Rheological and Injectability Evaluation of Sterilized Poloxamer-407-Based Hydrogels Containing Docetaxel-Loaded Lipid Nanoparticles.

Nanostructured lipid carriers (NLCs) have the potential to increase the bioavailability and reduce the side effects of docetaxel (DTX). However, only a small fraction of nanoparticles given intravenously can reach a solid tumor. In situ-forming gels combined with nanoparticles facilitate local administration and promote drug retention at the tumor site. Injectable hydrogels based on poloxamer 407 are excellent candidates for this hybrid nanoparticle-hydrogel system because of their thermoresponsive behavior and biocompatibility. Therefore, this work aimed to develop injectable poloxamer hydrogels containing NLCs for intratumoral delivery of DTX. To ensure sterility, the obtained hydrogels were autoclaved (121 °C for 15 min) after preparation. Then, the incorporation of NLCs into the poloxamer hydrogels and the impact of steam sterilization on the nanocomposite hydrogels were evaluated concerning sol-gel transition, injectability, and physicochemical stability. All formulations were extruded through the tested syringe-needle systems with acceptable force (2.2-13.4 N) and work (49.5-317.7 N·mm) of injection. Following steam sterilization, injection became easier in most cases, and the physicochemical properties of all hydrogels remained practically unchanged according to the spectroscopical and thermal analysis. The rheological evaluation revealed that the nanocomposite hydrogels were liquid at 25 °C and underwent rapid gelation at 37 °C. However, their sterilized counterparts gelled at 1-2 °C above body temperature, suggesting that the autoclaving conditions employed had rendered these nanocomposite hydrogels unsuitable for local drug delivery.

Prevalence and factors associated with anxiety, depression and burnout in gynecology and obstetrics residents during the COVID-19 pandemic.

Objective: To determine the prevalence of anxiety, depression and burnout in residents of Gynecology and Obstetrics during COVID-19 pandemic in Brazil and its associated factors. Methods: Cross-sectional study involving all regions of Brazil, through the application of a sociodemographic questionnaire, the Hospital Anxiety and Depression Scale (HAD) and the Maslach Burnout Inventory (MBI-HSS) instrument. Multivariate analysis was performed after adjusting the Poisson model. Results: Among the 719 participating medical residents, screening was positive for anxiety in 75.7% and for depression in 49.8% of cases. Burnout syndrome was evidenced in 41.3% of the physicians studied. Those with depression are more likely to have anxiety (OR 0.797; 95%CI 0.687 - 0.925) and burnout syndrome (OR 0.847 95%CI 0.74 - 0.97). Residents with anxiety (OR 0.805; 95%CI 0.699 - 0.928) and burnout (OR 0.841; 95%CI 0.734 - 0.963) are more likely to have depression. Conclusion: High prevalence of anxiety, depression and burnout were found in residents of Gynecology and Obstetrics in Brazil, in addition to important correlations between anxiety-depression and depression-burnout.

Analytical Techniques for Characterizing Tumor-Targeted Antibody-Functionalized Nanoparticles.

The specific interaction between cell surface receptors and corresponding antibodies has driven opportunities for developing targeted cancer therapies using nanoparticle systems. It is challenging to design and develop such targeted nanomedicines using antibody ligands, as the final nanoconjugate's specificity hinges on the cohesive functioning of its components. The multicomponent nature of antibody-conjugated nanoparticles also complicates the characterization process. Regardless of the type of nanoparticle, it is essential to perform physicochemical characterization to establish a solid foundation of knowledge and develop suitable preclinical studies. A meaningful physicochemical evaluation of antibody-conjugated nanoparticles should include determining the quantity and orientation of the antibodies, confirming the antibodies' integrity following attachment, and assessing the immunoreactivity of the obtained nanoconjugates. In this review, the authors describe the various techniques (electrophoresis, spectroscopy, colorimetric assays, immunoassays, etc.) used to analyze the physicochemical properties of nanoparticles functionalized with antibodies and discuss the main results.

