Clinical and molecular characterization of limb-girdle muscular dystrophy 2G/R7 in a large cohort of Brazilian patients.

Limb-girdle muscular dystrophy type 2G/R7 (LGMD2G/R7) is an ultra-rare condition initially identified within the Brazilian population. We aimed to expand clinical and genetic information about this disease, including its worldwide distribution. A multicenter historical cohort study was performed at 13 centers in Brazil in which data from index cases and their affected relatives from consecutive families with LGMD2G/R7 were reviewed from July 2017 to August 2023. Additionally, a systematic literature review was conducted to identify case reports and series of the disease worldwide. Forty-one LGMD2G/R7 cases were described in the Brazilian cohort, being all subjects homozygous for the c.157C>T/(p.Gln53*) variant in TCAP. Survival curves showed that the median disease duration before individuals required walking aids was 21 years. Notably, women exhibited a slower disease progression, requiring walking aids 13 years later than men. LGMD2G/R7 was frequently reported not only in Brazil but also in China and Bulgaria, with 119 cases identified globally, with possible founder effects in the Brazilian, Eastern European, and Asian populations. These findings are pivotal in raising awareness of LGMD2G/R7, understanding its progression, and identifying potential modifiers. This can significantly contribute to the development of future natural history studies and clinical trials for this disease.

In vitro antimicrobial, antibiofilm photodynamic activity, and molecular dynamic simulations of tetra-cationic porphyrinmembrane interactions against foodborne microorganisms.

This manuscript presents a new report on the in vitro antimicrobial photo-inactivation of foodborne microorganisms (Salmonella spp. and Listeria monocytogenes) using tetra-cationic porphyrins. Isomeric tetra-cationic porphyrins (3MeTPyP, 4MeTPyP, 3PtTPyP, and 4PtTPyP) were tested, and antimicrobial activity assays were performed at specific photosensitizer concentrations under dark and white-light LED irradiation conditions. Among the tested bacterial strains, 4MeTPyP exhibited the highest efficiency, inhibiting bacterial growth within just 60 min at low concentrations (17.5 µM). The minimal inhibitory concentration of 4MeTPyP increased when reactive oxygen species scavengers were present, indicating the significant involvement of singlet oxygen species in the photooxidation mechanism. Furthermore, the checkerboard assay testing the association of 4MeTPyP showed an indifferent effect. Atomic force microscopy analyses and dynamic simulations were conducted to enhance our understanding of the interaction between this porphyrin and the strain's membrane.

Referred pain is associated with greater odontogenic spontaneous pain and a heightened pain sensitivity in patients with symptomatic irreversible pulpitis.

BACKGROUND: Symptomatic irreversible pulpitis often results in heightened reactions to thermal stimuli such as pain evoked by a cold stimulus, and spontaneous odontogenic pain (unprovoked pain). OBJECTIVE: This study primarily compared the clinical manifestations of odontogenic spontaneous pain and pain provoked by cold stimulus specifically focusing on their sensory discriminative characteristics (intensity, duration and quality) between symptomatic irreversible pulpitis patients with and without referred pain. METHODS: Twenty-three patients with symptomatic irreversible pulpitis with referred pain and 12 patients without referred pain were included in this cross-sectional study. The following outcomes were assessed: odontogenic spontaneous pain and its descriptors; pain evoked by cold stimulus and qualitative sensory testing before and after local anaesthesia; referred pain location; use of analgesic medication; complementary anaesthesia efficacy. T-test, chi-squared and McNemar tests were applied to the data (p < .50). RESULTS: Patients with referred pain presented a greater odontogenic pain intensity (p < .05) when considered the average of the last 24 h. These patients also showed higher pain rating and pain descriptors (p < .05). Intensity and duration of the pain evoked by cold stimulus in the non-affected contralateral tooth at baseline were higher for patients with referred pain (p < .05). CONCLUSION: Symptomatic irreversible pulpitis patients with referred pain present greater odontogenic spontaneous pain and a heightened pain sensitivity. Therefore, patients with referred pain seem more complex from a pain severity perspective, supporting the clinical utility of discriminating symptomatic irreversible pulpitis with and without referred pain.

