Persistent Organic Pollutants in Tagus Estuary Salt Marshes: Patterns of Contamination and Plant Uptake.

The presence of anthropogenic compounds, including organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs), was studied in three salt marshes within the Tagus estuary, Portugal, along an anthropogenic pressure gradient. Results revealed differences in OCPs and PCBs among the marshes, with differing concentration levels. Specifically, one marsh, with surrounding agricultural activity, showed the highest OCP concentrations, while another, with a historical industrial past, exhibited elevated PCB levels. In contrast, a third marsh, part of a natural reserve, displayed comparatively lower concentrations of both substances. Sediment concentrations, likely influenced by agricultural practices, were found to be comparable to or higher than those observed in other Portuguese estuaries. The halophyte Spartina maritima was found to absorb OCPs, particularly in its aboveground tissues, suggesting bioaccumulation within the plant. Additionally, PCB levels appeared to be influenced by industrial history, with one marsh displaying notably higher concentrations. In conclusion, the persistence of organochlorine compounds in the salt marsh ecosystems notwithstanding the regulatory prohibitions implemented in the 1990s highlights the need for continuous monitoring and study of such sites and the necessity of remediation practices, which are imperative to mitigate ecological and health risks in these polluted salt marshes.

Biosocial Technical Systems: An Emerging Approach to Analyse Responses to Novel Diseases.

The COVID-19 pandemic emerged in the context of a parallel epidemic of information, namely an infodemic. With the development of vaccines occurring in record time, a disinformation campaign ensued rendering the infodemic ever more troubling. As COVID-19 had to be curbed with vaccines opinion pools and surveys indicated that a minority, but relevant, part of the general public had weakened trust in public health policies and also on governmental responses to the pandemic in general. This dissent in public opinion on pandemic response is interpreted in this chapter as a controversy related to the efficacy and risks associated to vaccines. Such controversy gained momentum partly because traditional scientific communication has been largely unidirectional rather than bi-directional. We propose to apply a novel biosocial technical perspective to examine the COVID-19 pandemic controversy and communication, articulating social, biological and technical issues. The interaction between COVID-19 and vaccines, i.e. artefact-biological interactions, resulted in vaccine development in record time. However, the interaction between social systems and vaccine as artefacts was plagued by partial public reluctance in their acceptance. This rendered communication efforts ever more relevant, bringing lessons related to the importance of a more fluid bi-directional communication in future disease epidemics.

Hemoplasmas in wild rodents and marsupials from the Caatinga Biome, Brazil.

A total of 231 blood samples from wild mammals belonging to the orders Rodentia (n = 142) and Didelphimorphia (n = 89) were screened by real-time PCR assay (qPCR), being six Rhipidomys sp., 118 Thrichomys laurentius, nine Rattus rattus, four Kerodon rupestris, five Necromys lasiurus, 42 Didelphis albiventris and 47 Monodelphis domestica. Results using qPCR showed that 32 of the total 231 (13.85 %) samples were positive for hemoplasma sequences of the 16S rRNA gene. Sequences from two D. albiventris showed 99.77-99.89 % identity with 'Candidatus Mycoplasma haemoalbiventris' and 99.09 % with 'Candidatus Mycoplasma haemodidelphidis', respectively. Furthermore, one M. domestica and five T. laurentius showed 99.72-99.77 % identity with Mycoplasma sp., and one K. rupestris showed 98.13 % identity with 'Candidatus Mycoplasma haematohydrochaerus'; and from two Rattus rattus showed 99.65-99.89 % identity with Mycoplasma sp. and 'Candidatus Mycoplasma haemomuris'. The 23S rRNA gene sequences obtained from the two D. albiventris showed 100 % identity with 'Ca. M. haemoalbiventris' whereas the sequences from the R. rattus showed only 85.31 % identity with 'Candidatus Mycoplasma haematohydrochaerus'. Two T. laurentius and one K. rupestris showed 84.66-92.97 % identity with 'Candidatus Mycoplasma haemosphiggurus'. Based on phylogenetic and Neighbor-Net network analyses of the 16S and 23S rRNA genes, potential novel species are described. In addition, 'Ca. M. haemoalbiventris' was detected in Didelphis albiventris, and Mycoplasma sp. was detected in Rattus sp. rodents from the Caatinga biome, Brazil.

