Turbulent blood flow in a cerebral artery with an aneurysm.

Unruptured intracranial aneurysms are common in the general population, and many uncertainties remain when predicting rupture risks and treatment outcomes. One of the cutting-edge tools used to investigate this condition is computational fluid dynamics (CFD). However, CFD is not yet mature enough to guide the clinical management of this disease. In addition, recent studies have reported significant flow instabilities when refined numerical methods are used. Questions remain as to how to properly simulate and evaluate this flow, and whether these instabilities are really turbulence. The purpose of the present study is to evaluate the impact of the simulation setup on the results and investigate the occurrence of turbulence in a cerebral artery with an aneurysm. For this purpose, direct numerical simulations were performed with up to 200 cardiac cycles and with data sampling rates of up to 100,000 times per cardiac cycle. Through phase-averaging or triple decomposition, the contributions of turbulence and of laminar pulsatile waves to the velocity, pressure and wall shear stress fluctuations were distinguished. For example, the commonly used oscillatory shear index was found to be closely related to the laminar waves introduced at the inlet, rather than turbulence. The turbulence energy cascade was evaluated through energy spectrum estimates, revealing that, despite the low flow rates and Reynolds number, the flow is turbulent near the aneurysm. Phase-averaging was shown to be an approach that can help researchers better understand this flow, although the results are highly dependent on simulation setup and post-processing choices.

Effects of Resistance Training Techniques on Metabolic Responses in Trained Males.

This study investigated the effects of biset, drop-set and traditional resistance training (RT) techniques on metabolic responses in resistance-trained males. Fifteen trained males (age 29.7 ± 6.1 years; body mass 83.4 ± 7.6 kg; RT experience 11.4 ± 6.7 years; one-repetition maximum (1RM) barbell bench press: body mass ratio 1.4 ± 0.1 a.u.) were assigned to three experimental conditions, in a randomized crossover design. The experimental conditions were bi-set (3×10 repetitions at 70%1RM in barbell bench press followed by 10 repetitions at 60%1RM in incline bench press), drop-set (3×10 repetitions at 70%1RM followed by 10 repetitions at 50%1RM in barbell bench press) and traditional RT (3×20 at 60%1RM in barbell bench press). A portable gas analyzer was used to assess energy expenditure and maximal oxygen uptake during the experimental protocols. Blood lactate levels were assessed at baseline and 1, 3, and 5 minutes after the training session. There were no differences for total training volume (p = 0.999). Post hoc comparisons revealed that bi-set elicited higher aerobic energy expenditure (p = 0.003 vs. drop-set; p < 0.001 vs. traditional RT) and aerobic oxygen consumption (p = 0.034 vs. drop-set; p < 0.001 vs. traditional RT) than other RT schemes. There were no differences regarding anaerobic EE between-conditions (p > 0.05). There was a main effect of time and condition for blood lactate levels (p < 0.001). Post hoc comparisons revealed that drop-set training elicited higher blood lactate levels than traditional RT (p = 0.009). The results suggest that RT techniques may have a potential role in optimizing metabolic responses in resistance-trained males.

Pulsation artifact removal from intra-operatively recorded local field potentials using sparse signal processing and data-specific dictionary.

Neural recordings frequently get contaminated by ECG or pulsation artifacts. These large amplitude components can mask the neural patterns of interest and make the visual inspection process difficult. The current study describes a sparse signal representation strategy that targets to denoise pulsation artifacts in local field potentials (LFPs) recorded intraoperatively. To estimate the morphology of the artifact, we first detect the QRS-peaks from the simultaneously recorded ECG trace as an anchor point. After the LFP data has been epoched with respect to each beat, a pool of raw data segments of a specific length is generated. Using the K-singular value decomposition (K-SVD) algorithm, we constructed a data-specific dictionary to represent each contaminated LFP epoch in a sparse fashion. Since LFP is aligned to each QRS complex and the background neural activity is uncorrelated to the anchor points, we assumed that constructed dictionary will be formed to mainly represent the pulsation artifact. In this scheme, we performed an orthogonal matching pursuit to represent each LFP epoch as a linear combination of the dictionary atoms. The denoised LFP data is thus obtained by calculating the residual between the raw LFP and its approximation. We discuss and demonstrate the improvements in denoised data and compare the results with respect to principal component analysis (PCA). We noted that there is a comparable change in the signal for visual inspection to observe various oscillating patterns in the alpha and beta bands. We also see a noticeable compression of signal strength in the lower frequency band (<13 Hz), which was masked by the pulsation artifact, and a strong increase in the signal-to-noise ratio (SNR) in the denoised data.Clinical Relevance- Pulsation artifact can mask relevant neural activity patterns and make their visual inspection difficult. Using sparse signal representation, we established a new approach to reconstruct the quasiperiodic pulsation template and computed the residue signal to achieve noise-free neural activity.