Repurposing approved drugs as potential efflux pump inhibitors in multidrug-resistant Pseudomonas aeruginosa.

Aim: To evaluate the action of promethazine, fluoxetine and carbonyl cyanide 3-chlorophenylhydrazone as efflux pump inhibitors (EPIs) against multidrug-resistant Pseudomonas aeruginosa. Methods: The effect of the compounds was evaluated in planktonic cells and bacterial biofilms. Accumulation tests were performed with ethidium bromide to prove their action as EPIs. Then, they were associated with antimicrobials. Results: Effect on planktonic cells and biofilms was found. Assays with ethidium bromide indicate their action as EPIs. Significant reductions in the metabolic activity of biofilms were observed after the association with the antimicrobials, especially for meropenem. Conclusion: It is possible to prove the action of these compounds as EPIs for P. aeruginosa and demonstrate the relevance of efflux pumps in antimicrobial resistance.

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Effects of bioconversion by Battus polydamas on the chemical composition of Aristolochia spp. and evaluation of antimicrobial activity and biocompatibility.

Aristolochia plants are emblematic from an ethnopharmacological viewpoint and are know to possess numerous biological properties, including antiseptic. However, the medicinal potential of these species is debatable because of their representative chemical constituents, aristolochic acids (AAs) and aristolactams (ALs), which are associated, for instance, with nephropathy and cancer. These contrasting issues have stimulated the development of approaches intended to detoxification of aristoloquiaceous biomasses, among which is included the bioconversion method using larvae of the specialist phytophagous insect Battus polydamas, previously shown to be viable for chemical diversification and to reduce toxicity. Thus, eleven Aristolochia spp. were bioconverted, and the antimicrobial activities of the plant methanolic extracts and its respective bioconversion products were evaluated. The best results were found for Aristolochia esperanzae, Aristolochia gibertii, and Aristolochia ringens against Bacillus cereus, with MIC ranging from 7.8 to 31.25 µg/mL. These three species were selected for chemical, antioxidant, cytotoxic, hemolytic, and mutagenic analyses. Chemical analysis revealed 65 compounds, 21 of them possible bioconversion products. The extracts showed potential to inhibit the formation and degradation of B. cereus biofilms. Extracts of A. gibertii and its bioconverted biomass showed antioxidant activity comparable to dibutylhydroxytoluene (BHT) standard. Bioconversion decreased the hemolytic activity of A. esperanzae and the cytotoxicities of A. esperanzae and A. gibertii. None of the extracts was found to be mutagenic. The bioactivities of the fecal extracts were maintained, and biocompatibility was improved. Therefore, the results obtained in this study reveal positive expectations about the natural detoxification process of the Aristolochia species.

Bending of a lipid membrane edge by annexin A5 trimers.

Plasma membrane damage occurs in healthy cells and more frequently in cancer cells where high growth rates and metastasis result in frequent membrane damage. The annexin family of proteins plays a key role in membrane repair. Annexins are recruited at the membrane injury site by Ca+2 and repair the damaged membrane in concert with several other proteins. Annexin A4 (ANXA4) and ANXA5 form trimers at the bilayer surface, and previous simulations show that the trimers induce high local negative membrane curvature on a flat bilayer. The membrane-curvature-inducing property of ANXA5 is presumed to be vital to the membrane repair mechanism. A previously proposed descriptive model hypothesizes that ANXA5-mediated curvature force is utilized at the free edge of the membrane at a wound site to pull the wound edges together, resulting in the formation of a "neck"-shaped structure, which, when combined with a constriction force exerted by ANXA6, leads to membrane repair. The molecular details and mechanisms of repair remain unknown, in part because the membrane edge is a transient structure that is difficult to investigate both experimentally and computationally. For the first time, we investigate the impact of ANXA5 near a membrane edge, which is modeled by a bicelle under periodic boundary conditions. ANXA5 trimers induce local curvature on the membrane leading to global bending of the bicelle. The global curvature depends on the density of annexins on the bicelle, and the curvature increases with the ANXA5 concentration until it reaches a plateau. The simulations suggest that not only do annexins induce local membrane curvature, but they can change the overall shape of a free-standing membrane. We also demonstrate that ANXA5 trimers reduce the rate of phosphatidylserine lipid diffusion from the cytoplasmic to the exoplasmic leaflet along the edge of the bicelle. In this way, membrane-bound annexins can potentially delay the apoptotic signal triggered by the presence of phosphatidylserine lipids in the outer leaflet, thus biding time for repair of the membrane hole. Our findings provide new insights into the role of ANXA5 at the edges of the membrane (the injury site) and support the curvature-constriction model of membrane repair.

