Liquid-liquid phase separation and fibrillation of the prion protein modulated by a high-affinity DNA aptamer.

Structural conversion of cellular prion protein (PrPC) into scrapie PrP (PrPSc) and subsequent aggregation are key events associated with the onset of transmissible spongiform encephalopathies (TSEs). Experimental evidence supports the role of nucleic acids (NAs) in assisting this conversion. Here, we asked whether PrP undergoes liquid-liquid phase separation (LLPS) and if this process is modulated by NAs. To this end, two 25-mer DNA aptamers, A1 and A2, were selected against the globular domain of recombinant murine PrP (rPrP90-231) using SELEX methodology. Multiparametric structural analysis of these aptamers revealed that A1 adopts a hairpin conformation. Aptamer binding caused partial unfolding of rPrP90-231 and modulated its ability to undergo LLPS and fibrillate. In fact, although free rPrP90-231 phase separated into large droplets, aptamer binding increased the number of droplets but noticeably reduced their size. Strikingly, a modified A1 aptamer that does not adopt a hairpin structure induced formation of amyloid fibrils on the surface of the droplets. We show here that PrP undergoes LLPS, and that the PrP interaction with NAs modulates phase separation and promotes PrP fibrillation in a NA structure and concentration-dependent manner. These results shed new light on the roles of NAs in PrP misfolding and TSEs.

Effect of pH on the Complex Coacervation and on the Formation of Layers of Sodium Alginate and PDADMAC.

In this study, we investigated the thermodynamic features of a system based on oppositely charged polyelectrolytes, sodium alginate, and poly(diallyldimethylammonium chloride) (PDADMAC) at different pH values. Additionally, a comparison of the effects of the thermodynamic parameters on the growth of the layers based on the same polymers is presented. For this investigation, different techniques were combined to compare results from the association in solution and coassembled layers at the silicon surface. Dynamic light scattering (DLS) and isothermal titration calorimetry (ITC) were used for studies in solution, and the layer-by-layer technique was employed for the preparation of the polymer layers. Ellipsometry and atomic force microscopy (AFM) were used to characterize the layer thickness growth as a function of the solution pH, and interferometric confocal microscopy was employed to analyze the topography and roughness of the films. The titration of both polyelectrolytes in two different sequences of additions confirmed the mechanism; it involved a two-step process that was monitored by varying the enthalpy, as determined by ITC experiments, and the structural evolution of the aggregates into coacervates, according to DLS. The primary process is aggregation to form polyelectrolyte complexes having a smaller hydrodynamic diameter, which abruptly transit toward a secondary process because of the formation of coacervate particles that have a larger hydrodynamic diameter. Independent of pH and the sequence of addition, for the first process, both directions are entropically driven. However, the binding enthalpy (ΔHb) decreased with a decrease in the pH of the solution. The layers grown for the PDADMAC/sodium alginate system demonstrated pH sensitivity with either linear or exponential behavior, depending on the pH values of the polyelectrolyte solutions. The more endothermic process at pH 10 afforded layers with a smaller thickness and with linear growth according to the increase in the number of layers from 5 to 20. Decreases in the pH of the solution resulted in the layers growing exponentially; additionally, a decrease in the ΔHb of the association in the solution was observed. The layer thicknesses measured using ellipsometry and AFM data were in good agreement. Additionally, the influence of pH on the roughness and topography of the films was observed. Films from basic dipping solutions resulted in surfaces that were more homogeneous with less roughness; in contrast, films with more layers and those formed in a low-pH dipping solution were rougher and less homogeneous.

Social and economic value of Portuguese community pharmacies in health care.