Entropy Production in a Fractal System with Diffusive Dynamics.

We study the entropy production in a fractal system composed of two subsystems, each of which is subjected to an external force. This is achieved by using the H-theorem on the nonlinear Fokker-Planck equations (NFEs) characterizing the diffusing dynamics of each subsystem. In particular, we write a general NFE in terms of Hausdorff derivatives to take into account the metric of each system. We have also investigated some solutions from the analytical and numerical point of view. We demonstrate that each subsystem affects the total entropy and how the diffusive process is anomalous when the fractal nature of the system is considered.

Results for Nonlinear Diffusion Equations with Stochastic Resetting.

In this study, we investigate a nonlinear diffusion process in which particles stochastically reset to their initial positions at a constant rate. The nonlinear diffusion process is modeled using the porous media equation and its extensions, which are nonlinear diffusion equations. We use analytical and numerical calculations to obtain and interpret the probability distribution of the position of the particles and the mean square displacement. These results are further compared and shown to agree with the results of numerical simulations. Our findings show that a system of this kind exhibits non-Gaussian distributions, transient anomalous diffusion (subdiffusion and superdiffusion), and stationary states that simultaneously depend on the nonlinearity and resetting rate.

Construction of semi-supervised spatial projections to identify the source of beta- and high frequency oscillations in Parkinson's disease.

Traditional deep brain stimulation (DBS) treatment for Parkinson's disease (PD) targets the placement of DBS leads into subthalamic nucleus (STN). Extraction of neurobiomarkers from STN local field potential activity can be used for the optimization of DBS. Beta (12-30 Hz) and high frequency oscillations (200-450 Hz, HFO) of STN and their phase-amplitude coupling have been previously correlated with symptom severity in PD. The typical approach is to take bipolar derivations of electrode contacts in order to enhance recordings of local brain activity and suppress noise levels. This approach can often cancel the signals in correlated neighboring contacts and create ambiguity in which monopolar contact to select for the identification of the main source of the oscillatory signal. To improve local specificity and help identify the source of beta and HFO in terms of electrode contact, we propose a semi supervised blind-source separation method. This approach presents a novel perspective to investigate electrophysiology by projecting the recorded channels into a subspace of virtual channels. We show the contribution of each channel to the identified source and correlate the spatial information with imaging and postoperative programming parameters. We anticipate such a source identification strategy can be used in the future to investigate the distribution of beta and HFO on individual contacts of the DBS lead and can improve the interpretation of these signals.

Regulation of brain cognitive states through auditory, gustatory, and olfactory stimulation with wearable monitoring.

Inspired by advances in wearable technologies, we design and perform human-subject experiments. We aim to investigate the effects of applying safe actuation (i.e., auditory, gustatory, and olfactory) for the purpose of regulating cognitive arousal and enhancing the performance states. In two proposed experiments, subjects are asked to perform a working memory experiment called n-back tasks. Next, we incorporate listening to different types of music, drinking coffee, and smelling perfume as safe actuators. We employ signal processing methods to seamlessly infer participants' brain cognitive states. The results demonstrate the effectiveness of the proposed safe actuation in regulating the arousal state and enhancing performance levels. Employing only wearable devices for human monitoring and using safe actuation intervention are the key components of the proposed experiments. Our dataset fills the existing gap of the lack of publicly available datasets for the self-management of internal brain states using wearable devices and safe everyday actuators. This dataset enables further machine learning and system identification investigations to facilitate future smart work environments. This would lead us to the ultimate idea of developing practical automated personalized closed-loop architectures for managing internal brain states and enhancing the quality of life.

Radiation-Induced Lumbosacral Plexopathy.

