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Abstract Background The purpose of this paper is to describe and analyze the professional expectations of medical students during the 2007-2008 academic year at the public medical schools of Angola, Guinea-Bissau and Mozambique, and to identify their social and geographical origins, their professional expectations and difficulties relating to their education and professional future. Methods Data were collected through a standardised questionnaire applied to all medical students registered during the 2007-2008 academic year. Results Students decide to study medicine at an early age. Relatives and friends seem to have an especially important influence in encouraging, reinforcing and promoting the desire to be a doctor. The degree of feminization of the student population differs among the different countries. Although most medical students are from outside the capital cities, expectations of getting into medical school are already associated with migration from the periphery to the capital city, even before entering medical education. Academic performance is poor. This seems to be related to difficulties in accessing materials, finances and insufficient high school preparation. Medical students recognize the public sector demand but their expectations are to combine public sector practice with private work, in order to improve their earnings. Salary expectations of students vary between the three countries. Approximately 75% want to train as hospital specialists and to follow a hospital-based career. A significant proportion is unsure about their future area of specialization, which for many students is equated with migration to study abroad. Conclusions Medical education is an important national investment, but the returns obtained are not as efficient as expected. Investments in high-school preparation, tutoring, and infrastructure are likely to have a significant impact on the success rate of medical schools. Special attention should be given to the socialization of students and the role model status of their teachers. In countries with scarce medical resources, the hospital orientation of students' expectations is understandable, although it should be associated with the development of skills to coordinate hospital work with the network of peripheral facilities. Developing a local postgraduate training capacity for doctors might be an important strategy to help retain medical doctors in the home country.

The diversity of non-biting midges (Chironomidae, Diptera) remains an unresolved topic, with estimates of species numbers ranging from 6000 to 15,000 according to various authors. To assess Chironomidae diversity in Lithuania, we evaluate the effectiveness of COI gene-based species delimitation methods for providing rapid diversity estimates. Nevertheless, differences between tree-based and distance-based approaches can result in varying group classifications, which may cause species numbers to be overestimated or underestimated. For our study, we analyzed a dataset of 109 specimens sampled from six Lithuanian streams. By applying multiple methods, such as Assemble Species by Automatic Partitioning (ASAP), Automatic Barcode Gap Discovery (ABGD), the generalized mixed Yule-coalescent (GMYC) model, and the Bayesian implementation of the Poisson Tree Processes (bPTP) model, we found that species estimates ranged from 28 to 58. Among these methods, ASAP proved to be the most effective for our dataset, identifying 58 putative species. These results reinforce our assumption that the current understanding of Chironomidae species diversity is incomplete.

Watersheds are highly complex aquatic systems crucial to human and ecological functions. Land cover impacts watersheds, and assessing the landscape structure and configuration is essential for effective management and planning, mainly when using indicators of anthropogenic disturbances such as distance to nature (D 2 N). This study aimed to assess the temporal change in land cover and anthropogenic influence in the Piracicaba Capivari Jundiaí Watershed (WPCJ) and the Sorocaba Médio Tietê Watershed (WSMT) in southeastern Brazil from 2010 to 2020, providing valuable information for decision-making. In both watersheds, agricultural activities predominated, covering more than 60% of the area, with forestry being dominant in 2010 and sugarcane in 2020. However, forest areas have increased in both study regions. The D 2 N analysis indicated a predominance of urbanized or intensively cultivated landscapes (> 60%). There was an improvement in 2020 compared to 2010, attributed to increased natural areas. Regarding the temporal change in D 2 N, more areas transitioned toward artificial rather than natural states in both WPCJ and WSMT, although a considerable portion remained unchanged (> 75%). This analysis is one of the few that applies D 2 N to watershed assessment. We emphasize the need for continued planning and conservation efforts in the WPCJ and WSMT and the potential for using this approach to assess anthropogenic influence in other watersheds., Competing Interests: Declarations: All authors have read, understood, and have complied as applicable with the statement on “Ethical responsibilities of Authors” as found in the Instructions for Authors. Ethical approval: This article does not contain any studies with human participants or animals performed by any authors. Conflict of interest: The authors declare no competing interests., (© 2025. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

