SPTSSA variants alter sphingolipid synthesis and cause a complex hereditary spastic paraplegia.

Sphingolipids are a diverse family of lipids with critical structural and signalling functions in the mammalian nervous system, where they are abundant in myelin membranes. Serine palmitoyltransferase, the enzyme that catalyses the rate-limiting reaction of sphingolipid synthesis, is composed of multiple subunits including an activating subunit, SPTSSA. Sphingolipids are both essential and cytotoxic and their synthesis must therefore be tightly regulated. Key to the homeostatic regulation are the ORMDL proteins that are bound to serine palmitoyltransferase and mediate feedback inhibition of enzymatic activity when sphingolipid levels become excessive. Exome sequencing identified potential disease-causing variants in SPTSSA in three children presenting with a complex form of hereditary spastic paraplegia. The effect of these variants on the catalytic activity and homeostatic regulation of serine palmitoyltransferase was investigated in human embryonic kidney cells, patient fibroblasts and Drosophila. Our results showed that two different pathogenic variants in SPTSSA caused a hereditary spastic paraplegia resulting in progressive motor disturbance with variable sensorineural hearing loss and language/cognitive dysfunction in three individuals. The variants in SPTSSA impaired the negative regulation of serine palmitoyltransferase by ORMDLs leading to excessive sphingolipid synthesis based on biochemical studies and in vivo studies in Drosophila. These findings support the pathogenicity of the SPTSSA variants and point to excessive sphingolipid synthesis due to impaired homeostatic regulation of serine palmitoyltransferase as responsible for defects in early brain development and function.

The experience of children and family in pediatric home hospitalization: A systematic review of qualitative evidence.

OBJECTIVE: To synthesize the experiences of children and parents/caregivers in the process of pediatric home hospitalization (PHH). INTRODUCTION: The practice of home hospitalization (HH), while not a new concept has expanded in recent years. This model of care consists of continuous care at home for children with acute illness or acute chronic disease and presents itself as an alternative to conventional hospitalization (Middel, 2007; Parab et al., 2013). Excellence in pediatric healthcare is fundamental and this review provides a necessary understanding towards the experiences of children and their families in HH. METHODS: Research was carried out in three phases and included both published and grey literature in the CINAHL, MEDLINE, MedicLatina, PubMed, Cochrane Library, Psychology and Behavioral Sciences Collection, and OpenAIRE databases to find relevant articles. Studies published in Portuguese, English, Spanish, and French with no time limit were considered. RESULTS: Findings were aggregated into five categories: communication and care experiences, parental dynamics and role carers, benefits and challenges for parents and children, relationship between parents, children's, and healthcare professionals and enhancing continuity of care and family support. According to ConQual the confidence level of the results was moderate in all articles. CONCLUSIONS: Through PHH, it is possible to avoid the impact of a conventional hospitalization since it promotes family union, increases the affective bond, the feeling of security, comfort, tranquillity, relief, confidence, and autonomy, reducing stressors such as anxiety, fear, nervousness, uncertainty, and fear.

Detecting, quantifying, and discriminating the mechanism of mosaic chromosomal aneuploidies using MAD-seq.

Current approaches to detect and characterize mosaic chromosomal aneuploidy are limited by sensitivity, efficiency, cost, or the need to culture cells. We describe the mosaic aneuploidy detection by massively parallel sequencing (MAD-seq) capture assay and the MADSEQ analytical approach that allow low (<10%) levels of mosaicism for chromosomal aneuploidy or regional loss of heterozygosity to be detected, assigned to a meiotic or mitotic origin, and quantified as a proportion of the cells in the sample. We show results from a multi-ethnic MAD-seq (meMAD-seq) capture design that works equally well in populations of diverse racial and ethnic origins and how the MADSEQ analytical approach can be applied to exome or whole-genome sequencing data, revealing previously unrecognized aneuploidy or copy number neutral loss of heterozygosity in samples studied by the 1000 Genomes Project, cell lines from public repositories, and one of the Illumina Platinum Genomes samples. We have made the meMAD-seq capture design and MADSEQ analytical software open for unrestricted use, with the goal that they can be applied in clinical samples to allow new insights into the unrecognized prevalence of mosaic chromosomal aneuploidy in humans and its phenotypic associations.