Lumbosacral plexopathy (LSP) encompasses a group of disorders affecting post-ganglionic fibers derived from the L1-S4 roots. The differential diagnosis is challenging and includes other neuropathies of medullary, radicular, or peripheral origin. Defining the etiology is equally crucial, as LSP management relies on its cause. A thorough clinical history should address potential neoplastic disease (new-onset, progression, or relapse), diabetes mellitus, lumbar or pelvic trauma, and previous exposure to radiation. This is the case of a 78-year-old male, with a history of prostatic adenocarcinoma, treated with image-guided radiation therapy and hormone therapy five years before, with no evidence of relapse on follow-up. The patient presented with bilateral weakness, numbness, and paresthesia of lower limbs, gradually progressing over a three-month period, and followed by an acute worsening with inability to stand or walk. He also referred to distal mild edema, episodic hematuria, and urinary incontinence. Physical examination revealed paraparesis affecting proximal and distal leg muscles, along with bilateral hypoesthesia, impaired deep tendon reflexes, and proprioception below knee level. Pelvic, dorsal, and lumbosacral MRI excluded neoplastic lesions but identified somatic fracture of L5 without medullary or conus medullaris compromise. These findings did not explain the clinical picture. Further neurophysiologic studies characterized sensory-motor deficits as post-ganglionic, with specific spontaneous discharges of the muscle fibers, known as myokymia. These findings were consistent with radiation-induced LSP and were supported by MRI. Radiation-induced cystitis was also documented in pelvic MRI and urethral cystoscopy. This case highlights the clinical picture and differential diagnosis of radiation-induced LSP. Despite more typical symptoms and course, a neoplastic origin should always be carefully investigated and excluded. Radiation protocol should be carefully accessed, and its complications should not be overlooked, as they might cause severe morbidity.

Micro-mechanical fingerprints of the rat bladder change in actinic cystitis and tumor presence.

Tissue mechanics determines tissue homeostasis, disease development and progression. Bladder strongly relies on its mechanical properties to perform its physiological function, but these are poorly unveiled under normal and pathological conditions. Here we characterize the mechanical fingerprints at the micro-scale level of the three tissue layers which compose the healthy bladder wall, and identify modifications associated with the onset and progression of pathological conditions (i.e., actinic cystitis and bladder cancer). We use two indentation-based instruments (an Atomic Force Microscope and a nanoindenter) and compare the micromechanical maps with a comprehensive histological analysis. We find that the healthy bladder wall is a mechanically inhomogeneous tissue, with a gradient of increasing stiffness from the urothelium to the lamina propria, which gradually decreases when reaching the muscle outer layer. Stiffening in fibrotic tissues correlate with increased deposition of dense extracellular matrix in the lamina propria. An increase in tissue compliance is observed before the onset and invasion of the tumor. By providing high resolution micromechanical investigation of each tissue layer of the bladder, we depict the intrinsic mechanical heterogeneity of the layers of a healthy bladder as compared with the mechanical properties alterations associated with either actinic cystitis or bladder tumor.

Cannabidiol effect in pentylenetetrazole-induced seizures depends on PI3K.

BACKGROUND: The phytocannabinoid cannabidiol (CBD) has previously shown to have anticonvulsant effects in preclinical and clinical studies. Recently, CBD has been approved to treat certain types of drug-resistant epileptic syndromes. However, the underlying mechanism of action remains unclear. The phosphatidylinositol 3-kinase (PI3K) signaling pathway has been proposed to modulate seizures and might be recruited by CBD. Thus, we tested the hypothesis that the anticonvulsant effect of CBD involves PI3K in a seizure model induced by pentylenetetrazole (PTZ). METHODS: We employed pharmacological and genetic approaches to inhibit PI3K and quantified its effects on seizure duration, latency, and number. RESULTS: PI3K genetic ablation increased the duration and number of seizures. CBD inhibited PTZ-induced seizures in mice. Genetic deletion of PI3K or pretreatment with the selective inhibitor LY294002 prevented CBD effects. CONCLUSION: Our data strengthen the hypothesis that the CBD anticonvulsant effect requires the PI3K signaling pathway.

Proteomic Signatures of Microbial Adaptation to the Highest Ultraviolet-Irradiation on Earth: Lessons From a Soil Actinobacterium.