In soybean breeding programs, a great deal of time is devoted to the use of methods that perform selection of individual plants during the initial generations. Our hypothesis is that BLUPIS (simulated individual BLUP) can be efficient when applied in the initial stages of soybean breeding programs. This study aimed to explore the potential of BLUPIS in the early generations of a soybean breeding program, as well as to assess the viability of the strategy of dividing the useful area of experimental plots for estimating genotypic effects and plant selection. The experiment involved 84 segregating populations and 15 soybean parents in the F 2 and F 3 generations. Yield data was collected from the 2019/2020 and 2020/2021 cropping seasons. In the F 2 generation, different data exploration methods were applied to determine the most suitable adaptation to be used in the F 3 generation. The individual BLUP (BLUPI) was compared with BLUPIS using information from different replications and/or equal to the information used in BLUPI. The selection conducted by BLUPIS and BLUPI showed high concordance regarding the selected plants. In the F 3 generation, segregating populations were selected based on positive genotypic effects, and individual plants within these populations were further selected according to the number of plants determined by BLUPIS. The division of the plot area was an efficient strategy for selecting segregating populations and individual plants within superior populations in the F 3 generation, resulting in genetic gains of approximately 1.56 g per plant. When combined with the strategy of advancing generations in the off-season, the BLUPIS approach reduces the time required to achieve a high level of homozygosity. Therefore, BLUPIS proved to be a powerful statistical tool for early selection based on grain yield in soybeans., Competing Interests: Declarations. Ethical approval and consent to participate: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

The agouti (Dasyprocta prymnolopha) is a medium-sized, wild rodent that is highly rustic and docile. Its size and ease of management make it a viable candidate for an alternative animal model to traditional murine subjects. However, data on the epidermal strata of agoutis are lacking, with significant uncertainties persisting regarding their skin's characterization. This study aimed to describe and quantify the epidermal strata of skin biopsies from male and female agoutis raised in captivity, to further validate the species as a model for dermatological research. Ultrastructural evaluations through atomic force microscopy (AFM) and stereological analyses were conducted, revealing significant differences between the layers of the skin; notably, the dermis exhibited a greater total volume than the epidermis. The findings suggest that the epidermal strata are well-defined, with the volume likely correlating to the size and cellular density of the keratinocytes. Corneodesmosomes and tonofilaments were identified across all epidermal layers, indicating the probable maintenance of anchoring protein activity, even post-cornification of these cells. These results suggest that the agouti may serve as a promising model for dermatological studies, owing to the homogeneity of its cutaneous tissue across different body regions and the distinct volume and morphology of its epithelial stratification, which could enhance the applicability of systematic investigative methods in the future., Competing Interests: Declaration of Competing Interest Authors declare no conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: The etiology of preeclampsia (PE) may be associated with the increased of production of reactive species and decreased antioxidant activity of enzymes. Inadequate intake of Zn can affect gestational health due to its biological functions, such as its role in the antioxidant defense system. The study aimed to assess the nutritional status of Zn and antioxidant enzymes in postpartum women and its correlation with neonatal outcomes., Methods: A cross-sectional analytical study was carried out at a referral gynecology and obstetrics hospital. A total of 119 women (PE = 58, HP = 61) participated in the study. A quantitative food-frequency questionnaire was used to assess food consumption and further analyze the dietary Zn levels. Zinc levels in plasma and erythrocytes samples were analyzed by flame atomic absorption spectrometry, catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) levels were determined by UV-Vis spectrophotometry., Results: Plasma and dietary intake Zn results were considered adequate and without statistical difference between groups. SOD levels were significantly higher in the HP group (p = 0.011), and CAT levels were higher in the PE group (p = 0.050). There was a positive correlation between SOD activity in women with PE and the weight of their newborns (r = 0.336, p=0.021)., Conclusion: The results showed adequate Zn levels (consumption and serum levels) in the groups studied, although with a reduction of plasma Zn in the PE group compared to the PH group. Zinc in plasma fractions and erythrocytes are important markers for oxidative stress, in particular, plasma Zn seems to be related to the rapid response to preeclampsia. The activity of antioxidant enzymes was elevated in the groups studied. Better SOD activity improves birth weight in children of pregnant women with preeclampsia., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Carla Soraya Costa Maia reports financial support was provided by Coordination of Higher Education Personnel Improvement. Thaynan C. Dias reports financial support was provided by National Council for Scientific and Technological Development. Daniela V. Azevedo reports financial support was provided by Foundation for Scientific and Technological Development and Support of Ceará. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (Copyright © 2024 Elsevier GmbH. All rights reserved.)

