Development of a new affinity maturation protocol for the construction of an internalizing anti-nucleolin antibody library.

Over the last decades, monoclonal antibodies have substantially improved the treatment of several conditions. The continuous search for novel therapeutic targets and improvements in antibody's structure, demands for a constant optimization of their development. In this regard, modulation of an antibody's affinity to its target has been largely explored and culminated in the discovery and optimization of a variety of molecules. It involves the creation of antibody libraries and selection against the target of interest. In this work, we aimed at developing a novel protocol to be used for the affinity maturation of an antibody previously developed by our group. An antibody library was constructed using an in vivo random mutagenesis approach that, to our knowledge, has not been used before for antibody development. Then, a cell-based phage display selection protocol was designed to allow the fast and simple screening of antibody clones capable of being internalized by target cells. Next generation sequencing coupled with computer analysis provided an extensive characterization of the created library and post-selection pool, that can be used as a guide for future antibody development. With a single selection step, an enrichment in the mutated antibody library, given by a decrease in almost 50% in sequence diversity, was achieved, and structural information useful in the study of the antibody-target interaction in the future was obtained.

A versatile high-throughput assay based on 3D ring-shaped cardiac tissues generated from human induced pluripotent stem cell-derived cardiomyocytes.

We developed a 96-well plate assay which allows fast, reproducible, and high-throughput generation of 3D cardiac rings around a deformable optically transparent hydrogel (polyethylene glycol [PEG]) pillar of known stiffness. Human induced pluripotent stem cell-derived cardiomyocytes, mixed with normal human adult dermal fibroblasts in an optimized 3:1 ratio, self-organized to form ring-shaped cardiac constructs. Immunostaining showed that the fibroblasts form a basal layer in contact with the glass, stabilizing the muscular fiber above. Tissues started contracting around the pillar at D1 and their fractional shortening increased until D7, reaching a plateau at 25±1%, that was maintained up to 14 days. The average stress, calculated from the compaction of the central pillar during contractions, was 1.4±0.4 mN/mm2. The cardiac constructs recapitulated expected inotropic responses to calcium and various drugs (isoproterenol, verapamil) as well as the arrhythmogenic effects of dofetilide. This versatile high-throughput assay allows multiple in situ mechanical and structural readouts.

A Case Report of Sheehan Syndrome: A Rare Cause of Hypopituitarism.

Sheehan syndrome is a rare cause of hypopituitarism characterized by pituitary gland necrosis after postpartum hemorrhage. It is a pertinent cause of maternal morbidity and mortality in developing countries with deficient obstetrical care but is frequently overlooked in developed countries where its occurrence is uncommon. We present the case of a 66-year-old female diagnosed with Sheehan syndrome more than 30 years after her last delivery complicated by postpartum hemorrhage. Although the patient presented several symptoms and signs of pituitary hormonal deficiencies, a diagnosis had never been established before. The complete laboratory analysis revealed a deficiency in the pituitary axis, and the cranial imaging showed an empty sella turcica. Hormonal replacement therapy resolved several impairments in terms of general energy, physical capacity, temperature regulation, skin characteristics, and sexual function. It also improved her cardiovascular and osteoporosis risk, and, most importantly, prevented a potential life-threatening adrenal crisis. This report highlights the subtle manifestations of Sheehan syndrome to help clinicians establish a prompt diagnosis. Even in developed countries such as Portugal, this condition should be regarded as a potential cause of hypopituitarism.

Incorporating environmental heterogeneity and observation effort to predict host distribution and viral spillover from a bat reservoir.

Predicting the spatial occurrence of wildlife is a major challenge for ecology and management. In Latin America, limited knowledge of the number and locations of vampire bat roosts precludes informed allocation of measures intended to prevent rabies spillover to humans and livestock. We inferred the spatial distribution of vampire bat roosts while accounting for observation effort and environmental effects by fitting a log Gaussian Cox process model to the locations of 563 roosts in three regions of Peru. Our model explained 45% of the variance in the observed roost distribution and identified environmental drivers of roost establishment. When correcting for uneven observation effort, our model estimated a total of 2340 roosts, indicating that undetected roosts (76%) exceed known roosts (24%) by threefold. Predicted hotspots of undetected roosts in rabies-free areas revealed high-risk areas for future viral incursions. Using the predicted roost distribution to inform a spatial model of rabies spillover to livestock identified areas with disproportionate underreporting and indicated a higher rabies burden than previously recognized. We provide a transferrable approach to infer the distribution of a mostly unobserved bat reservoir that can inform strategies to prevent the re-emergence of an important zoonosis.

