Incidence of Compartment Syndrome Following Peri-Pandemic Intervention for Non-traumatic Acute Limb Ischemia.

OBJECTIVE: During the pandemic, our institution anecdotally observed a significant proportion of acute limb ischemia (ALI) patients developing compartment syndrome (CS) following revascularization compared to pre-pandemic rates. To determine whether this perceived increase was occurring globally, we utilized the TriNetX database to evaluate the incidence of CS secondary to ALI intervention in both the pre-pandemic (2017-2019) and pandemic eras (2020-2022). METHODS: We conducted a multicenter query using the TriNetX global research network for ALI patients receiving treatment. Incidence of CS diagnosis within 1 calendar day of ALI intervention was calculated for each era. Demographics and comorbidities were then compared between CS and non-CS patients within each era. Risk of adverse outcomes within 30 days of CS diagnosis was also determined for each era, including mortality, major amputation, and re-intervention. RESULTS: The pre-pandemic cohort contained 7736 patients while the pandemic era cohort included 8,306, for 16,042 total patients. A significant increase in CS incidence (risk ratio (RR) = 1.23, P = 0.0026) was demonstrated within the pandemic era. An increased prevalence of comorbidities such as dyslipidemia (pre-pandemic: P = 0.0022; pandemic: P = 0.0026) and peripheral vascular disease (P < 0.0001, both eras) was observed in the non-CS cohort within both eras. 30-day mortality was significantly increased in CS patients (pre-pandemic: RR = 3.057; pandemic: RR = 2.710; P < 0.0001 both eras) compared to non-CS patients. CS patients were more likely to receive major amputation (pre-pandemic: RR = 3.734; pandemic: RR = 2.809; P < 0.0001 both eras) and/or re-intervention within 30 days (pre-pandemic: RR = 1.871, P < 0.0001; pandemic: RR = 1.370, P = 0.0218) over non-CS patients. CONCLUSIONS: The incidence of CS following revascularization for ALI rose worldwide during the pandemic. Patients who developed CS are younger with fewer comorbidities than non-CS patients. Despite a more favorable comorbid profile, CS patients demonstrate significantly higher rates of adverse outcomes. Further investigation is necessary to determine the specific underlying mechanisms driving this increased incidence in CS among ALI patients.

Burnout is not associated with trainee performance on the Vascular Surgery In-Training Exam.

OBJECTIVE: The Vascular Surgery In-Training Examination (VSITE) is a yearly exam evaluating vascular trainees' knowledge base. Although multiple studies have evaluated variables associated with exam outcomes, few have incorporated training program-specific metrics. The purpose of this study is to evaluate the impact of the learning environment and burnout on VSITE performance. METHODS: Data was collected from a confidential, voluntary survey administered after the 2020 to 2022 VSITE as part of the SECOND Trial. VSITE scores were calculated as percent correct then standardized per the American Board of Surgery. Generalized estimating equations with robust standard errors and an independent correlation structure were used to evaluate trainee and program factors associated with exam outcomes. Analyses were further stratified by integrated and independent training paradigms. RESULTS: A total of 1385 trainee responses with burnout data were collected over 3 years (408 in 2020, 459 in 2021, 498 in 2022). On average, 46% of responses reported at least weekly burnout symptoms. On unadjusted analysis, burnout symptoms correlated with a 14 point drop in VSITE score (95% confidence interval [CI], -24 to -4; P = .006). However, burnout was no longer significant after adjusted analysis. Instead, higher postgraduate year level, being in a relationship, identifying as male gender with or without kids, identifying as non-Hispanic white, larger programs, and having a sense of belonging within a program were associated with higher VSITE scores. CONCLUSIONS: Despite high rates of burnout, trainees generally demonstrate resilience in gaining the medical knowledge necessary to pass the VSITE. Performance on standardized exams is associated with trainee and program characteristics, including availability of support systems and program belongingness.

Lymphocyte recovery after bendamustine therapy in patients with mantle cell lymphoma. Results of a retrospective analysis and prognostic impact in the CAR-T era.