BACKGROUND: Community pharmacies are major contributors to health care systems across the world. Several studies have been conducted to evaluate community pharmacies services in health care. The purpose of this study was to estimate the social and economic benefits of current and potential future community pharmacies services provided by pharmacists in health care in Portugal. METHODS: The social and economic value of community pharmacies services was estimated through a decision-model. Model inputs included effectiveness data, quality of life (QoL) and health resource consumption, obtained though literature review and adapted to Portuguese reality by an expert panel. The estimated economic value was the result of non-remunerated pharmaceutical services plus health resource consumption potentially avoided. Social and economic value of community pharmacies services derives from the comparison of two scenarios: "with service" versus "without service". RESULTS: It is estimated that current community pharmacies services in Portugal provide a gain in QoL of 8.3% and an economic value of 879.6 million euros (M€), including 342.1 M€ in non-remunerated pharmaceutical services and 448.1 M€ in avoided expense with health resource consumption. Potential future community pharmacies services may provide an additional increase of 6.9% in QoL and be associated with an economic value of 144.8 M€: 120.3 M€ in non-remunerated services and 24.5 M€ in potential savings with health resource consumption. CONCLUSIONS: Community pharmacies services provide considerable benefit in QoL and economic value. An increase range of services including a greater integration in primary and secondary care, among other transversal services, may add further social and economic value to the society.

Unraveling Prion Protein Interactions with Aptamers and Other PrP-Binding Nucleic Acids.

Transmissible spongiform encephalopathies (TSEs) are a group of neurodegenerative disorders that affect humans and other mammals. The etiologic agents common to these diseases are misfolded conformations of the prion protein (PrP). The molecular mechanisms that trigger the structural conversion of the normal cellular PrP (PrPC) into the pathogenic conformer (PrPSc) are still poorly understood. It is proposed that a molecular cofactor would act as a catalyst, lowering the activation energy of the conversion process, therefore favoring the transition of PrPC to PrPSc. Several in vitro studies have described physical interactions between PrP and different classes of molecules, which might play a role in either PrP physiology or pathology. Among these molecules, nucleic acids (NAs) are highlighted as potential PrP molecular partners. In this context, the SELEX (Systematic Evolution of Ligands by Exponential Enrichment) methodology has proven extremely valuable to investigate PrP-NA interactions, due to its ability to select small nucleic acids, also termed aptamers, that bind PrP with high affinity and specificity. Aptamers are single-stranded DNA or RNA oligonucleotides that can be folded into a wide range of structures (from harpins to G-quadruplexes). They are selected from a nucleic acid pool containing a large number (1014-1016) of random sequences of the same size (~20-100 bases). Aptamers stand out because of their potential ability to bind with different affinities to distinct conformations of the same protein target. Therefore, the identification of high-affinity and selective PrP ligands may aid the development of new therapies and diagnostic tools for TSEs. This review will focus on the selection of aptamers targeted against either full-length or truncated forms of PrP, discussing the implications that result from interactions of PrP with NAs, and their potential advances in the studies of prions. We will also provide a critical evaluation, assuming the advantages and drawbacks of the SELEX (Systematic Evolution of Ligands by Exponential Enrichment) technique in the general field of amyloidogenic proteins.

Mammalian prion protein (PrP) forms conformationally different amyloid intracellular aggregates in bacteria.

BACKGROUND: An increasing number of proteins are being shown to assemble into amyloid structures that lead to pathological states. Among them, mammalian prions outstand due to their ability to transmit the pathogenic conformation, becoming thus infectious. The structural conversion of the cellular prion protein (PrP(C)), into its misfolded pathogenic form (PrP(Sc)) is the central event of prion-driven pathologies. The study of the structural properties of intracellular amyloid aggregates in general and of prion-like ones in particular is a challenging task. In this context, the evidence that the inclusion bodies formed by amyloid proteins in bacteria display amyloid-like structural and functional properties make them a privileged system to model intracellular amyloid aggregation. RESULTS: Here we provide the first demonstration that recombinant murine PrP and its C-terminal domain (90-231) attain amyloid conformations inside bacteria. Moreover, the inclusions formed by these two PrP proteins display conformational diversity, since they differ in fibril morphology, binding affinity to amyloid dyes, stability, resistance to proteinase K digestion and neurotoxicity. CONCLUSIONS: Overall, our results suggest that modelling PrP amyloid formation in microbial cell factories might open an avenue for a better understanding of the structural features modulating the pathogenic impact of this intriguing protein.

Biophysical and morphological studies on the dual interaction of non-octarepeat prion protein peptides with copper and nucleic acids.

