Mapping possible interaction sites for crotoxin in CNF, a gamma PLA2 inhibitor from Crotalus durissus terrificus rattle snake, using SPOT synthesis.

Phospholipases A2 (PLA2s) are main components of snake venoms. Several snake species possess endogenous PLA2 inhibitors in their circulating blood, which are generally known as sbPLIs (an acronym for snake blood phospholipase A2inhibitors). The sbPLIs are categorized in three classes (alpha, beta or gamma) depending on the existence of distinguishing protein domains in their structure. The Crotalus durrissus terrificus venom has a highly neurotoxic PLA2 - crotoxin (CTX) - in its composition and the self-protection of the snake is mainly ensured by a sbγPLI named CNF (standing for Crotalusneutralizing factor). In an attempt to find smaller molecules able to inhibit the catalytic activity of CTX, in the present study we used linear peptide arrays to identify CNF segments possibly involved in the interaction with the toxin. Five reacting segments were identified as possible interacting regions. The target peptides were synthesized and located in the in silico CNF structure. Although all of them are exposed to the solvent, high concentrations were needed to inhibit the PLA2 activity of the whole venom or CTX. Limitations of the methodology employed and particular characteristics of CTX inhibition by CNF are discussed.

Antiaging effects of a skin care formulation containing nanoencapsulated antioxidants: A clinical, in vitro, and ex vivo study.

BACKGROUND: The development of effective cosmetic products for the reduction of the signs of skin aging is a complex process which requires an optimized combination of ingredients and specialized systems to deliver the actives to the skin layers. AIM: To evaluate the tolerance and antiaging clinical efficacy of a cosmetic formulation containing a blend of nanoencapsulated antioxidants: ascorbyl palmitate, resveratrol, tocopherol, caffeine, carnosine, and niacinamide. METHODS: Clinical efficacy was determined by subjective and instrumental analyses of collagen synthesis by fluorescence spectroscopy, by three-dimensional imaging analysis of suborbital edema, and by analysis of skin hydration and sebum content by biophysical techniques-Corneometer® and Sebumeter®. RESULTS: The studied formulation was safe and effective for the improvement of skin appearance by increasing collagen synthesis and skin moisturizing and by reducing facial blemishes, swelling, and oiliness. A preclinical exploratory approach using an experimental model of human cell and skin cultures agreed with the observed antiaging effects, identifying mechanisms related to the containment of oxidative stress, reduction of melanin production, increased synthesis of type I procollagen, and regulation of the epidermal cohesion protein filaggrin. CONCLUSIONS: The skin benefits obtained resulted from the combination of the ingredients in the formulation and the nanoencapsulation-based delivery system, which favors the solubility, safety, efficacy, and bioavailability of the preparation to the skin.

CD8+ tissue-resident memory T-cell development depends on infection-matching regulatory T-cell types.

Immunological memory is critical for immune protection, particularly at epithelial sites, which are under constant risk of pathogen invasions. To counter invading pathogens, CD8+ memory T cells develop at the location of infection: tissue-resident memory T cells (TRM). CD8+ T-cell responses are associated with type-1 infections and type-1 regulatory T cells (TREG) are important for CD8+ T-cell development, however, if CD8+ TRM cells develop under other infection types and require immune type-specific TREG cells is unknown. We used three distinct lung infection models, to show that type-2 helminth infection does not establish CD8+ TRM cells. Intracellular (type-1) and extracellular (type-3) infections do and rely on the recruitment of response type-matching TREG population contributing transforming growth factor-ß. Nevertheless, type-1 TREG cells remain the most important population for TRM cell development. Once established, TRM cells maintain their immune type profile. These results may have implications in the development of vaccines inducing CD8+ TRM cells.

Morphological and molecular study of Syllinae (Annelida, Syllidae) from Bermuda, with the description of five new species.