Bendamustine in combination with rituximab (BR) or with rituximab and cytarabine (R-BAC) is the standard first-line immunochemotherapy in mantle cell lymphoma (MCL) for elderly patients and patients ineligible for intensive regimens or autologous transplantation. As bendamustine causes prolonged lymphopenia and the literature lacks evidence of its persistence in patients with MCL, this retrospective analysis aims to estimate the lymphocyte recovery time, also in view of potential immunotherapy with CAR-T cells. Data were collected from 44 consecutive MCL patients who received bendamustine (BR or R-BAC) as first-line therapy at the Hematology Unit of Sapienza University Hospital between May 2011 and April 2022. Twenty patients (45%) were treated with R-BAC and 24 (55%) with BR. At baseline, the median lymphocyte count was 1795/µl (range: 370-11730/µL). One month after the end of therapy, it was 450/µl (range: 50-3300/µl) and 3 months after 768/µl (range: 260-1650/µl). After 6 and 9 months, we observed a gradual increase in median lymphocyte count of 900/µl (range: 370-2560/µl and 130-2770/µl, respectively). After 12 months median lymphocyte count was 1256/µl (range: 240-4140/µl). Median lymphocyte count at 1, 3, 6, and 9 months post-treatment was significantly lower than baseline but showed recovery by the 12 months. This finding is crucial for MCL patients considering CAR-T cell therapy, suggesting a minimum 9-month interval between bendamustine administration and leukapheresis.

Paradigm Shift of Interventional Strategies and Outcomes for Acute Limb Ischemia Post-Pandemic.

PURPOSE: We performed a large-scale comparison of patients treated for acute limb ischemia (ALI) in the pre-COVID (2017-2019) and COVID (2020-2022) eras to evaluate changes in interventional strategies and compare factors associated with adverse outcomes. We sought to characterize patient outcomes in an evolving ALI treatment algorithm in response to pandemic-associated presentation delays and rapid technological advancements in mechanical thrombectomy (MT). METHODS: Using the TriNetX global research network, we conducted a multicenter query across 80 health care organizations (HCOs) spanning 4 countries for patients treated for ALI. Propensity score matching was performed to account for comorbidities. Risk of adverse outcomes within 30 days was calculated for each era, including re-intervention (RI30), major/minor amputation, and death. Patients were then stratified by initial intervention: open revascularization (OR), MT, or catheter-directed thrombolysis and adjunctive endovascular procedures alone (CDT/EP). Risk of adverse outcomes was compared between treatment groups of the same era. RESULTS: After propensity score matching, the pre-COVID era and COVID era cohorts included 7344 patients each. COVID era patients experienced a statistically significant higher risk of 30-day mortality (RR=1.211, p=0.027). Mechanical thrombectomy interventions were performed more frequently in the COVID era (RR=1.314, p<0.0001). Comparing outcomes between treatment groups, MT patients required RI30 more than OR patients (pre-COVID: RR=2.074, p=0.006; COVID: RR=1.600, p=0.025). Open revascularization patients had higher 30-day mortality (pre-COVID: RR=2.368, p<0.0001; COVID: RR=2.013, p<0.0001) and major amputations (pre-COVID: RR=2.432, p<0.0001; COVID: RR=2.176, p<0.0001) than CDT/EP. Pre-COVID CDT/EP patients were at higher risk for RI30 (RR=1.449, p=0.005) and minor amputations (RR=1.500, p=0.010) than OR. The MT group had higher major amputation rates than CDT/EP (pre-COVID: RR=2.043, p=0.019; COVID: RR=1.914, p=0.007). COVID-era MT patients had greater 30-day mortality (RR=1.706, p=0.031) and RI30 (RR=1.544, p=0.029) than CDT/EP. CONCLUSION: Significant shifts toward an MT-based approach have been observed in the last 3 years. Although MT required more RI30 than OR, there was no associated consequence of mortality and limb salvage. The increased mortality seen among COVID-era patients could be explained by delayed presentation, as well as poorly understood pro-thrombogenic or pro-inflammatory mechanisms related to the first waves of COVID. More research is necessary to determine an optimal treatment algorithm. CLINICAL IMPACT: Comorbid risk factors and severity of ischemia must be carefully considered before selecting an interventional strategy to prevent adverse outcomes and maximize limb salvage. Open revascularization strategies are associated with increased mortality and limb loss compared to less-invasive thrombolytic therapy alone. Mechanical thrombectomy (MT)-based approaches have been increasingly used in the last 3 years. Patients receiving MT are more likely to require reintervention within 30 days.

Study of the influence of tributyrin-supplemented diets on the gut bacterial communities of rainbow trout (Oncorhynchus mykiss).

