Synthesis and structural characterization of new macrocyclic inhibitors of the Zika virus NS2B-NS3 protease.

Three new series of macrocyclic active site-directed inhibitors of the Zika virus (ZIKV) NS2B-NS3 protease were synthesized. First, attempts were made to replace the basic P3 lysine residue of our previously described inhibitors with uncharged and more hydrophobic residues. This provided numerous compounds with inhibition constants between 30 and 50 nM. A stronger reduction of the inhibitory potency was observed when the P2 lysine was replaced by neutral residues, all of these inhibitors possess Ki values >1 µM. However, it is possible to replace the P2 lysine with the less basic 3-aminomethylphenylalanine, which provides a similarly potent inhibitor of the ZIKV protease (Ki = 2.69 nM). Crystal structure investigations showed that the P2 benzylamine structure forms comparable interactions with the protease as lysine. Twelve additional structures of these inhibitors in complex with the protease were determined, which explain many, but not all, SAR data obtained in this study. All individual modifications in the P2 or P3 position resulted in inhibitors with low antiviral efficacy in cell culture. Therefore, a third inhibitor series with combined modifications was synthesized; all of them contain a more hydrophobic  d-cyclohexylalanine in the linker segment. At a concentration of 40 µM, two of these compounds possess similar antiviral potency as ribavirin at 100 µM. Due to their reliable crystallization in complex with the ZIKV protease, these cyclic compounds are very well suited for a rational structure-based development of improved inhibitors.

ACE2 acts as a novel regulator of TMPRSS2-catalyzed proteolytic activation of influenza A virus in airway cells.

The transmembrane serine protease 2 (TMPRSS2) activates the outer structural proteins of a number of respiratory viruses including influenza A virus (IAV), parainfluenza viruses, and various coronaviruses for membrane fusion. Previous studies showed that TMPRSS2 interacts with the carboxypeptidase angiotensin-converting enzyme 2 (ACE2), a cell surface protein that serves as an entry receptor for some coronaviruses. Here, by using protease activity assays, we determine that ACE2 increases the enzymatic activity of TMPRSS2 in a non-catalytic manner. Furthermore, we demonstrate that ACE2 knockdown inhibits TMPRSS2-mediated cleavage of IAV hemagglutinin (HA) in Calu-3 human airway cells and suppresses virus titers 100- to 1.000-fold. Transient expression of ACE2 in ACE2-deficient cells increased TMPRSS2-mediated HA cleavage and IAV replication. ACE2 knockdown also reduced titers of MERS-CoV and prevented S cleavage by TMPRSS2 in Calu-3 cells. By contrast, proteolytic activation and multicycle replication of IAV with multibasic HA cleavage site typically cleaved by furin were not affected by ACE2 knockdown. Co-immunoprecipitation analysis revealed that ACE2-TMPRSS2 interaction requires the enzymatic activity of TMPRSS2 and the carboxypeptidase domain of ACE2. Together, our data identify ACE2 as a new co-factor or stabilizer of TMPRSS2 activity and as a novel host cell factor involved in proteolytic activation and spread of IAV in human airway cells. Furthermore, our data indicate that ACE2 is involved in the TMPRSS2-catalyzed activation of additional respiratory viruses including MERS-CoV.IMPORTANCEProteolytic cleavage of viral envelope proteins by host cell proteases is essential for the infectivity of many viruses and relevant proteases provide promising drug targets. The transmembrane serine protease 2 (TMPRSS2) has been identified as a major activating protease of several respiratory viruses, including influenza A virus. TMPRSS2 was previously shown to interact with angiotensin-converting enzyme 2 (ACE2). Here, we report the mechanistic details of this interaction. We demonstrate that ACE2 increases or stabilizes the enzymatic activity of TMPRSS2. Furthermore, we describe ACE2 involvement in TMPRSS2-catalyzed cleavage of the influenza A virus hemagglutinin and MERS-CoV spike protein in human airway cells. These findings expand our knowledge of the activation of respiratory viruses by TMPRSS2 and the host cell factors involved. In addition, our results could help to elucidate a physiological role for TMPRSS2.