Although polychaetes from the Bermuda Archipelago have been studied since the beginning of the twentieth century, syllids have been particularly neglected in this area, which is surprising considering this family is usually a dominant group in marine benthic ecosystems. To fill this knowledge gap, we have carried out an extensive analysis of Bermudan Syllidae, combining morphological and molecular data including four nuclear and mitochondrial markers (cytochrome c oxidase subunit I, 18S rRNA, 16S rRNA and 28S rRNA). We have identified and established the phylogenetic position of five new species, Haplosyllis anitae n. sp., Haplosyllis guillei n. sp., Haplosyllis larsi n. sp., Haplosyllis vassiae n. sp. and Syllis laiae n. sp., together with Haplosyllis cf. cephalata. Overall, our results extend the knowledge on the diversity of Syllidae in Bermuda, increasing the number of species present in the area to 25. Our results also recover Opisthosyllis and Syllis as non-monophyletic genera, for which traditional diagnostic morphological features do not accurately reflect their evolutionary histories, and thus we propose that these groups should be reorganized based on molecular characters.

Long-term neurological complications in COVID-19 survivors: study protocol of a prospective cohort study (NeurodegCoV-19).

BACKGROUND: Evidence suggests an association between SARS-CoV-2 infection and worse performance on cognitive tests, and a higher risk of Parkinson's disease (PD) and dementia up to 6 and 12 months after infection, respectively. Longer follow-ups with comparison groups are needed to clarify the potentially increased risk of neurodegenerative diseases in COVID-19 survivors, namely those infected before mass vaccination. METHODS: A prospective study started in July 2022 with four cohorts of 150 individuals each, defined according to SARS-CoV-2 infection and hospitalisation status between March 2020 and February 2021: cohort 1-hospitalised due to SARS-CoV-2 infection; cohort 2-hospitalised, COVID-19-free; cohort 3-infected, not hospitalised; cohort 4-not infected, not hospitalised. Cohort 2 will be matched to cohort 1 according to age, sex, level of hospitalisation care and length of stay; cohort 4 will be age-matched and sex-matched to cohort 3. Baseline, 1-year and 2-year follow-up evaluations will include: cognitive performance assessed with the Montreal Cognitive Assessment (MoCA) and neuropsychological tests; the assessment of prodromal markers of PD with Rapid Eye Movement Sleep Behaviour Disorder single-question Screen and self-reported olfactory and gustative alterations; screening of PD with the 9-item PD screening questionnaire; gait evaluation with Timed Up&Go test. Suspected cases of cognitive impairment and PD will undergo a clinical evaluation by a neurologist. Frequency measures of neurological complications, prodromal markers and diagnoses of dementia and PD, will be presented. The occurrence of cognitive decline-the difference between baseline and 1-year MoCA scores 1.5 SD below the mean of the distribution of the variation-will be compared between cohorts 1 and 2, and cohorts 3 and 4 with OR estimated using multivariate logistic regression. ETHICS AND DISSEMINATION: This study received ethics approval from the Ethics Committees of the health units Unidade Local de Saúde de Matosinhos and Centro Hospitalar de Entre Douro e Vouga, and informed consent is signed for participating. Results will be disseminated among the scientific community and the public.

On the hormonal control of posterior regeneration in the annelid Platynereis dumerilii.

Regeneration is the process by which many animals are able to restore lost or injured body parts. After amputation of the posterior part of its body, the annelid Platynereis dumerilii is able to regenerate the pygidium, the posteriormost part of its body that bears the anus, and a subterminal growth zone containing stem cells that allows the subsequent addition of new segments. The ability to regenerate their posterior part (posterior regeneration) is promoted, in juvenile worms, by a hormone produced by the brain and is lost when this hormonal activity becomes low at the time the worms undergo their sexual maturation. By characterizing posterior regeneration at the morphological and molecular levels in worms that have been decapitated, we show that the presence of the head is essential for multiple aspects of posterior regeneration, as well as for the subsequent production of new segments. We also show that methylfarnesoate, the molecule proposed to be the brain hormone, can partially rescue the posterior regeneration defects observed in decapitated worms. Our results are therefore consistent with a key role of brain hormonal activity in the control of regeneration and growth in P. dumerilii, and support the hypothesis of the involvement of methylfarnesoate in this control.

Annelid adult cell type diversity and their pluripotent cellular origins.