Does the recall of caregiver eating messages exacerbate the pathogenic impact of shame on eating and weight-related difficulties?

PURPOSE: The central role of caregiver eating messages has been recognized on later individual's eating behaviour, body image and weight status. Additionally, shame is a painful emotion also associated with the development and maintenance of body image and eating-related difficulties. The main aim of the present study was to explore the moderator effect of the recall of caregiver eating messages on the associations between external shame, body weight and shape-related concerns, and the adoption of inflexible eating rules. METHODS: The sample comprised 479 women from the general population, aged from 18 to 60 years, who completed self-report measures. RESULTS: Results of the correlation analyses showed that early caregiver restrictive/critical eating messages were positively associated with external shame, inflexible eating and overvaluation of body weight and shape. Path analysis results demonstrated a moderator effect of early caregiver critical eating messages on the relationship between external shame and both weight and shape-related concerns and inflexible eating. These results revealed that caregiver restrictive/critical eating messages exacerbated the impact of shame on these psychopathological indices, with the tested model accounting for 19% and 38% of the variance of inflexible eating rules and body weight and shape concerns, respectively. CONCLUSIONS: These findings seem to suggest important research and clinical implications, contributing to the understanding of disordered eating patterns, and appear to represent a new avenue for the development of prevention and intervention programs. Particularly, these findings support the relevance of targeting caregiver eating-related attitudes and messages in prevention interventions for eating psychopathology. LEVEL OF EVIDENCE: Level V, cross-sectional descriptive study.

Job insecurity among nurses, nursing technicians and nursing aides in public hospitals.

OBJECTIVE: To identify the contribution of all the dimensions involved in job insecurity among nursing professionals in Brazilian state public hospitals. METHOD: This was a quantitative descriptive study conducted between March 2015 and February 2016, with nurses, nursing technicians and nursing aides. The data were analyzed using exploratory factor analysis. RESULTS: The sample consisted of 265 nurses (n=161 with job security and n=104 outsourced) and 810 nursing technicians and nursing aides (n=597 with job security and n=213 outsourced). Among the nurses, "Working conditions" accounted for 46.8% of their job insecurity. Among nursing technicians and nursing aides "intensity of work due to work process organization" best explained the construct of job insecurity (51.2%). CONCLUSION: Job insecurity is expressed differently depending on the type of nursing professional. The difference in the distribution of the dimensions showed that job insecurity is related to the hierarchical position of each nursing professional in the technical division of nursing work and the role of each professional in the work process.

UHPLC-MS-based HDAC Assay Applied to Bio-guided Microfractionation of Fungal Extracts.

INTRODUCTION: Histone deacetylases (HDAC) are considered as promising targets for cancer treatment. Today, four HDAC inhibitors, vorinostat, romidepsin, belinostat, and panobinostat, have been approved by the Food and Drug Administration (FDA) for cancer treatment, while others are in clinical trials. Among them, several are naturally occurring fungal metabolites. OBJECTIVE: To develop and optimise an enzyme assay for bio-guided identification of HDAC inhibitors in fungal strains. METHODS: Fluorescence and MS-based HDAC enzymatic assays were compared during the bio-guided fractionation of Penicillium griseofulvum. The MS-based approach was then optimised to evaluate HDAC selectivity using the human recombinant class I isoform HDAC1 and the class II isoform HDAC6. RESULTS: Fluorescence-based assays have several drawbacks when used for bio-guided fractionation because of the native fluorescence and the trypsin inhibitory ability of compounds present in many extracts. The MS-based method led to the isolation of gliocladride C, which is selective for HDAC1 and salirepol, which showed an HDAC6 selectivity. Their activity and presence in P. griseofulvum is described here for the first time. CONCLUSION: The UHPLC-ESI-MS/MS-based method using specific HDAC isoforms is suitable to isolate selective HDAC inhibitors by bio-guided fractionation of fungal strains. Also, it decreases potential interferences with natural products compared to the fluorescence-based assay.

Synthesis of a selective HDAC6 inhibitor active in neuroblasts.