An economical in-class sticker microfluidic activity develops student expertise in microscale physics and device manufacturing.

Educating new students in miniaturization science remains challenging due to the non-intuitive behavior of microscale objects and specialized layer-by-layer assembly approaches. In our analysis of the existing literature, we noted that it remains difficult to have low cost activities that elicit deep learning. Furthermore, few activities have stated learning goals and measurements of effectiveness. To that end, we created a new educational activity that enables students to build and test microfluidic mixers, valves, and bubble generators in the classroom setting with inexpensive, widely-available materials. Although undergraduate and graduate engineering students are able to successfully construct the devices, our activity is unique in that the focus is not on successfully building and operating each device. Instead, it is to gain understanding about miniaturization science, device design, and construction so as to be able to do so independently. Our data show that the activity is appropriate for developing the conceptual understanding of graduate and advanced undergraduate students (n = 57), as well as makes a lasting impression on the students. We also report on observations related to student patterns of misunderstanding and how miniaturization science provides a unique opportunity for educational researchers to elicit and study misconceptions. More broadly, since this activity teaches participants a viable approach to creating microsystems and can be implemented in nearly any global setting, our work democratizes the education of miniaturization science. Noting the broad potential of point-of-care technologies in the global setting, such an activity could empower local experts to address their needs.

Memory support training and lifestyle modifications to promote healthy aging in persons at risk for Alzheimer's disease: a digital application supported intervention (Brain Boosters).

BACKGROUND: Evidence-based interventions to protect against cognitive decline among older adults at risk for Alzheimer's disease and related dementias (ADRD) are urgently needed. Rehabilitation approaches to support memory and behavioral/lifestyle interventions are recognized as promising strategies for preserving or improving cognitive health, although few previous interventions have combined both approaches. This paper describes the protocol of the Brain Boosters intervention, which synergistically combines training in compensatory and healthy lifestyle behaviors and supports implementation and tracking of new behaviors with a digital application. METHODS: The study utilizes a single-site, single-blinded, randomized controlled design to compare a structured lifestyle and compensatory aid intervention to an education-only self-guided intervention. We plan to enroll 225 community-dwelling adults (25% from underrepresented groups) aged 65 + who endorse subjective cognitive decline (SCD) and low baseline levels of healthy lifestyle behaviors. Both interventions will be administered in group format, consisting of 15 two-hour classes that occur weekly for ten weeks and taper to bi-monthly and monthly, for an intervention duration of 6 months. Participants in both interventions will receive education about a variety of memory support strategies and healthy lifestyle behaviors, focusing on physical and cognitive activity and stress management. The structured intervention will also receive support in adopting new behaviors and tracking set goals aided by the Electronic Memory and Management Aid (EMMA) digital application. Primary outcomes include global cognition (composite of memory, attention, and executive function tests) and everyday function (Everyday Cognition Questionnaire). Data will be collected at baseline and outcome visits, at approximately 6, 12, and 18 months. Qualitative interviews, self-report surveys (e.g., indicators of self-determination, health literacy) and EMMA data metrics will also be used to identify what components of the intervention are most effective and for whom they work. DISCUSSION: Successful project completion will provide valuable information about how individuals with SCD respond to a compensation and preventative lifestyle intervention assisted by a digital application, including an understanding of factors that may impact outcomes, treatment uptake, and adherence. The work will also inform development, scaling, and personalization of future interventions that can delay disability in individuals at risk for ADRD. TRIAL REGISTRATION: ClinicalTrials.gov. (NCT05027789, posted 8/30/2021).

Repeated toluene and cyclohexane inhalation produces differential effects on HPA and HPT axes in adolescent male rats.

