m6A landscape is more pervasive when Trypanosoma brucei exits the cell cycle.

N6-methyladenosine (m6A) is a mRNA modification with important roles in gene expression. In African trypanosomes, this post-transcriptional modification is detected in hundreds of transcripts and it affects the stability of the variant surface glycoprotein (VSG) transcript in the proliferating blood stream form. However, how the m6A landscape varies across the life cycle remains poorly defined. Using full-length, non-fragmented RNA, we immunoprecipitated and sequenced m6A-modified transcripts across three life cycle stages of Trypanosoma brucei - slender (proliferative), stumpy (quiescent), and procyclic forms (proliferative). We found that 1037 transcripts are methylated in at least one of these three life cycle stages. While 21% of methylated transcripts are common in the three stages of the life cycle, globally each stage has a distinct methylome. Interestingly, 47% of methylated transcripts are detected in the quiescent stumpy form only, suggesting a critical role for m6A when parasites exit the cell cycle and prepare for transmission by the Tsetse fly. In this stage, we found that a significant proportion of methylated transcripts encodes for proteins involved in RNA metabolism, which is consistent with their reduced transcription and translation. Moreover, we found that not all major surface proteins are regulated by m6A, as procyclins are not methylated, and that, within the VSG repertoire, not all VSG transcripts are demethylated upon parasite differentiation to procyclic form. This study reveals that the m6A regulatory landscape is specific to each life cycle stage, becoming more pervasive as T. brucei exits the cell cycle.

A CRISPRi-based genetic resource to study essential Staphylococcus aureus genes.

IMPORTANCE: Staphylococcus aureus is an important clinical pathogen that causes a high number of antibiotic-resistant infections. The study of S. aureus biology, and particularly of the function of essential proteins, is of particular importance to develop new approaches to combat this pathogen. We have optimized a clustered regularly interspaced short palindromic repeat interference (CRISPRi) system that allows efficient targeting of essential S. aureus genes. Furthermore, we have used that system to construct a library comprising 261 strains, which allows the depletion of essential proteins encoded by 200 genes/operons. This library, which we have named Lisbon CRISPRi Mutant Library, should facilitate the study of S. aureus pathogenesis and biology.

Validity and reliability of a new incremental step test for people with chronic obstructive pulmonary disease.

BACKGROUND: Incremental step tests (IST) can be used to assess exercise capacity in people with chronic obstructive pulmonary disease (COPD). The development of a new step test based on the characteristics of the incremental shuttle walk test (ISWT) is an important study to explore. We aimed to develop a new IST based on the ISWT in people with COPD, and assess its validity (construct validity) and reliability, according to Consensus-based Standards for the selection of health status Measurement Instruments (COSMIN) recommendations. METHODS: A cross-sectional study was conducted in participants recruited from hospitals/clinics. During the recruitment, the participants who presented a 6-minute walk test (6MWT) report in the previous month were also identified and the respective data was collected. Subsequently, participants attended two sessions at their homes. IST was conducted on the first visit, along with the 1 min sit-to-stand (1MSTS) test. IST was repeated on a second visit, performed 5-7 days after the first one. Spearman's correlations were used for construct validity, by comparing the IST with the 6MWT and the 1MSTS. Intraclass correlation coefficient (ICC2,1), SE of measurement (SEM) and minimal detectable change at 95% CI (MDC95) were used for reliability. The learning effect was explored with the Wilcoxon signed-rank test. RESULTS: 50 participants (70.8±7.5 years) were enrolled. IST was significant and moderate correlated with the 6MWT (ρ=0.50, p=0.020), and with the 1MSTS (ρ=0.46, p=0.001). IST presented an ICC2,1=0.96, SEM=10.1 (16.6%) and MDC95=27.9 (45.8%) for the number of steps. There was a statistically significant difference between the two attempts of the IST (p=0.030). CONCLUSION: Despite the significant and moderate correlations with the 6MWT and 1MSTS, the inability to full compliance with the COSMIN recommendations does not yet allow the IST to be considered valid in people with COPD. On the other hand, the IST is a reliable test based on its high ICC, but a learning effect and an 'indeterminate' measurement error were shown. TRIAL REGISTRATION NUMBER: NCT04715659.

N6-methyladenosine in poly(A) tails stabilize VSG transcripts.