In recent years, the role of HDAC6 in neurodegeneration has been partially elucidated, which led some authors to propose HDAC6 inhibitors as a therapeutic strategy to treat neurodegenerative diseases. In an effort to develop a selective HDAC6 inhibitor which can cross the blood brain barrier (BBB), a modified hydroxamate derivative (compound 3) was designed and synthetized. This compound was predicted to have potential for BBB penetration based on in silico and in vitro evaluation of passive permeability. When tested for its HDAC inhibitory activity, the IC50 value of compound 3 towards HDAC6 was in the nM range in both enzymatic and cell-based assays. Compound 3 showed a cell-based selectivity profile close to that of tubastatin A in SH-SY5Y human neuroblastoma cells, and a good BBB permeability profile.

Cross metathesis with hydroxamate and benzamide BOC-protected alkenes to access HDAC inhibitors and their biological evaluation highlighted intrinsic activity of BOC-protected dihydroxamates.

Conditions for the metathesis of alkenes in the convergent synthesis of HDAC inhibitors have been improved by continuous catalyst flow injection in the reaction media. Intermediate and target compounds obtained were tested for their ability to induce HDAC inhibition and tubulin acetylation, revealing the key role of the tert-butyloxycarbonyl (BOC) group for more HDAC6 selectivity. Molecular modelling added rationale for this BOC effect.

Cell-based multi-substrate assay coupled to UHPLC-ESI-MS/MS for a quick identification of class-specific HDAC inhibitors.

Histone deacetylases (HDAC) are involved in several diseases including cancer, cardiovascular and neurodegenerative disorders, and the search for inhibitors is a current topic in drug discovery. Four HDAC inhibitors have already been approved by the FDA for cancer therapy and others are under clinical studies. However, the clinical utility of some of them is limited because of unfavorable toxicities associated with their broad range of HDAC inhibitory effects. Toxicity could be decreased by using HDAC inhibitors with improved specificity. To date, the most popular screening assays are based on fluorescence-labeled substrates incubated with an enzymatic source (cells extracts or recombinant isoforms). Here, we describe a high-throughput cell-based UHPLC-ESI-MS/MS assay able to rapidly predict activity against HDAC1 and HDAC6 in a cell environment. This method is predicted to be a useful tool to accelerate the search for class-selective HDAC inhibitors in drug discovery.

Prevention of False-Positive Results: Development of an HPTLC Autographic Assay for the Detection of Natural Tyrosinase Inhibitors.

A simple and rapid high-performance thin-layer chromatography-based autographic assay was established to screen plant extracts for the presence of tyrosinase-inhibiting substances. Three mobile phases were selected for the chromatographic separation of different types of extracts. After development, the plate was sprayed with the substrate solution Levodopa followed by a solution of the enzyme tyrosinase. Several known tyrosinase inhibitors were tested simultaneously as positive controls. They were detected as white spots with white light in remission from the plate as well as with white light transmitted through the plate. Some of the investigated extracts included spots showing a different behaviour; some lipophilic substances appeared as white spots in white light remission but were black in white light transmission. This behaviour, which could lead to false-positive results, was due to poor wettability of the corresponding spots. False-positive results were eliminated by adding Triton X-100 to the Levodopa solution and drying the plate after 10 minutes incubation with a molecular sieve. Tyrosinase inhibitors can be clearly identified as white spots against a dark background in white light remission as well as in white light transmitted through the plate. The established high-performance thin-layer chromatography autographic assay was validated and can be used as a standard method for the detection of tyrosinase inhibitors in plant extracts without causing false-positive results.

Alkaloids from psychotria target sirtuins: in silico and in vitro interaction studies.

