Conductive Polypropylene Additive Manufacturing Feedstock: Application to Aqueous Electroanalysis and Unlocking Nonaqueous Electrochemistry and Electrosynthesis.

Additive manufacturing electrochemistry is an ever-expanding field; however, it is limited to aqueous environments due to the conductive filaments currently available. Herein, the production of a conductive poly(propylene) filament, which unlocks the door to organic electrochemistry and electrosynthesis, is reported. A filament with 40 wt % carbon black possessed enhanced thermal stability, excellent low-temperature flexibility, and high conductivity. The filament produced highly reproducible additive manufactured electrodes that were electrochemically characterized, showing a k0 of 2.00 ± 0.04 × 10-3 cm s-1. This material was then applied to three separate electrochemical applications. First, the electroanalytical sensing of colchicine within environmental waters, where a limit of detection of 10 nM was achieved before being applied to tap, bottled, and river water. Second, the electrodes were stable in organic solvents for 100 cyclic voltammograms and 15 days. Finally, these were applied toward an electrosynthetic reaction of chlorpromazine, where the electrodes were stable for 24-h experiments, outperforming a glassy carbon electrode, and were able to be reused while maintaining a good electrochemical performance. This material can revolutionize the field of additive manufacturing electrochemistry and expand research into a variety of new fields.

Exploring the Relationship between Telomere Length and Cognitive Changes in Post-COVID-19 Subjects.

BACKGROUND/OBJECTIVES: Emerging evidence suggests that patients suffering from COVID-19 may experience neurocognitive symptoms. Furthermore, other studies indicate a probable association between leukocyte telomere length (LTL) and neurocognitive changes in subjects with post-COVID-19 condition. Our study was designed to determine the correlation between telomere length and cognitive changes in post-COVID-19 subjects. METHODS: This study included 256 subjects, categorized based on SARS-CoV-2 infection from 2020 to 2023. In addition, subjects with a psychiatric diagnosis were considered. Moreover, the MoCA and MMSE scales were applied. Telomere length was determined using a polymerase chain reaction, and statistical analysis was employed using ANOVA and X2 tests. RESULTS: We identified a decrease in LTL in individuals with post-COVID-19 conditions compared to those without SARS-CoV-2 infection (p ≤ 0.05). However, no association was found between LTL and cognitive impairment in the subjects post-COVID-19. CONCLUSIONS: The findings suggest that LTL is affected by SARS-CoV-2 infection. Nonetheless, this important finding requires further research by monitoring neurological changes in subjects with post-COVID condition.

Adaptation to Noise in Spectrotemporal Modulation Detection and Word Recognition.

Noise adaptation is the improvement in auditory function as the signal of interest is delayed in the noise. Here, we investigated if noise adaptation occurs in spectral, temporal, and spectrotemporal modulation detection as well as in speech recognition. Eighteen normal-hearing adults participated in the experiments. In the modulation detection tasks, the signal was a 200ms spectrally and/or temporally modulated ripple noise. The spectral modulation rate was two cycles per octave, the temporal modulation rate was 10 Hz, and the spectrotemporal modulations combined these two modulations, which resulted in a downward-moving ripple. A control experiment was performed to determine if the results generalized to upward-moving ripples. In the speech recognition task, the signal consisted of disyllabic words unprocessed or vocoded to maintain only envelope cues. Modulation detection thresholds at 0 dB signal-to-noise ratio and speech reception thresholds were measured in quiet and in white noise (at 60 dB SPL) for noise-signal onset delays of 50 ms (early condition) and 800 ms (late condition). Adaptation was calculated as the threshold difference between the early and late conditions. Adaptation in word recognition was statistically significant for vocoded words (2.1 dB) but not for natural words (0.6 dB). Adaptation was found to be statistically significant in spectral (2.1 dB) and temporal (2.2 dB) modulation detection but not in spectrotemporal modulation detection (downward ripple: 0.0 dB, upward ripple: -0.4 dB). Findings suggest that noise adaptation in speech recognition is unrelated to improvements in the encoding of spectrotemporal modulation cues.

Effects of ipsilateral, contralateral, and bilateral noise precursors on psychoacoustical tuning curves in humans.