Annelids are a broadly distributed, highly diverse, economically and environmentally important group of animals. Most species can regenerate missing body parts, and many are able to reproduce asexually. Therefore, many annelids can generate all adult cell types in adult stages. However, the putative adult stem cell populations involved in these processes, as well as the diversity of adult cell types generated by them, are still unknown. Here, we recover 75,218 single cell transcriptomes of Pristina leidyi , a highly regenerative and asexually-reproducing freshwater annelid. We characterise all major annelid adult cell types, and validate many of our observations by HCR in situ hybridisation. Our results uncover complex patterns of regionally expressed genes in the annelid gut, as well as neuronal, muscle and epidermal specific genes. We also characterise annelid-specific cell types such as the chaetal sacs and globin + cells, and novel cell types of enigmatic affinity, including a vigilin + cell type, a lumbrokinase + cell type, and a diverse set of metabolic cells. Moreover, we characterise transcription factors and gene networks that are expressed specifically in these populations. Finally, we uncover a broadly abundant cluster of putative stem cells with a pluripotent signature. This population expresses well-known stem cell markers such as vasa, piwi and nanos homologues, but also shows heterogeneous expression of differentiated cell markers and their transcription factors. In these piwi + cells, we also find conserved expression of pluripotency regulators, including multiple chromatin remodelling and epigenetic factors. Finally, lineage reconstruction analyses reveal the existence of differentiation trajectories from piwi + cells to diverse adult types. Our data reveal the cell type diversity of adult annelids for the first time and serve as a resource for studying annelid cell types and their evolution. On the other hand, our characterisation of a piwi+ cell population with a pluripotent stem cell signature will serve as a platform for the study of annelid stem cells and their role in regeneration.

Development of Photoprotective Formulations: Influence of Formulation Composition on the SPF and Mechanical Properties.

The evaluation of the vehicle formulation is important during the development of sunscreens, as it influences their efficacy. In this context, the aim of the present study was to develop photoprotective formulations and evaluate the influence of the formulation components in the sun protection factor (SPF) and physical-mechanical and sensory properties of the formulations. We evaluated four sunscreens through a 22 full factorial design in terms of concentration and emulsifier type. The design of experiments (DOE) parameters were SPF, thixotropy, and work of shear. After the screening of the formulations by DOE, the SPF values, mechanical and sensory properties, and stability were evaluated. All study formulations showed non-Newtonian pseudoplastic behavior, compatible with sunscreens, and presented SPF values above 30. The factors evaluated in DOE had significant interactions for all the analyzed parameters. The concentration of the phosphate-based emulsifier influenced the SPF parameter. The work of shear was influenced by the concentration of polyglyceryl-based emulsifier. The concentration and the type of emulsifier influenced the thixotropy. Finally, effective sunscreens were developed, and the type and concentration of emulsifiers had an influence on the SPF of the formulations. In addition, the formulations chosen by DOE were stable and showed good sensory properties.

Evaluation of Morphological and Structural Skin Alterations on Diabetic Subjects by Biophysical and Imaging Techniques.

Diabetes causes increased production of advanced glycation end products (AGEs), which may lead to irreversible damage to collagen fibers, and early and more accentuated signs of skin aging. Thus, the objective of this study was to evaluate diabetic skin's mechanical and morphological characteristics and compare these to healthy skin. Twenty-eight female participants aged between 39 and 55 years were enrolled: half had type 2 diabetes, and the others were healthy. Wrinkles, transepidermal water loss (TEWL), stratum corneum water content, skin color, elasticity, morphological and structural characteristics of epidermis and dermis echogenicity were evaluated using biophysical and skin imaging techniques. Higher TEWL values were observed in participants with diabetes, who also showed lower skin elasticity and wrinkles with greater volume, area, and depth. In addition, the Reflectance Confocal Microscopy (RCM) imaging analysis showed that all participants with diabetes presented polycyclic papillae and deformed and amorphous collagen fibers. The obtained data showed significant differences between healthy and diabetic skin and could help develop more specific topical treatments to improve the treatment of skin conditions in people with diabetes. Finally, RCM is an advanced imaging technique that allows for a more profound analysis of diabetic skin, which could assist in the evaluation of dermocosmetic treatments to improve the skin alterations caused by this disease.

Development of multifunctional sunscreens: Evaluation of physico-mechanical and film-forming properties.