Tadalafil, a potent phosphodiesterase inhibitor 5 (PDE-5), is commonly used for the management of erectile dysfunction. However, its therapeutic potential extends beyond this indication. This study aimed to investigate the impact of tadalafil on the recovery of testicular parenchyma in male Wistar rats exposed to testicular thermal stress. Fifty-four Wistar rats were subjected to testicular thermal stress and randomly assigned to receive either tadalafil treatment (TAD) or no treatment (control). TAD was administered intraperitoneally at a dose of either 0.9 mg/kg or 1.8 mg/kg. Biometric parameters, histopathological assessment of the testis, serum testosterone levels, oxidative stress, and interleukin levels were evaluated on days 7, 15, and 30 after thermal shock. The animals were euthanized at the end of each experimental period, and samples were collected. TAD treatment maintained testicular weight and reduced the testicular degenerative process up to day 7 post-injury. However, despite TAD therapy, serum testosterone levels were decreased in the treated groups at days 7 and 15 post-thermal stress. TAD also decreased TNF-α and NO levels at different doses but had no effect on IL-6. The treatment with TAD after heat shock demonstrated anti-inflammatory and antioxidant properties but did not prevent the aggravation of testicular lesions in subsequent periods, even with the systematic reduction in TNF-α and NO levels. Therefore, this selective PDE-5 inhibitor, at the dosages used, did not have a positive impact on testosterone levels during the post-thermal stress period, which could compromise the resumption of the spermatogenic process., Competing Interests: Conflicts of interest: The authors have no conflict of interest to declare., (Copyright © The Author(s).)

South America, particularly within its tropical belt, is renowned for its unparalleled high levels of species richness, surpassing other major biomes. Certain neotropical areas harbor fragmented knowledge of insect diversity and face imminent threats from biodiversity loss and climate change. Hence, there is an urgent need for rapid estimation methods to complement slower traditional taxonomic approaches. A variety of algorithms for delimiting species through single-locus DNA barcodes have been developed and applied for rapid species diversity estimates across diverse taxa. However, tree-based and distance-based methods may yield different group assignments, leading to potential overestimation or underestimation of putative species. Here, we investigate the performance of different DNA-based species delimitation approaches to rapidly estimate the diversity of Polypedilum (Chironomidae, Diptera) in South America. Additionally, we test the hypothesis that significant differences exist in the community structure of Polypedilum fauna between South America and its neighboring regions, particularly the Nearctic. Our analysis encompasses a dataset of 1492 specimens from 598 locations worldwide, with a specific focus on South America. Within this region, we analyzed a subset of 247 specimens reported from 37 locations. Using various methods including the Barcode Index Number (BIN), Bayesian Poisson tree processes (bPTP), multi-rate Poisson tree processes (mPTP), single-rate Poisson tree processes (sPTP), and generalized mixed Yule coalescent (sGMYC), we identify molecular operational taxonomic units (MOTUs) ranging from 267 to 520. Our results indicate that the sGMYC method is the most suitable for estimating putative species in our dataset, resulting in the identification of 75 species in the Neotropical region, particularly in South America. Notably, this region exhibited higher species richness in comparison to the Palearctic and Oriental realms. Additionally, our findings suggest potential differences in species composition of Polypedilum fauna between the Neotropical and the adjacent Nearctic realms, highlighting high levels of endemism and species richness in the first. These results support our hypothesis that there are substantial differences exist in species composition between the Polypedilum fauna in South America and the neighboring regions., Competing Interests: The authors declare no conflict of interest., (© 2023 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Ethnopharmacological Relevance: Aloysia gratissima leaves are popularly used to treat respiratory, digestive, and nervous system disorders. Several studies have been carried out to determine the biological activity of A. gratissima, such as its antibacterial and anti-edematogenic activities, but despite the beneficial uses of A. gratissima, few studies have examined the toxicological profile of this plant., Aim of the Study: This study aimed to determine the chemical composition, cytotoxic, genotoxic, mutagenic potential, and antioxidant activity of an aqueous extract of A. gratissima leaves (AG-AEL)., Material and Methods: The phytochemical constitution of AG-AEL was assessed by colorimetric analyses and High-performance liquid chromatography (HPLC). The inorganic elements were detected by Particle-Induced X-ray Emission (PIXE). The antioxidant, cytotoxicity, genotoxic, and mutagenic activities were evaluated in vitro by Di(phenyl)-(2,4,6-trinitrophenyl)iminoazanium (DPPH), Sulforhodamine B (SRB) assay, comet assay, and Salmonella/microsome assays., Results: AG-AEL indicated the presence of terpenoids, flavonoids, and phenolic acids. HPLC detected rutin at 2.41 ± 0.33 mg/100 mg. PIXE analysis indicated the presence of Mg, Si, P, S, K, Ca, Mn, and Zn. The 50% inhibitory concentration was 84.17 ± 3.17 μg/mL in the DPPH assay. Genotoxic effects were observed using the Comet assay in neuroblastoma (SH-SY5Y) cells and mutations were observed in TA102 and TA97a strains. The extract showed cytotoxic activities against ovarian (OVCAR-3), glioblastoma (U87MG), and colon (HT-29) cancer cell lines., Conclusions: In conclusion, AG-AEL increased DNA damage, induced frameshift, and oxidative mutations, and showed cytotoxic activities against different cancer cells. The in vitro toxicological effects observed suggest that this plant preparation should be used with caution, despite its pharmacological potential., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