Epigenetic enzymes such as histone deacetylases play a crucial role in the development of ageing-related diseases. Among the 18 histone deacetylase isoforms found in humans, class III histone deacetylases, also known as sirtuins, seem to be promising targets for treating neurodegenerative conditions. Recently, Psychotria alkaloids, mainly monoterpene indoles, have been reported for their inhibitory properties against central nervous system cholinesterase and monoamine oxidase proteins. Given the multifunctional profile of these alkaloids in the central nervous system, and the fact that the indole scaffold has been previously associated with sirtuin inhibition, we hypothesized that these indole derivatives could also interact with sirtuins. In the present study, alkaloids previously isolated from Psychotria spp. were evaluated for their potential interaction with human sirtuin 1 and sirtuin 2 by molecular docking and molecular dynamics simulation approaches. The in silico results allowed for the selection of five potentially active compounds, namely, prunifoleine, 14-oxoprunifoleine, E-vallesiachotamine, Z-vallesiachotamine, and vallesiachotamine lactone. The sirtuin inhibition of these compounds was confirmed in vitro in a dose-response manner, with preliminary information on their pharmacokinetics properties.

Aurones as histone deacetylase inhibitors: identification of key features.

In this study, a total of 22 flavonoids were tested for their HDAC inhibitory activity using fluorimetric and BRET-based assays. Four aurones were found to be active in both assays and showed IC50 values below 20 µM in the enzymatic assay. Molecular modelling revealed that the presence of hydroxyl groups was responsible for good compound orientation within the isoenzyme catalytic site and zinc chelation.

Anti-Mycobacterium tuberculosis activity and cytotoxicity of Calophyllum brasiliense Cambess (Clusiaceae).

We evaluated the in vitro anti-Mycobacterium tuberculosis activity and the cytotoxicity of dichloromethane extract and pure compounds from the leaves of Calophyllum brasiliense. Purification of the dichloromethane extract yielded the pure compounds (-) mammea A/BB (1), (-) mammea B/BB (2) and amentoflavone (3). The compound structures were elucidated on the basis of spectroscopic and spectrometric data. The contents of bioactive compounds in the extracts were quantified using high performance liquid chromatography coupled to an ultraviolet detector. The anti-M. tuberculosis activity of the extracts and the pure compounds was evaluated using a resazurin microtitre assay plate. The cytotoxicity assay was performed in J774G.8 macrophages using the 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide colourimetric method. The quantification of the dichloromethane extract showed (1) and (2) at concentrations of 31.86 ± 2.6 and 8.24 ± 1.1 µg/mg of extract, respectively. The dichloromethane and aqueous extracts showed anti-M. tuberculosis H37Rv activity of 62.5 and 125 µg/mL, respectively. Coumarins (1) and (2) showed minimal inhibitory concentration ranges of 31.2 and 62.5 µg/mL against M. tuberculosis H37Rv and clinical isolates. Compound (3) showed no activity against M. tuberculosis H37Rv. The selectivity index ranged from 0.59-1.06. We report the activity of the extracts and coumarins from the leaves of C. brasiliense against M. tuberculosis.

Study of leaf metabolome modifications induced by UV-C radiations in representative Vitis, Cissus and Cannabis species by LC-MS based metabolomics and antioxidant assays.

UV-C radiation is known to induce metabolic modifications in plants, particularly to secondary metabolite biosynthesis. To assess these modifications from a global and untargeted perspective, the effects of the UV-C radiation of the leaves of three different model plant species, Cissus antarctica Vent. (Vitaceae), Vitis vinifera L. (Vitaceae) and Cannabis sativa L. (Cannabaceae), were evaluated by an LC-HRMS-based metabolomic approach. The approach enabled the detection of significant metabolite modifications in the three species studied. For all species, clear modifications of phenylpropanoid metabolism were detected that led to an increased level of stilbene derivatives. Interestingly, resveratrol and piceid levels were strongly induced by the UV-C treatment of C. antarctica leaves. In contrast, both flavonoids and stilbene polymers were upregulated in UV-C-treated Vitis leaves. In Cannabis, important changes in cinnamic acid amides and stilbene-related compounds were also detected. Overall, our results highlighted phytoalexin induction upon UV-C radiation. To evaluate whether UV-C stress radiation could enhance the biosynthesis of bioactive compounds, the antioxidant activity of extracts from control and UV-C-treated leaves was measured. The results showed increased antioxidant activity in UV-C-treated V. vinifera extracts.

Clinical and sociodemographic factors associated with time spent sitting in military police.