Cochlear tuning and hence auditory frequency selectivity are thought to change in noisy environments by activation of the medial olivocochlear reflex (MOCR). In humans, auditory frequency selectivity is often assessed using psychoacoustical tuning curves (PTCs), a plot of the level required for pure-tone maskers to just mask a fixed-level pure-tone probe as a function of masker frequency. Sometimes, however, the stimuli used to measure a PTC are long enough that they can activate the MOCR by themselves and thus affect the PTC. Here, PTCs for probe frequencies of 500 Hz and 4 kHz were measured in forward masking using short maskers (30 ms) and probes (10 ms) to minimize the activation of the MOCR by the maskers or the probes. PTCs were also measured in the presence of long (300 ms) ipsilateral, contralateral, and bilateral broadband noise precursors to investigate the effect of the ipsilateral, contralateral, and bilateral MOCR on PTC tuning. Four listeners with normal hearing participated in the experiments. At 500 Hz, ipsilateral and bilateral precursors sharpened the PTCs by decreasing the thresholds for maskers with frequencies at or near the probe frequency with minimal effects on thresholds for maskers remote in frequency from the probe. At 4 kHz, by contrast, ipsilateral and bilateral precursors barely affected thresholds for maskers near the probe frequency but broadened PTCs by reducing thresholds for maskers far from the probe. Contralateral precursors barely affected PTCs. An existing computational model was used to interpret the results. The model suggested that despite the apparent differences, the pattern of results is consistent with the ipsilateral and bilateral MOCR inhibiting the cochlear gain similarly at the two probe frequencies and more strongly than the contralateral MOCR.

Phenome-Wide Association Study of Latent Autoimmune Diabetes from a Southern Mexican Population Implicates rs7305229 with Plasmatic Anti-Glutamic Acid Decarboxylase Autoantibody (GADA) Levels.

Latent autoimmune diabetes in adults (LADA) is characterized by the presence of glutamate decarboxylase autoantibodies (GADA). LADA has intermediate features between type 1 diabetes and type 2 diabetes. In addition, genetic risk factors for both types of diabetes are present in LADA. Nonetheless, evidence about the genetics of LADA in non-European populations is scarce. This study aims to perform a genome-wide association study with a phenome-wide association study of LADA in a southeastern Mexican population. We included 59 patients diagnosed with LADA from a previous study and 3121 individuals without diabetes from the MxGDAR/ENCODAT database. We utilized the GENESIS package in R to perform the genome-wide association study (GWAS) of LADA and PLINK for the phenome-wide association study (PheWAS) of LADA features. Nine polymorphisms reach the nominal association level (1 × 10-5) in the GWAS. The PheWAS showed that rs7305229 is genome-wide and associated with serum GADA levels in our sample (p = 1.84 × 10-8). rs7305229 is located downstream of the FAIM2 gene; previous reports associate FAIM2 variants with childhood obesity, body mass index, body adiposity measures, lymphocyte CD8+ activity, and anti-thyroid peroxidase antibodies. Our findings reveal that rs7305229 affects the GADA levels in patients with LADA from southeastern Mexico. More studies are needed to determine if this risk genotype exists in other populations with LADA.

Click editing enables programmable genome writing using DNA polymerases and HUH endonucleases.

Genome editing technologies based on DNA-dependent polymerases (DDPs) could offer several benefits compared with other types of editors to install diverse edits. Here, we develop click editing, a genome writing platform that couples the advantageous properties of DDPs with RNA-programmable nickases to permit the installation of a range of edits, including substitutions, insertions and deletions. Click editors (CEs) leverage the 'click'-like bioconjugation ability of HUH endonucleases with single-stranded DNA substrates to covalently tether 'click DNA' (clkDNA) templates encoding user-specifiable edits at targeted genomic loci. Through iterative optimization of the modular components of CEs and their clkDNAs, we demonstrate the ability to install precise genome edits with minimal indels in diverse immortalized human cell types and primary fibroblasts with precise editing efficiencies of up to ~30%. Editing efficiency can be improved by rapidly screening clkDNA oligonucleotides with various modifications, including repair-evading substitutions. Click editing is a precise and versatile genome editing approach for diverse biological applications.