Unveiling the Mystery: A Case Report on Oropharyngeal Synovial Sarcoma.

Synovial sarcoma, originating from mesenchymal cells, represents a rare and aggressive sarcoma subtype. This case report describes a rare occurrence of synovial sarcoma in the soft palate, with only a few cases described in the literature. A 38-year-old male presented with a painless mass on the soft palate, which raised suspicion of an abscess and emphasized the importance of considering malignancy in persistent or progressive soft tissue masses, even in atypical anatomical locations. The diagnostic workup, including imaging modalities such as maxillofacial computed tomography (CT) scan, magnetic resonance imaging (MRI), and positron emission tomography-fluorodeoxyglucose (PET-FDG) scan, played a crucial role in confirming the diagnosis and assessing disease extension.  The standard treatment is the complete excision of the tumor. Nevertheless, when it comes to tumors located in the head and neck region, defining standardized margins proves to be a challenge. Radiotherapy can play an important role, particularly in those with tumors larger than 5 cm or positive margins. While chemotherapy offers certain advantages, its application remains a subject of controversy despite its potential benefits. Timely referral and multidisciplinary management are essential in optimizing patient outcomes. Although synovial sarcoma poses diagnostic and therapeutic challenges, advances in diagnostic techniques and personalized medicine offer hope for improved outcomes.

Association between food environments and fetal growth in pregnant Brazilian women.

INTRODUCTION: Birth weight is described as one of the main determinants of newborns' chances of survival. Among the associated causes, or risk factors, the mother's nutritional status strongly influences fetal growth and birth weight outcomes of the concept. This study evaluates the association between food deserts, small for gestational age (SGA), large for gestational age (LGA) and low birth weight (LBW) newborns. DESIGN: This is a cross-sectional population study, resulting from individual data from the Live Birth Information System (SINASC), and commune data from mapping food deserts (CAISAN) in Brazil. The newborn's size was defined as follows: appropriate for gestational age (between 10 and 90th percentile), SGA (< 10th percentile), LGA (> 90th percentile), and low birth weight < 2,500 g. To characterize food environments, we used tertiles of the density of establishments which sell in natura and ultra-processed foods. Logistic regression modeling was conducted to investigate the associations of interest. RESULTS: We analyzed 2,632,314 live births in Brazil in 2016, after appropriate adjustments, women living in municipalities with limited availability of fresh foods had a higher chance of having newborns with SGA [OR2nd tertile: 1.06 (1.05-1.07)] and LBW [OR2nd tertile: 1.11 (1.09-1.12)]. Conversely, municipalities with greater availability of ultra-processed foods had a higher chance of having newborns with SGA [OR3rd tertile: 1.04 (1.02-1.06)] and LBW [OR2nd tertile: 1.13 (1.11-1.16)]. Stratification by race showed that Black and Mixed/Brown women had a higher chance of having newborns with SGA [OR3rd tertile: 1.09 (1.01-1.18)] and [OR3rd tertile: 1.06 (1.04-1.09)], respectively, while Mixed-race women also had a higher chance of having newborns with LBW [OR3rd tertile: 1.17 (1.14-1.20)]. Indigenous women were associated with LGA [OR3rd tertile: 1.20 (1.01-1.45)]. CONCLUSION: The study found that living in areas with limited access to healthy foods was associated with an increased risk of SGA and low birth weight among newborns, particularly among Black and Mixed/Brown women. Therefore, urgent initiatives aimed at reducing social inequalities and mitigating the impact of poor food environments are needed in Brazil.

Pancreas Rejection in the Artificial Intelligence Era: New Tool for Signal Patients at Risk.