Dynamic enhancer landscapes in human craniofacial development.

The genetic basis of human facial variation and craniofacial birth defects remains poorly understood. Distant-acting transcriptional enhancers control the fine-tuned spatiotemporal expression of genes during critical stages of craniofacial development. However, a lack of accurate maps of the genomic locations and cell type-resolved activities of craniofacial enhancers prevents their systematic exploration in human genetics studies. Here, we combine histone modification, chromatin accessibility, and gene expression profiling of human craniofacial development with single-cell analyses of the developing mouse face to define the regulatory landscape of facial development at tissue- and single cell-resolution. We provide temporal activity profiles for 14,000 human developmental craniofacial enhancers. We find that 56% of human craniofacial enhancers share chromatin accessibility in the mouse and we provide cell population- and embryonic stage-resolved predictions of their in vivo activity. Taken together, our data provide an expansive resource for genetic and developmental studies of human craniofacial development.

Detection of plasma anti-lipopolysaccharide (LPS)antibodies against enterohemorrhagic Escherichia coli(EHEC) in asymptomatic kindergarten teachers fromBuenos Aires province

Resumen En Argentina, el síndrome urémico hemolítico causado por Escherichia coli enterohemorrágica (EHEC) tiene la más alta incidencia del mundo. Las infecciones por EHEC tienen un comportamiento endemoepidémico y causan del 20 al 30% de los síndromes de diarrea sanguinolenta en niños menores de 5 años. En el período 2016-2020, se notificaron 272 nuevos casos por año al Sistema de Vigilancia de Salud Nacional. Múltiples factores son responsables de la alta incidencia de SUH en Argentina, incluyendo la transmisión persona-persona. Con el objetivo de detectar posibles portadores asintomáticos de EHEC, realizamos un estudio preliminar de la frecuencia de anticuerpos antilipopolisacáridos contra los serotipos de EHEC más prevalentes en Argentina. El estudio se realizó con muestras de plasma obtenidas de 61 maestras y maestros de jardines de infantes de 26 instituciones del distrito de José C. Paz, localizado en el área suburbana de la provincia de Buenos Aires, Argentina. El 51% de las muestras presentaron anticuerpos contra los serotipos de lipopolisacáridos O157, O145, O121 y O103; el 6,4% de las muestras positivas tuvieron el isotipo IgM (n=2), el 61,3% el isotipo IgG (n=19) y el 32,3% los isotipos IgM e IgG (n=10). Dado que los anticuerpos antilipopolisacáridos presentan usualmente una duración corta, la detección de IgM específica podría indicar una infección reciente. Además, el alto porcentaje de muestras positivas hallado podría indicar una exposición frecuente a las cepas de EHEC en la cohorte estudiada. Asimismo, la gran población de adultos portadores asintomáticos de estas cepas patógenas podría contribuir al comportamiento endémico, a través de la transmisión persona-persona. El perfeccionamiento de programas educacionales continuos en jardines de infantes podría constituir una medida importante para reducir los casos de síndrome urémico hemolítico, no solo en Argentina, sino también en el mundo.

Abstract In Argentina, hemolytic uremic syndrome (HUS) caused by EHEC has the highest incidence in the world. EHEC infection has an endemo-epidemic behavior, causing 20-30% of acute bloody diarrhea syndrome in children under 5 years old. In the period 2016-2020, 272 new cases per year were notified to the National Health Surveillance System. Multiple factors are responsible for HUS incidence in Argentina including person-to-person transmission. In order to detect possible EHEC carriers, we carried out a preliminary study of the frequency of kindergarten teachers with anti-LPS antibodies against the most prevalent EHEC serotypes in Argentina. We analyzed 61 kindergarten teachers from 26 institutions from José C. Paz district, located in the suburban area of Buenos Aires province, Argentina. Fifty-one percent of the plasma samples had antibodies against O157, O145, O121 and O103 LPS: 6.4% of the positive samples had IgM isotype (n=2), 61.3% IgG isotype (n=19) and 32.3% IgM and IgG (n=10). Given that antibodies against LPS antigens are usually short-lived specific IgM detection may indicate a recent infection. In addition, the high percentage of positive samples may indicate a frequent exposure to EHEC strains in the cohort studied, as well as the existence of a large non-symptomatic population of adults carrying pathogenic strains that could contribute to the endemic behavior through person-to-person transmission. The improvement of continuous educational programs in kindergarten institutions could be a mandatory measure to reduce HUS cases not only in Argentina but also globally.