OBJECTIVE: To verify the association between clinical and sociodemographic factors and time spent sitting in military police. METHOD: This is a cross-sectional study, with 432 military police officers from Eastern Regional Policing Command units of the Military Police of Bahia de Feira de Santana. Data collection took place from August to December 2022 through Google Forms using the International Physical Activity Questionnaire. RESULTS: Men predominated (82.35%), race/color was black (87.04%), the head of the family had completed higher education (47.69%) and police officers with a partner (81.94%). The risk of time spent sitting ≥ 180 minutes per day was lower in males (IRR < 1). Increasing age was associated with a lower risk of time spent sitting ≥ 180 minutes per day (IRR < 1). CONCLUSION: Male police officers with more years of experience were less exposed to sedentary behavior. Specific interventions and health policies aimed at combating sedentary behavior become relevant, aiming to promote health and prevent diseases.

Educational intervention on perceived stress among adults with type 2 diabetes and metabolic syndrome: a non-randomized clinical trial.

Objective: To assess the effectiveness of an educational intervention on perceived stress and metabolic syndrome parameters among adults with type 2 diabetes mellitus. Method: Fifty-one adults (aged 48.73±7.84; 86.3% of women) were included in a non-randomized clinical trial performed in a healthcare unit for six months (Brazilian Clinical Trial Registry: RBR-43K52N). All participants were diagnosed with type 2 diabetes mellitus and metabolic syndrome (intervention group, n=26; control group, n=25). The intervention consisted of a nurse-led educational health-promoting program with a multidisciplinary approach organized in seven workshops. The primary outcome was decreased perceived stress, and the secondary outcome was improvement in metabolic syndrome parameters according to perceived stress levels. These outcomes were assessed at two points in time, at the baseline and follow-up. Results: Participation in the intervention program resulted in a significant decrease in perceived stress (p=0.028). The stressed participants in the intervention group experienced a significant decrease in blood glucose levels (p=0.001) and a significant increase in high-density lipoprotein-cholesterol (p=0.003) concentrations after the six-month intervention. Conclusion: The nurse-led educational health-promoting program decreased perceived stress among adults with type 2 diabetes mellitus and metabolic syndrome, improving fasting blood glucose and high-density lipoprotein cholesterol among the stressed participants in the intervention group.

The spectrum of neurological presentation in individuals affected by TBL1XR1 gene defects.

BACKGROUND: TBL1XR1 encodes a F-box-like/WD40 repeat-containing protein that plays a role in transcription mediated by nuclear receptors and is a known genetic cause of neurodevelopmental disease of childhood (OMIM# 608628). Yet the developmental trajectory and progression of neurologic symptoms over time remains poorly understood. METHODS: We developed and distributed a survey to two closed Facebook groups devoted to families of patients with TBL1XR1-related disorder. The survey consisted of 14 subsections focused upon the developmental trajectories of cognitive, behavioral, motor, and other neurological abnormalities. Data were collected and managed using REDCap electronic data capture tools. RESULTS: Caregivers of 41 patients with a TBL1XR1-related disorder completed the cross-sectional survey. All reported variants affecting a single amino acid, including missense mutations and in-frame deletions, were found in the WD40 repeat regions of Tbl1xr1. These are domains considered important for protein-protein interactions that may plausibly underlie disease pathology. The majority of patients were diagnosed with a neurologic condition before they received their genetic diagnosis. Language appeared most significantly affected with only a minority of the cohort achieving more advanced milestones in this domain. CONCLUSION: TBL1XR1-related disorder encompasses a spectrum of clinical presentations, marked by early developmental delay ranging in severity, with a subset of patients experiencing developmental regression in later childhood.

Exploring men's health in medium and high complexity care in Brazil: A deductive thematic analysis of social determinants.