INTRODUCTION: Pancreas transplantation is currently the only treatment that can re-establish normal endocrine pancreatic function. Despite all efforts, pancreas allograft survival and rejection remain major clinical problems. The purpose of this study was to identify features that could signal patients at risk of pancreas allograft rejection. METHODS: We collected 74 features from 79 patients who underwent simultaneous pancreas-kidney transplantation (SPK) and used two widely-applicable classification methods, the Naive Bayesian Classifier and Support Vector Machine, to build predictive models. We used the area under the receiver operating characteristic curve and classification accuracy to evaluate the predictive performance via leave-one-out cross-validation. RESULTS: Rejection events were identified in 13 SPK patients (17.8%). In feature selection approach, it was possible to identify 10 features, namely: previous treatment for diabetes mellitus with long-term Insulin (U/I/day), type of dialysis (peritoneal dialysis, hemodialysis, or pre-emptive), de novo DSA, vPRA_Pre-Transplant (%), donor blood glucose, pancreas donor risk index (pDRI), recipient height, dialysis time (days), warm ischemia (minutes), recipient of intensive care (days). The results showed that the Naive Bayes and Support Vector Machine classifiers prediction performed very well, with an AUROC and classification accuracy of 0.97 and 0.87, respectively, in the first model and 0.96 and 0.94 in the second model. CONCLUSION: Our results indicated that it is feasible to develop successful classifiers for the prediction of graft rejection. The Naive Bayesian generated nomogram can be used for rejection probability prediction, thus supporting clinical decision making.

The Effectiveness of Nursing Rehabilitation Interventions on Self-Care for Older Adults with Respiratory Disorders: A Systematic Review with Meta-Analysis.

Background: This research work aimed to summarize the rehabilitation nursing interventions published in the scientific literature that most contribute to effective adherence to self-care in older adults with respiratory diseases. Methods: A systematic literature review with meta-analysis was conducted by searching the EBSCOhost platform (CINAHL Complete, MEDLINE Complete, Cochrane, and MedicLatina) using the PRISMA methodology. Five articles were selected for final analysis. Meta-analysis was carried out using Comprehensive Meta-Analysis (CMA) software, and the results were presented in a forest plot. Results: Thirty-one self-promoting rehabilitation nursing interventions were identified, with the most effective being those related to the assessment of progress in physical capacity/activity tolerance (functional status category/domain) and the assessment of the increase in health-related quality of life (health-related quality of life category/domain). Conclusions: Rehabilitation nursing interventions such as self-management programs led by nurses, community-based and home-based rehabilitation programs, and inspiratory muscle training can effectively reduce and enable the effective control of symptoms associated with respiratory disorders, boosting older adults' empowerment to engage in self-care.

An Online Anomaly Detection Approach for Fault Detection on Fire Alarm Systems.

The early detection of fire is of utmost importance since it is related to devastating threats regarding human lives and economic losses. Unfortunately, fire alarm sensory systems are known to be prone to failures and frequent false alarms, putting people and buildings at risk. In this sense, it is essential to guarantee smoke detectors' correct functioning. Traditionally, these systems have been subject to periodic maintenance plans, which do not consider the state of the fire alarm sensors and are, therefore, sometimes carried out not when necessary but according to a predefined conservative schedule. Intending to contribute to designing a predictive maintenance plan, we propose an online data-driven anomaly detection of smoke sensors that model the behaviour of these systems over time and detect abnormal patterns that can indicate a potential failure. Our approach was applied to data collected from independent fire alarm sensory systems installed with four customers, from which about three years of data are available. For one of the customers, the obtained results were promising, with a precision score of 1 with no false positives for 3 out of 4 possible faults. Analysis of the remaining customers' results highlighted possible reasons and potential improvements to address this problem better. These findings can provide valuable insights for future research in this area.

How to choose the appropriate cannulation technique for vascular access in hemodialysis patients.

The cannulation technique is important for the survival of the arteriovenous access. Choosing the appropriate technique is a complex decision. Such choice must be customized to patients, considering their characteristics, the type of arteriovenous access and the experience of the hemodialysis team. We describe seven items that can help nurses to identify the appropriate cannulation technique: type of arteriovenous access; drainage vein; hygienic self-care profile; experience of the nursing staff in the cannulation technique and nurse-to-patient ratio; hemodialysis treatment method; patient's condition; and patient's decision. Such items can help nurses in decision-making on the "ideal" cannulation technique for each patient.