SARS-CoV-2 remains a health threat with the continuous emergence of new variants. This work aims to expand the knowledge about the SARS-CoV-2 receptor-binding domain (RBD) interactions with cell receptors and monoclonal antibodies (mAbs). By using constant-pH Monte Carlo simulations, the free energy of interactions between the RBD from different variants and several partners (Angiotensin-Converting Enzyme-2 (ACE2) polymorphisms and various mAbs) were predicted. Computed RBD-ACE2-binding affinities were higher for two ACE2 polymorphisms (rs142984500 and rs4646116) typically found in Europeans which indicates a genetic susceptibility. This is amplified for Omicron (BA.1) and its sublineages BA.2 and BA.3. The antibody landscape was computationally investigated with the largest set of mAbs so far in the literature. From the 32 studied binders, groups of mAbs were identified from weak to strong binding affinities (e.g. S2K146). These mAbs with strong binding capacity and especially their combination are amenable to experimentation and clinical trials because of their high predicted binding affinities and possible neutralization potential for current known virus mutations and a universal coronavirus.Communicated by Ramaswamy H. Sarma.

The aim in this research paper was to investigate the effect of using calcium monophosphate (MCP) and MCP mixed with commercial phosphates salts, in total or partial replacement of calcium chloride (CaCl 2 ) in the manufacture of Minas Frescal cheese. Initially, model cheeses were made to perform the rheological analysis during the coagulation process. Of these, the five best treatments were chosen to carry out the production of Minas Frescal cheese, used only CaCl 2 and MCP, and partial replacements of MCP + polyphosphate, MCP + potassium monophosphate (MKP) and MCP. The cheeses showed no significant difference in physicochemical composition, yield and syneresis, however, the cheese with partial replacement of CaCl 2 by MCP + polyphosphate and MCP + MKP showed the highest hardness values, like the control. This demonstrates that it is possible to replace calcium chloride without significant changes in the physicochemical characteristics and yield of Minas Frescal cheese, and it is still possible to modulate the hardness of the cheese produced according to the type of calcium/phosphate source used. This allows the industry to replace the source of calcium in the manufacture of Minas Frescal cheese according to the desired hardness.

Zika virus (ZIKV) from Uganda (UG) expresses a phenotype related to fetal loss, whereas the variant from Brazil (BR) induces microcephaly in neonates. The differential virulence has a direct relation to biomolecular mechanisms that make one strain more aggressive than the other. The nonstructural protein 1 (NS1) is a key viral toxin to comprehend these viral discrepancies because of its versatility in many processes of the virus life cycle. Here, we aim to examine through coarse-grained models and molecular dynamics simulations the protein-membrane interactions for both NS1 ZIKV-UG and NS1 ZIKV-BR dimers. A first evaluation allowed us to establish that the NS1 proteins, in the membrane presence, explore new conformational spaces when compared to systems simulated without a lipid bilayer. These events derive from both differential coupling patterns and discrepant binding affinities to the membrane. The N-terminal domain, intertwined loop, and greasy finger proposed previously as binding membrane regions were also computationally confirmed by us. The anchoring sites have aromatic and ionizable residues that manage the assembly of NS1 toward the membrane, especially for the Ugandan variant. Furthermore, in the presence of the membrane, the difference in the dynamic cross-correlation of residues between the two strains is particularly pronounced in the putative epitope regions. This suggests that the protein-membrane interaction induces changes in the distal part related to putative epitopes. Taken together, these results open up new strategies for the treatment of flaviviruses that would specifically target these dynamic differences.