Dietary supplementation with triglyceride tributyrin (TBT), a butyrate precursor, has been associated with beneficial effects on fish health and improvements in the ability of carnivorous fish to tolerate higher levels of plant-based protein. In this study, we aimed to investigate the effects of a plant-based diet supplemented with TBT on the structural diversity and putative function of the digesta-associated bacterial communities of rainbow trout (Oncorhynchus mykiss). In addition to this, we also assessed the response of fish gut digestive enzyme activities and chyme metabolic profile in response to TBT supplementation. Our results indicated that TBT had no significant effects on the overall fish gut bacterial communities, digestive enzyme activities or metabolic profile when compared with non-supplemented controls. However, a more in-depth analysis into the most abundant taxa showed that diets at the highest TBT concentrations (0.2% and 0.4%) selectively inhibited members of the Enterobacterales order and reduced the relative abundance of a bacterial population related to Klebsiella pneumoniae, a potential fish pathogen. Furthermore, the predicted functional analysis of the bacterial communities indicated that increased levels of TBT were associated with depleted KEGG pathways related to pathogenesis. The specific effects of TBT on gut bacterial communities observed here are intriguing and encourage further studies to investigate the potential of this triglyceride to promote pathogen suppression in the fish gut environment, namely in the context of aquaculture.

[Utility of the medial region of pro-adrenomodulin for the detection of true bacteremia in elderly patients treated in the emergency department for suspected infection]. / Utilidad de la región medial de la pro-adrenomodulina para la detección de bacteriemia verdadera en pacientes mayores atendidos en urgencias por sospecha de infección.

OBJECTIVE: The prediction of bacteremia in the emergency department (ER) is important for initial decision-making. The elderly population is a diagnosis challenge. The objective was to evaluate the accuracy of mid regional pro-adrenomedullin (MR-proADM) to identify true bacteremia (BV) in elderly patients attended in 3 hospital emergency departments. METHODS: Observational study including patients ≥75 years of age or older attended in the ER for suspected infection in whom a blood culture (BC) was extracted. Sociodemographic, comorbidity, hemodynamic and analytical variables, biomarkers [MR-proADM, procalcitonin (PCT), C-reactive protein (CRP) and lactate] and final diagnosis were collected. The primary outcome was a true positive on a blood culture. RESULTS: A total of 109 patients with a mean age of 83 (SD: 5.5) years were included. A final diagnosis of BV was obtained in 22 patients (20.2%). The independent variables to predict it were PCT (OR: 13.9; CI95%: 2.702-71.703; p=0.002), MR-proADM (OR: 4.081; CI95%: 1.026-16.225; p=0.046) and temperature (OR: 2.171; CI95%: 1.109-4.248; p=0.024). Considering the cut-off point for MR-proADM (2.13 mg/dl), a sensitivity (Se) of 73%, specificity (E) of 71%, a positive predictive value (PPV) of 39%, a negative predictive value (NPV) of 91%, a positive likelihood ratio (LHR+) of 2.53 and a negative likelihood ratio (LHR-) of 0.38; for PCT (0.76 mg/dl) a Se of 90%, E of 65%, PPV of 40%, NPV of 96%, LHR+ 2,64 and a LHR- of 0.14 were obtained. When combining both, a Se of 69%, E of 84%, PPV of 52%, NPV of 91%, LHR+ of 4.24 and LHR- of 0.38 were observed. CONCLUSIONS: Elevated levels of PCT and MR-proADM were independently associated with an increased risk of BV and the combination of both improves the accuracy to identify these patients.

Sex-Related Disparities in Acute Limb Ischemia Treatment Outcomes.