Brazilian Physicians Mental Health: A Cross-Sectional Nationwide Study exploring factors associated with prevalence of suicide planning and attempts.

OBJECTIVES: To report suicide planning and attempts' in a lifetime among Brazilian physicians and to explore associated risk factors. METHODS: A nation-wide, online survey based on the Tool for the Assessment of Suicide Risk and Satisfaction with Life Scale was conducted among Brazilian physicians (January 2018 - January 2019). Multivariate explored associations of demographics, psychological, and work-related factors on suicide planning and attempts reports. RESULTS: Among 4,148 respondents, 1,946 (53.5%) were male, 2,527 (60.9%) were 30-60 years old, 2,675 (64.5%) had 2-4 work-contracts and 1,725 (41.6%) reported a weekly workload of 40-60 hours. Overall prevalence of suicide plans was 8.8% (n=364) and suicide attempts were reported by 3.2% (n=133) of respondents. Daily (AdjOR=7.857;95%CI 2.282-27.051, p=0.002) or weekly emotional exhaustion (AdjOR=7.953; 95%CI 2.403-26.324, p=0.001), daily frustration with work (AdjOR=3.093;95%CI 1.711-5.588, p<0.001), and being bisexual (AdjOR=5.083;95%CI 2.544-10.158, p<0.001) were significantly associated with higher odds of reports. Among extremely dissatisfied professionals 38.3% reported having made suicide planning and attempts, while among extremely satisfied only 2.8% reported it (p<0.001). CONCLUSIONS: Brazilian physicians with a lifetime history of suicide planning and attempts presented a higher association with emotional exhaustion and frustration with work. Urgent actions are needed to promote professional protection policies and resilience.

Photodynamic Therapy Directed to Melanoma Skin Cancer by Thermosensitive Hydrogel Containing Chlorophyll A.

Melanoma, a severe form of skin cancer intricately linked to genetic and environmental factors, is predicted to reach 100,000 new cases worldwide by 2040, underscoring the need for effective and safe treatment options. In this study, we assessed the efficacy of a photosensitizer called Chlorophyll A (Chl-A) incorporated into hydrogels (HGs) made of chitosan (CS) and poloxamer 407 (P407) for Photodynamic Therapy (PDT) against the murine melanoma cell line B16-F10. The HG was evaluated through various tests, including rheological studies, SEM, and ATR-FTIR, along with cell viability assays. The CS- and P407-based HGs effectively released Chl-A and possessed the necessary properties for topical application. The photodynamic activity of the HG containing Chl-A was evaluated in vitro, demonstrating high therapeutic potential, with an IC50 of 25.99 µM-an appealing result when compared to studies in the literature reporting an IC50 of 173.8 µM for cisplatin, used as a positive control drug. The developed formulation of CS and P407-based HG, serving as a thermosensitive system for topical applications, successfully controlled the release of Chl-A. In vitro cell studies associated with PDT exhibited potential against the melanoma cell line.

White matter abnormalities in 15 subjects with SPG76.