Objectives: To investigate the repercussions of the COVID-19 pandemic on lymphedema patients from an endemic area of lymphatic filariasis. Methods: The study descriptive compared sociodemographic and clinical aspects, risk of falling and quality of life, prior and during the COVID-19 pandemic in 28 lymphedema patients, older than 18 years old and under investigation of filarial infection. For the evaluation of functional mobility, the Time Up and Go test and The Medical Outcome Study Short Form-36 Health for quality of life, was used. Results: An increase in interdigital and dermal lesions, a higher frequency of acute dermatolymphangioadenitis crises and risk of falling, worsening of quality of life in the domains of physical functioning, general health, vitality, and mental health during the pandemic was observed. Conclusion: Our findings of clinical worsening and quality of life of patients during the COVID-19 pandemic indicate the need to reinforce the goal of the Lymphatic Filariasis Program regarding the follow-up of these patients in the actions of the Global Program for the Elimination of Lymphatic Filariasis, due to the discontinuity in the care during the pandemic., Competing Interests: The authors declare that they do not have any conflicts of interest., (Copyright © 2023 Aquino, Aguiar-Santos, Rocha, Coutinho, Nascimento, Moretti, Silva, Lima, Bonfim and Medeiros.)

A highly efficient and robust multiple scales in silico protocol, consisting of atomistic Molecular Dynamics (MD), coarse-grain (CG) MD, and constant-pH CG Monte Carlo (MC), has been developed and used to study the binding affinities of selected antigen-binding fragments of the monoclonal antibody (mAbs) CR3022 and several of its here optimized versions against 11 SARS-CoV-2 variants including the wild type. Totally 235,000 mAbs structures were initially generated using the RosettaAntibodyDesign software, resulting in top 10 scored CR3022-like-RBD complexes with critical mutations and compared to the native one, all having the potential to block virus-host cell interaction. Of these 10 finalists, two candidates were further identified in the CG simulations to be the best against all SARS-CoV-2 variants. Surprisingly, all 10 candidates and the native CR3022 exhibited a higher affinity for the Omicron variant despite its highest number of mutations. The multiscale protocol gives us a powerful rational tool to design efficient mAbs. The electrostatic interactions play a crucial role and appear to be controlling the affinity and complex building. Studied mAbs carrying a more negative total net charge show a higher affinity. Structural determinants could be identified in atomistic simulations and their roles are discussed in detail to further hint at a strategy for designing the best RBD binder. Although the SARS-CoV-2 was specifically targeted in this work, our approach is generally suitable for many diseases and viral and bacterial pathogens, leukemia, cancer, multiple sclerosis, rheumatoid, arthritis, lupus, and more., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Some insects produce venoms to defend against predators and directly interact with opioid receptors. In the present study, it was identified two alkaloids in the wasp venom species Hymenoepimecis bicolor . It was demonstrated that these could act as potential inhibitors of opioid receptors through their robust affinity to the receptors. The interaction profile was given to opioid receptors (μOR), with 60% of targets similar to alkaloid 1, with 0.25 probability, and 46.7% of targets similar to alkaloid 2, with a probability 0.17 of affinity as a target, which is considered signaling macromolecules and can mediate the most potent analgesic and addictive properties of opiate alkaloids. Notably, both alkaloids showed -7.6 kcal/mol affinity to the morphine agonies through six residues, Gly124, Asp147, Trp293, Ile296, Ile322, and Tyr326. These observations suggest further research on opioid receptors using in vitro studies of possible therapeutic applications.Communicated by Ramaswamy H. Sarma.