BACKGROUND: Although a substantial impetus behind disparities research in healthcare exists, those that are sex-related within vascular surgery outcomes are largely unexplored. Consequently, published guidelines lack specificity when it comes to treating male and female patients with vascular disease. Disparities related to patients suffering from chronic limb-threatening ischemia have been broached, although no extensive studies assessing disparities in acute limb ischemia treatment outcomes have come to the forefront. In this study, our aim is to identify and quantify sex-related disparities as they pertain to interventions for acute limb ischemia. METHODS: Using the TriNetX global research network, we conducted a multicenter query across 48 healthcare organizations spanning 5 countries for patients treated for acute limb ischemia. We determined the number of male and female patients that received one of the following interventions: open revascularization, percutaneous mechanical thrombectomy, or catheter-directed thrombolysis and/or adjunctive endovascular procedures. Propensity score matching was performed to account for comorbidities. Risk of adverse outcomes within 30 days was calculated for each sex, including reintervention, major amputation, and death. Risk of adverse outcomes was then compared between treatment groups of the same sex and between sexes. Type-I errors were reduced through utilization of the Holm-Bonferroni method to correct P values. RESULTS: Within our study, we noted several important findings. Females were more likely to receive catheter-directed thrombolysis and/or adjunctive endovascular procedures (P = 0.001) than males. There were no significant differences in the rates of open revascularization or percutaneous mechanical thrombectomy between males and females. Overall, females were more likely to die within 30 days (P < 0.0001) and greater number of males required reintervention within 30 days (P < 0.0001). Analyzing outcomes within individual treatment groups, females undergoing open revascularization or catheter-directed thrombolysis and/or adjunctive endovascular intervention demonstrated a profound increase in mortality within 30 days of intervention (P = 0.0072 and P = 0.0206, respectively), but these differences were not reflected in the percutaneous mechanical thrombectomy group. Limb salvage rates in females were higher than males overall although there were no significant sex differences within any treatment groups specifically. CONCLUSIONS: In conclusion, there was a significantly higher risk of death in females across all treatment groups in the studied timeframe. Limb salvage rates were higher for females in the open revascularization (OR) treatment group, while males were more likely to require a reintervention across all treatment groups. By evaluating these disparities, we can provide greater insight into personalized treatment for patients presenting with acute limb ischemia.

Effect of Grape Pomace Flour in Savory Crackers: Technological, Nutritional and Sensory Properties.

The wine industry generates large amounts of by-products that are usually destined as waste. Grape pomace is the residue of the winemaking process and is rich in compounds with functional properties, such as dietary fiber and phenolic compounds. The aim of this research was to study the influence of white and red grape pomace flour (GPF) addition in the enhancement of functional properties of savory crackers. Different levels of incorporation were tested (5%, 10% and 15% (w/w)). Analysis of physical properties, nutritional composition and sensory acceptability were conducted to evaluate the effect of GPF incorporation. GPF cracker stability throughout a four-week period was achieved with regard to firmness and color. These products presented distinctive and appealing colors, ranging from a violet (GPF of Touriga Nacional variety) to a brown hue (GPF of Arinto variety). Concerning nutritional composition, both crackers incorporated with 10% GPF of Arinto or Touriga Nacional varieties could be considered "high in fiber", as per the Regulation (EC) No. 1924/2006, suggesting a functional food. GPF crackers demonstrated an overall great acceptance of this kind of innovative foods, with the majority indicating that they would certainly/probably buy them. Moreover, the cracker with 10% Arinto GPF achieved the most balanced and overall preference.

Medical and Personal Characteristics Can Predict the Risk of Lung Metastasis.

AIMS: Understanding the correlations between underlying medical and personal characteristics of a patient with cancer and the risk of lung metastasis may improve clinical management and outcomes. We used machine learning methodologies to predict the risk of lung metastasis using readily available predictors. MATERIALS AND METHODS: We retrospectively analysed a cohort of 11 164 oncological patients, with clinical records gathered between 2000 and 2020. The input data consisted of 94 parameters, including age, body mass index (BMI), sex, social history, 81 primary cancer types, underlying lung disease and diabetes mellitus. The strongest underlying predictors were discovered with the analysis of the highest performing method among four distinct machine learning methods. RESULTS: Lung metastasis was present in 958 of 11 164 oncological patients. The median age and BMI of the study population were 63 (±19) and 25.12 (±5.66), respectively. The random forest method had the most robust performance among the machine learning methods. Feature importance analysis revealed high BMI as the strongest predictor. Advanced age, smoking, male gender, alcohol dependence, chronic obstructive pulmonary disease and diabetes were also strongly associated with lung metastasis. Among primary cancers, melanoma and renal cancer had the strongest correlation. CONCLUSIONS: Using a machine learning-based approach, we revealed new correlations between personal and medical characteristics of patients with cancer and lung metastasis. This study highlights the previously unknown impact of predictors such as obesity, advanced age and underlying lung disease on the occurrence of lung metastasis. This prediction model can assist physicians with preventive risk factor control and treatment strategies.

Managing False Lumen Patency in Chronic Thoracic Aortic Dissecting Aneurysms With Coil Embolization.

