Entire Protocol Research Articles

Testing the limits of SPDM: Authentication of intermittently connected devices

The Security Protocol and Data Model (SPDM) is an open standard for authentication, attestation, and key exchange among hardware units, such as CPUs and peripheral components. In principle, SPDM was designed to operate over a somewhat stable communication channel, meaning that connection losses usually require the re-execution of the entire protocol. This puts into question SPDM’s suitability for battery-powered devices, which may keep only intermittent communications aiming to save energy. To address this question, we evaluate different authentication approaches that build upon and extend SPDM’s native key bootstrapping capabilities to handle intermittent authentication. In particular, we show that the combination of SPDM and a Time-based One-Time Password (TOTP) protocol is a promising solution for this scenario. We analyze the performance of the proposed authentication schemes using a proof-of-concept virtual device. The TOTP-based scheme was shown to be the fastest, the reconnection step being at least twice and up to 900× faster than possible straightforward applications of SPDM. Also, our scheme requires less memory to operate. Finally, we discuss the possibility of integrating intermittent authentication capabilities into the SPDM standard itself.

Beyond bedwetting: How successful treatment is observed in sleep macrostructure

IntroductionSleep enuresis (SE), commonly known as bedwetting, refers to involuntary urination during sleep. It is a prevalent condition affecting approximately 15 % of children at age 5, 10 % by age 7, and 5 % by age 10. This condition can significantly impact both children and their families. The pathophysiology of SE is complex and not yet fully understood. There are several established treatment methods, but limited information on their sleep dynamics. ObjectiveThis study aimed to evaluate differences in sleep structure before and after treatment in patients with monosymptomatic SE (MSE), focusing on alarm therapy, desmopressin, and a combination of both. The analysis compared pre- and post-treatment differences within each treatment arm. The analysis was conducted for both successful and unsuccessful treatment outcomes. Methods/resultsThis was a prospective study with MSE patients, aged 6–16 years, diagnosed by multidisciplinary assessment. Of the 140 initial applicants 75 were initially included in the study and randomized for therapeutic intervention in three treatment arms, namely: alarm, desmopressin and alarm + desmopressin. Therapeutic response was evaluated 12 months after treatment discontinuation. Polysomnographic evaluation pre and post treatment was carried out. 51/75 completed the entire protocol. 42/51 were successfully treated and had a median age of 10 [8–12] and the non-success, 8 [7–10]. Among the successful patients, the percentage of N2 sleep decreased from a median of 55.7 %–48.5 % (p = 0.0004) in the alarm arm, from 58.8 % to 50 % (p = 0.002) in the desmopressin arm, and from 54.7 % to 50.9 % (p = 0.044) in the combined arm. The percentage of N3 sleep increased from 25.7 % to 30.1 % (p = 0.004) in the alarm arm, from 21.6 % to 26 % (p = 0.032) in the desmopressin arm, and from 23.7 % to 28.3 % (p = 0.014) in the combined arm. The arousal index significantly increased from pre-to post-treatment in all arms: in the alarm arm, from 1.25 to 2.8 (p = 0.002); in the desmopressin arm, from 1.3 to 2.7 (p = 0.019); and in the combined treatment arm, from 1 to 4 (p = 0.003). No significant differences were observed in the non-successful arm or among those who experienced complete resolution of the enuresis without treatment. ConclusionThe observed increase in N3 sleep and arousal and decrease in N2 sleep following successful treatment, regardless of the specific interventions, underscores the role of sleep in the pathophysiology of enuresis. Conversely, the lack of sleep differences in the non-successful arm further highlights the importance of sleep, beyond developmental factors, in influencing the clinical outcomes of enuresis, especially since all children were assessed 12 months after the start of treatment.

Effect of cold beverages on whole-body heat exchange in young and older males during intermittent exercise in the heat.