The exposome consists of several factors such as solar radiation and pollution, which can provoke skin damage and lead to premature skin aging. Thus, the use of multifunctional sunscreens is critical in order to prevent this damage. In addition, film formation is very important to reach the expected SPF. Within this context, the objective of the present study was to develop and evaluate the in vivo SPF, sensory, physico-mechanical, and film-forming properties of sunscreens containing a biopolymer from Tara and red algae. A clinical study of the film-forming effect and of skin hydration was performed by instrumental measurements and by biophysical and skin imaging techniques. The SPF of both formulations, with or without the biopolymer, was 45.6. This result was 10.09% higher than expected. higher than expected. However, the sunscreen added to the biopolymer showed better sensory and texture properties, significantly increased skin hydration and reduced transepidermal water loss. The film-forming property was observed by the analysis of Reflectance Confocal Microscopy images 2, 4, and 6 h after formulation application, and this result was more pronounced for the sunscreen added to the biopolymer. Thus, the film-forming property of the biopolymer was important for prolonging the skin barrier function due to film formation and to obtain more effective and multifunctional sunscreens that provide longer protection.

Reliability and validity of the Patient Generated Index (PGI) in patients with chronic kidney disease (CKD) on dialysis, a new approach to quality of life.

PURPOSE: To evaluate the test-retest reliability and validity of the Patient Generated Index (PGI) in individuals with Chronic Kidney Disease (CDK) undergoing hemodialysis. METHODS: Through a non-experimental study with repeated measures, PGI was applied twice to assess internal consistency and test-retest reliability. Correlations with the Kidney Disease Quality of Life Short Form (KDQOL-SF), the Human Activity Profile (HAP) questionnaire, the Social Participation Scale, and the Glittre ADL Test were used. RESULTS: 91 individuals with CKD were evaluated. There was high reliability for the PGI (ICC= 0.97) PGI correlated with KQDOL - SF in Functional Capacity r = 0.38 (p < 0.001), Emotional Well-Being r = 0.31 (p = 0.003), Social Aspect r = 0.22 (p = 0.036), Emotional Function r = 0.22 (p = 0.038) and Effect of Kidney Disease r = 0.21 (p = 0.042), and Physical scores r = 0.24 (p = 0.021)), Mental r = 0.21 (p = 0.05) and General r = 0.22 (p = 0.037) summarized. There was a significant correlation between PGI and HAP r = 0.40 (p < 0.001) and the Social Participation Scale r = -0.36 (p < 0.001). There was no correlation between the PGI and Glittre ADL scores r = 0.12 (p = 0.247). CONCLUSION: In adults receiving hemodialysis, the PGI proved to be an accurate and reliable instrument for the assessment of the quality of life from the perspective of the patient.IMPLICATIONS FOR REHABILITATIONAlthough hemodialysis treatment is associated with increased survival and symptom control, there is a significant change in the patient's lifestyle.In order to provide a more focused view of the individual, the Patient Generated Index (PGI) was created to evaluate the quality of life.PGI is reliable and correlates with KQDOL - SF and the Social Participation Scale in this population.

Film-Forming Properties of Topical Formulations for Skin and Hair: In Vivo and In Vitro Studies Using Biophysical and Imaging Techniques.

The film-forming effect is an important property of formulations for skin improvement and hair protection. Reflectance confocal microscopy, an advanced imaging technique, is an important tool for its evaluation. Thus, the aim of this study was to evaluate the film-forming properties of cosmetic formulations based on starches and containing a Spirulina maxima extract after their application to skin and hair, using sensorial analysis and instrumental measurements, with emphasis on reflectance confocal microscopy. Two formulations based on starches and PEG-75 lanolin containing or not (Vehicle) a Spirulina maxima dry extract were developed. The rheological behavior and sensorial properties of both formulations were evaluated. The film-forming property on the skin was evaluated in terms of skin hydration by RCM imaging analysis and transepidermal water loss (TEWL). The hair tresses were evaluated in terms of mechanical properties and RCM imaging analysis. The formulations showed pseudoplastic behavior and a low hysteresis area. In addition, the presence of Spirulina in the formulation did not interfere in the rheological parameters. Both formulations reduced TEWL and maintained the stratum corneum water content. The protective effect was also observed in the hair, since the application of the formulation with Spirulina showed a better performance in the tensile test when compared to vehicle. Thus, the proposed formulation showed film formation on the skin and hair surface that brought immediate benefits such as a reduction of TEWL. Finally, the formulations were shown to have film-forming effects and their use can be suggested for skin and hair protection against daily damage.