Misused volatile solvents typically contain toluene (TOL) as the main psychoactive ingredient. Cyclohexane (CHX) can also be present and is considered a safer alternative. Solvent misuse often occurs at early stages of life, leading to permanent neurobehavioral impairment and growth retardation. However, a comprehensive examination of the effects of TOL and CHX on stress regulation and energy balance is lacking. Here, we compared the effect of a binge-pattern exposure to TOL or CHX (4,000 or 8,000 ppm) on body weight, food intake, the hypothalamus-pituitary-adrenal (HPA) and hypothalamus-pituitary-thyroid (HPT) axes in male adolescent Wistar rats. At 8,000 ppm, TOL decreased body weight gain without affecting food intake. In addition, TOL and CHX altered the HPA and HPT axes' function in a solvent- and concentration-dependent manner. The highest TOL concentration produced HPA axis hyperactivation in animals not subjected to stress, which was evidenced by increased corticotropin-releasing-factor (CRF) release from the median eminence (ME), elevated adrenocorticotropin hormone (ACTH) and corticosterone serum levels, and decreased CRF mRNA levels in the hypothalamic paraventricular nucleus (PVN). TOL (8,000 ppm) also increased triiodothyronine (T3) serum levels, decreased pro-thyrotropin-releasing-hormone (pro-TRH) mRNA transcription in the PVN, pro-TRH content in the ME, and serum thyroid stimulating hormone (TSH) levels. CHX did not affect the HPA axis. We propose that the increased HPT axis activity induced by TOL can be related to the impaired body weight gain associated with inhalant misuse. These findings may contribute to a better understanding of the effects of the misused solvents TOL and CHX.

In vitro testing of host-targeting small molecule antiviral matriptase/TMPRSS2 inhibitors in 2D and 3D cell-based assays.

The outbreak of coronavirus disease 2019 (COVID-19) pandemic strongly stimulated the development of small molecule antivirals selectively targeting type II transmembrane serine proteases (TTSP), required for the host-cell entry of numerous viruses. A set of 3-amidinophenylalanine derivatives (MI-21, MI-472, MI-477, MI-485, MI-1903 and MI-1904), which inhibit the cleavage of certain viral glycoproteins was characterized in 2D and 3D primary human hepatocyte models on collagen- and Matrigel-coating using a CCK-8 assay to evaluate their cytotoxicity, a resorufin-based method to detect redox imbalances, fluorescence and ultrafiltration experiments to evaluate their interactions with human serum albumin (HSA) and α-acidic glycoprotein (AGP), and luminescence measurement to assess CYP3A4 modulation. For elucidation of selectivity of the applied compounds towards matriptase, transmembrane serine protease 2 (TMPRRS2), thrombin and factor Xa (FXa) Ki values were determined. It was proven that cell viability was only deteriorated by inhibitor MI-1903, and redox status was not influenced by administration of the selected inhibitors at 50 µM for 24 h. MI-472 and MI-477 formed relatively stable complexes with AGP. CYP3A4 inhibition was found to be strong in PHHs exposed to all inhibitors with the exception of MI-21, which seems to be a promising drug candidate also due to its better selectivity towards matriptase and TMPRSS2 over the blood clotting proteases thrombin and FXa. Our in vitro pharmacokinetic screening with these inhibitors helps to select the compounds with the best selectivity and safety profile suitable for a further preclinical characterization without animal sacrifice.

Reducción cerrada y fijación intramedular de fracturas del cuello del quinto metacarpiano: técnica retrógrada vs. anterógrada. Estudio prospectivo / Closed reduction and intramedullary fixation of fifth metacarpal neck fractures: Retrograde vs. anterograde technique. Prospective study