To mitigate health risks associated with occupational heat stress, workers are advised to adhere to a work-rest regimen, and hydrate regularly. However, it remains unclear if beverage temperature influences whole-body heat exchange during work-rest cycles, and if responses differ in older workers who have a blunted heat loss capacity. Ten young (mean [SD]: 22 [3] years) and 10 older (60 [4] years) males performed four 15-min bouts of moderate-intensity cycling at a fixed rate of metabolic heat production (200 W·m-2), each interspersed by 15-min rest in dry heat (40°C, ~12% relative humidity). On separate days, participants consumed either ice-slurry (~0°C), standardized to provide a heat transfer capacity of 75 kJ·m-2, or an identical mass of warm fluid (37.5°C) before the first and third exercise bouts. Evaporative and dry heat exchange (direct calorimetry) and metabolic heat production (indirect calorimetry) were measured continuously to determine cumulative heat storage (summation of heat loss and heat gain) over the entire protocol. Rectal temperature was also measured continuously. Relative to warm fluid, ice-slurry ingestion reduced cumulative heat storage in young (69 [181] vs. 216 [94] kJ) and older males (90 [104] vs. 254 [140] kJ, main effect: p < 0.01), but was unaffected by age (p = 0.49). However, rectal temperature was unaffected by beverage temperature in both groups (all p ≥ 0.15). We show that cold fluid ingestion is an appropriate administrative control for both young and older males as it can mitigate increases in body heat content during moderate-intensity work-rest cycles in dry heat.

Interfacial property determination from dynamic pendant-drop characterizations.

The properties of the interface between materials have practical implications in various fields, encompassing capillary action, foam and emulsion stability, adhesion properties of materials and mass and heat transfer processes. Studying the dynamics of interfaces is also fundamental for understanding intermolecular interactions, change of molecular conformations and molecular aggregations. Pendant-drop tensiometry and its extension, the oscillating drop method, are simple, versatile methods used to measure surface tension, interfacial tension and interfacial rheological properties. These methods can, however, generate unreliable results because of inadequate material preparation, an incorrect calibration method, inappropriate selection of data for analysis, neglect of optical influences or operating the system outside the linear viscoelastic regime. In addition, many studies fail to report accurate uncertainties. This protocol addresses all these critical points and provides detailed descriptions of some operation tips relating to purifying methods for different kinds of material, the time frame for analyzing measurement data, the correction method for optical effects, implementation of the oscillating method with a common programmable pump and remedies for some common problems encountered during the measurement. Examples of interfacial tension measurements for two- and three-phase systems, as well as interfacial dilational modulus measurements for N2 and surfactant solutions, are provided to illustrate procedural details and results. A single measurement takes minutes to hours to complete, while the entire protocol, including the leak test, cleaning, repeated measurements and data analysis, may take several days.

AN AGAROSE GEL-BASED APPROACH FOR THE REMOVAL OF METALLIC-BASE INK STAIN FROM THE PAPER SURFACES OF A 17TH CENTURY BOOK

Removing ink stains from the pages of ancient and valuable books is a very challenging and sometimes impossible task. Only in some circumstances, restorers are able to improve the appearance of an ink-stained page by lightening the spots using an ink eradicator. This paper reports on an approach for the removal of ink stains from a 17th century book using a combination of two commercial houseware products, the rust remover for textile and bleach. The products were chosen on the basis of their active ingredients such as hydrofluoric acid (HF), which facilitates the removal of metal ions contained in the inks (such as iron and chromium) and sodium hypochlorite (NaClO), known for its whitening abilities. The products were first tested by direct application to one of the several stained pages as a control experiment. The ink stain was successfully removed from the book, but by SEM analysis the page results in a clear detachment of the cellulose fibers from the paper pulp, which reflect a significant change in the paper fine structure. Moreover, the mixing of chemicals occurring during this procedure generated toxic vapors and the entire protocol could only be performed under a chemical fume hood. A new procedure was developed to mitigate these negative effects. Each of the two commercial products was dispersed in a 2% agarose solution shortly before its gelification and the two gels thus obtained were applied consecutively on the surface of the paper to be cleaned. The results showed an effective removal of the ink stain from treated page, avoiding irreparable damage to the paper structure and improving the working conditions of the restorers. Hence, the proposed gel cleaning procedure offers three main advantages: i) a simple and practical tool for effective removal of metallic ink stains from the paper surface; ii) reduction of damage caused by aggressive reagents (HF and NaClO) contained in cleaning products for a better conservation of paper fibers; iii) a great decrease of the risk of accidentally exposing the operators to harmful toxic vapor (produced by accidental mixing of aforementioned chemical reagents).