Phenolic Compounds Characterization of Caryocar brasiliense Peel with Potential Antioxidant Activity.

The pequi (Caryocar brasiliense) fruit peel, despite being frequently discarded, has a high content of bioactive compounds, and therefore has a high nutritional value. The present study aimed to explore the bioactivities in the pequi peel, particularly their potential health benefits at the level of antioxidant activity. The exploitation of this fruit could also present significant economic benefits and applications of pequi by-products would represent a reduction in waste, having a positive impact on the environment. Phenolic compounds present in the pequi exocarp and external mesocarp were identified by paper spray mass spectrometry (PS-MS) and quantified by HPLC. The total phenolic content (TPC) along with the amount of 2,2-diphenyl-1-picrylhydrazyl (DPPH), Ferric Reducing Antioxidant Power (FRAP), and the amount of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) were also determined in peel extracts. Epicatechin was the most abundant phenolic compound found, followed by the caffeic, salicylic, and gallic acids. In addition, fingerprinting revealed compounds related to several beneficial health effects. In short, the results obtained were encouraging for potential applications of pequi peel in the field of functional foods.

Extracellular Vesicles: The Next Generation of Biomarkers and Treatment for Central Nervous System Diseases.

It has been widely established that the characterization of extracellular vesicles (EVs), particularly small EVs (sEVs), shed by different cell types into biofluids, helps to identify biomarkers and therapeutic targets in neurological and neurodegenerative diseases. Recent studies are also exploring the efficacy of mesenchymal stem cell-derived extracellular vesicles naturally enriched with therapeutic microRNAs and proteins for treating various diseases. In addition, EVs released by various neural cells play a crucial function in the modulation of signal transmission in the brain in physiological conditions. However, in pathological conditions, such EVs can facilitate the spread of pathological proteins from one brain region to the other. On the other hand, the analysis of EVs in biofluids can identify sensitive biomarkers for diagnosis, prognosis, and disease progression. This review discusses the potential therapeutic use of stem cell-derived EVs in several central nervous system diseases. It lists their differences and similarities and confers various studies exploring EVs as biomarkers. Further advances in EV research in the coming years will likely lead to the routine use of EVs in therapeutic settings.

The degree of bowel remission predicts phenotype progression in Crohn's disease.

BACKGROUND: Patients with Crohn's disease (CD) are at risk of progressing from inflammatory to stricturing and penetrating phenotypes. The influence of the depth of remission on the risk of progression has not been adequately evaluated. METHODS: A retrospective cohort study including surgically naïve CD patients with inflammatory phenotype evaluated concomitantly by magnetic resonance enterography and colonoscopy. The degree of remission was correlated with the risk of progressing to stricturing and penetrating phenotypes. RESULTS: Three hundred nineteen CD patients were included: 27.0% with transmural remission, 16.0% with isolated endoscopic remission, 14.4% with isolated radiologic remission, and 42.6% without remission. Patients with transmural remission presented the lowest rates of phenotype progression (1.2%), with a significant difference compared to isolated radiologic remission (10.9%, p = 0.019), to isolated endoscopic remission (19.6%, p ≤ 0.001), and to no remission (46.3%, p ≤ 0.001). In multivariate regression analysis, transmural remission (OR 0.017 95% CI 0.002-0.135, p < 0.001), isolated radiologic remission (OR 0.139 95% CI 0.049-0.396, p < 0.001), and isolated endoscopic remission (OR 0.301 95% CI 0.123-0.736, p = 0.008) resulted in lower rates of phenotype progression compared to no remission. No patient with transmural or isolated radiologic remission progressed to penetrating phenotypes. CONCLUSION: The degree of bowel remission correlates with the risk of phenotype progression. Patients with transmural remission are at the lowest risk of progressing to stricturing and penetrating phenotypes.

