No effect of ovarian stimulation and oocyte yield on euploidy and live birth rates: an analysis of 12 298 trophectoderm biopsies.

STUDY QUESTION: Does ovarian stimulation affect embryo euploidy rates or live birth rates (LBRs) after transfer of euploid embryos? SUMMARY ANSWER: Euploidy rates and LBRs after transfer of euploid embryos are not significantly influenced by gonadotropin dosage, duration of ovarian stimulation, estradiol level, follicle size at ovulation trigger or number of oocytes retrieved, regardless of a woman's age. WHAT IS KNOWN ALREADY: Aneuploidy rates increase steadily with age, reaching >80% in women >42 years old. The goal of ovarian stimulation is to overcome this high aneuploidy rate through the recruitment of several follicles, which increases the likelihood of obtaining a euploid embryo that results in a healthy conceptus. However, several studies have suggested that a high response to stimulation might be embryotoxic and/or increase aneuploidy rates by enhancing abnormal segregation of chromosomes during meiosis. Furthermore, a recent study demonstrated a remarkable difference in euploidy rates, ranging from 39.5 to 82.5%, among young oocyte donors in 42 fertility centres, potentially suggesting an iatrogenic etiology resulting from different stimulation methods. STUDY DESIGN, SIZE, DURATION: This is a retrospective cohort study that included 2230 in vitro fertilisation (IVF) with preimplantation genetic testing for aneuploidy (PGT-A) cycles and 930 frozen-thawed single euploid embryo transfer (FET) cycles, performed in our centre between 2013 and 2017. PARTICIPANTS/MATERIALS, SETTING, METHODS: A total of 12 298 embryos were analysed for ploidy status. Women were divided into five age groups (<35, 35-37, 38-40, 41-42 and >42 years old). Outcomes were compared between different durations of stimulation (<10, 10-12 and ≥13 days), total gonadotropin dosages (<4000, 4000-6000 and >6000 IU), numbers of oocytes retrieved (<10, 10-19 and ≥20 oocytes), peak estradiol levels (<2000, 2000-3000 and >3000 pg/mL), and sizes of the largest follicle on the day of trigger (<20 and ≥20 mm). MAIN RESULTS AND THE ROLE OF CHANCE: Within the same age group, both euploidy rates and LBRs were comparable between cycles regardless of their differences in total gonadotropin dosage, duration of stimulation, number of oocytes harvested, size of the largest follicles or peak estradiol levels. In the youngest group, (<35 years, n = 3469 embryos), euploidy rates were comparable between cycles with various total gonadotropin dosages (55.6% for <4000 IU, 52.9% for 4000-6000 IU and 62.3% for >6000 IU; P = 0.3), durations of stimulation (54.4% for <10 days, 55.2% for 10-12 days and 60.9% for >12 days; P = 0.2), number of oocytes harvested (59.4% for <10 oocytes, 55.2% for 10-19 oocytes and 53.4% for ≥20 oocytes; P = 0.2), peak estradiol levels (55.7% for E2 < 2000 pg/mL, 55.4% for E2 2000-3000 pg/mL and 54.8% for E2 > 3000 pg/mL; P = 0.9) and sizes of the largest follicle (55.6% for follicles <20 mm and 55.1% for follicles ≥20 mm; P = 0.8). Similarly, in the oldest group (>42 years, n = 1157 embryos), euploidy rates ranged from 8.7% for gonadotropins <4000 IU to 5.1% for gonadotropins >6000 IU (P = 0.3), from 10.8% for <10 days of stimulation to 8.5% for >12 days of stimulation (P = 0.3), from 7.3% for <10 oocytes to 7.4% for ≥20 oocytes (P = 0.4), from 8.8% for E2 < 2000 pg/mL to 7.5% for E2 > 3000 pg/mL (P = 0.8) and from 8.2% for the largest follicle <20 mm to 8.9% for ≥20 mm (P = 0.7). LBRs after single FET were also comparable between these groups. LIMITATIONS, REASONS FOR CAUTION: Although this large study (2230 IVF/PGT-A cycles, 12 298 embryos and 930 single FET cycles) demonstrates the safety of ovarian stimulation in terms of aneuploidy and implantation potential of euploid embryos, a multi-centre study may help to prove the generalisability of our single-centre data. WIDER IMPLICATIONS OF THE FINDINGS: These findings reassure providers and patients that gonadotropin dosage, duration of ovarian stimulation, estradiol level, follicle size at ovulation trigger and number of oocytes retrieved, within certain ranges, do not appear to significantly influence euploidy rates or LBRs, regardless of the woman's age. STUDY FUNDING/COMPETING INTEREST(S): No external funding was received and there are no competing interests to declare. TRIAL REGISTRATION NUMBER: N/A.