Feasibility and Safety of a Home-based Electroencephalogram Neurofeedback Intervention to Reduce Chronic Neuropathic Pain: A Cohort Clinical Trial

ObjectiveTo evaluate the feasibility, safety, and potential health benefits of an 8-week home-based neurofeedback intervention. DesignSingle-group preliminary study. SettingCommunity-based. ParticipantsNine community dwelling adults with chronic neuropathic pain, 6 women and 3 men, with an average age of 51.9 years (range, 19-78 years) and with a 7-day average minimum pain score of 4 of 10 on the visual analog pain scale. InterventionsA minimum of 5 neurofeedback sessions per week (40min/session) for 8 consecutive weeks was undertaken with a 12-week follow-up baseline electroencephalography recording period. Main Outcome MeasuresPrimary feasibility outcomes: accessibility, tolerability, safety (adverse events and resolution), and human and information technology (IT) resources required. Secondary outcomes: pain, sensitization, catastrophization, anxiety, depression, sleep, health-related quality of life, electroencephalographic activity, and simple participant feedback. ResultsOf the 23 people screened, 11 were eligible for recruitment. One withdrew and another completed insufficient sessions for analysis, which resulted in 9 datasets analyzed. Three participants withdrew from the follow-up baselines, leaving 6 who completed the entire trial protocol. Thirteen adverse events were recorded and resolved: 1 was treatment-related, 4 were equipment-related, and 8 were administrative-related (eg, courier communication issues). The human and IT resources necessary for trial implementation were identified. There were also significant improvements in pain levels, depression, and anxiety. Six of 9 participants perceived minimal improvement or no change in symptoms after the trial, and 5 of 9 participants were satisfied with the treatment received. ConclusionsIt is feasible and safe to conduct a home-based trial of a neurofeedback intervention for people with chronic neuropathic pain, when the human and IT resources are provided and relevant governance processes are followed. Improvements in secondary outcomes merit investigation with a randomized controlled trial.

Randomized Comparison of Progression of Atherosclerotic Plaques and Calcification of Coronary Artery in Atrial Fibrillation Patients Treated With Edoxaban Versus Warfarin (The REPRESENT-AF trial)

Although the adverse effects of long-term use of vitamin K oral anticoagulant (OAC), warfarin, on the coronary vasculature are well-established, it remains unknown whether nonvitamin K oral anticoagulants play a role in the attenuation of plaque progression and coronary calcification. This study aimed to compare the changes in atherosclerotic plaques and calcification of the coronary arteries in patients with atrial fibrillation (AF) treated with edoxaban and warfarin. A total of 150 OAC-naïve patients with AF and atherosclerotic lesions on coronary computed tomography angiography (CCTA) were enrolled and randomly assigned to the edoxaban or warfarin treatment groups. All enrolled patients received rosuvastatin 10 mg and 119 patients completed the entire study protocol. A total of 12 months after the assigned OAC treatment, follow-up CCTA was performed and changes in plaque and calcium volumes of the coronary arteries were analyzed. The baseline characteristics of the 2 groups were well-balanced. The percentage of time in therapeutic range in the warfarin group was 61.1%. Compared with the baseline CCTA, there was a significant reduction in plaque volume after 12 months of OAC and rosuvastatin administration in both groups, and the extent of regression did not differ significantly between the groups. The increase in calcium volume was greater in the warfarin group than in the edoxaban group; however, the difference was not significant. In OAC-naïve patients with AF and atherosclerotic coronary lesions who were treated with moderate-intensity statin, edoxaban use did not have a positive effect on atherosclerotic plaques and coronary calcification compared with warfarin use over a 12-month follow-up period.

A novel methodology for the diffusion coefficient determination in porous media by X-ray high-resolution radiographies

The first step in diagnosing a patient with cancer is the biopsy and the study of the tissue sample taken, suitably processed, is essential for a correct diagnosis. Tissue processors are machines that automatically perform the entire processing protocol, often relying on empirical timing related to chemical diffusions in the biological samples. The determination of the diffusion coefficient in porous materials, such as histological tissue, is still under study with multiple methods often resulting in inaccurate predictions and with few amounts of literature data for biological samples. The authors present an experimental technique, based on a multitude of X-ray radiographs acquired over time, where the intensity of the attenuation to the ionising beam is proportional to the concentration of the solvent that diffuses in the histological preparation. The paper so describes a proof-of-concept of a novel method to estimate the diffusion coefficient in a histological sample, based on the acquisition of time-dependent X-ray radiographs and the solution of an inverse diffusion problem.

Cryopreserving the intact intervertebral disc without compromising viability.