Background: Men's health is influenced by a complex interplay of social, economic, and cultural determinants. Understanding how these aspects affect the health of adult cisgender men in medium and high-complexity healthcare settings is essential for improving healthcare services and promoting better health outcomes. Objective: This study aimed to analyze the health status of adult cisgender men in medium and high-complexity healthcare settings based on social determinants and conditioners. Methods: This study employed a qualitative design involving 45 adult cisgender men receiving care in medium/high complexity services in Bahia, Brazil. Semi-structured interviews were conducted from July 2019 to February 2020, and data were interpreted based on Dahlgren and Whitehead's Model of Social Determinants of Health using deductive thematic analysis. Results: Proximal determinants included biological aspects, preventive behaviors, lifestyle/social life, and aging processes. Intermediate factors included work conditions, access/utilization of healthcare services/medications, and psychosocial factors. Macro determinants involved income distribution, power dynamics, resource allocation, health inequalities/iniquities, morbidity, culture, political decisions, environmental factors, and structural elements. Conclusion: The health status of men in medium/high complexity care was profoundly influenced by structural social determinants. These determinants impacted healthcare attention, service organization, cultural influences, the reproduction of hegemonic masculinity patterns, lifestyle, social support, and socioeconomic conditions necessary to realize the right to health. Nursing practices should conduct comprehensive assessments that extend beyond physical health indicators.

Monoamine oxidase inhibition by monoterpene indole alkaloids and fractions obtained from Psychotria suterella and Psychotria laciniata.

Alkaloid fractions of Psychotria suterella (SAE) and Psychotria laciniata (LAE) as well as two monoterpene indole alkaloids (MIAs) isolated from these fractions were evaluated against monoamine oxidases (MAO-A and -B) obtained from rat brain mitochondria. SAE and LAE were analysed by HPLC-PDA and UHPLC/HR-TOF-MS leading to the identification of the compounds 1, 2, 3 and 4, whose identity was confirmed by NMR analyses. Furthermore, SAE and LAE were submitted to the enzymatic assays, showing a strong activity against MAO-A, characterized by IC(50) values of 1.37 ± 1.05 and 2.02 ± 1.08 µg/mL, respectively. Both extracts were also able to inhibit MAO-B, but in higher concentrations. In a next step, SAE and LAE were fractionated by RP-MPLC affording three and four major fractions, respectively. The RP-MPLC fractions were subsequently tested against MAO-A and -B. The RP-MPLC fractions SAE-F3 and LAE-F4 displayed a strong inhibition against MAO-A with IC(50) values of 0.57 ± 1.12 and 1.05 ± 1.15 µg/mL, respectively. The MIAs 1 and 2 also inhibited MAO-A (IC(50) of 50.04 ± 1.09 and 132.5 ± 1.33 µg/mL, respectively) and -B (IC(50) of 306.6 ± 1.40 and 162.8 ± 1.26 µg/mL, respectively), but in higher concentrations when compared with the fractions. This is the first work describing the effects of MIAs found in neotropical species of Psychotria on MAO activity. The results suggest that species belonging to this genus could consist of an interesting source in the search for new MAO inhibitors.

Arrival time at a referral hospital and functional disability of people with stroke: a cohort study.

BACKGROUND: Stroke is a major cause of death and functional disability worldwide. Knowledge of the associated factors is essential for defining education, management, and healthcare strategies. OBJECTIVE: To analyze the association between arrival time at a neurology referral hospital (ATRH) and functional disability in patients with ischemic stroke 90 days after the event. DESIGN AND SETTING: Prospective cohort study conducted at a public institution of higher education in Brazil. METHODS: This study included 241 people aged ≥ 18 years who presented ischemic stroke. The exclusion criteria were death, inability to communicate without companions who could answer the research questions, and > 10 days since ictus. Disability was assessed using the Rankin score (mR). Variables for which associations showed a P value ≤ 0.20 in bivariate analysis were tested as modifiers between ATRH and disability. Significant interaction terms were used for multivariate analysis. Multivariate logistic regression analysis was performed with all variables, arriving at the complete model and adjusted beta measures. The confounding variables were included in the robust logistic regression model, and Akaike's Information Criterion was adopted to choose the final model. The Poisson model assumes a statistical significance of 5% and risk correction. RESULTS: Most participants (56.0%) arrived at the hospital within 4.5 hours of symptom onset, and 51.7% presented with mRs of 3 to 5 after 90 days of ictus. In the multivariate model, ATRH ≥ 4.5 hours and females were associated with more significant disability. CONCLUSIONS: Arrival at the referral hospital 4.5 hours after the onset of symptoms or wake-up stroke was an independent predictor of a high degree of functional disability.