Intraductal papillary neoplasms of the bile ducts: a comparative study of a rare disease in Europe and Nagoya, Japan.

BACKGROUND: Intraductal papillary neoplasm of the bile ducts (IPNB) is a rare disease in Western countries. The aim of this study was to compare tumor characteristics, management strategies, and outcomes between Western and Eastern patients who underwent surgical resection for IPNB. METHODS: A multi-institutional retrospective series of patients with IPNB undergoing surgery between January 2010 and December 2020 was gathered under the auspices of the European-African Hepato-Pancreato-Biliary Association (E-AHPBA), and at Nagoya University Hospital, Japan. RESULTS: A total of 85 patients (51% male; median age 66 years) from 28 E-AHPBA centers were compared to 91 patients (64% male; median age 71 years) from Nagoya. Patients in Europe had more multiple lesions (23% vs 2%, P < .001), less invasive carcinoma (42% vs 85%, P < .001), and more intrahepatic tumors (52% vs 24%, P < .001) than in Nagoya. Patients in Europe experienced less 90-day grade >3 Clavien-Dindo complications (33% vs 68%, P < .001), but higher 90-day mortality rate (7.0% vs 0%, P = .03). R0 resections (81% vs 82%) were similar. Overall survival, excluding 90-day postoperative deaths, was similar in both regions. DISCUSSION: Despite performing more extensive resections, the low perioperative mortality rate observed in Nagoya was probably influenced by a combination of patient-, tumor-, and surgery-related factors.

Detection of plasma anti-lipopolysaccharide (LPS) antibodies against enterohemorrhagic Escherichia coli (EHEC) in asymptomatic kindergarten teachers from Buenos Aires province.

In Argentina, hemolytic uremic syndrome (HUS) caused by EHEC has the highest incidence in the world. EHEC infection has an endemo-epidemic behavior, causing 20-30% of acute bloody diarrhea syndrome in children under 5 years old. In the period 2016-2020, 272 new cases per year were notified to the National Health Surveillance System. Multiple factors are responsible for HUS incidence in Argentina including person-to-person transmission. In order to detect possible EHEC carriers, we carried out a preliminary study of the frequency of kindergarten teachers with anti-LPS antibodies against the most prevalent EHEC serotypes in Argentina. We analyzed 61 kindergarten teachers from 26 institutions from José C. Paz district, located in the suburban area of Buenos Aires province, Argentina. Fifty-one percent of the plasma samples had antibodies against O157, O145, O121 and O103 LPS: 6.4% of the positive samples had IgM isotype (n=2), 61.3% IgG isotype (n=19) and 32.3% IgM and IgG (n=10). Given that antibodies against LPS antigens are usually short-lived specific IgM detection may indicate a recent infection. In addition, the high percentage of positive samples may indicate a frequent exposure to EHEC strains in the cohort studied, as well as the existence of a large non-symptomatic population of adults carrying pathogenic strains that could contribute to the endemic behavior through person-to-person transmission. The improvement of continuous educational programs in kindergarten institutions could be a mandatory measure to reduce HUS cases not only in Argentina but also globally.