Tissue cryopreservation requires saturation of the structure with cryoprotectants (CPAs) that are also toxic to cells within a short timeframe unless frozen. The race between CPA delivery and cell death is the main barrier to realizing transplantation banks that can indefinitely preserve tissues and organs. Unrealistic cost and urgency leaves less life-threatening ailments unable to capitalize on traditional organ transplantation systems that immediately match and transport unfrozen organs. For instance, human intervertebral discs (IVD) could be transplanted to treat back pain or used as ex vivo models for studying regenerative therapies, but both face logistical hurdles in organ acquisition and transport. Here we aimed to overcome those challenges by cryopreserving intact IVDs using compressive loading and swelling to accelerate CPA delivery. CPAs were tested on bovine nucleus pulposus cells to determine the least cytotoxic solution. Capitalizing on our CPAs Computed Tomography (CT) contrast enhancement, we imaged and quantified saturation time in intact bovine IVDs under different conditions in a bioreactor. Finally, the entire protocol was tested, including 1 week of frozen storage, to confirm tissue viability in multiple IVD regions after thawing. Results showed cryopreserving medium containing dimethyl sulfoxide and ethylene glycol gave over 7.5 h before cytotoxicity. While non-loaded IVDs required over 3 days to fully saturate, a dynamic loading protocol followed by CPA addition and free-swelling decreased saturation time to <5 h. After cryopreserving IVDs for 1 week with the optimized CPA and permeation method, all IVD regions had 85% cell viability, not significantly different from fresh unfrozen controls. This study created a novel solution to a roadblock in IVD research and development. Using post-compression swelling CPA can be delivered to an intact IVD over 20× more quickly than previous methods, enabling cryopreservation of the IVD with no detectable loss in cell viability.

High-resolution topographic surveying and change detection with the iPhone LiDAR.

This paper introduces a comprehensive protocol leveraging open-access techniques to create small- to medium-scale 3D representations of the environment by using iPhone and iPad light detection and ranging (LiDAR). The protocol focuses on two capabilities of the iPhone LiDAR. The first capability is 3D modeling: iPhone LiDAR rapidly generates detailed indoor and outdoor 3D models, providing insights into object size, volume and geometry. The second capability is change detection: the 3D models created by the LiDAR sensor can be used for precise measurement of changes over time. Compared to other 3D topographic surveying methods, this method is rapid, high resolution, low cost and easy to use. The protocol outlines iPhone LiDAR scanning practices, model export and change detection. The expected results after executing the protocol are (i) a detailed 3D model of a small- to medium-sized object or area of interest and (ii) a distance point cloud revealing change between two point clouds of the same object or area between different times. The entire protocol can be conducted within 2 h by anyone with an iPhone with the LiDAR sensor and a computer. This protocol empowers scientists, students and community members conducting research with a cheap, easy-to-use method for addressing a range of questions and challenges, thus benefiting experts and the broader community.

Feasibility of a Recumbent Stepper for Short-Interval, Low-Volume High-Intensity Interval Exercise in Stroke.

Studies investigating high-intensity interval exercise (HIIE) in stroke typically emphasize treadmill training. However, a literature review suggested that seated devices such as a recumbent stepper or cycle offer a promising alternative for HIIE since exercise can be prescribed using peak power output (PPO). Therefore, this would give health care professionals the ability to monitor and adapt power output for the target heart rate range. The purpose of this secondary analysis was to examine the feasibility of prescribing short-interval, low-volume HIIE using PPO in chronic stroke. We used several methods to test feasibility: (1)Acceptability: Measured by the percentage of participants who completed the entire HIIE protocol; (2) Implementation was assessed by the number of reported cardiac or serious adverse events during submaximal exercise testing and HIIE and the average percentage of participants reaching vigorous intensity, defined by the American College of Sports Medicine as at least 77% of age-predicted maximal heart rate (HRmax). Data were available for 28 participants who were 32.2 (17.2) months post-stroke and 61.4 (11.9) years of age. Twenty-eight participants completed HIIE per protocol. No cardiac or serious adverse events occurred during the submaximal exercise test or during HIIE. The rapid switching between HIIE and recovery showed no evidence of blood pressure reaching unsafe thresholds. Average intensity during HIIE reached 76.8% HRmax, which is slightly below the target of 77.0%. A single bout of short-interval, low-volume HIIE, prescribed using PPO, was feasible in chronic stroke.

Rapid and visual detection of Shiga-toxigenic Escherichia coli (STEC) in carabeef meat harnessing loop-mediated isothermal amplification (LAMP).

Shiga toxigenic E. coli are important foodborne zoonotic pathogens. The present study was envisaged to standardize loop-mediated isothermal amplification assays targeting stx1 and stx2 genes for rapid and visual detection of STEC and compare its sensitivity with PCR. The study also assessed the effect of short enrichment on the detection limit of LAMP and PCR. The developed LAMP assays were found to be highly specific. Analytical sensitivity of LAMP was 94fg/µLand 25.8fg/µL for stx-1 and stx-2 while LOD of 5 CFU/g of carabeef was measured after 6-12h enrichment. The study highlights the importance of short (6-12h) enrichment for improving the sensitivity of LAMP. The entire detection protocol could be performed within 9h yielding results on the same day. The developed LAMP assays proved to be a handy and cost-effective alternative for screening STEC contamination in meat.