Comparativo da especificidade das escalas de Tinetti e SPPB na avaliação de idosos saudáveis e com disfunções neuromotoras no sul de Goiás / Comparison of the specificity of the tinetti and sppb scales in the assessment of healthy elderly people and those with neuromotor dysfunctions in southern Goiás

Introdução: o aumento da população idosa no mundo e as alterações fisiológicas decorrentes desse processo refletem a necessidade de boas ferramentas de avaliação para a identificação precoce de possíveis declínios. Objetivo: comparar os achados da avaliação pelas escalas Short Physical Performance Battery (SPPB) e Índice de Equilíbrio e Marcha de Tinetti validadas no Brasil, verificando a especificidade de cada escala na avaliação de idosos saudáveis e com disfunções neuromotoras. Métodos: 76 indivíduos, 43 saudáveis e 33 com disfunção neuromotora, de ambos os sexos, com idade mínima de 60 anos, residentes nos municípios de Caldas Novas-GO, Itumbiara-GO e Goiatuba-GO, foram avaliados em dias alternados pelas escalas SPPB e Tinetti. Resultados: os grupos apresentaram pontuações maiores nas avaliações pelo Índice Tinetti, o que sugere maior sensibilidade da SPPB na avaliação da marcha e do equilíbrio desses indivíduos. Conclusão: SPPB mostrou-se mais específica que o Índice de Tinetti, sendo capaz de detectar alterações que o Índice Tinetti não foi capaz de encontrar.

Introduction: the increase in the elderly population in the world and the physiological changes resulting from this process reflect the need for good assessment tools for the early identification of possible declines. Objective: to compare the assessment findings by the Short Physical Performance Battery (SPPB) and Tinetti's Balance and Gait Index validated in Brazil, verifying the specificity of each scale in the assessment of healthy elderly people and those with neuromotor dysfunctions. Methods: 76 individuals, 43 healthy and 33 with neuromotor dysfunction, of both sexes, aged at least 60 years, living in the cities of Caldas Novas-GO, Itumbiara-GO and Goiatuba-GO, were evaluated on alternate days by the SPPB and Tinetti scales. Results: the groups presented higher scores in the evaluations by Tinetti, which suggests greater sensitivity of the SPPB in the evaluation of gait and balance of these individuals. Conclusion: SPPB was more specific than Tinetti, being able to detect changes that the Tinetti Index was not able to find.

Exploring Inflammatory Asthma Phenotypes: Proteomic Signatures in Serum and Induced Sputum.

Asthma drug responses may differ due to inflammatory mechanisms triggered by the immune cells in the pulmonary microenvironment. Thus, asthma phenotyping based on the local inflammatory profile may aid in treatment definition and the identification of new therapeutic targets. Here, we investigated protein profiles of induced sputum and serum from asthma patients classified into eosinophilic, neutrophilic, mixed granulocytic, and paucigranulocytic asthma, according to inflammatory phenotypes. Proteomic analyses were performed using an ultra-performance liquid chromatography (ultra-HPLC) system coupled to the Q Exactive Hybrid Quadrupole Orbitrap Mass Spectrometer. Fifty-two (52) proteins showed significant differences in induced sputum among the groups, while only 12 were altered in patients' sera. Five proteins in the induced sputum were able to discriminate all phenotypic groups, while four proteins in the serum could differentiate all except the neutrophilic from the paucigranulocytic inflammatory pattern. This is the first report on comparative proteomics of inflammatory asthma phenotypes in both sputum and serum samples. We have identified a potential five-biomarker panel that may be able to discriminate all four inflammatory phenotypes in sputum. These findings not only provide insights into potential therapeutic targets but also emphasize the potential for personalized treatment approaches in asthma management.

ISG15/USP18/STAT2 is a molecular hub regulating IFN I-mediated control of Dengue and Zika virus replication.

The establishment of a virus infection is the result of the pathogen's ability to replicate in a hostile environment generated by the host's immune system. Here, we found that ISG15 restricts Dengue and Zika viruses' replication through the stabilization of its binding partner USP18. ISG15 expression was necessary to control DV replication driven by both autocrine and paracrine type one interferon (IFN-I) signaling. Moreover, USP18 competes with NS5-mediated STAT2 degradation, a major mechanism for establishment of flavivirus infection. Strikingly, reconstitution of USP18 in ISG15-deficient cells was sufficient to restore the STAT2's stability and restrict virus growth, suggesting that the IFNAR-mediated ISG15 activity is also antiviral. Our results add a novel layer of complexity in the virus/host interaction interface and suggest that NS5 has a narrow window of opportunity to degrade STAT2, therefore suppressing host's IFN-I mediated response and promoting virus replication.

Synthesis of ionically imprinted Poly(Alylthiourea) in the presence of 1-(2-Pyridylazo)-2-Napthol (PAN) for preconcentration in magnetic dispersive solid phase of nickel ions in water and food samples.