BACKGROUND AND OBJECTIVES: Hereditary spastic paraplegias (HSPs) are heterogenous genetic disorders characterized by progressive pyramidal tract involvement. SPG76 is a recently identified form of HSP, caused by biallelic calpain-1 (CAPN1) variants. The most frequently described MRI abnormality in SPG76 is mild cerebellar atrophy and non-specific white matter abnormalities were reported in only one case. Following the identification of prominent white matter abnormalities in a subject with CAPN1 variants, which delayed the diagnosis, we aimed to verify the presence of MRI patterns of white matter involvement specific to this HSP. METHODS: We performed a retrospective radiological qualitative analysis of 15 subjects with SPG76 (4 previously unreported) initially screened for white matter involvement. Moreover, we performed quantitative analyses in our proband with available longitudinal studies. RESULTS: We observed bilateral, periventricular white matter involvement in 12 subjects (80%), associated with multifocal subcortical abnormalities in 5 of them (33.3%). Three subjects (20%) presented only multifocal subcortical involvement. Longitudinal quantitative analyses of our proband revealed increase in multifocal white matter lesion count and increased area of periventricular white matter involvement over time. DISCUSSION: SPG76 should be added to the list of HSPs with associated white matter abnormalities. We identified periventricular white matter involvement in subjects with SPG76, variably associated with multifocal subcortical white matter abnormalities. These findings, in the presence of progressive spastic paraparesis, can mislead the diagnostic process towards an acquired white matter disorder.

Films composed of white angico gum and chitosan containing chlorhexidine as an antimicrobial agent.

Anadenanthera colubrina, popularly known as white angico, is a species extensively cultivated in Brazil, mainly in the cerrado region, including the state of Piauí. This study examines the development of films composed of white angico gum (WAG) and chitosan (CHI) and containing chlorhexidine (CHX), an antimicrobial agent. The solvent casting method was used to prepare films. Different combinations and concentrations of WAG and CHI were used to obtain films with good physicochemical characteristics. Properties such as the in vitro swelling ratio, the disintegration time, folding endurance, and the drug content were determined. The selected formulations were characterised by scanning electron microscopy, Fourier-transform infrared spectroscopy, differential scanning calorimetry, thermogravimetric analysis, and X-ray diffraction, and the CHX release time and antimicrobial activity were evaluated. CHX showed a homogenous distribution in all CHI/WAG film formulations. The optimised films showed good physicochemical properties with 80% CHX release over 26 h, which is considered promising for local treatment of severe lesions in the mouth. Cytotoxicity tests of the films did not show toxicity. The antimicrobial and antifungal effects were very effective against the tested microorganisms.

Clinical and demographic features of minor salivary gland tumors: A collaborative study of 480 cases.

OBJECTIVE: To describe the frequency, clinical, and demographic features of minor salivary gland tumors and possible associated factors. MATERIALS AND METHODS: A cross-sectional study was conducted. Clinical and demographic data were collected from biopsy records of two oral pathology services. Chi-square test, Fisher's exact test, and descriptive statistical analysis were performed. RESULTS: A total of 480 (0.89%) minor salivary gland tumors were retrieved, 272 (56.7%) benign and 147 (30.7%) malignant. Sixty-one (12.6%) had no subtype specification. Most patients were women (307/64.0%), in sixth decade of life (80/16.7%), with a mean age of 45.32 years. Palate was the most common site (336/70.1%). Pleomorphic adenoma (PA; 245/51.1%), mucoepidermoid carcinoma (MEC; 70/14.6%), and adenoid cystic carcinoma (ACC; 43/8.9%) were the most frequent tumors. Symptomatic case, recurrence, and tobacco use were associated with malignancy (p < 0.05). PA and MEC were more frequent in palate (p < 0.05). No association between the three most frequent histological types and gender or age group was observed (p > 0.05). CONCLUSIONS: This represents one of the largest exclusive series of minor salivary gland tumors in Brazil and worldwide. PA, MEC, and ACC were the most frequent tumors. Clinical and demographic data are similar from Brazilian studies or from other countries.

Potential binding modes of the gut bacterial metabolite, 5-hydroxyindole, to the intestinal L-type calcium channels and its impact on the microbiota in rats.