Prevalence and factors associated with anxiety, depression and burnout in gynecology and obstetrics residents during the COVID-19 pandemic

Abstract Objective: To determine the prevalence of anxiety, depression and burnout in residents of Gynecology and Obstetrics during COVID-19 pandemic in Brazil and its associated factors. Methods: Cross-sectional study involving all regions of Brazil, through the application of a sociodemographic questionnaire, the Hospital Anxiety and Depression Scale (HAD) and the Maslach Burnout Inventory (MBI-HSS) instrument. Multivariate analysis was performed after adjusting the Poisson model. Results: Among the 719 participating medical residents, screening was positive for anxiety in 75.7% and for depression in 49.8% of cases. Burnout syndrome was evidenced in 41.3% of the physicians studied. Those with depression are more likely to have anxiety (OR 0.797; 95%CI 0.687 - 0.925) and burnout syndrome (OR 0.847 95%CI 0.74 - 0.97). Residents with anxiety (OR 0.805; 95%CI 0.699 - 0.928) and burnout (OR 0.841; 95%CI 0.734 - 0.963) are more likely to have depression. Conclusion: High prevalence of anxiety, depression and burnout were found in residents of Gynecology and Obstetrics in Brazil, in addition to important correlations between anxiety-depression and depression-burnout.

Single-cell analysis of chromatin accessibility in the adult mouse brain.

Recent advances in single-cell technologies have led to the discovery of thousands of brain cell types; however, our understanding of the gene regulatory programs in these cell types is far from complete1-4. Here we report a comprehensive atlas of candidate cis-regulatory DNA elements (cCREs) in the adult mouse brain, generated by analysing chromatin accessibility in 2.3 million individual brain cells from 117 anatomical dissections. The atlas includes approximately 1 million cCREs and their chromatin accessibility across 1,482 distinct brain cell populations, adding over 446,000 cCREs to the most recent such annotation in the mouse genome. The mouse brain cCREs are moderately conserved in the human brain. The mouse-specific cCREs-specifically, those identified from a subset of cortical excitatory neurons-are strongly enriched for transposable elements, suggesting a potential role for transposable elements in the emergence of new regulatory programs and neuronal diversity. Finally, we infer the gene regulatory networks in over 260 subclasses of mouse brain cells and develop deep-learning models to predict the activities of gene regulatory elements in different brain cell types from the DNA sequence alone. Our results provide a resource for the analysis of cell-type-specific gene regulation programs in both mouse and human brains.

Validation and reliability analysis of the Questionnaire on the Emotional Impact of Vertigo-CIEV version translated to the Brazilian Portuguese language. / Análise de validação e confiabilidade do Questionário Emocional da Vertigem-CIEV em versão traduzida para o Português-Brasileiro.

PURPOSE: To determine the internal consistency and reliability of the "Questionário de Impacto Emocional da Vertigem (CIEV)" and to validate the instrument with respect to the Dizziness Handicap Inventory (DHI) in a sample of individuals with balance disorders. METHODS: 38 subjects participated in the study, males and females, aged from 23 to 85 years, who presented dizziness, vertigo, and/or falls complaints and attended to the Vestibular Disorders clinic at the University Hospital. Individuals with hearing complaints and/or tinnitus unrelated to dizziness, previous psychiatric comorbidities, and/or cognitive impairments were excluded. We performed an anamnesis and collected complementary data from the medical records. After that, the self-perception questionnaires, DHI, and CIEV, were applied. Statistical analysis was performed in which the Cronbach's alpha verified the internal consistency of the CIEV. Reliability and validity of the CIEV related to the DHI were calculated using Intraclass Correlation Index (ICC) and Pearson's correlation test, respectively. RESULTS: There was a statistically significant correlation between the scores obtained, for both reliability and validation analysis (p<0.001). The mean ICC showed a moderate correlation between the total scores (0.695) and a strong correlation with the physical, emotional, and functional DHI domains (0.706 to 0.869), being the emotional aspect the highest degree (0.869). Pearson's correlation showed strong correlation between the total scores (r=0.820) and varied from moderate to strong, with strongest correlations to the DHI emotional domain (r=0.788). CONCLUSION: The outcomes illustrate important contribution to validation parameters to consider clinical use of the CIEV in the Brazilian population, aiming to identify emotional aspects in patients with balance disorders.