On-line monitoring of photosynthetic activity based on pH data to assess microalgae cultivation.

Microalgae performance of outdoor cultivation systems is influenced by environmental and operating dynamics. Monitoring and control systems are needed to maximise biomass productivity and nutrient recovery. The goal of this work was to corroborate that pH data could be used to monitor microalgae performance by means of data from an outdoor membrane photobioreactor (MPBR) plant. In this system, microalgae photosynthetic activity was favoured over other physical and biological processes, so that the pH data dynamics was theoretically related to the microalgae carbon uptake rate (CUR). Short- and long-term continuous operations were tested to corroborate the relationship between the first derivate of pH data dynamics (pH') and microalgae photosynthetic activity. Short-term operations showed a good correlation between gross pH' values and MPBR performance. An indicator of the maximum daily average microalgae activity was assessed by a combination of on-line pH' measurements obtained in the long-term and a microalgae growth kinetic model. Both indicators contributed to the development of advanced real-time monitoring and control systems to optimise microalgae cultivation technology.

Plant-wide modelling in wastewater treatment: showcasing experiences using the Biological Nutrient Removal Model.

Plant-wide modelling can be considered an appropriate approach to represent the current complexity in water resource recovery facilities, reproducing all known phenomena in the different process units. Nonetheless, novel processes and new treatment schemes are still being developed and need to be fully incorporated in these models. This work presents a short chronological overview of some of the most relevant plant-wide models for wastewater treatment, as well as the authors' experience in plant-wide modelling using the general model BNRM (Biological Nutrient Removal Model), illustrating the key role of general models (also known as supermodels) in the field of wastewater treatment, both for engineering and research.

Position paper - progress towards standards in integrated (aerobic) MBR modelling.

Membrane bioreactor (MBR) models are useful tools for both design and management. The system complexity is high due to the involved number of processes which can be clustered in biological and physical ones. Literature studies are present and need to be harmonized in order to gain insights from the different studies and allow system optimization by applying a control. This position paper aims at defining the current state of the art of the main integrated MBR models reported in the literature. On the basis of a modelling review, a standardized terminology is proposed to facilitate the further development and comparison of integrated membrane fouling models for aerobic MBRs.

B subset cells in patients with chronic granulomatous disease in a Mexican population.

INTRODUCTION: Chronic granulomatous disease (CGD) is a disorder of phagocyte function, characterized by pyogenic infections and granuloma formation caused by defects in NADPH oxidase complex activity. Although the effect of CGD mainly reflects the phagocytic compartment, B cell responses are also impaired in patients with CGD. MATERIALS AND METHODS: Flow cytometric analysis was performed on peripheral blood samples from 35 CGD patients age-matched with healthy controls (HC). The target cells of our study were the naive (IgD+/CD27-), memory (IgD-/CD27+), and B1a (CD5+) cells. Immunoglobulins (Igs) were also measured. This study was performed in a Latin American cohort. RESULTS: We found significantly higher levels of naive B cells and B1a cells, but lower levels of memory B cells were found in CGD patients compared to HC. There was no significant difference of cell percentages per inheritance type. DISCUSSION: Our findings suggest that the deficiency of NADPH oxidase components can affect the differentiation of naive B cells to memory B cells. Consequently, memory cells will be low, which also influenced the expression of CD27 in memory B cells and as a result, the percentage of naive cells increases. An altered phenotype of B lymphocytes in CGD patients may contribute to the opportunistic infections and autoimmune disorders that are seen in this disease.