Applying sunscreen SPF 50 with high antioxidant capacity during fifteen days improves the dermis echogenicity and reduces the reddish skin undertone.

BACKGROUND: Of the many effects induced by UV radiation on the skin, erythema is one of the most well-known features, which is a cutaneous inflammatory reaction correlated with acute photodamage. The utilization of sunscreen may reduce this process. AIMS: To evaluate the utilization of a sunscreen SPF50 with high antioxidant capacity during 15 days by young men without photoprotection habits. METHODS: For this, we evaluated erythema, skin hydration properties, and dermis echogenicity using skin imaging techniques. Forty male participants (aged between 18 and 28 years old), 36 without previous photoprotection habits, were recruited, and the erythema was evaluated using a visual score and skin colorimeter. Macroscopic images (VivaCam®) were also obtained. Dermis echogenicity was evaluated using high-frequency ultrasonography. All the participants received a sunscreen SPF 50 to use for 15 days. RESULTS: The visual score presented a strong correlation (r = 0.8657) with the colorimeter results. Visually and using the biophysical methodologies was possible to observe the reduction of the visual erythema. The dermis echogenicity also improved, probably correlated with the acute inflammation reduction. No alterations were observed in the skin hydration and skin barrier parameters. CONCLUSIONS: The utilization of complementary and correlated different skin biophysical and imaging techniques in this study allows a better comprehension regarding the skin early photoaging process due the direct sun exposure. The utilization with a SPF 50 sunscreen with high antioxidant potential allows for a reduction in the erythema after 15 days of usage, a quick result, however, did not improved the skin barrier or SC hydration.

Application of a Reflectance Confocal Microscopy Imaging Analysis Score for the Evaluation of Non-Melanogenic Changes in Male Photoaged Skin.

The photoaging process is characterized by skin changes due to ultraviolet radiation exposure and is the principal environmental factor affecting skin aging. Reflectance confocal microscopy permits noninvasive skin imaging to understand how the photoaging process may change skin. Since men do not habitually use sunscreen, the application of skin imaging techniques is important to understand the influence of sunlight on their skin health. The aim of this study was to develop a score based on RCM imaging analyses to evaluate the morphological and structural changes in the photoaged skin according to literature data. The score was applied in order to determine possible correlations between chronological aging and sunscreen use behavior among men. Thus, 40 men aged 18 to 50 years were recruited, images from the frontal region of their skin were obtained and the score was applied. It was observed that habits are more important than age for the skin photoaging process. Men with photoprotection habits showed overall better skin morphological and structural characteristics regardless of age, demonstrating that sun protection behavior is a major key factor in the understanding of photoaging, so that men should be encouraged to start the use of cosmetic products and perform self-care.

Intestinal tissue-resident T cell activation depends on metabolite availability.

The metabolic capacity of many cells is tightly regulated and can adapt to changes in metabolic resources according to environmental changes. Tissue-resident memory (TRM) CD8+ T cells are one of the most abundant T cell populations and offer rapid protection against invading pathogens, especially at the epithelia. TRM cells metabolically adapt to their tissue niche, such as the intestinal epithelial barrier. In the small intestine, the types of TRM cells are intraepithelial lymphocytes (IELs), which contain high levels of cytotoxic molecules and express activation markers, suggesting a heightened state of activation. We hypothesize that the tissue environment may determine IEL activity. We show that IEL activation, in line with its semiactive status, is metabolically faster than circulating CD8+ T cells. IEL glycolysis and oxidative phosphorylation (OXPHOS) are interdependently regulated and are dependent on rapid access to metabolites from the environment. IELs are restrained by local availability of metabolites, but, especially, glucose levels determine their activity. Importantly, this enables functional control of intestinal TRM cells by metabolic means within the fragile environment of the intestinal epithelial barrier.

