values for most protocols except for 2-day exercise stress. This study is the first to derive TIDr threshold values using a normal population defined by zero CAC and normal MPI. TIDr was found to vary depending on stress modality, protocol as well as the software used.

SENODAY: A New Perspective of Handling Breast Cancer

BackgroundTo evaluate the feasibility of a same day breast cancer diagnosis and management protocol, consequently decreasing time to treatment and immediately reassuring patients with benign diagnosis. Materials and MethodsA total of 60 women underwent breast exam during SENODAY in our cancer center between January 2020 and December 2022. Patients are first seen by a breast surgeon who mentions whether the patient's history and physical exam are suspicious of malignancy. Patients are then sent to the radiologist who performs a complete radiologic assessment, classifies the lesions, and performs a biopsy when necessary. The specimen is sent to the pathologist who uses the imprint cytology technique to obtain a preliminary diagnosis. Effective counseling is established in case of breast cancer diagnosis. ResultsAmong 60 women, 25 patients were reassured by breast imaging and 35 underwent histopathological analysis (17 patients with a 1-day protocol and 18 with the standard definitive technique). Clinical examination was found to have a sensitivity of 100% and a specificity of 89.47%. The positive predictive value was 80 % and the negative predictive value was 100%. However, we did not find a strong correlation between imaging and definitive pathology. Moreover, on imprint cytology, sensitivity, specificity, PPV, and NPV were 100%. Finally, the mean time-to-treat was 28.6 days. ConclusionsSENODAY reassured 68.3% of patients. It also offered effective counseling and a treatment plan to newly diagnosed breast cancer patients within 1 day. Same day histological diagnosis by imprint cytology is effective and feasible with an excellent accuracy.

Evaluating the effects of counterconditioning, novelty-facilitated, and standard extinction on the spontaneous recovery of threat expectancy and conditioned stimulus valence.

Extinction training has proved effective to diminish the expectancy of the aversive unconditioned stimulus (US). However, the negative valence of the conditioned stimulus (CS) may still stay intact. In fact, several studies have suggested that the CS negative valence may be a factor that promotes the return of fear. Our study focuses on the role of changes in the CS valence as a potential mechanism to reduce the spontaneous recovery of threat expectancies. To do that, we evaluated counterconditioning (CC), a technique aimed to reduce the CS negative valence by paring it with a positive stimulus and compared its efficacy to that of a novelty-facilitated extinction (NFE) and a standard extinction interventions. Using a 2-day protocol, participants first learned the relationship between a figure and an aversive sound, using a differential conditioning paradigm, and were then randomly assigned to one of three different groups. For the CC group, CS+ or cue A was paired with a positive US. The standard extinction group was exposed to cue A alone. For a third NFE group, cue A was followed by a neutral US. Finally, on the second day, spontaneous recovery was tested. Our findings did not provide evidence to suggest that CC could be more effective to prevent or reduce the return of threat expectancies or influence valence ratings when compared with NFE and standard extinction.

Myocardial perfusion SPECT in Germany from 2012 to 2021: insights into development and quality indicators

PurposeThis paper summarises the results of 4 national surveys on the numbers, utilisation and technique of myocardial perfusion SPECT (MPS) from 2012 to 2021.MethodsA one-page questionnaire for information on MPS in 2012, 2015, 2018 and 2021 was sent to German centres practising nuclear medicine. To check for representativeness, the numbers obtained were related to official annual data and furthermore to the numbers of invasive coronary angiography procedures (ICA).ResultsMPS examinations increased by > 40% from 2012 to 2021 and showed a centralisation with increasing MPS per centre. In 2020, a mild impact of the COVID-19 pandemic could be observed in the form of only a slight MPS increase, which was compensated in the following year. Outpatient care cardiologists represent the most important referrer (70%). Mostly, 2-day protocols were used. One-day protocols and stress-only protocols showed insignificant changes. The use of exercise stress decreased steadily. In 2021, exercise stress was replaced by pharmacological stress as the most frequent stress modality. Camera systems showed a shift to more SPECT-CT systems. The use of gated SPECT increased to almost 90%. Quantitative scoring showed an increasing acceptance. The ratio of invasive coronary angiographies (ICA) to MPS was between 3.9 and 4.5. A significant proportion of ICA in the context of CCS (chronic coronary syndrome) was performed without prior testing for ischaemia.ConclusionThe 2012 to 2021 MPS surveys reveal a continuously growing number of examinations with only a mild temporary effect of the COVID-19 pandemic and a centralisation with increasing numbers per centre. Performance and technical data reveal a high-grade adherence of MPS practice to the current ESC guideline. A large potential of non-invasive diagnostics remains for the future.