Optimising an outdoor membrane photobioreactor for tertiary sewage treatment.

The operation of an outdoor membrane photobioreactor plant which treated the effluent of an anaerobic membrane bioreactor was optimised. Biomass retention times of 4.5, 6, and 9 days were tested. At a biomass retention time of 4.5 days, maximum nitrogen recovery rate:light irradiance ratios, photosynthetic efficiencies and carbon biofixations of 51.7 ±â€¯14.3 mg N·mol-1, 4.4 ±â€¯1.6% and 0.50 ±â€¯0.05 kg CO2·m3influent, respectively, were attained. Minimum membrane fouling rates were achieved when operating at the shortest biomass retention time because of the lower solid concentration and the negligible amount of cyanobacteria and protozoa. Hydraulic retention times of 3.5, 2, and 1.5 days were tested at the optimum biomass retention times of 4.5 days under non-nutrient limited conditions, showing no significant differences in the nutrient recovery rates, photosynthetic efficiencies and membrane fouling rates. However, nitrogen recovery rate:light irradiance ratios and photosynthetic efficiency significantly decreased when hydraulic retention time was further shortened to 1 day, probably due to a rise in the substrate turbidity which reduced the light availability in the culture. Optimal carbon biofixations and theoretical energy recoveries from the biomass were obtained at hydraulic retention time of 3.5 days, which accounted for 0.55 ±â€¯0.05 kg CO2·m-3influent and 0.443 ±â€¯0.103 kWh·m-3influent, respectively.

Resource recovery from sulphate-rich sewage through an innovative anaerobic-based water resource recovery facility (WRRF).

This research work proposes an innovative water resource recovery facility (WRRF) for the recovery of energy, nutrients and reclaimed water from sewage, which represents a promising approach towards enhanced circular economy scenarios. To this aim, anaerobic technology, microalgae cultivation, and membrane technology were combined in a dedicated platform. The proposed platform produces a high-quality solid- and coliform-free effluent that can be directly discharged to receiving water bodies identified as sensitive areas. Specifically, the content of organic matter, nitrogen and phosphorus in the effluent was 45 mg COD·L-1, 14.9 mg N·L-1 and 0.5 mg P·L-1, respectively. Harvested solar energy and carbon dioxide biofixation in the form of microalgae biomass allowed remarkable methane yields (399 STP L CH4·kg-1 CODinf) to be achieved, equivalent to theoretical electricity productions of around 0.52 kWh per m3 of wastewater entering the WRRF. Furthermore, 26.6% of total nitrogen influent load was recovered as ammonium sulphate, while nitrogen and phosphorus were recovered in the biosolids produced (650 ± 77 mg N·L-1 and 121.0 ± 7.2 mg P·L-1).

Economic and environmental sustainability of submerged anaerobic MBR-based (AnMBR-based) technology as compared to aerobic-based technologies for moderate-/high-loaded urban wastewater treatment.

The objective of this study was to assess the economic and environmental sustainability of submerged anaerobic membrane bioreactors (AnMBRs) in comparison with aerobic-based technologies for moderate-/high-loaded urban wastewater (UWW) treatment. To this aim, a combined approach of steady-state performance modelling, life cycle analysis (LCA) and life cycle costing (LCC) was used, in which AnMBR (coupled with an aerobic-based post-treatment) was compared to aerobic membrane bioreactor (AeMBR) and conventional activated sludge (CAS). AnMBR with CAS-based post-treatment for nutrient removal was identified as a sustainable option for moderate-/high-loaded UWW treatment: low energy consumption and reduced sludge production could be obtained at given operating conditions. In addition, significant reductions can be achieved in different aspects of environmental impact (global warming potential (GWP), abiotic depletion, acidification, etc.) and LCC over existing UWW treatment technologies.