Conversion of prion protein (PrP) to an altered conformer, the scrapie PrP (PrP(Sc)), is a critical step in the development of transmissible spongiform encephalopathies. Both Cu(II) and nucleic acid molecules have been implicated in this conversion. Full-length PrP can bind up to six copper ions; four Cu(II) binding sites are located in the octarepeat domain (residues 60-91), and His-96 and His-111 coordinate two additional copper ions. Experimental evidence shows that PrP binds different molecules, resulting in diverse cellular signaling events. However, there is little information about the interaction of macromolecular ligands with Cu(II)-bound PrP. Both RNA and DNA sequences can bind PrP, and this interaction results in reciprocal conformational changes. Here, we investigated the interaction of Cu(II) and nucleic acids with amyloidogenic non-octarepeat PrP peptide models (comprising human PrP residues 106-126 and hamster PrP residues 109-149) that retain His-111 as the copper-anchoring residue. The effect of Cu(II) and DNA or RNA sequences in the aggregation, conformation, and toxicity of PrP domains was investigated at low and neutral pH. Circular dichroism and EPR spectroscopy data indicate that interaction of the PrP peptides with Cu(II) and DNA occurs at pH 7. This dual interaction induces conformational changes in the peptides, modulating their aggregation, and affecting the morphology of the aggregated species, resulting in different cytotoxic effects. These results provide new insights into the role of Cu(II) and nucleic acid sequences in the structural conversion and aggregation of PrP, which are both critical events related to prion pathogenesis.

MedGAN: optimized generative adversarial network with graph convolutional networks for novel molecule design.

Generative Artificial Intelligence can be an important asset in the drug discovery process to meet the demand for novel medicines. This work outlines the optimization and fine-tuning steps of MedGAN, a deep learning model based on Wasserstein Generative Adversarial Networks and Graph Convolutional Networks, developed to generate new quinoline-scaffold molecules from complex molecular graphs, including hyperparameter adjustments and evaluations of drug-likeness attributes such as pharmacokinetics, toxicity, and synthetic accessibility. The best model was capable of generating 25% valid molecules, 62% fully connected, from which 92% were quinolines, 93% were novel, and 95% unique, preserving chirality, atom charge, and favorable drug-like properties while generating 4831 novel quinolines. These results provide valuable insights into how activation functions, optimizers, learning rates, neuron units, molecule size and constitution, and scaffold structure affect the performance of generative models and their potential to create new molecular structures, enhancing deep learning applications in computational drug design.

Tele-Intensive Care Unit Program in Brazil: Implementation and Expansion.

In this scientific report, we aimed to describe the implementation and expansion of a Tele-Intensive Care Unit (Tele-ICU) program in Brazil, highlighting the pillars of success, improvements, and perspectives. Tele-ICU program emerged during the COVID-19 pandemic at the Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo (HCFMUSP), focusing on clinical case discussions and training of health practitioners in public hospitals of the state of São Paulo in Brazil, to support health care professionals for treating COVID-19 patients. The success of implementing this initiative endorsed the project expansion to other five hospitals from different macroregions of the country, leading to the Tele-ICU-Brazil. These projects assisted 40 hospitals, allowing more than 11,500 teleinterconsultations (exchange of medical information between health care professionals using a licensed online platform) and training more than 14,800 health care professionals, reducing mortality and length of hospitalized patients. A segment in telehealth for the obstetrics health care was implemented after detecting these were a susceptible group of patients to COVID-19 severity. As a perspective, this segment will be expanded to 27 hospitals in the country. The Tele-ICU projects reported here were the largest digital health ICU programs ever established in Brazilian National Health System until know. Their results were unprecedented and proved to be crucial for supporting health care professionals nationwide during the COVID-19 pandemic and guide future initiatives in digital health in Brazil's National Health System.

Nonspecific prion protein-nucleic acid interactions lead to different aggregates and cytotoxic species.