Effect of 2 Day vs. 3 Day Superstimulation Protocol on In Vivo Maturation of OPU derived oocytes in Buffaloes

The study was planned to evaluate effect of two different superstimulation protocols (2 Day vs. 3 Day) on in vivo maturation of oocytes in buffaloes. Buffaloes (n=8) were superstimulated twice using 2x2 design at an interval of 30-35 days using 200 mg and 300 mg FSH divided into four and six tapering doses (each dose of 50 mg) given over 2 and 3 days at 12 hourly intervals. Prostaglandin F2α analogue (500 μg Cloprostenol; i/m) at 72 and 84 hrs after start of superstimulatory treatment. Oocytes were retrieved from &gt; 6 mm follicles at 48 h after the last FSH injection by ovum pick up. Oocyte maturation was assessed by the degree of cumulus cell expansion and by Orcein staining of denuded oocytes. OPU of 60 and 91 follicles resulted in recovery of 38 and 50 cumulus oocyte complexes (COCs) in 2-Day and 3-Day group, respectively. Significantly higher proportion of expanded oocytes was recovered in 3-Day protocol compared to 2-Day protocol (36.0% vs. 13.2%) and higher number of compact COCs were recovered in 2-Day protocol. In 3-Day protocol, significantly higher percentage of oocytes retrieved was at Metaphase-II stage compared to 2-Day protocol (40% vs.15.8%, respectively). The preliminary trial demonstrated that 3-day superstimulatory protocol yielded higher percentage of in vivo matured oocytes than 2-Day protocol in buffaloes.

Improved detection of coronary artery disease by CZT regional coronary blood flow evaluation.

CZT cameras have enabled the noninvasive quantification of myocardial flow reserve (MFR), an important physiologic measure. This study aimed to compare myocardial perfusion SPECT (MPS) with or without MFR evaluation for the detection of obstructive coronary artery disease (CAD). 48 patients with CAD (>50% obstruction) detected at invasive coronary angiography or CT angiography underwent dipyridamole MPS and MFR evaluation within 30 days. A 1-day protocol (rest-stress) was used to quantify MFR. The acquisition of dynamic rest and stress images was initiated simultaneously to 99mTc sestamibi injection (370 and 1,110 MBq, respectively), both lasting for 11 min, followed by 5-min imaging. Pharmacologic stress with dipyridamole (0.56 mg/kg for 4 min) was performed with the patient positioned in the CZT camera. The images were processed and time-activity curves were generated, calculating global and regional MFR in a semiautomatic software. A global or regional MFR <2 was considered abnormal. MPS perfusion images were classified as normal or abnormal. The images were interpreted by experienced physicians blinded to the results of MFR and coronary angiography/CT. Mean age of the population was 61 ± 9 years, 54.2% female. Twenty patients (41.7%) had single-vessel CAD, 22 (45.8%) 2-vessel CAD and 6 (12.5%), triple-vessel CAD. Among the 82 vessels with obstruction, 48 had perfusion abnormalities in MPS and 60 had reduced MFR, while among the normal vessels, had 54 normal MPS and 52 had preserved MFR. The sensitivity of MFR (69%) was higher than that of MPS (55.2%), without significant changes in specificity (86 vs. 83.7%). MFR in the CZT camera is more sensitive for the detection of CAD than perfusion abnormalities in MPS, especially in patients with multivessel CAD.