Introducción: La fractura de quinto metacarpiano es una dolencia muy frecuente que puede ocasionar deformidad y afectación funcional para la prensión adecuada de la mano. La reinserción a las actividades cotidianas o laborales se relaciona con el tratamiento recibido y la rehabilitación. En fracturas de cuello de quinto metacarpiano la fijación interna con aguja Kirschner es un método de tratamiento convencional con variantes que afectan su desenlace. Objetivo: Comparar los resultados funcionales y clínicos del tratamiento de las fracturas de quinto metacarpiano con el uso de agujas Kirschner vía retrógrada versus anterógrada. Material y métodos: Estudio comparativo, longitudinal, prospectivo, realizado en un hospital de tercer nivel de Traumatología, en pacientes con fractura de cuello de quinto metacarpiano, con seguimiento clínico, radiográfico y con escala Quick DASH a la tercera, sexta y octava semana postoperatoria. Resultados: Se incluyeron 60 pacientes (58 hombres, 2 mujeres), con un promedio de edad de 29,63±10,15 años, con fractura de quinto metacarpiano, tratados mediante reducción cerrada y estabilización con aguja Kirschner. La vía anterógrada mostró un rango de flexión metacarpofalángica a las 8 semanas de 89,11 grados (p<0,001; IC95% [−26.81; −11,42]), un valor de la escala DASH de 18,17 (p<0,001; IC95% [23,45; 39,12]) y un promedio de 27,35 días de incapacidad laboral (p=0,002; IC95% [16,22; 62,14]), comparada con la vía retrógrada. Conclusión: La estabilización con aguja Kirschner vía anterógrada mostró superioridad en resultados funcionales y en amplitud de movimiento metacarpofalángica comparados con los operados por vía retrógrada a la octava semana del postoperatorio.(AU)

Introduction: The fifth metacarpal fracture is a frequent pathology that can cause deformity and functional impairment for the adequate grip of the hand. Reintegration into daily or working activities is related to the treatment received and rehabilitation. In fractures of the neck of the fifth metacarpal, internal fixation with a Kirschner's wire is a conventional treatment method with variants that affect its outcome. Aim of the study: To compare the functional and clinical results of the treatment of fifth metacarpal fracture with the use of retrograde versus antegrade Kirschner wires. Material and methods: Comparative, longitudinal, prospective study at a third-level trauma center in patients with a fifth metacarpal neck fracture, with clinical, radiographic and Quick DASH scale follow-up at the 3rd, 6th, and 8th postoperative week. Results: Sixty patients were included (58 men, 2 women), age of 29.63±10.15 years, with a fifth metacarpal fracture, treated by closed reduction and stabilization with a Kirschner wire. The antegrade approach showed a metacarpophalangeal flexion range at 8 weeks of 89.11° (p<0.001; 95% CI [−26.81; −11.42]), a DASH scale value of 18.17 (p<0.001; 95% CI [23.45; 39.12]), and an average of 27.35 days to return to work (p=0.002; 95% CI [16.22; 62.14]), compared with the retrograde approach. Conclusion: Stabilization with antegrade Kirschner wire showed superior functional results, and metacarpophalangeal range of motion, compared to those operated via retrograde approach.(AU)

[Translated article] Closed reduction and intramedullary fixation of fifth metacarpal neck fractures: Retrograde vs. anterograde technique. Prospective study / Reducción cerrada y fijación intramedular de fracturas del cuello del quinto metacarpiano: técnica retrógrada vs. anterógrada. Estudio prospectivo