In the present study, a magnetic ion-imprinted polymer based on n-allylthiourea in the presence of 1-(2-pyridylazo)-2-naphthol (MIIP-PAN) was synthesized, characterized, and applied in the preconcentration of nickel ions by dispersive magnetic solid phase extraction (DMSPE) with FAAS detection. For comparison, non-imprinted polymer (MNIP-PAN) and imprinted polymer without PAN were synthesized. The characterization of the polymers was performed by FT-IR, DRX, TEM, TGA, VSM, and BET. Selectivity studies were performed comparing the competitive adsorption of Ni2+ with other cations on MIIP-PAN and MNIP-PAN, achieving higher relative selectivity coefficients for MIIP-PAN than for MNIP-PAN and NIP. Under optimized conditions, the method provided a preconcentration factor of 76.70, detection limit of 0.25 µg/L and intra-day (2.06 - 2.33 %) and inter-day (1.82 - 4.90 %) precision. The developed method was applied to samples of water, teas, and chocolate powder, and its precision was evaluated through tests of recovery and analysis of certified materials.

Elbow septic arthritis in an infant: an unusual presentation of invasive pneumococcal disease.

Acute septic arthritis is a rare, potentially severe infection that requires immediate treatment to avoid long-term morbidity. Most common aetiological agents are commonly used for empirical treatment, but the choice of antibiotics may be influenced by other factors, such as the patient's age and the epidemiological context.We report an infant with elbow arthritis, whose treatment was changed after Streptococcus pneumoniae serotype 9N was isolated in the blood and synovial fluid. The child underwent arthrocentesis and received intravenous ampicillin followed by oral amoxicillin, with a favourable response and no sequelae at 1-year follow-up.We report an uncommon manifestation of invasive pneumococcal disease in a young immunised healthy infant caused by a non-vaccine serotype. The presence of S. pneumoniae should be considered in joint infections, especially in infants and those with a history of respiratory symptoms.

Tight control using fecal calprotectin and early disease intervention increase the rates of transmural remission in Crohn's disease.

BACKGROUND: Increasing evidence supports the use of transmural remission as a treatment target in Crohn's disease (CD), but it is seldom achieved in clinical practice. Tight monitoring of inflammation using fecal calprotectin with reactive treatment escalation may potentially improve these results. AIMS: To evaluate if treatment escalation based on fecal calprotectin can improve the rates of transmural remission in CD. The influence of the timing of intervention on this strategy was also evaluated. METHODS: Retrospective cohort study including 256 CD patients with 2 consecutive assessments by MRI-enterography and colonoscopy and with regular monitoring using fecal calprotectin. For each occurrence of an elevated fecal calprotectin (≥250 µg/g), we evaluated whether a reactive adjustment of medical treatment was performed. The ratio of treatment escalation/elevated fecal calprotectin was correlated with the chances of reaching transmural remission. Early disease was defined as disease duration <18 months without previous exposure to immunomodulators and biologics. RESULTS: After a median follow-up of 2 years (IQR 1-4), 61 patients (23.8%) reached transmural remission. Ratios of escalation ≥50% resulted in higher rates of transmural remission (34.2% vs. 15.1%, p < 0.001). The effect was more pronounced in patients with early disease (50.0% vs. 12.0%, p = 0.003). In multivariate analysis, a treatment escalation ratio ≥50% (OR 3.46, 95% CI 1.67-7.17, p = 0.001) and early disease intervention (OR 3.24, 95% CI 1.12-9.34, p = 0.030) were independent predictors of achieving transmural remission. CONCLUSION: Tight-monitoring and reactive treatment escalation increase the rates of transmural remission in CD. Intervention in early disease further improves these results.

Impact of the Covid-19 pandemic on the use of screens in early childhood.

OBJECTIVE: To identify the impact of the Covid-19 pandemic on the use of digital screens in early childhood, from the perspective of mothers and education professionals. METHOD: Qualitative study, based on Bronfenbrenner's Bioecological Theory. Nine mothers of children under three years of age, enrolled in daycare centers in a municipality in the state of Paraíba, Brazil, and six education professionals who worked in these daycare centers participated. The data were collected between July and October 2021 via virtual interviews and analyzed according to inductive thematic analysis. RESULTS: The restrictions imposed by the pandemic made screens the only available resource for children's educational activities, interaction, leisure, and distraction, causing an exponential increase in the time they spent in front of screens. CONCLUSION: The pandemic had a direct impact on the use of digital screens in early childhood, increasing the child's exposure time, according to parents and education professionals.