Recommender systems have become one of the main tools for personalized content filtering in the educational domain. Those who support teaching and learning activities, particularly, have gained increasing attention in the past years. This growing interest has motivated the emergence of new approaches and models in the field, in spite of it, there is a gap in literature about the current trends on how recommendations have been produced, how recommenders have been evaluated as well as what are the research limitations and opportunities for advancement in the field. In this regard, this paper reports the main findings of a systematic literature review covering these four dimensions. The study is based on the analysis of a set of primary studies ( N  = 16 out of 756, published from 2015 to 2020) included according to defined criteria. Results indicate that the hybrid approach has been the leading strategy for recommendation production. Concerning the purpose of the evaluation, the recommenders were evaluated mainly regarding the quality of accuracy and a reduced number of studies were found that investigated their pedagogical effectiveness. This evidence points to a potential research opportunity for the development of multidimensional evaluation frameworks that effectively support the verification of the impact of recommendations on the teaching and learning process. Also, we identify and discuss main limitations to clarify current difficulties that demand attention for future research., Supplementary Information: The online version contains supplementary material available at 10.1007/s10639-022-11341-9., Competing Interests: Competing interestsThe authors have no competing interests to declare that are relevant to the content of this article., (© The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2022, Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Main Conclusion: ScHINT1 was identified at sugarcane SAM using subtractive libraries. Here, by bioinformatic tools, two-hybrid approach, and biochemical assays, we proposed that its role might be associated to control redox homeostasis. Such control is important for plant development and flowering transition, and this is ensured with some protein partners such as PAL and SBT that interact with ScHINT1. The shoot apical meristem transition from vegetative to reproductive is a crucial step for plants. In sugarcane (Saccharum spp.), this process is not well known, and it has an important impact on production due to field reduction. In view of this, ScHINT1 (Sugarcane HISTIDINE TRIAD NUCLEOTIDE-BINDING PROTEIN) was identified previously by subtractive cDNA libraries using Shoot Apical Meristem (SAM) by our group. This protein is a member of the HIT superfamily that was composed of hydrolase with an AMP site ligation. To better understand the role of ScHINT1 in sugarcane flowering, here its function in SAM was characterized using different approaches such as bioinformatics, two-hybrid assays, transgenic plants, and biochemical assays. ScHINT1 was conserved in plants, and it was grouped into four clades (HINT1, HINT2, HINT3, and HINT4). The 3D model proposed that ScHINT1 might be active as it was able to ligate to AMP subtract. Moreover, the two-hybrid approach identified two protein interactions: subtilase and phenylalanine ammonia-lyase. The evolutionary tree highlighted the relationships that each sequence has with specific subfamilies and different proteins. The 3D models constructed reveal structure conservation when compared with other PDB-related crystals, which indicates probable functional activity for the sugarcane models assessed. The interactome analysis showed a connection to different proteins that have antioxidative functions in apical meristems. Lastly, the transgenic plants with 35S::ScHINT1_AS (anti-sense orientation) produced more flowers than wild-type or 35S::ScHINT1_S (sense). Alpha-tocopherol and antioxidant enzymes measurement showed that their levels were higher in 35S::ScHINT_S plants than in 35S::ScHINT1_AS or wild-type plants. These results proposed that ScHINT1 might have an important role with other proteins in orchestrating this complex network for plant development and flowering., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The reference material (RM) is a technical requirement for the quality assurance of analytical results and proficiency tests or interlaboratory comparisons. Microbiological RMs are most available in the dehydrated form, mainly by freeze-drying, and maintaining bacterial survival after preparation is a challenge. Thus, obtaining the most resistant cells is essential. Considering that bacteria present cross-response to dehydration after being submitted to an array of stress conditions, this study aimed to evaluate the influence of growth conditions on enterobacteria for the production of mixed microbiological RMs by freeze-drying in skim milk powder. Salmonella enterica serovar Enteritidis, Cronobacter sakazakii, Escherichia coli, and Citrobacter freundii were grown in a minimal medium with 0.5 M NaCl and 0 to 5.0 mM of manganese sulfate (MnSO 4 ) until stationary phase. Salmonella Enteritidis presented an increased resistance to dehydration in the presence of Mn, while C. sakazakii was the most resistant to freeze-drying and further storage for 90 days. Mixed microbiological RMs were produced by freeze-drying and containing Salmonella Enteritidis and coliforms in skim milk powder with 100 mM of trehalose and the Salmonella survival rate was 91.2 to 93.6%. The mixed RM was stable after 30 days at -20 °C, and Salmonella and coliforms were detected by different methods being, the Rambach Agar the best for the bacterial differentiation. The results showed that the culture conditions applied in this study resulted in bacterial cells being more resistant to dehydration, freeze-drying, and stabilization for the production of mixed microbiological RMs more stable and homogeneous., (© 2022. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

Electrostatic intermolecular interactions are important in many aspects of biology. We have studied the main electrostatic features involved in the interaction of the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein with the human receptor Angiotensin-converting enzyme 2 (ACE2). As the principal computational tool, we have used the FORTE approach, capable to model proton fluctuations and computing free energies for a very large number of protein-protein systems under different physical-chemical conditions, here focusing on the RBD-ACE2 interactions. Both the wild-type and all critical variants are included in this study. From our large ensemble of extensive simulations, we obtain, as a function of pH, the binding affinities, charges of the proteins, their charge regulation capacities, and their dipole moments. In addition, we have calculated the p K a s for all ionizable residues and mapped the electrostatic coupling between them. We are able to present a simple predictor for the RBD-ACE2 binding based on the data obtained for Alpha, Beta, Gamma, Delta, and Omicron variants, as a linear correlation between the total charge of the RBD and the corresponding binding affinity. This "RBD charge rule" should work as a quick test of the degree of severity of the coming SARS-CoV-2 variants in the future.