Economic and environmental sustainability of an AnMBR treating urban wastewater and organic fraction of municipal solid waste.

The objective of this study was to evaluate the economic and environmental sustainability of a submerged anaerobic membrane bioreactor (AnMBR) treating urban wastewater (UWW) and organic fraction of municipal solid waste (OFMSW) at ambient temperature in mild/hot climates. To this aim, power requirements, energy recovery from methane (biogas methane and methane dissolved in the effluent), consumption of reagents for membrane cleaning, and sludge handling (polyelectrolyte and energy consumption) and disposal (farmland, landfilling and incineration) were evaluated within different operating scenarios. Results showed that, for the operating conditions considered in this study, AnMBR technology is likely to be a net energy producer, resulting in considerable cost savings (up to €0.023 per m(3) of treated water) when treating low-sulphate influent. Life cycle analysis (LCA) results revealed that operating at high sludge retention times (70 days) and treating UWW jointly with OFMSW enhances the overall environmental performance of AnMBR technology.

Electrical conductivity as a state indicator for the start-up period of anaerobic fixed-bed reactors.

The aim of this work was to analyse the applicability of electrical conductivity sensors for on-line monitoring the start-up period of an anaerobic fixed-bed reactor. The evolution of bicarbonate concentration and methane production rate was analysed. Strong linear relationships between electrical conductivity and both bicarbonate concentration and methane production rate were observed. On-line estimations of the studied parameters were carried out in a new start-up period by applying simple linear regression models, which resulted in a good concordance between both observed and predicted values. Electrical conductivity sensors were therefore identified as an interesting method for monitoring the start-up period of anaerobic fixed-bed reactors due to its reliability, robustness, easy operation, low cost, and minimum maintenance compared with the currently used sensors.

A ghrelin-growth hormone axis drives stress-induced vulnerability to enhanced fear.

Hormones in the hypothalamus-pituitary-adrenal (HPA) axis mediate many of the bodily responses to stressors, yet there is no clear relationship between the levels of these hormones and stress-associated mental illnesses such as posttraumatic stress disorder (PTSD). Therefore, other hormones are likely to be involved in this effect of stress. Here we used a rodent model of PTSD in which rats repeatedly exposed to a stressor display heightened fear learning following auditory Pavlovian fear conditioning. Our results show that stress-related increases in circulating ghrelin, a peptide hormone, are necessary and sufficient for stress-associated vulnerability to exacerbated fear learning and these actions of ghrelin occur in the amygdala. Importantly, these actions are also independent of the classic HPA stress axis. Repeated systemic administration of a ghrelin receptor agonist enhanced fear memory but did not increase either corticotropin-releasing factor (CRF) or corticosterone. Repeated intraamygdala infusion of a ghrelin receptor agonist produced a similar enhancement of fear memory. Ghrelin receptor antagonism during repeated stress abolished stress-related enhancement of fear memory without blunting stress-induced corticosterone release. We also examined links between ghrelin and growth hormone (GH), a major downstream effector of the ghrelin receptor. GH protein was upregulated in the amygdala following chronic stress, and its release from amygdala neurons was enhanced by ghrelin receptor stimulation. Virus-mediated overexpression of GH in the amygdala was also sufficient to increase fear. Finally, virus-mediated overexpression of a GH receptor antagonist was sufficient to block the fear-enhancing effects of repeated ghrelin receptor stimulation. Thus, ghrelin requires GH in the amygdala to exert fear-enhancing effects. These results suggest that ghrelin mediates a novel branch of the stress response and highlight a previously unrecognized role for ghrelin and growth hormone in maladaptive changes following prolonged stress.