A misfolded form of the prion protein (PrP) is the primary culprit in mammalian prion diseases. It has been shown that nucleic acids catalyze the misfolding of cellular PrP into a scrapie-like conformer. It has also been observed that the interaction of PrP with nucleic acids is nonspecific and that the complex can be toxic to cultured cells. No direct correlation has yet been drawn between changes in PrP structure and toxicity due to nucleic acid binding. Here we asked whether different aggregation, stability, and toxicity effects are detected when nonrelated DNA sequences interact with recombinant PrP. Using spectroscopic techniques to analyze PrP tertiary and secondary structure and cellular assays to assess toxicity, we found that rPrP-DNA interactions lead to different aggregated species, depending on the sequence and size of the oligonucleotide tested. A 21-mer DNA sequence (D67) induced higher levels of aggregation and also dissimilar structural changes in rPrP, compared to binding to oligonucleotides with the same length and different nucleotide sequences or different GC contents. The rPrP-D67 complex induced significant cell dysfunction, which appears to be correlated with the biophysical properties of the complex. Although sequence specificity is not apparent for PrP-nucleic acid interactions, we believe that particular nucleic acid patterns, possibly related to GC content, oligonucleotide length, and structure, govern PrP recognition. Understanding the structural and cellular effects observed for PrP-nucleic acid complexes may shed light on the still mysterious pathology of the prion protein.

Simulation-based Assessment to Measure Proficiency in Mechanical Ventilation among Residents.

Background: Mechanical ventilation (MV) skills are essential for clinicians caring for critically ill patients, yet few training programs use structured curricula and appropriate assessments. Objective structured clinical exams (OSCEs) have been used to assess clinical competency in many areas, but there are no OSCE models focused on MV. Objective: To develop and validate a simulation-based assessment (SBA) with an OSCE structure to assess baseline MV competence among residents and identify knowledge gaps. Methods: We developed an SBA using a lung simulator and a mechanical ventilator, and an OSCE structure, with six clinical scenarios in MV. We included internal medicine residents at the beginning of their rotation in the respiratory intensive care unit (ICU) of a university-affiliated hospital. A subset of residents was also evaluated with a validated multiple-choice exam (MCE) at the beginning and at the end of the ICU rotation. Scores on both assessments were normalized to range from 0 to 10. We used Cronbach's α coefficient to assess reliability and Spearman correlation to estimate the correlation between the SBA and the MCE. Results: We included 80 residents, of whom 42 also completed the MCE examinations. The final version of the SBA had 32 items, and the Cronbach's α coefficient was 0.72 (95% confidence interval [CI], 0.64-0.81). The average SBA score was 6.2 ± 1.3, and performance was variable across items, with 80% correctly adjusting initial ventilatory settings and only 12% correctly identifying asynchrony. The MCE had 24 questions, and the average score was 7.6 ± 2.4 at the beginning of the rotation and 8.2 ± 2.3 at the end of the rotation (increase of 0.6 points; 95% CI, 0.30-0.90; P < 0.001). There was moderate correlation between the SBA and the MCE (rho = 0.41; P = 0.002). Conclusion: We developed and validated an objective structured assessment on MV using a pulmonary simulator and a mechanical ventilator addressing the main competencies in MV. The performance of residents in the SBA at the beginning of an ICU rotation was lower than the performance in MCE, highlighting the need for greater emphasis on practical skills in MV during residency.

A multi-intervention protocol to improve sleep quality in a coronary care unit.

AIMS: Poor sleep is a frequent occurrence in the critical illness. Evaluate sleep quality and test the effect of a multi-intervention sleep care protocol in improving sleep quality in a coronary care unit (CCU). METHODS AND RESULTS: Quasi-experimental study, carried out in two phases. During the first phase, the control group (n = 58 patients) received usual care. Baseline sleep data were collected through the Richards-Campbell Sleep Questionnaire (RCSQ) and the Sleep in the Intensive Care Unit Questionnaire (SICUQ). During the second phase (n = 55 patients), a sleep care protocol was implemented. Interventions included actions to promote analgesia, reduce noise, brightness, and other general measures. Sleep data were collected again to assess the impact of these interventions. The intervention group had better scores in overall sleep depth [median (interquartile range)] [81 (65-96.7) vs. 69.7 (50-90); P = 0.046]; sleep fragmentation [90 (65-100) vs. 69 (42.2-92.7); P = 0.011]; return to sleep [90 (69.7-100) vs. 71.2 (40.7-96.5); P = 0.007]; sleep quality [85 (65-100) vs. 71.1 (49-98.1); P = 0.026]; and mean RCSQ score [83 (66-94) vs. 66.5 (45.7-87.2); P = 0.002] than the baseline group. The main barriers to sleep were pain [1 (1.0-5.5)], light [1 (1.0-5.0)], and noise [1 (1.0-5.0)]. The most rated sources of sleep-disturbing noise were heart monitor alarm [3 (1.0-5.25)], intravenous pump alarm [1.5 (1.0-5.00)]. and mechanical ventilator alarm [1 (1.0-5.0)]. All were significantly lower in the intervention group than in the baseline group. CONCLUSION: A multi-intervention protocol was feasible and effective in improving different sleep quality parameters and reducing some barriers to sleep in CCU patients.