The natural history of CVB3 myocarditis in C57BL/6J mice: an extended in-depth characterization

Background and aimsViral infections are the leading cause of myocarditis. Besides acute cardiac complications, late-stage sequelae such as myocardial fibrosis may develop, importantly impacting the prognosis. Coxsackievirus B3 (CVB)-induced myocarditis in mice is the most commonly used translational model to study viral myocarditis and has provided the majority of our current understanding of the disease pathophysiology. Nevertheless, the late stages of disease, encompassing fibrogenesis and arrhythmogenesis, have been underappreciated in viral myocarditis research to date. The present study investigated the natural history of CVB-induced myocarditis in C57BL/6J mice, expanding the focus beyond the acute phase of disease. In addition, we studied the impact of sex and inoculation dose on the disease course. Methods and resultsC57BL/6J mice (12 weeks old; n=154) received a single intraperitoneal injection with CVB to induce viral myocarditis, or vehicle (PBS) as control. Male mice (n=92) were injected with 5 × 105 (regular dose) (RD) or 5 × 106 (high dose) (HD) plaque-forming units of CVB, whereas female mice received the RD only. Animals were sacrificed 1, 2, 4, 8, and 11 weeks after CVB or PBS injection. Virally inoculated mice developed viral disease with a temporary decline in general condition and weight loss, which was less pronounced in female animals (P<.001). In male CVB mice, premature mortality occurred between days 8 and 23 after inoculation (RD: 21%, HD: 20%), whereas all female animals survived. Over the course of disease, cardiac inflammation progressively subsided, with faster resolution in female mice. There were no substantial group differences in the composition of the inflammatory cell infiltrates: predominance of cytotoxic T cells at day 7 and 14, and a switch from arginase1-reactive macrophages to iNOS-reactive macrophages from day 7 to 14 were the main findings. There was concomitant development and maturation of different patterns of myocardial fibrosis, with enhanced fibrogenesis in male mice. Virus was almost completely cleared from the heart by day 14. Serum biomarkers of cardiac damage and cardiac expression of remodeling genes were temporarily elevated during the acute phase of disease. Cardiac CTGF gene upregulation was less prolonged in female CVB animals. In vivo electrophysiology studies at weeks 8 and 11 demonstrated that under baseline conditions (i.e. in the absence of proarrhythmogenic drugs), ventricular arrhythmias could only be induced in CVB animals. The cumulative arrhythmia burden throughout the entire stimulation protocol was not significantly different between CVB and control groups. ConclusionCVB inoculation in C57BL/6J mice represents a model of acute self-limiting viral myocarditis, with progression to different patterns of myocardial fibrosis. Sex, but not inoculation dose, seems to modulate the course of disease.

Effect of foot core exercises vs ankle proprioceptive neuromuscular facilitation on pain, range of motion, and dynamic balance in individuals with plantar fasciitis: a comparative study

Plantar fasciitis is generally described as an inflammation of the plantar fascia and adjacent tissues around calcaneus tuberosity. Plantar fasciitis onset has been proposed to have an internal mechanical cause, even though this is inadequately recognized. Studies related to alternation in lower-extremity biomechanics that leads to reduced domes of the foot are hypothesized to cause tension in the underlying fascia. Similarly, despite a wealth of anecdotal data suggesting a direct connection between foot mechanism and disability. This condition typically leads to calf muscular stiffness, soreness in the bottom of the feet, decreased range and foot function, strength, and balance impairment. These results interfere with the normal biomechanics of ambulation. A total of (n=66) individuals with plantar fasciitis will be selected for the trial. Subjects will be allocated to Groups A and B at equal allocation with randomization. Group A will undergo foot core exercises, while Group B will undergo ankle proprioceptive neuromuscular facilitation, with both groups receiving conventional treatment. The regimen lasts for 40 minutes, 5 days, for 6 weeks. Foot core exercises are exercises to strengthen the intrinsic and extrinsic muscles of the foot, which results in the improvement in the alignment of the foot. Proprioceptive neuromuscular facilitation exercises are the facilitatory technique to improve the range and strength of the muscles, which results in the improvement in balance. Ultrasound and stretching techniques were added as the conventional therapy to reduce the pain. The outcome measures will be assessed on the Foot Functional Index, modified Star Excursion Balance Test (mSEBT), Visual Analogue Scale (VAS), and Weight Bearing Lunge Test (WBLT) will be assessed at the initiation and completion of the entire treatment protocol. Results will be analyzed prior to and after therapeutic intervention. Based on the comparison of the two treatments' effects on measuring outcomes in individuals with plantar fasciitis, an analysis will be conducted.