Histopathological and biochemical evidence of hepatopancreatic toxicity caused by cadmium in white shrimp, Palaemonetes argentinus.

Cadmium (Cd) is one of the most common pollutants in the environment and induces a range of tissue changes or damages and organ dysfunction. The histopathological effects of Cd and lipid peroxidation (LPO) on hepatopancreas of the freshwater shrimp, Palaemonetes argentinus, were studied. Shrimp were obtained from two lagoons with contrasting environmental quality, De los Padres (LP, impacted site) and Nahuel Rucá (NR, reference site), and were exposed to 3.06 and 12.24µgCdL(-1) for 3, 7, 10 and 15 days. The health status of both populations was also evaluated by histological analysis of control individuals. After exposure, shrimp were transferred to clean water for 28 days to evaluate the recuperation capacity of hepatopancreas. Control shrimp from NR exhibited a normal hepatopancreas structure; unlike control shrimp from LP which showed several alterations. These results were attributed to the different environmental quality of lagoons. The exposure to Cd resulted in several alterations in the histological structure of the hepatopancreas of both populations. The observed alterations included haemocytic and connective infiltrations in the intertubular space, erosioned microvilli, ripple of basal lamina, atrophied epithelium and necrosis, however, the latter was only observed in shrimp from LP. The exposure also caused an increase of LPO levels in both populations. P. argentinus was able to repair the hepatopancreas structure from the damage caused by Cd, evidenced by the histopathological results and LPO levels. Obtained results are indicating that the histological analysis of the hepatopancreas proved to be a highly sensitive method for evaluating water quality, in both environmental and laboratory conditions.

Diagnostic validity of the Alzheimer's disease functional assessment and change scale in mild cognitive impairment and mild to moderate Alzheimer's disease.

BACKGROUND: The Alzheimer's Disease Functional Assessment and Change Scale (ADFACS) is a functional assessment instrument widely used in clinical research. AIMS: To test the diagnostic and concurrent validity of the Spanish version of this scale and to describe the functional deficit pattern for mild cognitive impairment (MCI) and Alzheimer's disease (AD) dementia. METHODS: The ADFACS, the Interview for Deterioration in Daily Living Activities in Dementia (IDDD), and the Mini Mental State Examination (MMSE) were administered to 146 control subjects (CS) and 165 patients (67 MCI and 98 AD). Nonparametric tests were used to compare the diagnostic groups. Cronbach's α and correlations with the MMSE and the IDDD were calculated. Sensitivity, specificity and predictive values were studied. RESULTS: The ADFACS had a high internal consistency (α = 0.95). Three cutoff points of 1, 4, and 17 were provided to separate CS and MCI patients, MCI and mild AD patients, and mild AD and moderate AD patients, respectively. The ADFACS strongly correlated with functional (IDDD, 0.927) and cognitive (MMSE, 0.747) measures. A similar pattern of dysfunction, but in different grades, was found for the MCI and AD groups. CONCLUSION: The ADFACS is a reliable, valid, and sensitive instrument to assess functional abilities; it is useful in dementia assessment for elderly populations.

Anaerobic treatment of urban wastewater in membrane bioreactors: evaluation of seasonal temperature variations.

The objective of this study was to evaluate the effect of seasonal temperature variations on the anaerobic treatment of urban wastewater in membrane bioreactors (MBRs). To this aim, sludge production, energy recovery potential, chemical oxygen demand (COD) removal and membrane permeability were evaluated in a submerged anaerobic MBR fitted with industrial-scale membrane units. The plant was operated for 172 days, between summer and winter seasons. Sludge production increased and energy recovery potential decreased when temperature decreased. COD removal and membrane permeability remained nearby stable throughout the whole experimental period.

Assessment of thermohydraulic performance and entropy generation in an evacuated tube solar collector employing pure water and nanofluids as working fluids.