OBJETIVO: Determinar a consistência interna e confiabilidade do "Questionário de Impacto Emocional da Vertigem-CIEV" e validar o instrumento em relação ao Dizziness Handicap Inventory (DHI), em uma amostra de pacientes com distúrbio do equilíbrio corporal. MÉTODO: Participaram 38 sujeitos, idades entre 23 e 85 anos, ambos os sexos, com queixas relacionadas à tontura, desequilíbrios e/ou quedas, atendidos em um ambulatório de Otoneurologia do Hospital Universitário. Foram excluídos sujeitos com queixas auditivas e/ou zumbido sem tontura associada, comorbidades psiquiátricas prévias e/ou comprometimento cognitivo que impedisse a compreensão dos questionários. Foi realizada anamnese, levantamento de prontuário para caracterização da amostra e aplicados os questionários de autopercepção, DHI e CIEV. O alfa de Cronbach verificou a consistência interna do CIEV e a confiabilidade e validade do CIEV em relação ao DHI foram calculadas pelo Índice de Correlação intraclasse (ICC) e teste de Correlação de Pearson, respectivamente. RESULTADOS: Houve correlação estatisticamente significante entre os escores obtidos, tanto à análise de confiabilidade quanto de validação (p<0,001). O ICC médio demonstrou moderada correlação para o escore total (0,695) e forte correlação com os domínios físico, emocional e funcional do DHI (0,706 a 0,869) sendo o maior grau para o domínio emocional (0,869). A Correlação de Pearson demonstrou grau forte para o escore total (r=0,820) e variação de moderado a forte para os domínios, com melhor resultado também para o domínio emocional do DHI (r=0,788). CONCLUSÃO: Os achados representam parâmetros importantes de contribuição para a validação do CIEV para uso clínico na população brasileira, direcionado para a identificação de aspectos emocionais em pacientes com distúrbios do equilíbrio corporal.

Detection and characterization of circulating microvesicles containing Shiga toxin type 2 in a rat model of Hemolytic Uremic Syndrome.

Shiga toxin producing Escherichia coli (STEC) are foodborne pathogens that release Shiga toxin (Stx), virulence factor responsible for the development of Hemolytic Uremic Syndrome (HUS). Stx causes endothelial cell damage, which leads to platelets deposition and thrombi formation within the microvasculature. It has been described that Stx activates blood cells and induces the shedding of proinflammatory and prothrombotic microvesicles (MVs) containing the toxin. In this sense, it has been postulated that MVs containing Stx2 (MVs-Stx2+) can contribute to the physiopathology of HUS, allowing Stx2 to reach the target organs while evading the immune system. In this work, we propose that circulating MVs-Stx2+ can be a potential biomarker for the diagnosis and prognosis of STEC infections and HUS progression. We developed a rat HUS model by the intraperitoneal injection of a sublethal dose of Stx2 and observed: decrease in body weight, increase of creatinine and urea levels, decrease of creatinine clearance and histological renal damages. After characterization of renal damages, we investigated circulating total MVs and MVs-Stx2+ by flow cytometry at different times after Stx2 injection. Additionally, we evaluated the correlation of biochemical parameters such as creatinine and urea in plasma with MVs-Stx2+. As a result, we found a significant circulation of MVs-Stx2+ at 72 and 96 h after Stx2 injection, nevertheless no correlation with creatinine and urea plasma levels were detected. Our results suggest that MVs-Stx2+ may be an additional biomarker for the characterization and diagnosis of HUS progression. A further analysis is required in order to validate MVs-Stx2+ as biomarker of the disease.

Stimuli-Responsive Hydrogels for Protein Delivery.