Effect of stimulus duration on estimates of human cochlear tuning.

Auditory masking methods originally employed to assess behavioral frequency selectivity have evolved over the years to infer cochlear tuning. Behavioral forward masking thresholds for spectrally notched noise maskers and a fixed, low-level probe tone provide accurate estimates of cochlear tuning. Here, we use this method to investigate the effect of stimulus duration on human cochlear tuning at 500 Hz and 4 kHz. Probes were 20-ms sinusoids at 10 dB sensation level. Maskers were noises with a spectral notch symmetrically and asymmetrically placed around the probe frequency. For seven participants with normal hearing, masker levels at masking threshold were measured in forward masking for various notch widths and for masker durations of 30 and 400 ms. Measurements were fitted assuming rounded exponential filter shapes and the power spectrum model of masking, and equivalent rectangular bandwidths (ERBs) were inferred from the fits. At 4 kHz, masker thresholds were higher for the shorter maskers but ERBs were not significantly different for the two masker durations (ERB30ms=294 Hz vs. ERB400ms=277 Hz). At 500 Hz, by contrast, notched-noise curves were shallower for the 30-ms than the 400-ms masker, and ERBs were significantly broader for the shorter masker (ERB30ms=126 Hz vs. ERB400ms=55 Hz). We discuss possible factors that may underlay the duration effect at low frequencies and argue that it may not be possible to fully control for those factors. We conclude that tuning estimates are not affected by maker duration at high frequencies but should be measured and interpreted with caution at low frequencies.

Nonreciprocal feedback induces migrating oblique and horizontal banded vegetation patterns in hyperarid landscapes.

In hyperarid environments, vegetation is highly fragmented, with plant populations exhibiting non-random biphasic structures where regions of high biomass density are separated by bare soil. In the Atacama Desert of northern Chile, rainfall is virtually nonexistent, but fog pushed in from the interior sustains patches of vegetation in a barren environment. Tillandsia landbeckii, a plant with no functional roots, survives entirely on fog corridors as a water source. Their origin is attributed to interaction feedback among the ecosystem agents, which have different spatial scales, ultimately generating banded patterns as a self-organising response to resource scarcity. The interaction feedback between the plants can be nonreciprocal due to the fact that the fog flows in a well-defined direction. Using remote sensing analysis and mathematical modelling, we characterise the orientation angle of banded vegetation patterns with respect to fog direction and topographic slope gradient. We show that banded vegetation patterns can be either oblique or horizontal to the fog flow rather than topography. The initial and boundary conditions determine the type of the pattern. The bifurcation diagram for both patterns is established. The theoretical predictions are in agreement with observations from remote sensing image analysis.

Exploring the potential of cell-derived vesicles for transient delivery of gene editing payloads.

Gene editing technologies have the potential to correct genetic disorders by modifying, inserting, or deleting specific DNA sequences or genes, paving the way for a new class of genetic therapies. While gene editing tools continue to be improved to increase their precision and efficiency, the limited efficacy of in vivo delivery remains a major hurdle for clinical use. An ideal delivery vehicle should be able to target a sufficient number of diseased cells in a transient time window to maximize on-target editing and mitigate off-target events and immunogenicity. Here, we review major advances in novel delivery platforms based on cell-derived vesicles - extracellular vesicles and virus-like particles - for transient delivery of gene editing payloads. We discuss major findings regarding packaging, in vivo biodistribution, therapeutic efficacy, and safety concerns of cell-derived vesicles delivery of gene editing cargos and their potential for clinical translation.

Incidental diagnosis of cardiac fibroma in an infant with acute viral bronchiolitis.