This study conducts a numerical comparison of the thermal performance of three distinct working fluids (pure water, TiO2, and SiO2 water-based nanofluids) within an evacuated tube solar collector using Computational Fluid Dynamics. The study evaluates thermohydraulic performance alongside global and local entropy generation rates, while considering variations in solar radiation values and inlet mass flow rates. Results indicate that nanofluids demonstrate superior performance under low solar radiation, exhibiting higher outlet temperatures, velocities, thermal efficiency, and exergy efficiency compared to pure water. However, at the higher solar radiation level, the efficiency of SiO2 water-based nanofluid diminishes due to its impact on specific heat. Furthermore, the entropy generation analysis reveals significant reductions with TiO2 water-based nanofluid in all the phenomena considered (up to 79 %). The SiO2 nanofluid performance aligns closely with pure water under high radiation value. This investigation offers valuable insights into the utilization of nanofluids in solar collectors across diverse operating conditions, emphasizing their pivotal role in enhancing overall performance.

Relationship of aquatic environmental factors with the abundance of Vibrio cholerae, Vibrio parahaemolyticus, Vibrio mimicus and Vibrio vulnificus in the coastal area of Guaymas, Sonora, Mexico.

Members of the genus Vibrio are common in aquatic environments. Among them are V. cholerae, V. vulnificus, V. parahaemolyticus and V. mimicus. Several studies have shown that environmental factors, such as temperature, salinity, and dissolved oxygen, are involved in their epidemiology. Therefore, the main objective of this study is to determine if there is a correlation between the presence/amount of V. cholerae, V, vulnificus, V. parahaemolyticus and V. mimicus and the environmental conditions of the seawater off the coast of Guaymas, México. Quantification of all four pathogenic bacteria was performed using the most probable number method, and suspected colonies were identified by polymerase chain reaction (PCR). Correlations were found using principal component analysis. V. parahaemolyticus was the most abundant and widely distributed bacteria, followed by V. vulnificus, V. mimicus and V. cholerae. Positive correlations between V. parahaemolyticus, V. vulnificus and V. mimicus with temperature, salinity, electric conductivity, and total dissolved solids were found. The abundance of V. cholerae was mainly affected by the sampling site and not by physicochemical parameters.

Assigning F-words as ingredients of interventions for children with cerebral palsy functioning at GMFCS IV and V: A scoping review protocol.

Introduction: Children with Cerebral Palsy (CP) functioning at Gross Motor Function Classification System (GMFCS) levels IV and V require "on time" identification and intervention. Interventions offered continue to be a challenge, in high-, but even more so in middle-, and low-income countries. Aim: To describe the methods developed to explore the ingredients of published studies on early interventions in young children with cerebral palsy (CP) at highest risk of being non-ambulant based on the "F-words for child development framework" and the design of a scoping review exploring these ingredients. Method: An operational procedure was developed through expert panels to identify ingredients of published interventions and related F-words. After sufficient agreement among researchers was reached, a scoping review was designed. The review is registered in the Open Science Framework database. The "Population, Concept and Context" framework was used. Population: young children (0-5 years with CP and at highest risk for being non-ambulant (GMFCS levels IV or V); Concept: non-surgical and non-pharmacological early intervention services measuring outcomes from any ICF domain; Context: studies published from 2001 to 2021. After duplicated screening and selection, data will be extracted and quality will be assessed with the American Academy for Cerebral Palsy and Developmental Medicine (AACPDM) and Mixed Methods Appraisal (MMAT) tools. Results: We present the protocol to identify the explicit (directly measured outcomes and respective ICF domains) and implicit (intervention features not explicitly intended or measured) ingredients. Conclusion: Findings will support the implementation of the F-words in interventions for young children with non-ambulant CP.

Translational insights into mechanisms underlying residual venous obstruction and the role of factor XI, P-selectin and GPVI in recurrent venous thromboembolism.