In the Central Andean region in South America, high-altitude ecosystems (3500-6000 masl) are distributed across Argentina, Chile, Bolivia, and Peru, in which poly-extremophilic microbes thrive under extreme environmental conditions. In particular, in the Puna region, total solar irradiation and UV incidence are the highest on Earth, thus, restraining the physiology of individual microorganisms and the composition of microbial communities. UV-resistance of microbial strains thriving in High-Altitude Andean Lakes was demonstrated and their mechanisms were partially characterized by genomic analysis, biochemical and physiological assays. Then, the existence of a network of physiological and molecular mechanisms triggered by ultraviolet light exposure was hypothesized and called "UV-resistome". It includes some or all of the following subsystems: (i) UV sensing and effective response regulators, (ii) UV-avoidance and shielding strategies, (iii) damage tolerance and oxidative stress response, (iv) energy management and metabolic resetting, and (v) DNA damage repair. Genes involved in the described UV-resistome were recently described in the genome of Nesterenkonia sp. Act20, an actinobacterium which showed survival to high UV-B doses as well as efficient photorepairing capability. The aim of this work was to use a proteomic approach together with photoproduct measurements to help dissecting the molecular events involved in the adaptive response of a model High-Altitude Andean Lakes (HAAL) extremophilic actinobacterium, Nesterenkonia sp. Act20, under artificial UV-B radiation. Our results demonstrate that UV-B exposure induced over-abundance of a well-defined set of proteins while recovery treatments restored the proteomic profiles present before the UV-challenge. The proteins involved in this complex molecular network were categorized within the UV-resistome subsystems: damage tolerance and oxidative stress response, energy management and metabolic resetting, and DNA damage repair.

Closed-Loop Tracking and Regulation of Emotional Valence State From Facial Electromyogram Measurements.

Affective studies provide essential insights to address emotion recognition and tracking. In traditional open-loop structures, a lack of knowledge about the internal emotional state makes the system incapable of adjusting stimuli parameters and automatically responding to changes in the brain. To address this issue, we propose to use facial electromyogram measurements as biomarkers to infer the internal hidden brain state as feedback to close the loop. In this research, we develop a systematic way to track and control emotional valence, which codes emotions as being pleasant or obstructive. Hence, we conduct a simulation study by modeling and tracking the subject's emotional valence dynamics using state-space approaches. We employ Bayesian filtering to estimate the person-specific model parameters along with the hidden valence state, using continuous and binary features extracted from experimental electromyogram measurements. Moreover, we utilize a mixed-filter estimator to infer the secluded brain state in a real-time simulation environment. We close the loop with a fuzzy logic controller in two categories of regulation: inhibition and excitation. By designing a control action, we aim to automatically reflect any required adjustments within the simulation and reach the desired emotional state levels. Final results demonstrate that, by making use of physiological data, the proposed controller could effectively regulate the estimated valence state. Ultimately, we envision future outcomes of this research to support alternative forms of self-therapy by using wearable machine interface architectures capable of mitigating periods of pervasive emotions and maintaining daily well-being and welfare.

Sensing Apps and Public Data Sets for Digital Phenotyping of Mental Health: Systematic Review.

BACKGROUND: Mental disorders are normally diagnosed exclusively on the basis of symptoms, which are identified from patients' interviews and self-reported experiences. To make mental health diagnoses and monitoring more objective, different solutions have been proposed such as digital phenotyping of mental health (DPMH), which can expand the ability to identify and monitor health conditions based on the interactions of people with digital technologies. OBJECTIVE: This article aims to identify and characterize the sensing applications and public data sets for DPMH from a technical perspective. METHODS: We performed a systematic review of scientific literature and data sets. We searched 8 digital libraries and 20 data set repositories to find results that met the selection criteria. We conducted a data extraction process from the selected articles and data sets. For this purpose, a form was designed to extract relevant information, thus enabling us to answer the research questions and identify open issues and research trends. RESULTS: A total of 31 sensing apps and 8 data sets were identified and reviewed. Sensing apps explore different context data sources (eg, positioning, inertial, ambient) to support DPMH studies. These apps are designed to analyze and process collected data to classify (n=11) and predict (n=6) mental states/disorders, and also to investigate existing correlations between context data and mental states/disorders (n=6). Moreover, general-purpose sensing apps are developed to focus only on contextual data collection (n=9). The reviewed data sets contain context data that model different aspects of human behavior, such as sociability, mood, physical activity, sleep, with some also being multimodal. CONCLUSIONS: This systematic review provides in-depth analysis regarding solutions for DPMH. Results show growth in proposals for DPMH sensing apps in recent years, as opposed to a scarcity of public data sets. The review shows that there are features that can be measured on smart devices that can act as proxies for mental status and well-being; however, it should be noted that the combined evidence for high-quality features for mental states remains limited. DPMH presents a great perspective for future research, mainly to reach the needed maturity for applications in clinical settings.

Neurological development and iron supplementation in healthy late-preterm neonates: a randomized double-blind controlled trial.