OBJECTIVE: Pediatric cardiac tumors are rare and, among them, 90% are benign. Cardiac fibroma is the second most frequent tumor, after rhabdomyoma. The objective of this study is to report a case of cardiac fibroma diagnosed incidentally in a patient admitted with acute viral bronchiolitis. CASE DESCRIPTION: A 5-month-old male infant was admitted to the pediatric emergency department with acute viral bronchiolitis requiring hospitalization. He presented a detectable respiratory syncytial virus in oropharyngeal swab, blood test with lymphocytosis and a chest radiography revealed cardiomegaly. Further cardiologic testing was performed detecting elevation of cardiac biomarkers, an electrocardiogram with alteration of left ventricular repolarization and echocardiogram with a heterogeneous mass in the left ventricular, with areas of calcification. A chest angiotomography suggested rhabdomyosarcoma or cardiac fibroma and a magnetic resonance showed a mass, with characteristics suggesting fibroma. The final diagnosis was made after two cardiac catheterizations for biopsy of the lesion, confirming cardiac fibroma by anatomopathological examination. Because the patient had moderate to severe systolic dysfunction, he was submitted to heart transplant. COMMENTS: One third of cardiac fibromas are asymptomatic, generally diagnosed late through tests ordered for other reasons. The gold-standard test for definitive diagnosis is biopsy. Cardiac fibroma usually does not present spontaneous regression and, in most cases, partial or total surgical resection is necessary. When tumors are unresectable, heart transplantation should be indicated. It is essential to have detailed characterization of the cardiac mass to establish the most appropriate therapeutic approach for each patient.

Neuropsychiatric symptoms of post-COVID-19 condition in Mexican healthcare personnel / Síntomas neuropsiquiátricos post-COVID-19 en trabajadores de salud mexicanos

Abstract Objective: The aim of the present study was to identify anxiety and depression in health personnel who suffered COVID-19, and to associate them with blood inflammatory markers. Materials and methods: The design of this study was descriptive and cross-sectional. We evaluated 51 healthcare workers who survived COVID-19 disease with Hamilton scales for anxiety and depression, also we calculated inflammatory markers (systemic immune-inflammation index, SII; monocyte lymphocyte ratio, MLR; platelet lymphocyte ratio, PLR; and neutrophil lymphocyte ratio, NLR) using blood venous samples. This study was carried out from August 2021 to December 2022. Statistical analysis was performed using SPSS v. 26. Results: Our study included 51 healthcare personnel, females (n=29) and males (n=22). The mean age was 40.54 ± 11.00 years. The most frequent acute symptoms for COVID-19 presented were dysgeusia (n=20), anosmia (n=18), and headache (n=17). The most common comorbidities were overweight (n=24), obesity (n=22), and hypertension (n=11). According to the Hamilton Rating Scale for Anxiety (HARS) and Hamilton Rating Scale for Depression Rating (HRSD) we identify anxiety and depression in 72.5% (n=37) and 51% (n=26) within the health personnel, respectively. Conclusions: In our study, we observed a high frequency of anxiety and depression in healthcare workers with post COVID-19 condition. However, we did not observe an association between inflammatory markers (NLR, PLR, MLR, and SII) with anxiety and depression in health personnel postCOVID-19. We suggest follow-up assessments in healthcare personnel with post-COVID-19 condition, to evaluate if mixed emotional disorders persist.

Resumen Objetivo: Identificar ansiedad y depresión en el personal de salud que padeció COVID-19, y asociarlos con niveles de marcadores inflamatorios en sangre. Materiales y métodos: El estudio fue descriptivo y transversal. Evaluamos a 51 trabajadores del área de la salud con antecedente de COVID-19, se aplicó las escalas de ansiedad y depresión de Hamilton, y calculamos marcadores inflamatorios (índice de inmunidad/inflamación sistémica, índice monocito/linfocito, índice plaqueta/linfocito, índice neutrófilo/ linfocito) obtenidas de muestras de sangre venosa. El estudio se realizó durante el periodo de agosto del 2021 a diciembre del 2022. El análisis estadístico fue realizado en SPSS v. 26. Resultados: Nuestro estudio incluyó a 51 personas del área de salud, 29 mujeres y 22 hombres. La edad media fue 40.54 ± 11.00 años. Los síntomas agudos más frecuentes que presentaron los trabajadores fueron disgeusia (n=20), anosmia (n=18), y cefalea (n=17). Las comorbilidades más frecuentes fueron: sobrepeso (n= 24), obesidad (n=22), e hipertensión (n=11). Aplicando la Escala de Ansiedad de Hamilton y la Escala de Depresión de Hamilton, identificamos ansiedad y depresión en el personal de salud en 72.5% (n=37) y 51% (n=26), respectivamente. Conclusiones: En nuestro estudio observamos una alta frecuencia de ansiedad y depresión post COVID-19 en el personal de salud. No observamos asociación entre ansiedad, depresión y marcadores inflamatorios hematológicos en los trabajadores de salud en población mexicana que padeció COVID-19. Sugerimos realizar evaluaciones de seguimiento en el personal de salud en condición post COVID-19 para demostrar la persistencia de trastornos mixtos de las emociones.