Linking human tick bite risk with tick abundance in the environment: A novel approach to quantify tick bite risk using orienteers in Scotland.

The rate that people are bitten by ticks is critical in determining the risk of tick-borne infections but is rarely quantified accurately. Often tick abundance in the environment is used as a proxy for tick bite risk, but the relationship with risk is poorly understood. We used a novel citizen science approach to measure tick bite rate in orienteers, to assess the relationship between tick abundance and tick bite risk and to identify risk factors for tick bites. Eleven orienteering events were attended in Scotland between August 2018 and September 2019. The number of tick bites in orienteers, and the time and distance of activity were collected using an online questionnaire. Tick abundance in the same areas used for the orienteering events was estimated by surveying ticks on ground vegetation using blanket drags. Among orienteers, mean incidence was 409 tick bites per 1,000 person-hours. Tick abundance and tick bite rate were strongly correlated, indicating that data from questing tick surveys is a useful proxy for the risk of human tick bites. Tick bite rate was better explained by the activity duration than distance covered and was higher in orienteers that ran earlier in the day, exposed to higher temperatures and in woodland habitats. This study highlights the value of the citizen science approach used, which crucially included submission of activity reports both with and without ticks, to generate robust data on tick bite rate. Accurately measuring tick bite rate and understanding environmental factors that influence it are essential in mitigating the risk of tick-borne diseases.

The MetroPT dataset for predictive maintenance.

The paper describes the MetroPT data set, an outcome of a Predictive Maintenance project with an urban metro public transportation service in Porto, Portugal. The data was collected in 2022 to develop machine learning methods for online anomaly detection and failure prediction. Several analog sensor signals (pressure, temperature, current consumption), digital signals (control signals, discrete signals), and GPS information (latitude, longitude, and speed) provide a framework that can be easily used and help the development of new machine learning methods. This dataset contains some interesting characteristics and can be a good benchmark for predictive maintenance models.

Candida auris, a singular emergent pathogenic yeast: its resistance and new therapeutic alternatives.

Nowadays, fungal infections affect millions of people across the world. Candida auris, a new emergent yeast, is a worrisome pathogen because it associates with a high rate of incidence and prevalence, including in the nosocomial environment. The hard identification, the phenotypic plasticity, and the easy adaptation to stressful conditions are some of the C. auris traits that render this latest yeast singular challenging. C. auris infections have already been reported from more than 30 countries and are associated with high mortality rates. This is the result from rapid transmission and the difficulty of prevention, control, and eradication. There are several factors related to the high virulence of C. auris, such as the multidrug resistance, biofilm development, and the ability to escape the response of the innate immune system. So, C. auris infections are a serious and alarming problem, not only because of the high pathogenicity of the fungal agent but also because of the low effectiveness of the treatments available. Although new formulations have been developed against C. auris strains, a better understanding is essential to efficiently treat, prevent, and control C. auris infections.

Immunotherapy in triple-negative breast cancer: Insights into tumor immune landscape and therapeutic opportunities.

Triple-negative breast cancer (TNBC) is a clinically aggressive subtype of breast cancer that represents 15-20% of breast tumors and is more prevalent in young pre-menopausal women. It is the subtype of breast cancers with the highest metastatic potential and recurrence at the first 5 years after diagnosis. In addition, mortality increases when a complete pathological response is not achieved. As TNBC cells lack estrogen, progesterone, and HER2 receptors, patients do not respond well to hormone and anti-HER2 therapies, and conventional chemotherapy remains the standard treatment. Despite efforts to develop targeted therapies, this disease continues to have a high unmet medical need, and there is an urgent demand for customized diagnosis and therapeutics. As immunotherapy is changing the paradigm of anticancer treatment, it arises as an alternative treatment for TNBC patients. TNBC is classified as an immunogenic subtype of breast cancer due to its high levels of tumor mutational burden and presence of immune cell infiltrates. This review addresses the implications of these characteristics for the diagnosis, treatment, and prognosis of the disease. Herein, the role of immune gene signatures and tumor-infiltrating lymphocytes as biomarkers in TNBC is reviewed, identifying their application in patient diagnosis and stratification, as well as predictors of efficacy. The expression of PD-L1 expression is already considered to be predictive of response to checkpoint inhibitor therapy, but the challenges regarding its value as biomarker are described. Moreover, the rationales for different formats of immunotherapy against TNBC currently under clinical research are discussed, and major clinical trials are highlighted. Immune checkpoint inhibitors have demonstrated clinical benefit, particularly in early-stage tumors and when administered in combination with chemotherapy, with several regimens approved by the regulatory authorities. The success of antibody-drug conjugates and research on other emerging approaches, such as vaccines and cell therapies, will also be addressed. These advances give hope on the development of personalized, more effective, and safe treatments, which will improve the survival and quality of life of patients with TNBC.