Predicting cytogenetic risk in multiple myeloma using conventional whole-body MRI, spinal dynamic contrast-enhanced MRI, and spinal diffusion-weighted imaging

ObjectivesCytogenetic abnormalities are predictors of poor prognosis in multiple myeloma (MM). This paper aims to build and validate a multiparametric conventional and functional whole-body MRI-based prediction model for cytogenetic risk classification in newly diagnosed MM.MethodsPatients with newly diagnosed MM who underwent multiparametric conventional whole-body MRI, spinal dynamic contrast-enhanced (DCE-)MRI, spinal diffusion-weighted MRI (DWI) and had genetic analysis were retrospectively included (2011–2020/Ghent University Hospital/Belgium). Patients were stratified into standard versus intermediate/high cytogenetic risk groups. After segmentation, 303 MRI features were extracted. Univariate and model-based methods were evaluated for feature and model selection. Testing was performed using receiver operating characteristic (ROC) and precision-recall curves. Models comparing the performance for genetic risk classification of the entire MRI protocol and of all MRI sequences separately were evaluated, including all features. Four final models, including only the top three most predictive features, were evaluated.ResultsThirty-one patients were enrolled (mean age 66 ± 7 years, 15 men, 13 intermediate-/high-risk genetics). None of the univariate models and none of the models with all features included achieved good performance. The best performing model with only the three most predictive features and including all MRI sequences reached a ROC-area-under-the-curve of 0.80 and precision-recall-area-under-the-curve of 0.79. The highest statistical performance was reached when all three MRI sequences were combined (conventional whole-body MRI + DCE-MRI + DWI). Conventional MRI always outperformed the other sequences. DCE-MRI always outperformed DWI, except for specificity.ConclusionsA multiparametric MRI-based model has a better performance in the noninvasive prediction of high-risk cytogenetics in newly diagnosed MM than conventional MRI alone.Critical relevance statementAn elaborate multiparametric MRI-based model performs better than conventional MRI alone for the noninvasive prediction of high-risk cytogenetics in newly diagnosed multiple myeloma; this opens opportunities to assess genetic heterogeneity thus overcoming sampling bias.Key points• Standard genetic techniques in multiple myeloma patients suffer from sampling bias due to tumoral heterogeneity.• Multiparametric MRI noninvasively predicts genetic risk in multiple myeloma.• Combined conventional anatomical MRI, DCE-MRI, and DWI had the highest statistical performance to predict genetic risk.• Conventional MRI alone always outperformed DCE-MRI and DWI separately to predict genetic risk. DCE-MRI alone always outperformed DWI separately, except for the parameter specificity to predict genetic risk.• This multiparametric MRI-based genetic risk prediction model opens opportunities to noninvasively assess genetic heterogeneity thereby overcoming sampling bias in predicting genetic risk in multiple myeloma.Graphical

The Effect of Oral Care Foams and a Spray on Salivary pH Changes after Exposure to Acidic Beverages in Young Adults.

Soft drinks may have a deleterious effect on dental health due to a high titratable acidity and a low pH that could be sufficient to induce tooth demineralization. The use of oral care products immediately after acidic challenge may diminish the erosive potential of soft drinks. We assessed the effect of oral care foams and a spray on salivary pH changes after exposure to Coca-Cola® in young adults. Thirty-three consenting eligible patients were recruited in this double-blind, randomized, crossover study performed in six visits. Baseline examination included unstimulated salivary flow rate, stimulated salivary buffer capacity, and the simplified oral hygiene index (OHI-S) assessment. Salivary pH and time for pH recovery were registered after exposure to Coca-Cola® alone or that followed by the application of each of the studied products (an oral foam containing hydroxyapatite and probiotics, an oral foam containing amino fluoride, an alkaline oral spray, and tap water). Thirty-two patients completed the entire study protocol and were included in the final analysis. The mean minimum salivary pH and the mean oral clearance rate after rinsing with Coca-Cola® were 6.3 and 27 min, respectively. Further rinsing with any one of the tested solutions, including tap water, resulted in a significant improvement in these parameters. When the pH curves were plotted, the oral care products demonstrated a lower area under the curve that differed significantly from the area under the curve for Coca-Cola®; tap water did not differ significantly from Coca-Cola® and oral care products. Minimum salivary pH correlated positively with salivary buffer capacity and salivation rate, while salivary clearance correlated with OHI-S plaque scores. In conclusion, the effect of oral care foams and a spray on minimum salivary pH and salivary clearance after exposure to Coca-Cola® did not differ significantly among the tested products and tap water. Trial registration NCT06148662. Funding: none.