Thoracic endovascular aortic repair has become the preferred modality of treatment of complicated type B aortic dissections. However, persistent pressurization of the false lumen can lead to negative aortic remodeling with aneurysmal dilation. Described herein is the coil embolization technique that can be used to manage this complication and a review of the literature on the recent development of management options.

The use of patient-reported outcome and experience measures for health policy purposes: A scoping review in oncology.

The systematic use of patient-reported measures (PRMs) [i.e., patient-reported outcome measures (PROMs) and patient-reported experience measures (PREMs)] is advocated as an effective way to improve care practices. However, whether PRMs can lead to the performance assessment of healthcare organisations (HCOs) through valid quality indicators (QIs) for national purposes (i.e., public reporting and paying for performance) is open to debate. This study undertakes a scoping review to examine the use of PRMs as QIs for health policy purposes and to identify the challenges faced in the emblematic case of oncology. According to PRISMA guidelines, published papers, websites and reports published by national and international initiatives were analysed using five online databases (Web of Science, Scopus, PubMed, JSTOR and Google Advanced Search), and then studied using the same keywords. We selected 61 articles and 19 websites/reports and identified 29 PREMs and 48 PROMs from 14 countries and two international initiatives that routinely used them as QIs for HCOs' comparisons. Four types of barriers to this specific use were identified relating to the definition of a standard set, scientific soundness, data collection, and the actionability of such measures. Despite current developments, different barriers still must be overcome before PRMs can be used for health policy purposes in oncology. Future research is needed to ensure that valid QIs related to PRMs are applied at a national level.

Endovascular Aorto-Iliac Reconstruction vs. Aortobifemoral Bypass as First Choice for a Durable Revascularization for Aorto-Iliac Occlusive Disease.

INTRODUCTION: Aortoiliac occlusive disease (AIOD) is defined as atherosclerotic obstruction of the distal abdominal aorta and iliac arteries. Multiple options exist for management of AIOD including endovascular aortoiliac reconstruction and aortobifemoral bypass. As new technologies are developed, the best approach to manage this condition is evolving. METHODS: We performed a literature review to assess the current state of endovascular aortoiliac reconstruction and aortobifemoral bypass as options for revascularization of aortoiliac occlusive disease. CONCLUSION: Endovascular aortoiliac reconstruction and aortobifemoral bypass are both feasible and clinically effective options for management of aortoiliac occlusive disease. No randomized controlled trial has been performed to show one option to be more effective than the other. Recent literature demonstrates comparable long-term patency, limb salvage and survival among endovascular approaches to the treatment of AIOD with quicker recovery, lower costs and improved quality of life when compared to open aortobifemoral bypass (ABF) surgery. Selection of procedure should be tailored to the individual patient in order to develop an effective long-term successful strategy for management of aortoiliac occlusive disease. Further study is warranted to define durability of these endovascular approaches as well as patient specific characteristics that influence outcomes.

Spatially resolved diagnostics for optimization of large ion beam sources.

Giant negative ion sources for neutral beam injectors deliver huge negative ion currents, thanks to their multi-beamlet configuration. As the single-beamlet optics defines the transmission losses along the beamline, the extraction of a similar current for all beamlets is extremely desirable, in order to facilitate the beam source operation (i.e., around perveance match). This Review investigates the correlation between the vertical profile of beam intensity and the vertical profiles of plasma properties at the extraction region of the source, focusing on the influence of increasing cesium injection. Only by the combined use of all available source diagnostics, described in this Review, can beam features on the scale of the non-uniformities be investigated with a sufficient space resolution. At RF power of 50 kW/driver, with intermediate bias currents and a filter field of 2.4 mT, it is found that the central part of the four vertical beam segments exhibits comparable plasma density and beamlet currents; at the edges of the central segments, both the beam and electron density appear to decrease (probably maintaining fixed electron-to-ion ratio); at the bottom of the source, an increase of cesium injection can compensate for the vertical drifts that cause a much higher presence of electrons and a lower amount of negative ions.

Nitrogen dioxide pollution increases vulnerability to COVID-19 through altered immune function.