Intestinal microbiota and microbiota-derived metabolites play a key role in regulating the host physiology. Recently, we have identified a gut-bacterial metabolite, namely 5-hydroxyindole, as a potent stimulant of intestinal motility via its modulation of L-type voltage-gated calcium channels located on the intestinal smooth muscle cells. Dysregulation of L-type voltage-gated calcium channels is associated with various gastrointestinal motility disorders, including constipation, making L-type voltage-gated calcium channels an important target for drug development. Nonetheless, the majority of currently available drugs are associated with alteration of the gut microbiota. Using 16S rRNA sequencing this study shows that, when administered orally, 5-hydroxyindole has only marginal effects on the rat cecal microbiota. Molecular dynamics simulations propose potential-binding pockets of 5-hydroxyindole in the α1 subunit of the L-type voltage-gated calcium channels and when its stimulatory effect on the rat colonic contractility was compared to 16 different analogues, ex-vivo, 5-hydroxyindole stood as the most potent enhancer of the intestinal contractility. Overall, the present findings imply a potential role of microbiota-derived metabolites as candidate therapeutics for targeted treatment of slow intestinal motility-related disorders including constipation.

Centralidade do trabalho e saúde mental / Centrality of work for mental health

Partindo da noção de trabalho vivo, Christophe Dejours apresenta a tese da centralidade do trabalho para a subjetividade e a saúde mental. Para tanto, traz à baila a discussão sobre: a atividade, a insuficiência da tarefa e a experiência de fracasso diante do real; bem como a inteligência do corpo e a afetividade, marcando sua transformação à medida que realiza a atividade, em uma relação dinâmica entre o corpo, a subjetividade e a atividade. Não sendo o trabalho algo solipsista, tais elementos se entrecruzam em meio a outrem, o que requisita a cooperação e os aspectos complexos da formulação de regras para a própria atividade e demanda a confiança, a deliberação coletiva. É nesse coletivo, no viver juntos, que pode ocorrer o reconhecimento que, realizado sobre o que é feito, permite a realização de si no campo social, voltando-se para quem o faz, para uma identidade, que é o que protege a saúde mental. Na ausência da convivialidade e reconhecimento, o trabalho produz sofrimento patógeno. Desse modo, Dejours demonstra que nunca há neutralidade do trabalho no que diz respeito à subjetividade e à identidade, possibilitando a realização de si ou sua destruição

Based on the notion of living work, Christophe Dejours presents the thesis of the centrality of work for subjectivity and mental health. To do so, he discusses activity, the insufficiency of the task, the experience of failure in the face of reality, and the intelligence of the body and affectivity, marking its transformation as it performs the activity in a dynamic relationship between the body, subjectivity, and the activity. Rather than configuring a solipsistic text, such elements intersect each other, requiring cooperation and the complex aspects of formulating rules for the activity itself, which requires trust and collective deliberation. It is in this collective, in living together, that recognition can occur, which , based on what is done, enables self-realization in the social field, turning to those who do it and to an identity, which is what protects mental health. In the absence of conviviality and recognition, work produces pathogenic suffering. Thus, the author shows that work is never neutral toward subjectivity and identity, enabling self-realization or its own destruction

Vivências subjetivas de profissionais que realizam Vigilância em Saúde do Trabalhador / Subjective experiences of professionals who carry out Occupational Health Surveillance

Este artigo objetiva compreender a mobilização subjetiva que surge na atividade de trabalho de profissionais que realizam Vigilância em Saúde do Trabalhador (Visat). Participaram deste estudo cinco trabalhadores, de ambos os sexos, de um Centro de Referência em Saúde do Trabalhador (Cerest) da capital de um estado do Nordeste do Brasil, os quais foram entrevistados nos meses de janeiro e fevereiro de 2020. Os materiais produzidos foram analisados à luz da Psicodinâmica do Trabalho. Os resultados mostram que sua fonte de sofrimento está ligada ao que é vivido pelos trabalhadores das empresas que passam por Visat, cujos problemas relacionados à precarização muitas vezes ficam sem solução. Ao lado disso, aqueles que fazem Visat também se percebem muitas vezes precarizados em seus salários, vínculos e condições de trabalho. Finalmente, a falta de estrutura e investimento em saúde do trabalhador, assim como a legislação