Introducción: La fractura de quinto metacarpiano es una dolencia muy frecuente que puede ocasionar deformidad y afectación funcional para la prensión adecuada de la mano. La reinserción a las actividades cotidianas o laborales se relaciona con el tratamiento recibido y la rehabilitación. En fracturas de cuello de quinto metacarpiano la fijación interna con aguja Kirschner es un método de tratamiento convencional con variantes que afectan su desenlace. Objetivo: Comparar los resultados funcionales y clínicos del tratamiento de las fracturas de quinto metacarpiano con el uso de agujas Kirschner vía retrógrada versus anterógrada. Material y métodos: Estudio comparativo, longitudinal, prospectivo, realizado en un hospital de tercer nivel de Traumatología, en pacientes con fractura de cuello de quinto metacarpiano, con seguimiento clínico, radiográfico y con escala Quick DASH a la tercera, sexta y octava semana postoperatoria. Resultados: Se incluyeron 60 pacientes (58 hombres, 2 mujeres), con un promedio de edad de 29,63±10,15 años, con fractura de quinto metacarpiano, tratados mediante reducción cerrada y estabilización con aguja Kirschner. La vía anterógrada mostró un rango de flexión metacarpofalángica a las 8 semanas de 89,11 grados (p<0,001; IC95% [−26.81; −11,42]), un valor de la escala DASH de 18,17 (p<0,001; IC95% [23,45; 39,12]) y un promedio de 27,35 días de incapacidad laboral (p=0,002; IC95% [16,22; 62,14]), comparada con la vía retrógrada. Conclusión: La estabilización con aguja Kirschner vía anterógrada mostró superioridad en resultados funcionales y en amplitud de movimiento metacarpofalángica comparados con los operados por vía retrógrada a la octava semana del postoperatorio.(AU)

Introduction: The fifth metacarpal fracture is a frequent pathology that can cause deformity and functional impairment for the adequate grip of the hand. Reintegration into daily or working activities is related to the treatment received and rehabilitation. In fractures of the neck of the fifth metacarpal, internal fixation with a Kirschner's wire is a conventional treatment method with variants that affect its outcome. Aim of the study: To compare the functional and clinical results of the treatment of fifth metacarpal fracture with the use of retrograde versus antegrade Kirschner wires. Material and methods: Comparative, longitudinal, prospective study at a third-level trauma center in patients with a fifth metacarpal neck fracture, with clinical, radiographic and Quick DASH scale follow-up at the 3rd, 6th, and 8th postoperative week. Results: Sixty patients were included (58 men, 2 women), age of 29.63±10.15 years, with a fifth metacarpal fracture, treated by closed reduction and stabilization with a Kirschner wire. The antegrade approach showed a metacarpophalangeal flexion range at 8 weeks of 89.11° (p<0.001; 95% CI [−26.81; −11.42]), a DASH scale value of 18.17 (p<0.001; 95% CI [23.45; 39.12]), and an average of 27.35 days to return to work (p=0.002; 95% CI [16.22; 62.14]), compared with the retrograde approach. Conclusion: Stabilization with antegrade Kirschner wire showed superior functional results, and metacarpophalangeal range of motion, compared to those operated via retrograde approach.(AU)

Opportunities and considerations for studying liver disease with microphysiological systems on a chip.

Advances in microsystem engineering have enabled the development of highly controlled models of the liver that better recapitulate the unique in vivo biological conditions. In just a few short years, substantial progress has been made in creating complex mono- and multi-cellular models that mimic key metabolic, structural, and oxygen gradients crucial for liver function. Here we review: 1) the state-of-the-art in liver-centric microphysiological systems and 2) the array of liver diseases and pressing biological and therapeutic challenges which could be investigated with these systems. The engineering community has unique opportunities to innovate with new liver-on-a-chip devices and partner with biomedical researchers to usher in a new era of understanding of the molecular and cellular contributors to liver diseases and identify and test rational therapeutic modalities.

[Translated article] Closed reduction and intramedullary fixation of fifth metacarpal neck fractures: Retrograde vs. anterograde technique. Prospective study.