Essential Oils as a Good Weapon against Drug-Resistant Candida auris.

Candida auris is a recently found Candida species, mainly associated with nosocomial outbreaks in intensive care hospital settings, and unlike other Candida species, it can be transmitted through person-to-person or by contact with surfaces. C. auris is described as resistant to first-line antifungals and, consequently, associated with high mortality. Nowadays, essential oils (EOs) are known to be effective against fungal and bacterial infections. This work aimed to evaluate the effect of four EOs (tea tree, niaouli, white thyme and cajeput) against C. auris. The EO's effect on C. auris planktonic growth was evaluated by the minimum inhibitory concentration determination and by the agar disc diffusion method. Then, the same effect was evaluated on biofilm by colony-forming units' enumeration. The results showed that EOs were able to inhibit the C. auris planktonic growth, with an MIC50 between 0.78 and 1.56% and halos of 20-21 mm for white thyme and tea tree and 13-14 mm for cajeput and niaouli. In addition, the EOs were also able to completely inhibit biofilm formation. Moreover, white thyme and cajeput completely eradicate pre-formed biofilms, while tea tree and niaouli significantly reduce it. Thus, this work demonstrates that EOs are a possible therapeutic alternative and a future perspective for the hard fight against C. auris.

A comparative study between Fusarium solani and Neocosmospora vasinfecta revealed differential profile of fructooligosaccharide production.

Fructooligosaccharides (FOS) are fructose-based oligosaccharides employed as additives to improve the food's nutritional and technological properties. The rhizosphere of plants that accumulate fructopolysaccharides as inulin has been revealed as a source of filamentous fungi. These fungi can produce FOS either by inulin hydrolysis or by biosynthesis from sucrose, including unusual FOS with enhanced prebiotic properties. Here, we investigated the ability of Fusarium solani and Neocosmospora vasinfecta to produce FOS from different carbon sources. Fusarium solani and N. vasinfecta grew preferentially in inulin instead of sucrose, resulting in the FOS production as the result of endo-inulinase activities. N. vasinfecta was also able to produce the FOS 1-kestose and 6-kestose from sucrose, indicating transfructosylating activity, absent in F. solani. Moreover, the results showed how these carbon sources affected fungal cell wall composition and the expression of genes encoding for ß-1,3-glucan synthase and chitin synthase. Inulin and fructose promoted changes in fungal macroscopic characteristics partially explained by alterations in cell wall composition. However, these alterations were not directly correlated with the expression of genes related to cell wall synthesis. Altogether, the results pointed to the potential of both F. solani and N. vasinfecta to produce FOS at specific profiles.

Psychosocial risks factors among victim support workers during the COVID-19 pandemic: a study with the Copenhagen Psychosocial Questionnaire.