Proteins and peptides are potential therapeutic agents, but their physiochemical properties make their use as drug substances challenging. Hydrogels are hydrophilic polymeric networks that can swell and retain high amounts of water or biological fluids without being dissolved. Due to their biocompatibility, their porous structure, which enables the transport of various peptides and proteins, and their protective effect against degradation, hydrogels have gained prominence as ideal carriers for these molecules' delivery. Particularly, stimuli-responsive hydrogels exhibit physicochemical transitions in response to subtle modifications in the surrounding environment, leading to the controlled release of entrapped proteins or peptides. This review is focused on the application of these hydrogels in protein and peptide delivery, including a brief overview of therapeutic proteins and types of stimuli-responsive polymers.

Caloric restriction improves inflammation in different tissues of the Wistar rats with obesity and 2K1C renovascular hypertension.

Renovascular hypertension (RHV) is the cause of high blood pressure due to left renal ischemia, and obesity and hypertension cause an inflammatory response. This work analyzed the inflammatory and tissue repair profile in renal, hepatic, and cardiac tissues in an animal model of RVH associated with a high-fat diet and caloric restriction. The expressions of RORγ-t, IL-17, T-bet, and TNF-α decreased and IFN-γ increased in the right kidney. In relation to the left kidney, caloric restriction decreased the expression of IFN-γ. In the liver, caloric restriction decreased RORγ-t, IL-17, and T-bet. Hypertension associated with obesity decreased the expression of IFN-γ, while caloric restriction increased. In the right kidney, hypertension and obesity, associated or not with caloric restriction, increased the area of collagen fibers. In the heart and liver, caloric restriction reduced the area of collagen fibers. Caloric restriction increased vascular endothelial growth factor, reduced levels of growth transformation factor-ß1 (TGF-ß), and increased collagen I in the left kidney. Hypertension/obesity, submitted or not having caloric restriction, increased TGF-ß in liver. The results suggest that caloric restriction has beneficial effects in lowering blood pressure and regulating tissue proinflammatory cytokines. However, there was no change in the structure and composition of tissue repair markers.

The P300 Auditory Evoked Potential in Cochlear Implant Users: A Scoping Review

Abstract Introduction The P300 auditory evoked potential is a long-latency cortical potential evoked with auditory stimulation, which provides information on neural mechanisms underlying the central auditory processing. Objectives To identify and gather scientific evidence regarding the P300 in adult cochlear implant (CI) users. Data Synthesis A total of 87 articles, 20 of which were selected for this study, were identified and exported to the Rayyan search software. Those 20 articles did not propose a homogeneous methodology, which made comparison more difficult. Most articles (60%) in this review compare CI users with typical hearing people, showing prolonged P300 latency in CI users. Among the studies, 35% show that CI users present a smaller P300 amplitude. Another variable is the influence of the kind of stimulus used to elicit P300, which was prolonged in 30% of the studies that used pure tone stimuli, 10% of the studies that used pure tone and speech stimuli, and 60% of the studies that used speech stimuli. Conclusion This review has contributed with evidence that shows the importance of applying a controlled P300 protocol to diagnose and monitor CI users. Regardless of the stimuli used to elicit P300, we noticed a pattern in the increase in latency and decrease in amplitude in CI users. The user's experience with the CI speech processor over time and the speech test results seem to be related to the P300 latency and amplitude measurements.

AudBility: an online program for central auditory processing screening in school-aged children from 6 to 8 years old.

PURPOSE: To analyze the performance of students aged between in an auditory skills screening software program, considering the influence of biological determinants and the correlation of auditory tasks with the behavioral assessment tests of central auditory processing (PAC), as well as to present the cutoff points of the battery. METHODS: In the first stage, the sample consisted of 96 students with typical development, who underwent hearing screening at school. A self-perception questionnaire and the auditory tasks of sound localization (SL), temporal resolution (TR), temporal ordering of frequency (OT-F) and duration (OT-D), auditory closure (AC), dichotic digit- binaural integration (DD) and figure-ground (FG) were applied. Of these, 66 children participated in the second stage of the study, including basic and behavioral audiological assessment from PAC. RESULTS: The gender variable influenced the DD task to the right ear. Age influenced the outcome of five auditory tasks. The right ear performed better in the DD and OT-F tasks. At the age between 6 and 7 years, there was a correlation between screening and diagnosis in the tasks of AC, TR, DD, FG, and OT-F. At the age of 8 years, there was a correlation in the DD and OT-F tasks. The pass/fail criteria varied according to the task and biological determinants. CONCLUSION: There was a correlation between screening and diagnosis in a greater number of tasks in the age group between 6 and 7 years. The cut-off points for the auditory tasks should be analyzed according to age, sex and/or ear side.