Position paper of diagnosis and treatment of post-extubation laryngitis in children: a multidisciplinary expert-based opinion.

OBJECTIVES: To make recommendations on the diagnosis and treatment of post-extubation laryngitis (PEL) in children with or without other comorbidities. METHODS: A three-iterative modified Delphi method was applied. Specialists were recruited representing pediatric otolaryngologists, pediatric and neonatal intensivists. Questions and statements approached topics encompassing definition, diagnosis, endoscopic airway evaluation, risk factors, comorbidities, management, and follow-up. A consensus was defined as a supermajority >70%. RESULTS: Stridor was considered the most frequent symptom and airway endoscopy was recommended for definitive diagnosis. Gastroesophageal reflux and previous history of intubation were considered risk factors. Specific length of intubation did not achieve a consensus as a risk factor. Systemic corticosteroids should be part of the medical treatment and dexamethasone was the drug of choice. No consensus was achieved regarding dosage of corticosteroids, although endoscopic findings help defining dosage and length of treatment. Non-invasive ventilation, laryngeal rest, and use of comfort sedation scales were recommended. Indications for microlaryngoscopy and bronchoscopy under anesthesia were symptoms progression or failure to improve after the first 72-h of medical treatment post-extubation, after two failed extubations, and/or suspicion of severe lesions on flexible fiberoptic laryngoscopy. CONCLUSIONS: Management of post-extubation laryngitis is challenging and can be facilitated by a multidisciplinary approach. Airway endoscopy is mandatory and impacts decision-making, although there is no consensus regarding dosage and length of treatment.

Novel disposable and portable 3D-printed electrochemical apparatus for fast and selective screening of 25E-NBOH in forensic samples.

The advent of new psychoactive substances (NPS) has caused enormous difficulty for legal control since they are rapidly commercialized, and their chemical structures are routinely altered. In this aspect, derivatives phenethylamines, such as 25E-NBOH, have received great attention in the forensic scenario. Hence, we propose portable and cost-effective (U$ 5.00) 3D-printed devices for the electrochemical screening of 25E-NBOH for the first time. The cell and all electrodes were printed using acrylonitrile butadiene styrene filament (insulating material) and conductive filament (graphite embedded in a polylactic acid matrix), respectively, both by the fused deposition modeling (FDM) 3D printing technique. The electrochemical apparatus enables micro-volume analysis (50-2000 µL), especially important for low sample volumes. A mechanistic route for the electrochemical oxidation of 25E-NBOH is proposed based on cyclic voltammetric data, which showed two oxidation processes around +0.75 V and +1.00 V and a redox pair between +0.2 and -0.2 V (vs. graphite ink pseudo-reference). A fast and sensitive square-wave voltammetry method was developed, which exhibited a linear working range from 0.85 to 5.1 µmoL-1, detection limit of 0.2 µmol L-1, and good intra-electrode precision (n = 10, RSD <5.3 %). Inter-electrode measurements (n = 3, RSD <9.8 %) also attested that the electrode production process is reproducible. Interference tests in the presence of other drugs frequently found in blotting paper indicated high selectivity of the electrochemical method for screening of 25E-NBOH. Screening analysis of blotting paper confirmed the presence of 25E-NBOH in the seized samples. Moreover, a recovery percentage close to 100 % was found for a spiked saliva sample, suggesting the method's usefulness for quantitative purposes aimed at information on recent drug use.