BACKGROUND: Being a victim support worker (VSW) involves exposure to victims' suffering, pain, and traumatic events, which may trigger the risk of VSWs developing mental health problems. Psychosocial risks (PSR) and work-related stress are considered the most challenging issues in occupational safety and health, considering they impact individuals, organizations, and economies. METHODS: The purpose of the present study was to identify the PSR in a sample of 196 Portuguese victim support workers (VSW) (Mean age = 36.49; SD = 10.52). A questionnaire with socio-demographic characteristics, variables related to VSW's job, and the Portuguese medium version of the Copenhagen Psychosocial Questionnaire II (COPSOQ II) were used to assess these professionals' perception of PSR factors. RESULTS: The results reveal that although VSW recognizes some psychosocial factors favourable to their health and well-being, they also identify some PSR that place them at intermediate and severe risk, i.e., emotional and cognitive demands, which are the main areas of risk to the VSW. VSW over 38 years old scored higher in job insecurity, burnout, and offensive behaviours. CONCLUSIONS: These findings give important insights into the areas that must be enhanced in this context involving VSW. Additionally, the results highlight the relevance of encouraging a healthy and supportive work environment, preventing and promoting the health and well-being of VSW, particularly when considering the coronavirus disease (COVID-19) pandemic.

Public infrastructure for food and nutrition security in brazil: fufilling the constitutional commitment to the human right to adequate food.

Food insecurity is a critical global problem with social and public health consequences. In Brazil, access to adequate food is a fundamental human right guaranteed under the country's Constitution since 2010. As such, the State assumes the distinct and complementary obligations to respect, protect, promote and provide the Right to Adequate Food. The aim of this study is to present actions related to the "provision" dimension that have been developed as part of a network of public infrastructure strategies for food and nutrition security in Brazil. Through an exploratory, analytical literature review, the paper focuses on the operational designs and logistics of three main strategies: Food Banks, Community Kitchens, and Popular Restaurants. The Brazilian experience indicates that public actions are necessary, especially considering the urgency for those groups living with hunger and poverty. While similar programs can be found in other countries in South and North America, they are mostly offered by civil society organizations, and have not advanced toward public institutionalization. In fact, private programs are criticized for negating governments' obligation and responsibility in this area. Brazil's experience sheds light on public initiatives in meeting the State's obligations towards the Right to Adequate Food. Supplementary Information: The online version contains supplementary material available at 10.1007/s12571-022-01272-1.

Familial Amyloid Polyneuropathy Misdiagnosed as Systemic Sclerosis.

We present the case of a 53-year-old woman of Portuguese ancestry with a diagnosis of progressive systemic sclerosis (SSc), proposed for haematopoietic stem cell transplantation (HSCT). Clinical re-evaluation when assessing eligibility for the procedure led to the alternative diagnosis of familial amyloid polyneuropathy (FAP). We discuss the clinical presentations of FAP and SSc, focusing on their overlapping and distinguishing features. We emphasize the need for a high level of suspicion in order to establish an early diagnosis of FAP in the absence of a family history, and provide prognostic and genetic counselling. LEARNING POINTS: It is important to review diagnoses, especially when the clinical course is atypical.Cutaneous involvement is a commonly unrecognized feature of familial amyloid polyneuropathy.Hereditary conditions should be included in the differential diagnosis of multisystemic diseases, even in the absence of a family history.

Changes in cell wall composition and ultrastructure related to desiccation during the seed maturation of Paubrasilia echinata (brazilwood).

Paubrasilia echinata (brazilwood) is an endangered native tree from the Brazilian Atlantic Forest whose seeds tolerate maturation drying, but, unlike classic orthodox seeds, they quickly lose viability after shedding. This work analyzed the biochemical and ultrastructural changes during the maturation of brazilwood seeds, with particular attention to the cell walls and organization of the cellular components. The physiological seed maturity was accompanied by increased starch content and decreased soluble sugars. Arabinose increased considerably and was the predominant cell-wall sugar during maturation, suggesting a rise in arabinans that contribute to greater cell wall flexibility. This increase was consistent with the cell wall infolding observed in the hypocotyl axis and cotyledons during the maturation of brazilwood seeds. Ultrastructural analyses showed changes in the number and distribution of protein bodies and amyloplasts and the reorganization of lipid droplets into large drops or masses during seed desiccation. Our findings demonstrate that brazilwood seeds behave like other orthodox seeds during maturation, performing the cell wall and metabolic changes before the major decline in the seed water content. However, the high vacuolization and reorganization of lipid bodies observed at 65 DAA suggest that cell deterioration occurs to some extent at the end of the maturation period and could be responsible for reducing the longevity of the brazilwood dried seeds.