INTRODUCTION: The fifth metacarpal fracture is a frequent pathology that can cause deformity and functional impairment for the adequate grip of the hand. Reintegration into daily or working activities is related to the treatment received and rehabilitation. In fractures of the neck of the fifth metacarpal, internal fixation with a Kirschner's wire is a conventional treatment method with variants that affect its outcome. AIM OF THE STUDY: To compare the functional and clinical results of the treatment of fifth metacarpal fracture with the use of retrograde vs. antegrade Kirschner wires. MATERIAL AND METHODS: Comparative, longitudinal, prospective study at a third-level trauma centre in patients with a fifth metacarpal neck fracture, with clinical, radiographic and Quick DASH scale follow-up at the 3rd, 6th, and 8th postoperative week. RESULTS: Sixty patients were included (58 men, 2 women), age of 29.63±10.15 years, with a fifth metacarpal fracture, treated by closed reduction and stabilisation with a Kirschner wire. The antegrade approach showed a metacarpophalangeal flexion range at 8 weeks of 89.11° (p<0.001; 95% CI [-26.81; -11.42]), a DASH scale value of 18.17 (p<0.001; 95% CI [23.45; 39.12]), and an average of 27.35 days to return to work (p=0.002; 95% CI [16.22; 62.14]), compared with the retrograde approach. CONCLUSION: Stabilisation with antegrade Kirschner wire showed superior functional results, and metacarpophalangeal range of motion, compared to those operated via retrograde approach.

Closed reduction and intramedullary fixation of fifth metacarpal neck fractures: Retrograde vs. anterograde technique. Prospective study. / Reducción cerrada y fijación intramedular de fracturas del cuello del quinto metacarpiano: técnica retrógrada vs. anterógrada. Estudio prospectivo.

INTRODUCTION: The fifth metacarpal fracture is a frequent pathology that can cause deformity and functional impairment for the adequate grip of the hand. Reintegration into daily or working activities is related to the treatment received and rehabilitation. In fractures of the neck of the fifth metacarpal, internal fixation with a Kirschner's wire is a conventional treatment method with variants that affect its outcome. AIM OF THE STUDY: To compare the functional and clinical results of the treatment of fifth metacarpal fracture with the use of retrograde versus antegrade Kirschner wires. MATERIAL AND METHODS: Comparative, longitudinal, prospective study at a third-level trauma center in patients with a fifth metacarpal neck fracture, with clinical, radiographic and Quick DASH scale follow-up at the 3rd, 6th, and 8th postoperative week. RESULTS: Sixty patients were included (58 men, 2 women), age of 29.63±10.15 years, with a fifth metacarpal fracture, treated by closed reduction and stabilization with a Kirschner wire. The antegrade approach showed a metacarpophalangeal flexion range at 8 weeks of 89.11° (p<0.001; 95% CI [-26.81; -11.42]), a DASH scale value of 18.17 (p<0.001; 95% CI [23.45; 39.12]), and an average of 27.35 days to return to work (p=0.002; 95% CI [16.22; 62.14]), compared with the retrograde approach. CONCLUSION: Stabilization with antegrade Kirschner wire showed superior functional results, and metacarpophalangeal range of motion, compared to those operated via retrograde approach.

Universal pre-mixing dry-film stickers capable of retrofitting existing microfluidics.

Integrating microfluidic mixers into lab-on-a-chip devices remains challenging yet important for numerous applications including dilutions, extractions, addition of reagents or drugs, and particle synthesis. High-efficiency mixers utilize large or intricate geometries that are difficult to manufacture and co-implement with lab-on-a-chip processes, leading to cumbersome two-chip solutions. We present a universal dry-film microfluidic mixing sticker that can retrofit pre-existing microfluidics and maintain high mixing performance over a range of Reynolds numbers and input mixing ratios. To attach our pre-mixing sticker module, remove the backing material and press the sticker onto an existing microfluidic/substrate. Our innovation centers around the multilayer use of laser-cut commercially available silicone-adhesive-coated polymer sheets as microfluidic layers to create geometrically complex, easy to assemble designs that can be adhered to a variety of surfaces, namely, existing microfluidic devices. Our approach enabled us to assemble the traditional yet difficult to manufacture "F-mixer" in minutes and conceptually extend this design to create a novel space-saving spiral F-mixer. Computational fluid dynamic simulations and experimental results confirmed that both designs maintained high performance for 0.1 < Re < 10 and disparate input mixing ratios of 1:10. We tested the integration of our system by using the pre-mixer to fluorescently tag proteins encapsulated in an existing microfluidic. When integrated with another microfluidic, our pre-mixing sticker successfully combined primary and secondary antibodies to fluorescently tag micropatterned proteins with high spatial uniformity, unlike a traditional pre-mixing "T-mixer" sticker. Given the ease of this technology, we anticipate numerous applications for point-of-care devices, microphysiological-systems-on-a-chip, and microfluidic-based biomedical research.