Late-preterm infants (LPT) are at increased risk for long-term neurodevelopmental sequelae and iron deficiency. The aim of the study is to assess the positive effect of iron supplementation on psychomotor development in healthy LPT. We designed a randomized placebo-controlled double-blind trial dividing the newborns into two groups. Every patient was assessed using the Griffiths Mental Development Scales (GMDS)-II edition at 12-month post-conceptional age. The study was performed at the Neonatology Unit of our Hospital, in Italy. Sixty-six healthy LPT infants born between 340/7 and 366/7 weeks of gestational age were enrolled in the study. One group received martial prophylaxis from the third week of life to 6 months of post-conceptional age (2 mg/kg/day of iron pidolate), the other received placebo. Fifty-two of the enrolled infants were assessed using the GMDS at 12-month of post-conceptional age. Statistical analysis of the mean scores of the Griffiths subscales was performed. There was a difference in the mean developmental quotient (DQ) (p < 0.01) between the two groups: iron group mean DQ 121.45 ± 10.53 vs placebo group mean DQ 113.25 ± 9.70. Moreover, mean scores of the Griffiths subscales A, B, and D showed significant differences between the two groups (scale A p < 0.05, scale B p < 0.02, scale D p < 0.01, respectively).Conclusions: We recommend that all LPT neonates receive iron supplementation during the first 6 months of life in order to improve their 1-year neurodevelopmental quotient. What is Known: • Late-preterm infants (LPT) are at increased risk for long-term neurodevelopmental sequelae and also for iron deficiency. • Iron deficiency is an independent risk factor for adverse neurological outcomes. What is New: • Healthy late-preterm who received iron supplementation during the first 6 months of life achieved better neurological outcomes at 12-month post-conceptional age than LPT who received placebo. • Our study strongly supports the need for the implementation of martial prophylaxis in LPT neonates.

Smart Coatings Prepared via MAPLE Deposition of Polymer Nanocapsules for Light-Induced Release.

Herein, smart coatings based on photo-responsive polymer nanocapsules (NC) and deposited by laser evaporation are presented. These systems combine remotely controllable release and high encapsulation efficiency of nanoparticles with the easy handling and safety of macroscopic substrates. In particular, azobenzene-based NC loaded with active molecules (thyme oil and coumarin 6) were deposited through Matrix-Assisted Pulsed Laser Evaporation (MAPLE) on flat inorganic (KBr) and organic (polyethylene, PE) and 3D (acrylate-based micro-needle array) substrates. SEM analyses highlighted the versatility and performance of MAPLE in the fabrication of the designed smart coatings. DLS analyses, performed on both MAPLE- and drop casting-deposited NC, demonstrated the remarkable adhesion achieved with MAPLE. Finally, thyme oil and coumarin 6 release experiments further demonstrated that MAPLE is a promising technique for the realization of photo-responsive coatings on various substrates.

Influence of BMI, Age, and Gender on the Thickness of Most Common Thinned Flaps.

BACKGROUND: Since the description of superficial fascia flap harvesting, a new window of opportunity has been open in obese patients, where a higher subcutaneous thickness of tissue is present. To our knowledge, the impact of body mass index on superficial fascial flaps has not been reported. METHODS: We recruited 122 patients from April 2019 to January 2020. From these patients, the 3 most common thinned flaps were selected: the superficial circumflex iliac perforator flap; the anterolateral thigh flap at the perforator A, B, and C; and the thoracodorsal flap. Two vertical measures were registered: the distance from the skin to the superficial fascia, and from this point to the deep fascia. RESULTS: The average flap measurement presented here was within the range, as previous clinical studies. The superficial fatty layer thickness in the superficial circumflex iliac perforator and anterolateral thigh flap was somewhat similar in thickness between overweight and obese patients, showing a minimal increase with higher body mass index. The anterolateral thigh flap was found thicker among women, and no statistical difference was shown between age groups in any of the flaps. CONCLUSIONS: A better understanding of the fat layers' thickness will result in better planning, minimizing secondary debulking procedures, decreasing operative time, and reducing general complications among obese patients. Thus, a better understanding of flap structure and physiology in obese patients will lower complications and give more predictable results.

Effectiveness of Stepped-Care Intervention in Overweight and Obese Patients With Medial Tibiofemoral Osteoarthritis: A Randomized Controlled Trial.