Flaviviruses comprise a large group of arboviral species that are distributed in several countries of the tropics, neotropics, and some temperate zones. Since they can produce neurological pathologies or vascular damage, there has been intense research seeking better diagnosis and treatments for their infections in the last decades. The flavivirus NS1 protein is a relevant clinical target because it is involved in viral replication, immune evasion, and virulence. Being a key factor in endothelial and tissue-specific modulation, NS1 has been largely studied to understand the molecular mechanisms exploited by the virus to reprogram host cells. A central part of the viral maturation processes is the NS1 oligomerization because many stages rely on these protein-protein assemblies. In the present study, the self-associations of NS1 proteins from Zika, Dengue, and West Nile viruses are examined through constant-pH coarse-grained biophysical simulations. Free energies of interactions were estimated for different oligomeric states and pH conditions. Our results show that these proteins can form both dimers and tetramers under conditions near physiological pH even without the presence of lipids. Moreover, pH plays an important role mainly controlling the regimes where van der Waals interactions govern their association. Finally, despite the similarity at the sequence level, we found that each flavivirus has a well-characteristic protein-protein interaction profile. These specific features can provide new hints for the development of binders both for better diagnostic tools and the formulation of new therapeutic drugs., (Copyright © 2022. Published by Elsevier B.V.)

Background: Leukotriene B 4 (LTB 4 ) is a potent lipid mediator that stimulate the immune response. Because dental pulp inflammation and dentin repair are intrinsically related responses, the aim of this research was to investigate the potential of LTB 4 in inducing differentiation of dental pulp stem cells., Methods: Microspheres (MS) loaded with LTB 4 were prepared using an oil emulsion solvent extraction evaporation process and sterility, characterization, efficiency of LTB 4 encapsulation and in vitro LTB 4 release assay were investigated. Mouse dental pulp stem cells (OD-21) were stimulated with soluble LTB 4 or MS loaded with LTB 4 (0.01 and 0.1 μM). Cytotoxicity and cell viability was determined by lactate dehydrogenase and methylthiazol tetrazolium assays. Gene expression were measured by quantitative reverse transcription polymerase chain reaction after 3, 6, 24, 48 and 72 h. Mineralized nodule formation was assessed after 28 days of OD-21 cell stimulation with LTB 4  in mineralized media or not. Groups were compared using one-way ANOVA test followed by Dunnett's post-test (α = 0.05)., Results: Treatment with LTB 4 or MS loaded with LTB 4 (0.01 and 0.1 µm-μM) were not cytotoxic to OD-21 cells. Treatment with LTB 4 modulated the expression of the Ibsp (integrin binding sialoprotein) and Runx2 (runt-related transcription factor 2) genes differently depending on the experimental period analyzed. Interestingly LTB 4 loaded in microspheres (0.1 μM) allowed long term dental pulp cell differentiation and biomineralization., Conclusion: LTB 4 , soluble or loaded in MS, were not cytotoxic and modulated the expression of the Ibsp and Runx2 genes in cultured OD-21 cells. When LTB 4 was incorporated into MS, odontoblast differentiation and mineralization was induced in long term culture., (© 2022. The Author(s).)

The mechanism of protein-polyelectrolyte complexation on the wrong side of the isoelectric point has long puzzled researchers. Two alternative explanations have been proposed in the literature: (a) the charge-patch (CP) mechanism, based on the inhomogeneous distribution of charges on the protein, and (b) the charge-regulation (CR) mechanism, based on the variable charge of weak acid and base groups, which may invert the protein charge in the presence of another highly charged object. To discern these two mechanisms, we simulated artificially constructed short peptides, containing acidic and basic residues, arranged in a blocklike or alternating sequence. Our simulations of these peptides, interacting with polyelectrolytes, showed that charge patch and charge regulation alone can both lead to adsorption on the wrong side of the pI value. Their simultaneous presence enhances adsorption, whereas their absence prevents adsorption. Our simulation results were rationalized by following the variation of the charge regulation capacity and dipole moments of these peptides with the pH. Specifically for lysozyme, we found that charge patch prevails at physiological pH, whereas charge regulation prevails near the pI, thereby explaining seemingly contradicting conclusions in the literature. By applying the same approach to other proteins, we developed a general framework for assessing the role of the CP and CR mechanisms in existing case studies and for predicting how various proteins interact with polyelectrolytes at different pH values.