Telehealth for geriatric post-emergency department visits to promote age-friendly care.

OBJECTIVE: To describe a feasibility pilot study for older adults that addresses the digital divide, unmet health care needs, and the 4Ms of Age-Friendly Health Systems via the emergency department (ED) follow-up home visits supported by telehealth. DATA SOURCES AND STUDY SETTING: Data sources were a pre-implementation site survey and pilot phase individual-level patient data from six US Department of Veterans Affairs (VA) EDs. STUDY DESIGN: A pre-implementation survey assessed existing geriatric ED processes. In the pilot called SCOUTS (Supporting Community Outpatient, Urgent care & Telehealth Services), sites identified high-risk patients during an ED visit. After ED discharge, Intermediate Care Technicians (ICTs, former military medics), performed follow-up telephone, or home visits. During the follow-up visit, ICTs identified "what matters," performed geriatric screens aligned with Age-Friendly Health Systems, observed home safety risks, assisted with video telehealth check-ins with ED providers, and provided care coordination. SCOUTS visit data were recorded in the patient's electronic medical record using a standardized template. DATA COLLECTION/EXTRACTION METHODS: Sites were surveyed via electronic form. Administrative pilot data extracted from VA Corporate Data Warehouse, May-October 2021. PRINCIPLE FINDINGS: Site surveys showed none of the EDs had a formalized way of identifying the 4 M "what matters." During the pilot, ICT performed 56 telephone and 247 home visits. All home visits included a telehealth visit with an ED provider (n = 244) or geriatrician (n = 3). ICTs identified 44 modifiable home fall risks and 99 unmet care needs, recommended 80 pieces of medical equipment, placed 36 specialty care consults, and connected 180 patients to a Patient Aligned Care Team member for follow-up. CONCLUSIONS: A post-ED follow-up program in which former military medics perform geriatric screens and care coordination is feasible. Combining telehealth and home visits allows providers to address what matters and unmet care needs.

Boroleucine-Derived Covalent Inhibitors of the ZIKV Protease.

The Zika virus (ZIKV) remains a potential threat to the public health due to the lack of both an approved vaccination or a specific treatment. In this work, a series of peptidic inhibitors of the ZIKV protease with boroleucine as P1 residue was synthesized. The highest affinities with Ki values down to 8 nM were observed for compounds with basic residues in both P2 and P3 position and at the N-terminus. The low potency of reference compounds containing leucine, leucine-amide or isopentylamide as P1 residue suggested a covalent binding mode of the boroleucine-derived inhibitors. This was finally proven by crystal structure determination of the most potent inhibitor from this series in complex with the ZIKV protease.

Structure-Based Optimization and Characterization of Macrocyclic Zika Virus NS2B-NS3 Protease Inhibitors.

Zika virus (ZIKV) is a human pathogenic arbovirus. So far, neither a specific treatment nor a vaccination against ZIKV infections has been approved. Starting from our previously described lead structure, a series of 29 new macrocyclic inhibitors of the Zika virus protease containing different linker motifs have been synthesized. By selecting hydrophobic d-amino acids as part of the linker, numerous inhibitors with Ki values < 5 nM were obtained. For 12 inhibitors, crystal structures in complex with the ZIKV protease up to 1.30 Å resolution were determined, which contribute to the understanding of the observed structure-activity relationship (SAR). In immunofluorescence assays, an antiviral effect was observed for compound 26 containing a d-homocyclohexylalanine residue in its linker segment. Due to its excellent selectivity profile and low cytotoxicity, this inhibitor scaffold could be a suitable starting point for the development of peptidic drugs against the Zika virus and related flaviviruses.