Position paper of diagnosis and treatment of post-extubation laryngitis in children: a multidisciplinary expert-based opinion

Abstract Objectives To make recommendations on the diagnosis and treatment of post-extubation laryngitis (PEL) in children with or without other comorbidities. Methods A three-iterative modified Delphi method was applied. Specialists were recruited representing pediatric otolaryngologists, pediatric and neonatal intensivists. Questions and statements approached topics encompassing definition, diagnosis, endoscopic airway evaluation, risk factors, comorbidities, management, and follow-up. A consensus was defined as a supermajority >70%. Results Stridor was considered the most frequent symptom and airway endoscopy was recommended for definitive diagnosis. Gastroesophageal reflux and previous history of intubation were considered risk factors. Specific length of intubation did not achieve a consensus as a risk factor. Systemic corticosteroids should be part of the medical treatment and dexamethasone was the drug of choice. No consensus was achieved regarding dosage of corticosteroids, although endoscopic findings help defining dosage and length of treatment. Non-invasive ventilation, laryngeal rest, and use of comfort sedation scales were recommended. Indications for microlaryngoscopy and bronchoscopy under anesthesia were symptoms progression or failure to improve after the first 72-h of medical treatment post-extubation, after two failed extubations, and/or suspicion of severe lesions on flexible fiberoptic laryngoscopy. Conclusions Management of post-extubation laryngitis is challenging and can be facilitated by a multidisciplinary approach. Airway endoscopy is mandatory and impacts decision-making, although there is no consensus regarding dosage and length of treatment.

Incidental diagnosis of cardiac fibroma in an infant with acute viral bronchiolitis / Diagnóstico incidental de fibroma cardíaco em um lactente com bronquiolite viral aguda

ABSTRACT Objective: Pediatric cardiac tumors are rare and, among them, 90% are benign. Cardiac fibroma is the second most frequent tumor, after rhabdomyoma. The objective of this study is to report a case of cardiac fibroma diagnosed incidentally in a patient admitted with acute viral bronchiolitis. Case description: A 5-month-old male infant was admitted to the pediatric emergency department with acute viral bronchiolitis requiring hospitalization. He presented a detectable respiratory syncytial virus in oropharyngeal swab, blood test with lymphocytosis and a chest radiography revealed cardiomegaly. Further cardiologic testing was performed detecting elevation of cardiac biomarkers, an electrocardiogram with alteration of left ventricular repolarization and echocardiogram with a heterogeneous mass in the left ventricular, with areas of calcification. A chest angiotomography suggested rhabdomyosarcoma or cardiac fibroma and a magnetic resonance showed a mass, with characteristics suggesting fibroma. The final diagnosis was made after two cardiac catheterizations for biopsy of the lesion, confirming cardiac fibroma by anatomopathological examination. Because the patient had moderate to severe systolic dysfunction, he was submitted to heart transplant. Comments: One third of cardiac fibromas are asymptomatic, generally diagnosed late through tests ordered for other reasons. The gold-standard test for definitive diagnosis is biopsy. Cardiac fibroma usually does not present spontaneous regression and, in most cases, partial or total surgical resection is necessary. When tumors are unresectable, heart transplantation should be indicated. It is essential to have detailed characterization of the cardiac mass to establish the most appropriate therapeutic approach for each patient.

RESUMO Objetivo: Tumores cardíacos pediátricos são raros e, entre eles, 90% são benignos. O fibroma cardíaco é o segundo mais frequente, após o rabdomioma. O objetivo deste trabalho é relatar um caso de fibroma cardíaco, cujo diagnóstico foi incidental em um paciente internado por bronquiolite viral aguda. Descrição do caso: Lactente do sexo masculino com cinco meses foi atendido em um pronto atendimento pediátrico com quadro de bronquiolite viral aguda, necessitando de hospitalização. Em exames complementares, apresentou vírus sincicial respiratório detectável, hemograma com linfocitose, além de radiografia de tórax com cardiomegalia. Em investigação cardiológica, as enzimas cardíacas encontravam-se elevadas, o eletrocardiograma apresentava alteração de repolarização de parede lateral de ventrículo esquerdo e o ecocardiograma demonstrou massa heterogênea em ventrículo esquerdo, com áreas de calcificação. A angiotomografia de tórax sugeriu rabdomiossarcoma ou fibroma cardíaco e a ressonância magnética demonstrou massa cuja localização e características sugeriram fibroma. O diagnóstico foi conclusivo após dois cateterismos para biópsia da lesão, confirmando fibroma cardíaco pelo anatomopatológico. Em razão da disfunção sistólica moderada a grave, o paciente foi submetido a transplante cardíaco. Comentários: Um terço dos fibromas cardíacos é assintomático, geralmente diagnosticado tardiamente por meio de exames solicitados por outro motivo. O padrão-ouro para o diagnóstico definitivo é a biópsia. O fibroma cardíaco não costuma apresentar regressão espontânea, sendo submetido, na maioria dos casos, a ressecção cirúrgica parcial ou total. Nos tumores irressecáveis, o transplante cardíaco deve ser indicado. A caracterização detalhada da massa cardíaca é fundamental para traçar a conduta terapêutica mais adequada para cada paciente.