This study aims to understand the subjective mobilization that arises in the work activity of professionals who perform Worker Health Surveillance (WHS). In total, five workers of all genders from a Worker Health Reference Center (WHRC) in a capital city of a Northeastern state in Brazil participated in this study. They were interviewed in January and February 2020. The materials produced were analyzed in light of Work Psychodynamics. Results show that their source of suffering is linked to what they experience in companies undergoing WHS, whose precarization-related problems often remain unresolved. In addition to this, those who perform WHS also often perceive themselves as in precarious situations regarding their salaries, job security, and working conditions. Finally, the lack of structure and investment in workers' health and legislation (particularly at the municipal level) pose significant challenges for them to carry out their work

A multinational study of acute and long-term outcomes of Type 1 galactosemia patients who carry the S135L (c.404C > T) variant of GALT.

Patients with galactosemia who carry the S135L (c.404C > T) variant of galactose-1-P uridylyltransferase (GALT), documented to encode low-level residual GALT activity, have been under-represented in most prior studies of outcomes in Type 1 galactosemia. What is known about the acute and long-term outcomes of these patients, therefore, is based on very limited data. Here, we present a study comparing acute and long-term outcomes of 12 patients homozygous for S135L, 25 patients compound heterozygous for S135L, and 105 patients homozygous for two GALT-null (G) alleles. This is the largest cohort of S135L patients characterized to date. Acute disease following milk exposure in the newborn period was common among patients in all 3 comparison groups in our study, as were long-term complications in the domains of speech, cognition, and motor outcomes. In contrast, while at least 80% of both GALT-null and S135L compound heterozygous girls and women showed evidence of an adverse ovarian outcome, prevalence was only 25% among S135L homozygotes. Further, all young women in this study with even one copy of S135L achieved spontaneous menarche; this is true for only about 33% of women with classic galactosemia. Overall, we observed that while most long-term outcomes trended milder among groups of patients with even one copy of S135L, many individual patients, either homozygous or compound heterozygous for S135L, nonetheless experienced long-term outcomes that were not mild. This was true despite detection by newborn screening and both early and life-long dietary restriction of galactose. This information should empower more evidence-based counseling for galactosemia patients with S135L.

Effect of low-level light therapy before radiotherapy in oral squamous cell carcinoma: An in vitro study.

Radiation therapy for head and neck squamous cell carcinoma (HNSCC) is associated with several complications. Although photobiomodulation (PBM) has radioprotective effects in normal tissue, it could also enhance the growth of neoplastic cells. Thus, the present study aimed to investigate the cellular response of oral squamous cell carcinoma with pre-exposure to low-level phototherapy before radiotherapy. SCC9, Cal-27, A431, and HaCaT cell lines were subjected to low-level light therapy and radiotherapy. The cells were treated with a single energy density (300 J/cm2) of a light-emitting diode (660 nm) prior to ionizing radiation at different doses (0, 2, 4, and 6 Gy). After 24 h, wound scratch, proliferation, clonogenic cell survival, cell death, and reactive oxygen species (ROS) analyses were performed to evaluate cell response. The cell lines pre-exposed to PBM at the analyzed dosage were radiosensitive. The treatment significantly reduced cell proliferation and clonogenic cell survival. Migration and cell death assays also revealed positive results, with the treatment group showing lower rate of migration and higher cell death than did the control group. Moreover, PBM effectively increased the intracellular levels of ROS. PBM at 300 J/cm2 is a promising radiosensitizing modality to reduce the radiation dose and avoid the intolerable side effects of radiotherapy for HNSCC, thus increasing the probability of successful treatment. However, further studies are needed to support and confirm the results.