Previous ecological studies suggest the existence of possible interplays between the exposure to air pollutants and SARS-CoV-2 infection. Confirmations at individual level, however, are lacking. To explore the relationships between previous exposure to particulate matter < 10 µm (PM10) and nitrogen dioxide (NO2), the clinical outcome following hospital admittance, and lymphocyte subsets in COVID-19 patients with pneumonia. In 147 geocoded patients, we assessed the individual exposure to PM10 and NO2 in the 2 weeks before hospital admittance. We divided subjects according to the clinical outcome (i.e., discharge at home vs in-hospital death), and explored the lymphocyte-related immune function as an index possibly affecting individual vulnerability to the infection. As compared with discharged subjects, patients who underwent in-hospital death presented neutrophilia, lymphopenia, lower number of T CD45, CD3, CD4, CD16/56 + CD3 + , and B CD19 + cells, and higher previous exposure to NO2, but not PM10. Age and previous NO2 exposure were independent predictors for mortality. NO2 concentrations were also negatively related with the number of CD45, CD3, and CD4 cells. Previous NO2 exposure is a co-factor independently affecting the mortality risk in infected individuals, through negative immune effects. Lymphopenia and altered lymphocyte subsets might precede viral infection due to nonmodifiable (i.e., age) and external (i.e., air pollution) factors. Thus, decreasing the burden of air pollutants should be a valuable primary prevention measure to reduce individual susceptibility to SARS-CoV-2 infection and mortality.

C-reactive protein but not procalcitonin may predict antibiotic response and outcome in infections following major abdominal surgery.

We aimed to evaluate the usefulness of C-reactive protein (CRP) and procalcitonin (PCT) as markers of infection, sepsis and as predictors of antibiotic response after non-emergency major abdominal surgery. We enrolled, from June 2015 to June 2019, all patients who underwent surgery due to abdominal infection (peritoneal abscess, peritonitis) or having sepsis episode after surgical procedures (i.e. hepatectomy, bowel perforation, pancreaticoduodenectomy (PD), segmental resection of the duodenum (SRD) or biliary reconstruction in a Tertiary Care Hospital. Serum CRP (cut-off value < 5 mg/L) and PCT (cut-off value < 0.1mcg/L) were measured in the day when fever was present or within 24 h after abdominal surgery. Both markers were assessed every 48 h to follow-up antibiotic response and disease evolution up to disease resolution. We enrolled a total of 260 patients underwent non-emergency major abdominal surgery and being infected or developing infection after surgical procedure with one or more microbes (55% mixed Gram-negative infection including Klebsiella KPC, 35% Gram-positive infection, 10% with Candida infection), 58% of patients had ICU admission for at least 96 h, 42% of patients had fast track ICU (48 h). In our group of patients, we found that PCT had a trend to increase after surgical procedure; particularly, those undergoing liver surgery had higher PCT than those underwent different abdominal surgery (U Mann-Whitney p < 0.05). CRP rapidly increase after surgery in those developing infection and showed a statistical significant decrease within 48 h in those subject being responsive to antibiotic treatment and having a clinical response within 10 days independently form the pathogens (bacterial or fungal). Further we found that those having CRP higher than 250 mg/L had a reduced percentage of success treatment at 10 days compared to those < 250 mg/mL (U Mann-Whitney p < 0.05). PCT did not show any variation according to treatment response. CRP in our cohort seems to be a useful marker to predict antibiotic response in those undergoing non-emergency abdominal surgery, while PCT seem to be increased in those having major liver surgery, probably due to hepatic production of cytokines.

Single molecule DNA origami nanoarrays with controlled protein orientation.

The nanoscale organization of functional (bio)molecules on solid substrates with nanoscale spatial resolution and single-molecule control-in both position and orientation-is of great interest for the development of next-generation (bio)molecular devices and assays. Herein, we report the fabrication of nanoarrays of individual proteins (and dyes) via the selective organization of DNA origami on nanopatterned surfaces and with controlled protein orientation. Nanoapertures in metal-coated glass substrates were patterned using focused ion beam lithography; 88% of the nanoapertures allowed immobilization of functionalized DNA origami structures. Photobleaching experiments of dye-functionalized DNA nanostructures indicated that 85% of the nanoapertures contain a single origami unit, with only 3% exhibiting double occupancy. Using a reprogrammed genetic code to engineer into a protein new chemistry to allow residue-specific linkage to an addressable ssDNA unit, we assembled orientation-controlled proteins functionalized to DNA origami structures; these were then organized in the arrays and exhibited single molecule traces. This strategy is of general applicability for the investigation of biomolecular events with single-molecule resolution in defined nanoarrays configurations and with orientational control of the (bio)molecule of interest.