Click editing enables programmable genome writing using DNA polymerases and HUH endonucleases.

Genome editing technologies that install diverse edits can widely enable genetic studies and new therapeutics. Here we develop click editing, a genome writing platform that couples the advantageous properties of DNA-dependent DNA polymerases with RNA-programmable nickases (e.g. CRISPR-Cas) to permit the installation of a range of edits including substitutions, insertions, and deletions. Click editors (CEs) leverage the "click"-like bioconjugation ability of HUH endonucleases (HUHes) with single stranded DNA substrates to covalently tether "click DNA" (clkDNA) templates encoding user-specifiable edits at targeted genomic loci. Through iterative optimization of the modular components of CEs (DNA polymerase and HUHe orthologs, architectural modifications, etc.) and their clkDNAs (template configurations, repair evading substitutions, etc.), we demonstrate the ability to install precise genome edits with minimal indels and no unwanted byproduct insertions. Since clkDNAs can be ordered as simple DNA oligonucleotides for cents per base, it is possible to screen many different clkDNA parameters rapidly and inexpensively to maximize edit efficiency. Together, click editing is a precise and highly versatile platform for modifying genomes with a simple workflow and broad utility across diverse biological applications.

Knowledge regarding extracorporeal membrane oxygenation management among Brazilian pediatric intensivists: a cross-sectional survey.

OBJECTIVE: To assess Brazilian pediatric intensivists' general knowledge of extracorporeal membrane oxygenation, including evidence for its use, the national funding model, indications, and complications. METHODS: This was a multicenter cross-sectional survey including 45 Brazilian pediatric intensive care units. A convenience sample of 654 intensivists was surveyed regarding their knowledge on managing patients on extracorporeal membrane oxygenation, its indications, complications, funding, and literature evidence. RESULTS: The survey addressed questions regarding the knowledge and experience of pediatric intensivists with extracorporeal membrane oxygenation, including two clinical cases and 6 optional questions about the management of patients on extracorporeal membrane oxygenation. Of the 45 invited centers, 42 (91%) participated in the study, and 412 of 654 (63%) pediatric intensivists responded to the survey. Most pediatric intensive care units were from the Southeast region of Brazil (59.5%), and private/for-profit hospitals represented 28.6% of the participating centers. The average age of respondents was 41.4 (standard deviation 9.1) years, and the majority (77%) were women. Only 12.4% of respondents had taken an extracorporeal membrane oxygenation course. Only 19% of surveyed hospitals have an extracorporeal membrane oxygenation program, and only 27% of intensivists reported having already managed patients on extracorporeal membrane oxygenation. Specific extracorporeal membrane oxygenation management questions were responded to by only 64 physicians (15.5%), who had a fair/good correct response rate (median 63.4%; range 32.8% to 91.9%). CONCLUSION: Most Brazilian pediatric intensivists demonstrated limited knowledge regarding extracorporeal membrane oxygenation, including its indications and complications. Extracorporeal membrane oxygenation is not yet widely available in Brazil, with few intensivists prepared to manage patients on extracorporeal membrane oxygenation and even fewer intensivists recognizing when to refer patients to extracorporeal membrane oxygenation centers.

A novel 3D-printed graphite/polylactic acid sensor for the electrochemical determination of 2,4,6-trinitrotoluene residues in environmental waters.