Prion protein complexed to a DNA aptamer induce behavioral and synapse dysfunction in mice.

Conversion of the cellular prion protein (PrPC) into the scrapie form (PrPSc) is the leading step to the development of transmissible spongiform encephalopathies (TSEs), still incurable neurodegenerative disorders. Interaction of PrPC with cellular and synthetic ligands that induce formation of scrapie-like conformations has been deeply investigated in vitro. Different nucleic acid (NA) sequences bind PrP and convert it to ß-sheet-rich or unfolded species; among such NAs, a 21-mer double-stranded DNA, D67, was shown to induce formation of PrP aggregates that were cytotoxic. However, in vivo effects of these PrP-DNA complexes were not explored. Herein, aggregates of recombinant full-length PrP (rPrP23-231) induced by interaction with the D67 aptamer were inoculated into the lateral ventricle of Swiss mice and acute effects were investigated. The aggregates had no influence on emotional, locomotor and motor behavior of mice. In contrast, mice developed cognitive impairment and hippocampal synapse loss, which was accompanied by intense activation of glial cells in this brain region. Our results suggest that the i.c.v. injection of rPrP:D67 aggregates is an interesting model to study the neurotoxicity of aggregated PrP in vivo, and that glial cell activation may be an important step for behavioral and cognitive dysfunction in prion diseases.

The interplay between a GC-rich oligonucleotide and copper ions on prion protein conformational and phase transitions.

The prion protein (PrP) misfolding to its infectious form is critical to the development of prion diseases, whereby various ligands are suggested to participate, such as copper and nucleic acids (NA). The PrP globular domain was shown to undergo NA-driven liquid-liquid phase separation (LLPS); this latter may precede pathological aggregation. Since Cu(II) is a physiological ligand of PrP, we argue whether it modulates phase separation altogether with nucleic acids. Using recombinant PrP, we investigate the effects of Cu(II) (at 6 M equivalents) and a previously described PrP-binding GC-rich DNA (equimolarly to protein) on PrP conformation, oligomerization, and phase transitions using a range of biophysical techniques. Raman spectroscopy data reveals the formation of the ternary complex. Microscopy suggests that phase separation is mainly driven by DNA, whereas Cu(II) has no influence. Our results show that DNA can be an adjuvant, leading to the structural conversion of PrP, even in the presence of an endogenous ligand, copper. These results provide new insights into the role of Cu(II) and NA on the phase separation, structural conversion, and aggregation of PrP, which are critical events leading to neurodegeneration.

Implementation of Tele-ICU during the COVID-19 pandemic.

OBJECTIVE: To describe the implementation of a Tele-ICU program during the COVID-19 pandemic, as well as to describe and analyze the results of the first four months of operation of the program. METHODS: This was a descriptive observational study of the implementation of a Tele-ICU program, followed by a retrospective analysis of clinical data of patients with COVID-19 admitted to ICUs between April and July of 2020. RESULTS: The Tele-ICU program was implemented over a four-week period and proved to be feasible during the pandemic. Participants were trained remotely, and the program had an evidence-based design, the objective being to standardize care for patients with COVID-19. More than 100,000 views were recorded on the free online platforms and the mobile application. During the study period, the cases of 326 patients with COVID-19 were evaluated through the program. The median age was 60 years (IQR, 49-68 years). There was a predominance of males (56%). There was also a high prevalence of hypertension (49.1%) and diabetes mellitus (38.4%). At ICU admission, 83.7% of patients were on invasive mechanical ventilation, with a median PaO2/FiO2 ratio < 150. It was possible to use lung-protective ventilation in 75% of the patients. Overall, in-hospital mortality was 68%, and ICU mortality was 65%. CONCLUSIONS: Our Tele-ICU program provided multidisciplinary training to health care professionals and clinical follow-up for hundreds of critically ill patients. This public health care network initiative was unprecedented and proved to be feasible during the COVID-19 pandemic, encouraging the creation of similar projects that combine evidence-based practices, training, and Tele-ICU.