Profiling native pulmonary basement membrane stiffness using atomic force microscopy.

Mammalian cells sense and react to the mechanics of their immediate microenvironment. Therefore, the characterization of the biomechanical properties of tissues with high spatial resolution provides valuable insights into a broad variety of developmental, homeostatic and pathological processes within living organisms. The biomechanical properties of the basement membrane (BM), an extracellular matrix (ECM) substructure measuring only ∼100-400 nm across, are, among other things, pivotal to tumor progression and metastasis formation. Although the precise assignment of the Young's modulus E of such a thin ECM substructure especially in between two cell layers is still challenging, biomechanical data of the BM can provide information of eminent diagnostic potential. Here we present a detailed protocol to quantify the elastic modulus of the BM in murine and human lung tissue, which is one of the major organs prone to metastasis. This protocol describes a streamlined workflow to determine the Young's modulus E of the BM between the endothelial and epithelial cell layers shaping the alveolar wall in lung tissues using atomic force microscopy (AFM). Our step-by-step protocol provides instructions for murine and human lung tissue extraction, inflation of these tissues with cryogenic cutting medium, freezing and cryosectioning of the tissue samples, and AFM force-map recording. In addition, it guides the reader through a semi-automatic data analysis procedure to identify the pulmonary BM and extract its Young's modulus E using an in-house tailored user-friendly AFM data analysis software, the Center for Applied Tissue Engineering and Regenerative Medicine processing toolbox, which enables automatic loading of the recorded force maps, conversion of the force versus piezo-extension curves to force versus indentation curves, calculation of Young's moduli and generation of Young's modulus maps, where the pulmonary BM can be identified using a semi-automatic spatial filtering tool. The entire protocol takes 1-2 d.

Safety and feasibility of a functional electrical stimulation cycling-based muscular dysfunction diagnostic method in mechanically ventilated patients.

A nonvolitional diagnostic method based on FES-Cycling technology has recently been demonstrated for mechanically ventilated patients. This method presents good sensitivity and specificity for detecting muscle dysfunction and survival prognosis, even in unconscious patients. As the clinical relevance of this method has already been reported, we aimed to evaluate its safety and feasibility. An observational prospective study was carried out with 20 critically ill, mechanically ventilated patients. The FES-cycling equipment was set in a specific diagnostic mode. For safety determination, hemodynamic parameters and peripheral oxygen saturation were measured before and immediately after the diagnostic protocol, as well as venous oxygen saturation and blood lactate. The creatine phosphokinase level (CPK) was measured before and 24, 48, and 72 h after the test. The time taken to carry out the entire diagnostic protocol and the number of patients with visible muscle contraction (capacity of perceptive muscular recruitment) were recorded to assess feasibility. Heart rate [91 ± 23 vs. 94 ± 23 bpm (p = 0.0837)], systolic [122 ± 19 vs. 124 ± 19 mm Hg (p = 0.4261)] and diastolic blood pressure [68 ± 13 vs. 70 ± 15 mm Hg (p = 0.3462)], and peripheral [98 (96-99) vs. 98 (95-99) % (p = 0.6353)] and venous oxygen saturation [71 ± 14 vs. 69 ± 14% (p = 0.1317)] did not change after the diagnostic protocol. Moreover, blood lactate [1.48 ± 0.65 vs. 1.53 ± 0.71 mmol/L (p = 0.2320)] did not change. CPK did not change up to 72 h after the test [99 (59-422) vs. 125 (66-674) (p = 0.2799) vs. 161 (66-352) (p > 0.999) vs. 100 (33-409) (p = 0.5901)]. The time taken to perform the diagnostic assessment was 11.3 ± 1.1 min. In addition, 75% of the patients presented very visible muscle contractions, and 25% of them presented barely visible muscle contractions. The FES cycling-based muscular dysfunction diagnostic method is safe and feasible. Hemodynamic parameters, peripheral oxygen saturation, venous oxygen saturation, and blood lactate did not change after the diagnostic protocol. The muscle damage marker (CPK) did not increase up to 72 h after the diagnostic protocol.