Here, lab-made graphite and polylactic acid (Gpt-PLA) biocomposite materials were used to additively manufacture electrodes via the fused deposition modeling (FDM) technique for subsequent determination of the explosive 2,4,6-trinitrotoluene (TNT, considered a persistent organic pollutant). The surface of the 3D-printed material was characterized by SEM and Raman, which revealed high roughness and the presence of defects in the graphite structure, which enhanced the electrochemical response of TNT. The 3D-printed Gpt-PLA electrode coupled to square wave voltammetry (SWV) showed suitable performance for fastly determining the explosive residues (around 7 s). Two reduction processes at around -0.22 V and -0.36 V were selected for TNT detection, with linear ranges between 1.0 and 10.0 µM. Moreover, detection limits of 0.52 and 0.66 µM were achieved for both reduction steps. The proposed method was applied to determine TNT in different environmental water samples (tap water, river water, and seawater) without a dilution step (direct analysis). Recovery values between 98 and 106% confirmed the accuracy of the analyses. Additionally, adequate selectivity was achieved even in the presence of other explosives commonly used by military agencies, metallic ions commonly found in water, and also some electroactive camouflage species. Such results indicate that the proposed device is promising to quantify TNT residues in environmental samples, a viable on-site analysis strategy.

Hepatic levels of S-adenosylmethionine regulate the adaptive response to fasting.

There has been an intense focus to uncover the molecular mechanisms by which fasting triggers the adaptive cellular responses in the major organs of the body. Here, we show that in mice, hepatic S-adenosylmethionine (SAMe)-the principal methyl donor-acts as a metabolic sensor of nutrition to fine-tune the catabolic-fasting response by modulating phosphatidylethanolamine N-methyltransferase (PEMT) activity, endoplasmic reticulum-mitochondria contacts, ß-oxidation, and ATP production in the liver, together with FGF21-mediated lipolysis and thermogenesis in adipose tissues. Notably, we show that glucagon induces the expression of the hepatic SAMe-synthesizing enzyme methionine adenosyltransferase α1 (MAT1A), which translocates to mitochondria-associated membranes. This leads to the production of this metabolite at these sites, which acts as a brake to prevent excessive ß-oxidation and mitochondrial ATP synthesis and thereby endoplasmic reticulum stress and liver injury. This work provides important insights into the previously undescribed function of SAMe as a new arm of the metabolic adaptation to fasting.

Natural Retinol Analogs Potentiate the Effects of Retinal on Aged and Photodamaged Skin: Results from In Vitro to Clinical Studies.

INTRODUCTION: Plants are a source of natural ingredients with retinol-like properties that can deliver anti-aging benefits without the side effects typically associated with retinoid use. We hypothesized that by combining two such analogs, bakuchiol (BAK) and Vigna aconitifolia extract (VAE), with the potent retinoid retinal (RAL), the anti-photoaging potential of RAL could be enhanced without compromising its skin irritation profile. The purpose of this study was to demonstrate that BAK and VAE potentiate the anti-photoaging activity of RAL. METHODS: Gene expression profiling of full-thickness reconstructed skin was first used to examine the impact of BAK or VAE in combination with RAL on skin biology. Next, the irritative potential of this combination, and its capacity to reverse key signs of photoaging in an ex vivo model was assessed. Finally, a proof-of-concept open label clinical study was performed to evaluate the anti-photoaging capacity and skin compatibility of a cosmetic formulation (tri-retinoid complex; 3RC) containing this complex in combination with other well characterized anti-photoaging ingredients. RESULTS: In vitro profiling suggested that combining 0.1% RAL with BAK or VAE potentiates the effect of RAL on keratinocyte differentiation and skin barrier function without affecting its skin irritation profile. When formulated with other anti-photoaging ingredients, such as niacinamide and melatonin, 3RC reversed ultraviolet radiation-induced deficits in structural components of the dermal extracellular matrix, including hyaluronic acid and collagen. In vivo, it led to a reversal of clinical signs of age and photodamage, with statistically significant improvement to skin firmness (+5.6%), skin elasticity (+13.9%), wrinkle count (-43.2%), and skin tone homogeneity (+7.0%), observed within 28 days of once nightly use. Notably, the number of crow's feet wrinkles was reduced in 100% of subjects. Furthermore, 3RC was very well tolerated. CONCLUSION: These data suggest that 3RC is a highly effective and well-tolerated treatment for photoaging.