Cosmetic Formulations with Melaleuca alternifolia Essential Oil for the Improvement of Photoaged Skin: A Double-Blind, Randomized, Placebo-Controlled Clinical Study.

This aim of this study was to evaluate the penetration depth, antioxidant capacity and the clinical efficacy of Melaleuca alternifolia pure essential oil and in a nanoemulsion to prevent skin photoaging. For this, 2% of pure essential oil or 2% of this essential oil in a nanoemulsion were vehiculated in a formulation. The skin penetration was evaluated using confocal Raman microspectroscopy. The radical protection factor was evaluated using electron paramagnetic resonance spectroscopy. For a clinical study, 40 male participants, aged 18-28 years, were enrolled, being divided into three groups: vehicle formulation, M. alternifolia pure essential oil and M. alternifolia Nanoemulsion. All the participants also received a sunscreen SPF 50 to use during the day. Before and after 90 days of study, skin hydrolipidics and morphological characteristics were performed by skin imaging and biophysical techniques. The nanoemulsion presented a lower antioxidant capacity and a higher penetration through the stratum corneum, reaching the viable epidermis, improving the stratum granulosum morphology. The groups presented an increase in the papillary depth, improving in the dermis echogenicity and the collagen fibers. Melaleuca alternifolia essential provides the potential to improve photoaged skin, being the application of nanoemulsion able to reach deeper skin layers.

Distinct patterns of gene expression during regeneration and asexual reproduction in the annelid Pristina leidyi.

Regeneration, the ability to replace lost body parts, is a widespread phenomenon in the animal kingdom often connected to asexual reproduction or fission, since the only difference between the two appears to be the stimulus that triggers them. Both developmental processes have largely been characterized; however, the molecular toolkit and genetic mechanisms underlying these events remain poorly unexplored. Annelids, in particular the oligochaete Pristina leidyi, provide a good model system to investigate these processes as they show diverse ways to regenerate, and can reproduce asexually through fission under laboratory conditions. Here, we used a comparative transcriptomics approach based on RNA-sequencing and differential gene expression analyses to understand the molecular mechanisms involved in anterior regeneration and asexual reproduction. We found 291 genes upregulated during anterior regeneration, including several regeneration-related genes previously reported in other annelids such as frizzled, paics, and vdra. On the other hand, during asexual reproduction, 130 genes were found upregulated, and unexpectedly, many of them were related to germline development during sexual reproduction. We also found important differences between anterior regeneration and asexual reproduction, with the latter showing a gene expression profile more similar to that of control individuals. Nevertheless, we identified 35 genes that were upregulated in both conditions, many of them related to cell pluripotency, stem cells, and cell proliferation. Overall, our results shed light on the molecular mechanisms that control anterior regeneration and asexual reproduction in annelids and reveal similarities with other animals, suggesting that the genetic machinery controlling these processes is conserved across metazoans.

Impacts of sun protection and skin care habits in the biophysical and morphological properties of young men skin.

BACKGROUND: Previous studies have demonstrated that UVB radiation may cause changes in the epidermal permeability barrier and in the stratum corneum hydration. It is also well known that sun exposure causes erythema, skin cancer, and other physiologic alterations. Furthermore, because of the cultural barrier, men usually apply less sunscreen. AIMS: The objective of this study was to evaluate the cosmetic use and sun protection habits of young men, correlating their lifestyle with the biophysical and morphological skin characteristics, as well as to study how skin unprotected sun exposure can affect these features even in young people. PATIENTS/METHODS: 60 participants between 18 and 28 years old were divided into two groups: with photoprotection habits (PP habits) and without photoprotection habits (No PP habits). They were questioned about their sunscreen and cosmetic products use. The skin parameters were evaluated using biophysical and skin imaging techniques. RESULTS AND CONCLUSION: 60% of the participants did not apply sunscreen often, and 80% did not consume other skin care products. No PP habits group presented an increase in the transepidermal water loss-TEWL and sebum level, as well as a reduction in the stratum corneum water content and dermis echogenicity. The skin characteristics evaluated in this study were correlated with the sun protection habits and skin care behavior showing that men face some sociocultural barrier to start the use of cosmetic products. Thus, it is important to consider this information for the dermatological clinical and development of cosmetic products for men's skin.