RNA modifications are important regulators of gene expression1. In Trypanosoma brucei, transcription is polycistronic and thus most regulation happens post-transcriptionally2. N6-methyladenosine (m6A) has been detected in this parasite, but its function remains unknown3. Here we found that m6A is enriched in 342 transcripts using RNA immunoprecipitation, with an enrichment in transcripts encoding variant surface glycoproteins (VSGs). Approximately 50% of the m6A is located in the poly(A) tail of the actively expressed VSG transcripts. m6A residues are removed from the VSG poly(A) tail before deadenylation and mRNA degradation. Computational analysis revealed an association between m6A in the poly(A) tail and a 16-mer motif in the 3' untranslated region of VSG genes. Using genetic tools, we show that the 16-mer motif acts as a cis-acting motif that is required for inclusion of m6A in the poly(A) tail. Removal of this motif from the 3' untranslated region of VSG genes results in poly(A) tails lacking m6A, rapid deadenylation and mRNA degradation. To our knowledge, this is the first identification of an RNA modification in the poly(A) tail of any eukaryote, uncovering a post-transcriptional mechanism of gene regulation.

Effects of a Home-Based Pulmonary Rehabilitation Program in Patients with Chronic Obstructive Pulmonary Disease in GOLD B Group: A Pilot Study.

Patients with chronic obstructive pulmonary disease (COPD) in the Global Initiative for Chronic Obstructive Lung Disease (GOLD) B group can be included in pulmonary rehabilitation (PR) settings outside the hospitals. This study aimed to explore the feasibility of a home-based pulmonary rehabilitation (HBPR) program and assess its impact on patients with COPD in the GOLD B group. A real-world, pre-post intervention study was conducted with 12 weeks of HBPR (presential home visits and phone calls) using the self-management program Living Well with COPD. The 1-min sit-to-stand test (1MSTS), modified Medical Research Council Questionnaire (mMRC), COPD Assessment Test (CAT), Hospital Anxiety and Depression Scale (HADS), and London Chest Activity of Daily Living (LCADL) were used to assess the impact. Pre-post differences and correlations between changes in outcomes were calculated. In 30 patients (71.6 years, FEV1 (%) 52.8), significant improvements (p < 0.05) were observed on 1MSTS (Pre 17.2, Post 21.2), mMRC (Pre 2.0, Post 1.0), CAT (Pre 16.3, Post 9.9), HADS (Pre 14.4, Post 9.6), and LCADL (Pre 21.0, Post 15.8), with no adverse events reported. When significant, correlations between changes in outcomes were moderate or strong (0.48 ≤ ρ ≤ 0.66). HBPR can be feasible and safe, and it shows the potential to significantly improve outcomes of patients with COPD in the GOLD B group.

[Community life for all: mental health, participation and autonomy. SESPAS Report 2020]. / Vida comunitaria para todas: salud mental, participación y autonomía. Informe SESPAS 2020.

The objective of this article is to critically reflect on the participation in community of people with a diagnosis of mental disorder from a human rights and community health perspective. Firstly, we review basic concepts such as community mental health and the meaning of participation, which is understood as an end and not as a mean. It is important to increase the participation of people with a diagnosis in community spaces beyond the classic circuit of mental health care. This implies to create and share knowledge in a collective, horizontal and consensual way among all the people involved, especially the diagnosed people themselves. Secondly, the experience of the group of women of Radio Nikosia is narrated in first person by the participants themselves. The main highlights of the group are its horizontality, flexibility, and that is a self-organized space outside the health system where it is possible to express oneself without fear of being judged. Processes of trust, recognition, joy, social support and health are generated. Members of the group meet fortnightly and discuss on different topics chosen by them, and take part in political actions for women's own rights such as participation in the media, in feminist calls, in training and talks. We exemplify that other ways of doing community mental health are possible. The challenges are to make them visible, to systemize them and to be able to assess the impact they have on the health not only of the participants, but also the community.

Metabolic cross-feeding in imbalanced diets allows gut microbes to improve reproduction and alter host behaviour.

The impact of commensal bacteria on the host arises from complex microbial-diet-host interactions. Mapping metabolic interactions in gut microbial communities is therefore key to understand how the microbiome influences the host. Here we use an interdisciplinary approach including isotope-resolved metabolomics to show that in Drosophila melanogaster, Acetobacter pomorum (Ap) and Lactobacillus plantarum (Lp) a syntrophic relationship is established to overcome detrimental host diets and identify Ap as the bacterium altering the host's feeding decisions. Specifically, we show that Ap uses the lactate produced by Lp to supply amino acids that are essential to Lp, allowing it to grow in imbalanced diets. Lactate is also necessary and sufficient for Ap to alter the fly's protein appetite. Our data show that gut bacterial communities use metabolic interactions to become resilient to detrimental host diets. These interactions also ensure the constant flow of metabolites used by the microbiome to alter reproduction and host behaviour.