OBJECTIVE: To test the effectiveness of a 32-week, stepped-care intervention on disease remission rates in overweight and obese patients with medial tibiofemoral osteoarthritis (OA) compared to controls. METHODS: In this randomized controlled trial, eligible participants were ≥50 years of age with a body mass index of ≥28 kg/m2 and radiographic evidence of medial tibiofemoral OA. Participants were randomized to stepped-care (n = 87) or control group (n = 84). The stepped-care group received a 2-step intervention. The first step consisted of an 18-week diet and exercise program. The second step consisted of 4 treatment subgroups: 1) diet and exercise maintenance; 2) cognitive-behavioral therapy; 3) unloader knee brace; and 4) muscle strengthening exercises. Allocation into subgroups was based on disease remission state and clinical characteristics. The primary end point was the disease remission rate (yes/no) at 32 weeks, which was reached when participants achieved the Patient Acceptable Symptom State cutoff value for pain and for the patient global assessment of disease activity and/or functional impairment. RESULTS: Disease remission at 32 weeks was achieved by 18 of 68 (26%) in the control group and 32 of 82 (39%) in the stepped-care group (difference 12.6% [95% confidence interval -2.3, 27.4], P = 0.10). The stepped-care group showed an improvement in pain and function between baseline and 20 weeks. While functional improvement was maintained at 32 weeks, pain levels tended to get worse between weeks 20 and 32. CONCLUSION: The proposed intervention did not promote a significant difference in the rate of disease remission in comparison to the control group for overweight or obese patients with medial tibiofemoral OA.

Synergistic effect of a spinel ferrite on the adsorption capacity of nano bio-silica for the removal of methylene blue.

The synergistic effect of CoFe2O4 on the capacity of bio-silica extracted from rice husk for the removal of methylene blue (MB) was investigated. The novel composite of cobalt ferrite/nano bio-silica was prepared by dispersing cobalt and iron salt in ratio 1:2 in a solution containing bio-silica, calcined at 700°C and characterized. The adsorption capacity of the composite (253.6 mg g-1) was higher than that of bio-silica (52.6 mg g-1), and the process was exothermic and spontaneous. Langmuir and Freundlich models were applicable to explain the adsorption isotherm, while pseudo-second-order and Elovich are best applicable for the kinetics mechanism. The amount of MB that was removed, increased with an increase in ionic strength due to dimerization of MB. Regeneration and reusability of the adsorbents showed that they are economically viable. Energy-filtered transmission electron microscopy (EFTEM) and Fourier transformed infrared (FTIR) analysis of MB-loaded adsorbent confirmed the adsorption of MB.

Does the administration of preoperative pembrolizumab lead to sustained remission post-cystectomy? First survival outcomes from the PURE-01 study☆.

BACKGROUND: Initial studies of preoperative checkpoint inhibition before radical cystectomy (RC) have shown promising pathologic complete responses. We aimed to analyze the survival outcomes of patients enrolled in the PURE-01 study (NCT02736266). PATIENTS AND METHODS: We report the results of the secondary end points of PURE-01 in the final population of 143 patients. In particular, we report the event-free survival (EFS) outcomes, defined as the time from the first cycle of pembrolizumab to radiographic disease progression precluding RC, initiation of neoadjuvant chemotherapy (NAC), recurrence after RC, or death from any cause. Other end points were recurrence-free survival (RFS) and overall survival (OS). Subgroup analyses were carried out, including pathological response category, clinical complete responses (CR) assessed via multiparametric magnetic resonance imaging (mpMRI), and molecular subtyping. Cox regression analyses for EFS were also carried out. RESULTS: After a median [interquartile range (IQR)] follow-up of 23 (15-29) months, 12- and 24-month EFS were 84.5% [95% confidence interval (CI): 78.5-90.9] and 71.7% (62.7-82). The prognosis was favorable across all the different pathological response subgroups, with the exception of ypN+ (N = 21), showing a 24-month RFS (95% CI) of 39.3% (19.2% to 80.5%). A statistically significant EFS benefit was observed in patients with a clinical CR (P = 0.002). Programmed cell-death-ligand-1 combined positive score was significantly associated with longer EFS in multivariable analyses. Four patients refused RC after clinical evidence of CR, and none of them have recurred after a median follow-up of 10 months (IQR: 11-15). The claudin-low subtype displayed a numerically longer EFS after pembrolizumab and RC compared with the other subtypes. CONCLUSIONS: The EFS results from PURE-01 revealed that the immunotherapy effect was maintained post-RC in most patients. Pembrolizumab compared favorably with neoadjuvant chemotherapy, irrespective of the biomarker status. Molecular subtyping may be a useful tool to select the patients who are predicted to benefit the most from neoadjuvant pembrolizumab.