The P300 Auditory Evoked Potential in Cochlear Implant Users: A Scoping Review.

Introduction The P300 auditory evoked potential is a long-latency cortical potential evoked with auditory stimulation, which provides information on neural mechanisms underlying the central auditory processing. Objectives To identify and gather scientific evidence regarding the P300 in adult cochlear implant (CI) users. Data Synthesis A total of 87 articles, 20 of which were selected for this study, were identified and exported to the Rayyan search software. Those 20 articles did not propose a homogeneous methodology, which made comparison more difficult. Most articles (60%) in this review compare CI users with typical hearing people, showing prolonged P300 latency in CI users. Among the studies, 35% show that CI users present a smaller P300 amplitude. Another variable is the influence of the kind of stimulus used to elicit P300, which was prolonged in 30% of the studies that used pure tone stimuli, 10% of the studies that used pure tone and speech stimuli, and 60% of the studies that used speech stimuli. Conclusion This review has contributed with evidence that shows the importance of applying a controlled P300 protocol to diagnose and monitor CI users. Regardless of the stimuli used to elicit P300, we noticed a pattern in the increase in latency and decrease in amplitude in CI users. The user's experience with the CI speech processor over time and the speech test results seem to be related to the P300 latency and amplitude measurements.

Cell Type- and Tissue-specific Enhancers in Craniofacial Development.

The genetic basis of craniofacial birth defects and general variation in human facial shape remains poorly understood. Distant-acting transcriptional enhancers are a major category of non-coding genome function and have been shown to control the fine-tuned spatiotemporal expression of genes during critical stages of craniofacial development1-3. However, a lack of accurate maps of the genomic location and cell type-specific in vivo activities of all craniofacial enhancers prevents their systematic exploration in human genetics studies. Here, we combined histone modification and chromatin accessibility profiling from different stages of human craniofacial development with single-cell analyses of the developing mouse face to create a comprehensive catalogue of the regulatory landscape of facial development at tissue- and single cell-resolution. In total, we identified approximately 14,000 enhancers across seven developmental stages from weeks 4 through 8 of human embryonic face development. We used transgenic mouse reporter assays to determine the in vivo activity patterns of human face enhancers predicted from these data. Across 16 in vivo validated human enhancers, we observed a rich diversity of craniofacial subregions in which these enhancers are active in vivo. To annotate the cell type specificities of human-mouse conserved enhancers, we performed single-cell RNA-seq and single-nucleus ATAC-seq of mouse craniofacial tissues from embryonic days e11.5 to e15.5. By integrating these data across species, we find that the majority (56%) of human craniofacial enhancers are functionally conserved in mice, providing cell type- and embryonic stage-resolved predictions of their in vivo activity profiles. Using retrospective analysis of known craniofacial enhancers in combination with single cell-resolved transgenic reporter assays, we demonstrate the utility of these data for predicting the in vivo cell type specificity of enhancers. Taken together, our data provide an expansive resource for genetic and developmental studies of human craniofacial development.

Injectable Poloxamer Hydrogels for Local Cancer Therapy.