Limiting Overdistention or Collapse When Mechanically Ventilating Injured Lungs: A Randomized Study in a Porcine Model.

Rationale: It is unknown whether preventing overdistention or collapse is more important when titrating positive end-expiratory pressure (PEEP) in acute respiratory distress syndrome (ARDS). Objectives: To compare PEEP targeting minimal overdistention or minimal collapse or using a compromise between collapse and overdistention in a randomized trial and to assess the impact on respiratory mechanics, gas exchange, inflammation, and hemodynamics. Methods: In a porcine model of ARDS, lung collapse and overdistention were estimated using electrical impedance tomography during a decremental PEEP titration. Pigs were randomized to three groups and ventilated for 12 hours: PEEP set at ⩽3% of overdistention (low overdistention), ⩽3% of collapse (low collapse), and the crossing point of collapse and overdistention. Measurements and Main Results: Thirty-six pigs (12 per group) were included. Median (interquartile range) values of PEEP were 7 (6-8), 11 (10-11), and 15 (12-16) cm H2O in the three groups (P < 0.001). With low overdistension, 6 (50%) pigs died, whereas survival was 100% in both other groups. Cause of death was hemodynamic in nature, with high transpulmonary vascular gradient and high epinephrine requirements. Compared with the other groups, pigs surviving with low overdistension had worse respiratory mechanics and gas exchange during the entire protocol. Minimal differences existed between crossing-point and low-collapse animals in physiological parameters, but postmortem alveolar density was more homogeneous in the crossing-point group. Inflammatory markers were not significantly different. Conclusions: PEEP to minimize overdistention resulted in high mortality in an animal model of ARDS. Minimizing collapse or choosing a compromise between collapse and overdistention may result in less lung injury, with potential benefits of the compromise approach.

A phase II trial of induction systemic mFOLFIRINOX followed by hepatic arterial infusion of floxuridine and dexamethasone given concurrently with systemic mFOLFIRI as a first-line therapy in patients with unresectable liver-dominant intrahepatic cholangiocarcinoma (HELIX-1).

511 Background: Most patients with intrahepatic cholangiocarcinoma (ICC) have unresectable or multifocal liver-dominant disease. Survival after first-line systemic therapy remains poor, with median progression-free (mPFS) and overall survival of 7.2 and 12.8 months, respectively. Hepatic arterial infusion (HAI) therapy with floxuridine maximizes liver-specific treatment with minimal systemic toxicity and potentially offers improved disease control when combined with systemic therapy. Methods: HELIX-1 (NCT04251715) is an investigator-initiated, first-line, single-center, single-arm phase II clinical trial for patients with liver-dominant unresectable or multifocal ICC. The trial was designed with a patient safety run-in evaluating toxicity from the combined therapy. Pre-trial screening included laparoscopy, biopsies, and PET/CT. Eligible patients were treated with systemic mFOLFIRINOX for 4 cycles in order to select those likely to benefit from HAI. Patients with disease control on restaging proceeded to HAI pump placement and treatment with HAI floxuridine for 14 days followed by systemic mFOLFIRI on a 28-day cycle. The co-primary objectives were to assess safety of the combined therapeutic strategy and the disease control rate (DCR) at 6 months (RECIST v1.1) at end of trial (EOT). Results: A total of n=5 patients with liver-only ICC enrolled in the trial and completed the entire study protocol. The median age was 60 years (range 42-69) with a dominant lesion size of 9.8cm (range 8.4-14.5). All patients had both right and left hemiliver involvement with a median of 9 intrahepatic tumors (range 1-15). No patients experienced grade 3 or 4 adverse events, or hepatic dysfunction leading to cessation of HAI therapy. The DCR at 6 months was 100%, with a mPFS of 18.2 months. All five patients achieved partial radiographic response (PR) while receiving HAI therapy, and remain alive with liver-only disease at a median of 18.4 months (range 12.7-20) after study enrollment. After continuing treatment with HAI and systemic mFOLFIRI beyond the EOT, two patients transitioned to HAI treatment only and then to biochemical and radiographic surveillance without therapy after demonstrating PR and CA19-9 normalization. Conclusions: Integration of HAI floxuridine with mFOLFIRI following mFOLFIRINOX induction for patients with liver-only advanced ICC is well-tolerated and demonstrates longer DCR in comparison to historical controls. The combined regimen minimized systemic toxicity and allowed a large proportion of patients to transition to liver-only HAI treatment with maintained disease control. Future directions include combining HAI with new first-line systemic regimens for patients with advanced ICC. Clinical trial information: NCT04251715 .