[Characterization of patients with type 1 diabetes mellitus in southern Brazil: chronic complications and associated factors]. / Caracterização de pacientes com diabetes mellitus tipo 1 do sul do Brasil: complicações crônicas e fatores associados.

OBJECTIVE: To evaluate the prevalence of chronic vascular complications and associated factors in patients with type 1 diabetes mellitus (DM). METHODS: Cross sectional study with type 1 DM patients attending the Endocrine Division, Hospital de Clinicas de Porto Alegre. Patients were evaluated for presence of chronic vascular complications. RESULTS: We evaluated 573 patients, mean age of 33 years. The presence of diabetic retinopathy (DR) was observed in 43.3%, diabetes duration [OR: 1.07, 95% CI: 1.03 to 1.11, P <0001], the presence of diabetic nephropathy (DN) [OR: 3.40; CI 95%: 1.89 to 6.13, P <0001] and hypertension (HPT) [OR: 2.12, 95% CI: 1.16 to 3.87, P = 0014] were associated with DR. The DN was present in 34.5% and was associated with HPT [OR: 1.93, 95% CI: 1.16 to 3.21, P = 0001] and total cholesterol [OR: 1.0, 95% CI: 1.0-1.01, P = 0.05]. Seven patients had macrovascular disease. Only 22% achieved an A1c of <7.0%. HPT was 33% and 48% had levels <130/80 mm Hg and 45% of patients had values for LDL> 100 mg/dl. CONCLUSION: We observed a high prevalence of microvascular complications and HPT. Duration of DM, HPT and presence of DN were associated with DR. HPT and dyslipidemia were associated with DN. Most patients did not meet the desired glycemic control, blood pressure and lipid targets. Greater efforts are needed to intensify the pressure and metabolic control of patients with type 1 DM.

Characteristics and outcomes of patients with COVID-19 admitted to the ICU in a university hospital in São Paulo, Brazil - study protocol.

OBJECTIVES: We designed a cohort study to describe characteristics and outcomes of patients with coronavirus disease (COVID-19) admitted to the intensive care unit (ICU) in the largest public hospital in Sao Paulo, Brazil, as Latin America becomes the epicenter of the pandemic. METHODS: This is the protocol for a study being conducted at an academic hospital in Brazil with 300 adult ICU beds dedicated to COVID-19 patients. We will include adult patients admitted to the ICU with suspected or confirmed COVID-19 during the study period. The main outcome is ICU survival at 28 days. Data will be collected prospectively and retrospectively by trained investigators from the hospital's electronic medical records, using an electronic data capture tool. We will collect data on demographics, comorbidities, severity of disease, and laboratorial test results at admission. Information on the need for advanced life support and ventilator parameters will be collected during ICU stay. Patients will be followed up for 28 days in the ICU and 60 days in the hospital. We will plot Kaplan-Meier curves to estimate ICU and hospital survival and perform survival analysis using the Cox proportional hazards model to identify the main risk factors for mortality. ClinicalTrials.gov: NCT04378582. RESULTS: We expect to include a large sample of patients with COVID-19 admitted to the ICU and to be able to provide data on admission characteristics, use of advanced life support, ICU survival at 28 days, and hospital survival at 60 days. CONCLUSIONS: This study will provide epidemiological data about critically ill patients with COVID-19 in Brazil, which could inform health policy and resource allocation in low- and middle-income countries.

Performance of Noninvasive Ventilation Masks in a Lung Model of COPD Exacerbation.