COVID-19: isolamento social e suas consequências na vida de crianças com transtorno do espectro autista / COVID-19: social isolation and consequences in the lives of children with autistic spectrum disorder / COVID-19: el aislamiento social y sus consecuencias en la vida de los niños con trastorno del espectro autista

Objetivo: conhecer as percepções de profissionais de saúde acerca das consequências do isolamento social, causado pela pandemia da COVID-19, no desenvolvimento e na vida de crianças com Transtorno do Espectro Autista. Método: estudo exploratório, descritivo e de abordagem qualitativa, apoiado nos referenciais do Cuidado Centrado na Família e da Análise de Conteúdo de Bardin e aprovado pelo Comitê de Ética em Pesquisa. Desenvolvido em municípios do interior paulista, de setembro a outubro/2021, com participação de doze profissionais de saúde. Resultados: as experiências apreendidas estiveram representadas nas categorias temáticas: "Interrupção das terapêuticas e estratégias profissionais", "Implicações da pandemia no desenvolvimento da criança autista" e "Isolamento social da pandemia e a dinâmica familiar da criança com autismo". Conclusão: o isolamento social decorrente da pandemia do coronavírus interferiu na vida das crianças autistas e suas famílias, com consequências ao desenvolvimento social, motor, linguístico e pedagógico(AU)

Objective: to know the guidelines of health professionals about the consequences of social isolation, caused by the COVID-19 pandemic, on the development and lives of children with Autistic Spectrum Disorder. Method: field study, exploratory, descriptive and with a qualitative approach, supported by the references of Family-Centered Care and Bardin's Content Analysis and approved by the Research Ethics Committee. Developed in municipalities in the interior of São Paulo, from September to October/2021, with the participation of twelve health professionals. Results: the experiences learned followed instructions in the thematic categories: "Interruption of therapies and professional strategies", "Implications of the pandemic on the development of the autistic child" and "Social isolation of the pandemic and the family dynamics of the child with autism". Conclusion: the social isolation resulting from the coronavirus pandemic interfered in the lives of autistic children and their families, with consequences for social, motor, linguistic and pedagogical development(AU)

Objetivo: conocer las percepciones de los profesionales de la salud en cuanto a las consecuencias del aislamiento social, provocado por la pandemia de la COVID-19, en el desarrollo y la vida de los niños con Trastorno del Espectro Autista. Método: estudio exploratorio, descriptivo y con enfoque cualitativo, apoyado en los referenciales del Cuidado Centrado en la Familia y el Análisis de Contenido de Bardin y aprobado por el Comité de Ética en Investigación. El estudio tuvo lugar en municipios del interior de São Paulo, de septiembre a octubre/2021, contando con la participación de doce profesionales de la salud. Resultados: las experiencias aprendidas se representaron por las categorías temáticas: "Interrupción de terapias y estrategias profesionales", "Implicaciones de la pandemia en el desarrollo del niño autista" y "Aislamiento social de la pandemia y la dinámica familiar del niño con autismo". Conclusión: el aislamiento social resultante de la pandemia del coronavirus interfirió en la vida de los niños autistas y sus familias, con consecuencias para el desarrollo social, motor, lingüístico y pedagógico(AU)

Proactive Infliximab Monitoring Improves the Rates of Transmural Remission in Crohn's Disease: A Propensity Score-Matched Analysis.

BACKGROUND: Few patients can reach transmural remission in Crohn's disease (CD) with currently available therapies. Proactive optimization of infliximab (IFX) based on trough levels may potentially improve these results. METHODS: Retrospective cohort study including consecutive CD patients starting treatment with IFX. Rates of transmural remission were compared between patients with and without therapeutic drug monitoring (target level: 5-7 µg/mL). A propensity score-matched analysis was performed to adjust for potential confounders. RESULTS: A total of 195 CD patients were included, 57.9% receiving proactive therapeutic drug monitoring. The rates of transmural remission were higher in patients under proactive therapeutic drug monitoring (37.2% vs 18.3%; P = .004) with similar results in the propensity score-matched analysis (34.2% vs 17.1%; P = .025). In multivariate analysis, proactive therapeutic drug monitoring was independently associated with transmural remission (odds ratio, 2.95; 95% confidence interval, 1.44-6.06; P = .003). CONCLUSIONS: Proactive optimization of IFX based on trough levels increases the rates of transmural remission in CD.