Vida comunitaria para todas: salud mental, participación y autonomía. Informe SESPAS 2020 / Community life for all: mental health, participation and autonomy. SESPAS Report 2020

El objetivo de este artículo es realizar una reflexión crítica sobre la participación de las personas con un diagnóstico de trastorno mental en la comunidad desde una perspectiva de derechos y de construcción comunitaria de espacios colectivos. En la primera parte se realiza una revisión sobre conceptos básicos como salud mental comunitaria y el significado de participación, entendida como un fin y no como un medio. Se plantea la necesidad de aumentar la participación en espacios comunitarios más allá del circuito clásico de atención a la salud mental, procesos que implican crear y compartir conocimiento de manera colectiva, horizontal y consensuada entre todas las personas implicadas, en especial las propias personas diagnosticadas. En la segunda parte se presenta, como ejemplo, el grupo de mujeres de Radio Nikosia, narrado en primera persona por las propias participantes. Los aspectos destacados del grupo son su horizontalidad, flexibilidad y autoorganización en un espacio fuera del sistema sanitario donde es posible expresarse sin miedo a ser juzgadas. En este grupo se generan procesos de confianza, reconocimiento, alegría, apoyo social y salud. El grupo se reúne quincenalmente y reflexiona sobre diferentes temas escogidos. También se plantean acciones políticas por los propios derechos de las mujeres, como la participación en medios de comunicación, convocatorias feministas, formaciones y charlas. Se ejemplifica que otras formas de hacer salud mental comunitaria son posibles. Los retos son visibilizarlas, sistematizarlas y poder valorar el impacto que tienen en la salud de las personas y la comunidad

The objective of this article is to critically reflect on the participation in community of people with a diagnosis of mental disorder from a human rights and community health perspective. Firstly, we review basic concepts such as community mental health and the meaning of participation, which is understood as an end and not as a mean. It is important to increase the participation of people with a diagnosis in community spaces beyond the classic circuit of mental health care. This implies to create and share knowledge in a collective, horizontal and consensual way among all the people involved, especially the diagnosed people themselves. Secondly, the experience of the group of women of Radio Nikosia is narrated in first person by the participants themselves. The main highlights of the group are its horizontality, flexibility, and that is a self-organized space outside the health system where it is possible to express oneself without fear of being judged. Processes of trust, recognition, joy, social support and health are generated. Members of the group meet fortnightly and discuss on different topics chosen by them, and take part in political actions for women's own rights such as participation in the media, in feminist calls, in training and talks. We exemplify that other ways of doing community mental health are possible. The challenges are to make them visible, to systemize them and to be able to assess the impact they have on the health not only of the participants, but also the community

Combined Before-and-After Workplace Intervention to Promote Healthy Lifestyles in Healthcare Workers (STI-VI Study): Short-Term Assessment.

Health care workers (HCWs) are prone to a heavy psycho-physical workload. Health promotion programs can help prevent the onset of chronic and work-related diseases. The aim of the STI-VI 'before-and-after' study, with assessments scheduled at 6 and 12 months, was to improve the lifestyle of HCWs with at least one cardiovascular risk factor. A tailored motivational counseling intervention, focusing on dietary habits and physical activity (PA) was administered to 167 HCWs (53 males; 114 females). BMI, waist circumference, blood pressure, and cholesterol, triglyceride, and blood glucose levels were measured before and after the intervention. The 6-month results (total sample and by gender) showed a marked effect on lifestyle: PA improved (+121.2 MET, p = 0.01), and diets became more similar to the Mediterranean model (+0.8, p < 0.001). BMI dropped (-0.2, p < 0.03), and waist circumference improved even more (-2.5 cm; p < 0.001). Other variables improved significantly: total and LDL cholesterol (-12.8 and -9.4 mg/dL, p < 0.001); systolic and diastolic blood pressure (-4.4 and -2.5 mmHg, p < 0.001); blood glucose (-1.5 mg/dL, p = 0.05); and triglycerides (significant only in women), (-8.7 mg/dL, p = 0.008); but HDL cholesterol levels dropped too. If consolidated at 12 months, these results indicate that our intervention can help HCWs maintain a healthy lifestyle and work ability.