BACKGROUND: Noninvasive ventilation (NIV) reduces intubation and mortality in patients with COPD exacerbation who present with respiratory failure, and the type of mask may affect its success. Our objective was to compare the performance of 3 different NIV masks in a lung model. METHODS: We set the lung simulator mechanics and respiratory rate, and tested a small oronasal mask, a total face mask, and a large oronasal mask. We added CO2 at a constant rate into the system and monitored the end-tidal carbon dioxide. We used a mechanical ventilator to deliver NIV in 8 different combinations of inspiratory effort, pressure support, and expiratory positive airway pressure. We measured end-tidal carbon dioxide mask leakage, tidal volume, trigger time, time to achieve 90% of the inspiratory target during inspiration, and excess inspiratory time. RESULTS: We presented the mean ± SD of the 8 simulated conditions for each mask. The mean ± SD leakage was higher for the total face mask (51 ± 6 L/min) than for the small oronasal mask (37 ± 5 L/min) and for the large oronasal mask (21 ± 3 L/min), P < .001; but end-tidal carbon dioxide and tidal volume were similar. The mean ± SD 90% of the inspiratory target during inspiration was faster for the small oronasal mask (585 ± 49 ms) compared with the large oronasal (647 ± 107 ms) and total face mask (851 ± 105 ms), P < .001, all other variables were similar. CONCLUSIONS: In this model, we found that the type of mask had no impact on CO2 washout or on most synchrony variables.

Reliability of Thyroid Imaging Reporting and Data System (TI-RADS), and ultrasonographic classification of the American Thyroid Association (ATA) in differentiating benign from malignant thyroid nodules.

OBJECTIVE: Ultrasonography (US) is the best diagnostic tool for initial assessment of thyroid nodule. Recently, data reporting systems for thyroid lesions, such as the Thyroid Imaging Reporting and Data System (TI-RADS) and American Thyroid Association (ATA), which stratifies the risk for malignancy, have demonstrated good performance in differentiating malignant thyroid nodules. The purpose of this study is to determine the reliability of both data reporting systems in predicting thyroid malignancy in a tertiary care hospital. MATERIALS AND METHODS: We evaluated 195 thyroid nodules using modified TI-RADS and ATA risk stratification. The results were compared to the cyto-pathology analysis. Histopathological results were available for 45 cases after surgery, which is considered the golden standard for diagnosis of thyroid cancer. RESULTS: When compared with cytological results, sensitivity, specificity, negative predictive value (NPV), and accuracy were 100, 61.1, 100, and 63%, respectively, for TI-RADS; and 100, 75, 100, and 76%, respectively, for ATA. When compared with histopathological results, sensitivity, specificity, NPV, and accuracy were 90, 51.4, 94.7, and 60% respectively, for TI-RADS; and 100, 60, 100, and 68%, respectively, for ATA. All patients with malignant nodules were classified in the categories 4 or 5 of TI-RADS and in the intermediate or high suspicion risk according to the ATA system. CONCLUSION: Both TI-RADS and the ATA guidelines have high sensitivity and NPV for the diagnosis of thyroid carcinoma. These systems are feasible for clinical application, allowing to better select patients to undergo fine-needle aspiration biopsies.

Mammalian prion amyloid formation in bacteria.

Mammalian prion proteins (PrPs) that cause transmissible spongiform encephalopathies are misfolded conformations of the host cellular PrP. The misfolded form, the scrapie PrP (PrP(Sc)), can aggregate into amyloid fibrils that progressively accumulate in the brain, evolving to a pathological phenotype. A particular characteristic of PrP(Sc) is to be found as different strains, related to the diversity of conformational states it can adopt. Prion strains are responsible for the multiple phenotypes observed in prion diseases, presenting different incubation times and diverse deposition profiles in the brain. PrP biochemical properties are also strain-dependent, such as different digestion pattern after proteolysis and different stability. Although they have long been studied, strain formation is still a major unsolved issue in prion biology. The recreation of strain-specific conformational features is of fundamental importance to study this unique pathogenic phenomenon. In our recent paper, we described that murine PrP, when expressed in bacteria, forms amyloid inclusion bodies that possess different strain-like characteristics, depending on the PrP construct. Here, we present an extra-view of these data and propose that bacteria might become a successful model to generate preparative amounts of prion strain-specific assemblies for high-resolution structural analysis as well as for addressing the determinants of infectivity and transmissibility.