In the following study, we demonstrate that proactive optimization of infliximab using a trough level protocol (aim 5-7 µg/mL) results in higher rates of transmural remission compared with conventional infliximab treatment. These results remained significant in a propensity score­matched analysis.

Optimization of ultrasound-assisted extraction of sorghum phenolics and effect on the stability of 3-deoxyanthocyanins.

Sorghum 3-deoxyanthocyanins (3-DXAs) have greater stability when compared to other anthocyanins. However, the efficiency in extracting these phenolic compounds from cereals, using conventional methods, is low, because most of them are bound to the cell wall. Thus, the aim of this study was to optimize the ultrasound-assisted extraction (UAE) of anthocyanins and total phenolics from sorghum flour, and evaluate the stability of the 3-DXAs. Two frequencies (25 and 45 kHz) were applied in a Central Composite Rotational design to investigate the effect of the variables time (5-75 min) and temperature (30-65°C) using the UAE, with amplitude of the ultrasonic power set at 400 W. In addition, the stability of the 3-DXAs present in the extracts was evaluated. It was possible to successfully optimize the extraction of total anthocyanins (both frequencies) and phenolics (at 45 kHz), and then to obtain equations, to predict their concentrations, with high R2 . The efficiency of UAE was observed, increasing the yield of total anthocyanins, phenolics, and antioxidant capacity at the frequencies of 25 and 45 kHz by 30% and 27%, 10% and 5%, and 30% and 15%, respectively. The apigeninidin was the major 3-DXA found in the extracts, and the luteolinidin was the most stable over storage time. Overall, there was no difference in the 3-DXAs stability obtained by the UAE compared to the conventional method. Thus, ultrasound is an alternative to obtaining sorghum extracts rich in 3-DXAs and other phenolic compounds. PRACTICAL APPLICATION: The health benefits of sorghum 3-deoxyanthocyanins coupled with the growing interest of the food industry in producing healthier food products have motivated this study, because it is important to find ways to optimize 3-deoxyanthocyanins extraction. We have demonstrated that ultrasound-assisted extraction was efficient in extracting high amounts of 3-deoxyanthocyanins and other phenolics from sorghum flour. Moreover, some 3-deoxyanthocyanins have shown to be more stable than others after extraction. Thus, the ultrasound has great potential to produce sorghum phenolic extracts rich in 3-deoxyanthocyanins, which can be used as natural colorants and functional ingredients in foods.

HSP60 mimetic peptides from Mycobacterium leprae as new antigens for immunodiagnosis of Leprosy.

The early diagnosis of leprosy serves as an important tool to reduce the incidence of this disease in the world. Phage display (PD) technology can be used for mapping new antigens to the development of immunodiagnostic platforms. Our objective was to identify peptides that mimic Mycobacterium leprae proteins as serological markers using phage display technology. The phages were obtained in the biopanning using negative and positive serum from household contacts and leprosy patients, respectively. Then, the peptides were synthesized and validated in silico and in vitro for detection of IgG from patients and contacts. To characterize the native protein of M. leprae, scFv antibodies were selected against the synthetic peptides by PD. The scFv binding protein was obtained by immunocapture and confirmed using mass spectrometry. We selected two phase-fused peptides, MPML12 and MPML14, which mimic the HSP60 protein from M. leprae. The peptides MPML12 and MPML14 obtained 100% and 92.85% positivity in lepromatous patients. MPML12 and MPM14 detect IgG, especially in the multibacillary forms. The MPML12 and MPML14 peptides had positivity of 11.1% and 16.6% in household contacts, respectively. There was no cross-reaction in patient's samples with visceral leishmaniasis, tuberculosis and other mycobacteriosis for both peptides. Given these results and the easy obtainment of mimetic antigens, our peptides are promising markers for application in the diagnosis of leprosy, especially in endemic and hyperendemic regions.

Efficient genetic editing of human intestinal organoids using ribonucleoprotein-based CRISPR.