Impact of vaccination on the epidemiology and prognosis of pneumonia.

Adults with lung diseases, comorbidities, smokers, and elderly are at risk of lung infections and their consequences. Community-acquired pneumonia happen in more than 1% of people each year. Possible pathogens of community-acquired pneumonia include viruses, pneumococcus and atypicals. The CDC recommend vaccination throughout life to provide immunity, but vaccination rates in adults are poor. Tetravalent and trivalent influenza vaccine is designed annually during the previous summer for the next season. The available vaccines include inactivated, adjuvant, double dose, and attenuated vaccines. Their efficacy depends on the variant of viruses effectively responsible for the outbreak each year, and other reasons. Regarding the pneumococcal vaccine, there coexist the old polysaccharide 23-valent vaccine with the new conjugate 10-valent and 13-valent conjugate vaccines. Conjugate vaccines demonstrate their usefulness to reduce the incidence of pneumococcal pneumonia due to the serotypes present in the vaccine. Whooping cough is still present, with high morbidity and mortality rates in young infants. Adult's pertussis vaccine is available, it could contribute to the control of whooping cough in the most susceptible, but it is not present yet in the calendar of adults around the world. About 10 vaccines against SARS-CoV-2 have been developed in a short time, requiring emergency use authorization. A high rate of vaccination was observed in most of the countries. Booster doses became frequent after the loss of effectiveness against new variants. The future of this vaccine is yet to be written.

Skin Conductance Responses to a Discrete Threat in Virtual Reality: Associations with Psychopathy and Anxiety.

People with high levels of psychopathic traits are often described as fearless and lacking in emotional depth, particularly when evaluating threats in their environments. Skin conductance responsivity (SCR) to negative emotional stimuli represents a robust autonomic correlate of conduct problem behavior in children (Fanti et al., in Neuroscience and Biobehavioral Reviews, 100, 98-107, 2019). However, studies that have examined threat-related processing in youth with conduct problems have tended to use a variety of negative stimuli that might induce various and unspecific negative emotions. Few studies have taken in to account the moderating effects of anxiety on the relationship of distinct psychopathic traits (e.g., narcissism, callousness, impulsivity) with SCR to a fear inducing stimulus. In this study, we examined SCR to a virtual reality rollercoaster drop - that is, a discrete fear inducing event - in a sample of 75 youths (61 males; M = 14 years, SD = 1.4) enrolled in a non-mainstream school. The rollercoaster drop was used to more clearly examine an event-related response to a discrete threat, rather than examining SCR throughout the rollercoaster ride. We used the teacher-reported Antisocial Process Screening Device (Frick & Hare, in Antisocial process screening device: APSD. Toronto: Multi-Health Systems, 2001) to examine the relations of distinct psychopathic traits with SCR and self-reported anxiety. Lower anxiety was associated with higher callousness, but only in youths with low SCR to discrete threat. These findings suggest that fear and anxiety show complex and interactive relations with distinct psychopathic traits.

Borderline Personality Traits and Emotion Regulation Strategies in Adolescents: The Role of Implicit Theories.

Borderline Personality Disorder (BPD) is associated with emotion dysregulation. The emotion regulation strategies people adopt may depend on one's belief about emotion as either fixed or changeable (termed "implicit theory"). We test this experimentally by modifying beliefs about emotions using virtual reality (VR). A sample of 29 adolescents (ages 14-17 years) were recruited from two adolescent inpatient units for an uncontrolled pilot trial of a new brief intervention. We measured BPD traits, beliefs about emotion, treatment preference, cognitive reappraisal and rumination, before the VR game and 14-31 days later. Adolescents with higher levels of BPD traits endorsed fixed beliefs of emotion and reported higher levels of rumination and lower levels of cognitive reappraisal. After a one-time message delivered via VR, adolescents evidenced an increase in belief that their emotions were changeable. These findings suggest beliefs about emotions may have an important role in interventions for adolescents with BPD traits.