The widespread push to invest in local cancer therapies comes from the need to overcome the limitations of systemic treatment options. In contrast to intravenous administration, local treatments using intratumoral or peritumoral injections are independent of tumor vasculature and allow high concentrations of therapeutic agents to reach the tumor site with minimal systemic toxicity. Injectable biodegradable hydrogels offer a clear advantage over other delivery systems because the former requires no surgical procedures and promotes drug retention at the tumor site. More precisely, in situ gelling systems based on poloxamers have garnered considerable attention due to their thermoresponsive behavior, biocompatibility, ease of preparation, and possible incorporation of different anticancer agents. Therefore, this review focuses on the use of injectable thermoresponsive hydrogels based on poloxamers and their physicochemical and biological characterization. It also includes a summary of these hydrogel applications in local cancer therapies using chemotherapy, phototherapy, immunotherapy, and gene therapy.

Production and Characterization of k-Carrageenan Films Incorporating Cymbopogon winterianus Essential Oil as New Food Packaging Materials.

The global production of synthetic plastics from petroleum-based raw ingredients exceeds 150 million metric tons. The environment is threatened by tons of plastic waste, thus endangering wildlife and the public's health. These consequences increased the interest in biodegradable polymers as potential substitutes for traditional packaging materials. This study aimed to produce and characterize k-carrageenan films incorporating Cymbopogon winterianus essential oil, in which citronellal was determined to be the major compound (41.12%). This essential oil presented remarkable antioxidant activity, as measured through DPPH (IC50 = 0.06 ± 0.01%, v/v; AAI = 85.60 ± 13.42) and ß-carotene bleaching (IC50 = 3.16 ± 0.48%, v/v) methods. The essential oil also showed antibacterial properties against Listeria monocytogenes LMG 16779 (diameter of inhibition zone = 31.67 ± 5.16 mm and MIC = 8 µL/mL), which were also observed when incorporated in the k-carrageenan films. Moreover, scanning electron microscopy showed the reduction of the biofilms of this bacterium, and even its inactivation, due to visible destruction and loss of integrity when the biofilms were created directly on the developed k-carrageenan films. This study also revealed the quorum sensing inhibition potential of Cymbopogon winterianus essential oil (diameter of violacein production inhibition = 10.93 ± 0.81 mm), where it could impede intercellular communication and, hence, lower violacein synthesis. The produced k-carrageenan films were transparent (>90%) and slightly hydrophobic (water contact angle > 90°). This work demonstrated the viability of using Cymbopogon winterianus essential oil to produce k-carrageenan bioactive films as new food packaging materials. Future work should focus on the scale-up production of these films.

Effects of Steam Sterilization on the Properties of Stimuli-Responsive Polymer-Based Hydrogels.

Hydrogels based on stimuli-responsive polymers can change their characteristics in response to small variations in environmental conditions, such as temperature, pH, and ionic strength, among others. In the case of some routes of administration, such as ophthalmic and parenteral, the formulations must meet specific requirements, namely sterility. Therefore, it is essential to study the effect of the sterilization method on the integrity of smart gel systems. Thus, this work aimed to study the effect of steam sterilization (121 °C, 15 min) on the properties of hydrogels based on the following stimuli-responsive polymers: Carbopol® 940, Pluronic® F-127, and sodium alginate. The properties of the prepared hydrogels-pH, texture, rheological behavior, and sol-gel phase transition-were evaluated to compare and identify the differences between sterilized and non-sterilized hydrogels. The influence of steam sterilization on physicochemical stability was also investigated by Fourier-transform infrared spectroscopy and differential scanning calorimetry. The results of this study showed that the Carbopol® 940 hydrogel was the one that suffered fewer changes in the studied properties after sterilization. By contrast, sterilization was found to cause slight changes in the Pluronic® F-127 hydrogel regarding gelation temperature/time, as well as a considerable decrease in the viscosity of the sodium alginate hydrogel. There were no considerable differences in the chemical and physical characteristics of the hydrogels after steam sterilization. It is possible to conclude that steam sterilization is suitable for Carbopol® 940 hydrogels. Contrarily, this technique does not seem adequate for the sterilization of alginate or Pluronic® F-127 hydrogels, as it could considerably alter their properties.