Organoids, combined with genetic editing strategies, have the potential to offer rapid and efficient investigation of gene function in many models of human disease. However, to date, the editing efficiency of organoids with the use of non-viral electroporation methods has only been up to 30%, with implications for the subsequent need for selection, including turnaround time and exhaustion or adaptation of the organoid population. Here, we describe an efficient method for intestinal organoid editing using a ribonucleoprotein-based CRISPR approach. Editing efficiencies of up to 98% in target genes were robustly achieved across different gut anatomical locations and developmental timepoints from multiple patient samples with no observed off-target editing. The method allowed us to study the effect of loss of the tumour suppressor gene PTEN in normal human intestinal cells. Analysis of PTEN-deficient organoids defined phenotypes that likely relate to its tumour suppressive function in vivo, such as a proliferative advantage and increased organoid budding. Transcriptional profiling revealed differential expression of genes in pathways commonly known to be associated with PTEN loss, including mTORC1 activation.

The utilisation of primary health care system concepts positively impacts the assistance of patients with rare diseases despite limited knowledge and experience by health care professionals: A qualitative synopsis of the evidence including approximately 78 000 individuals.

Background: Individuals with rare diseases (RD) have been historically understudied. Previous publications reported that existing primary health care (PHC) workforces and associated infrastructure had been shown to improve their access and health-related outcomes in low- and middle-income countries (LMICs). As current evidence about the impact of PHC on patients diagnosed with RD is yet highly dispersed, this scoping review aimed to collate available evidence of the impact of PHC on patients with RD and summarize published information from multiple stakeholders about the perceived usefulness and barriers to effective use of the PHC system. Methods: We searched Embase, Health System Evidence, PubMed, LILACS / BVS, and The Cochrane Library, from inception to September 1, 2022, for publications providing clear expert- or experience-based insights or data from patients living with RD at the PHC level of care. We included publications highlighting barriers to integrated care of patients with RD, reported by multiple social actors involved in caring for patients with RD. Two investigators screened publications, extracted data, and clustered information among records deemed eligible for inclusion. Data synthesis was performed using narrative and thematic-based analysis. Major findings identified and coded through a semantic-driven analysis were processed in vosViewer software and reported using descriptive statistics. Findings: Eighty publications were included in this review. Quali-quantitative analyses evidenced that the PHC level is essential for approaching patients with RD, mainly due to its longitudinal, multidisciplinary, and coordinated care delivery. In addition, several publications highlighted that the medical curriculum is inappropriate for preparing health care providers to deal with patients presenting unusual signs and symptoms and being diagnosed with RD. PHC teams are essential in orienting patients and families on emergency events. Technology-related concepts were reported in 19 publications, emphasizing their effectiveness on early diagnosis, optimal treatment definition, improvement of quality of life, and long-lasting follow-up. Conclusions: We provided valuable information on the effectiveness of the PHC in fostering a creative, integrative, and supportive environment for patients living with RD. Our results can be helpful to several stakeholders in deciding what actions are still pending to achieve a solid and positive experience for patients with RD in the PHC. Registration: PROSPERO (CRD42022332347).

A highly sensitive nanobody-based immunoassay detecting SARS-CoV-2 nucleocapsid protein using all-recombinant reagents.

Antigen tests have been crucial for managing the COVID-19 pandemic by identifying individuals infected with SARS-CoV-2. This remains true even after immunity has been widely attained through natural infection and vaccination, since it only provides moderate protection against transmission and is highly permeable to the emergence of new virus variants. For this reason, the widespread availability of diagnostic methods is essential for health systems to manage outbreaks effectively. In this work, we generated nanobodies to the virus nucleocapsid protein (NP) and after an affinity-guided selection identified a nanobody pair that allowed the detection of NP at sub-ng/mL levels in a colorimetric two-site ELISA, demonstrating high diagnostic value with clinical samples. We further modified the assay by using a nanobody-NanoLuc luciferase chimeric tracer, resulting in increased sensitivity (detection limit = 61 pg/mL) and remarkable improvement in diagnostic performance. The luminescent assay was finally evaluated using 115 nasopharyngeal swab samples. Receiver Operating Characteristic (ROC) curve analysis revealed a sensitivity of 78.7% (95% confidence interval: 64.3%-89.3%) and specificity of 100.0% (95% confidence interval: 94.7%-100.0%). The test allows the parallel analysis of a large number of untreated samples, and fulfills our goal of producing a recombinant reagent-based test that can be reproduced at low cost by other laboratories with recombinant expression capabilities, aiding to build diagnostic capacity.