BACKGROUND: Residual venous obstruction (RVO) after deep vein thrombosis (DVT) is considered a risk factor of recurrent venous thromboembolism (VTE), arterial events and post-thrombotic syndrome (PTS). We hypothesized thrombo-inflammatory markers might be associated with RVO and clinical outcomes. MATERIALS AND METHODS: In a DVT cohort with routine RVO-assessment and 5-year follow-up, patients were invited for blood withdrawal after stopping anticoagulants. Thrombin generation potential, coagulation enzyme:inhibitor complexes, soluble platelet markers and clinical markers were measured in platelet-poor plasma. Associations were represented as odds ratio (OR) or hazard ratio (HR) per standard deviation. RESULTS: Patients with RVO (102/306, 33 %) had higher rates of PTS (24 vs. 12 %, p = 0.008), but similar rates of recurrence (16 vs. 15 %, p = 0.91) and arterial events (7 vs. 4 %, p = 0.26). RVO was associated with thrombin peak height (OR 1.40 [1.04-1.88]), endogenous thrombin potential (ETP, OR 1.35 [1.02-1.79]), and CRP (OR 1.74 [1.10-2.75]). Recurrent VTE was associated with ETP (HR 1.36 [1.03-1.81]), FXIa:C1-inhibitor (HR 1.34 [1.04-1.72]), thrombin:antithrombin (HR 1.36 [1.16-1.59]), soluble P-selectin (HR 2.30 [1.69-3.11]), soluble glycoprotein VI (sGPVI, HR 1.30 [1.01-1.69]), D-dimer (HR 1.56 [1.31-1.86]), and factor VIII (HR 1.44 [1.15-1.82]). Arterial events were associated with sGPVI (HR 1.80 [1.25-2.59]). PTS was not associated with any marker. CONCLUSIONS: Our findings indicate RVO was associated with thrombo-inflammation, but this did not predict clinical outcomes in this setting. Importantly, we found recurrent VTE was associated with ongoing coagulation and platelet activation in patients well beyond the acute phase of DVT. Furthermore, sGPVI indicated an increased risk of arterial events, highlighting the role of platelets in arterial thrombosis following DVT.

Plasma protein signatures for high on-treatment platelet reactivity to aspirin and clopidogrel in peripheral artery disease.

BACKGROUND: A significant proportion of patients with peripheral artery disease (PAD) displays a poor response to aspirin and/or the platelet P2Y12 receptor antagonist clopidogrel. This phenomenon is reflected by high on-treatment platelet reactivity (HTPR) in platelet function assays in vitro and is associated with an increased risk of adverse cardiovascular events. OBJECTIVE: This study aimed to elucidate specific plasma protein signatures associated with HTPR to aspirin and clopidogrel in PAD patients. METHODS AND RESULTS: Based on targeted plasma proteomics, 184 proteins from two cardiovascular Olink panels were measured in 105 PAD patients. VerifyNow ASPI- and P2Y12-test values were transformed to a continuous variable representing HTPR as a spectrum instead of cut-off level-defined HTPR. Using the Boruta random forest algorithm, the importance of 3 plasma proteins for HTPR in the aspirin, six in clopidogrel and 10 in the pooled group (clopidogrel or aspirin) was confirmed. Network analysis demonstrated clusters with CD84, SLAMF7, IL1RN and THBD for clopidogrel and with F2R, SELPLG, HAVCR1, THBD, PECAM1, TNFRSF10B, MERTK and ADM for the pooled group. F2R, TNFRSF10B and ADM were higher expressed in Fontaine III patients compared to Fontaine II, suggesting their relation with PAD severity. CONCLUSIONS: A plasma protein signature, including eight targets involved in proatherogenic dysfunction of blood cell-vasculature interaction, coagulation and cell death, is associated with HTPR (aspirin and/or clopidogrel) in PAD. This may serve as important systems-based determinants of poor platelet responsiveness to aspirin and/or clopidogrel in PAD and other cardiovascular diseases and may contribute to identify novel treatment strategies.