Carnauba (Copernicia prunifera) is a Brazilian palm tree used for wax production, which usually generates a large amount of waste. This work evaluated the carnauba waste for cellulase and xylanase production using Trichoderma reesei CCT2768 through a solid-state fermentation (SSF). Carnauba waste was used in its crude form (C-IN), pretreated (C-P) with alkaline hydrogen peroxide (AHP), and also recycled after the SSF process (C-PR). C-IN, C-P, and C-PR were characterized by XRD, FTIR, and SEM. Cellulase and xylanase production was performed by SSF for 72 h, and the enzymatic extracts obtained were mixed each other in different concentrations. FPase, CMCase, and xylanase activities were determined. Trichoderma reesei CCT-2768 has shown high performance to produce cellulases and xylanases. Total cellulase, CMCase, and β-glycosidase presented a highest activity when C-PPR1 (25% of C-PR and 75% of C-P) was used as a carbon source, with yield of 2.85 U/g, 41.21 U/g, and 2.80 U/g, respectively. The highest xylanase production was achieved when only the pretreated carnauba waste (C-P) was used, with an enzyme activity of 224.93 U/g. Carnauba has shown a promising carbon source capacity to induce the production of cellulolytic and xylanolytic enzymes by using T. reesei CCT2768, promoting the circular and ecofriendly economy, as well as a cost reduction, of the production process of these enzymes., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: Obesity is one of the most common nutritional disorders in dogs and cats and is related to the development metabolic comorbidities. Weight loss is the recommended treatment, but success is difficult due to the poor satiety control. Yeast beta-glucans are known as biological modifiers because of their innumerable functions reported in studies with mice and humans, but only one study with dogs was found. This study aimed to evaluate the effects of a diet supplemented with 0.1% beta-glucan on glucose, lipid homeostasis, inflammatory cytokines and satiety parameters in obese dogs. Fourteen dogs composed three experimental groups: Obese group (OG) with seven dogs with body condition score (BCS) 8 or 9; Lean group (LG) included seven non-obese dogs with a BCS of 5; and Supplemented Obese group (SOG) was the OG dogs after 90 days of consumption of the experimental diet., Results: Compared to OG, SOG had lower plasma basal glycemic values (p = 0.05) and reduced serum cholesterol and triglyceride levels. TNF-α was lower in SOG than in OG (p = 0.05), and GLP-1 was increased in SOG compared to OG and LG (p = 0.02)., Conclusion: These results are novel and important for recognizing the possibility of using beta-glucan in obesity prevention and treatment., (© 2021. The Author(s).)

Brazil stands out worldwide in the production of coffee. The observed increases in its productivity and morpho agronomic traits are the results of the improvement of several methodologies applied in obtaining improved cultivars, among which the predictive methods of genetic value stand out. These contribute significantly to the selection of higher genotypes, increasing the genetic gain per unit time. In this context, genomic-wide selection (GWS) is a tool that stands out, since it allows predicting the future phenotype of an individual based only on molecular information. Performing joint selection of traits is the interest of most breeding programs, and factor analysis (FA) has been used to assist in this end. The aim of this study was to evaluate the use of FA in the context of GWS, in genotypes of Coffea canephora . It was found that FA was efficient to elucidate the relationships between the traits and generate new variables. The factors formed can assist in the selection, as in addition to allowing joint interpretations, they present good estimates of predictive capacity, heritability and accuracy. Furthermore, high agreement was observed between the individuals selected based on the factors and those selected considering the individual traits. Additionally, it was observed agreement between the top 10% individuals selected based on the "vigor factor" and each variable individually. However, the selection based on "vigor factor" presented individuals with more suitable size from the phytotechnical point of view., Competing Interests: Conflict of interestThe authors have no conflict of interest to declare that are relevant to the content of this article., (© The Author(s), under exclusive licence to Springer Nature B.V. 2022.)

The use of natural products in research on tick control for Rhipicephalus microplus is increasing year by year, with promising results. In this regard, the aim of the present study was to phytochemically characterize the essential oils (EOs) of Leptospermum scoparium, Origanum vulgare and Litsea cubeba, and to evaluate the acaricidal activity of these EOs in solutions prepared using ethanol, dimethylsulfoxide (DMSO) and Tween 80 on larvae and females of R. microplus. In addition, three L. scoparium fractions were also isolated and their acaricidal activity on these larvae and adult females was tested. Gas chromatography and mass spectrometry results showed that cis-calamenene (29.82 %), carvacrol (64.85 %) and geranial (42.44 %) were the majority compounds of L. scoparium, O. vulgare and L. cubeba, respectively. Three fractions were isolated from L. scoparium: A1, rich in sesquiterpene hydrocarbons, and A2 and A3, rich in β-triketones. Bioassays on unfed larvae (immersion test) were performed using all the EOs at concentrations from 2.5 to 10.0 mg/mL; and using the three fractions obtained from L. scoparium EO at concentrations from 0.625 to 10 mg/mL. We observed 100 % mortality of larvae in all treatments with L. scoparium EO at all concentrations (diluted both in DMSO and in ethanol), and in treatments with O. vulgare EO diluted in DMSO. However, L. cubeba EO only gave rise to more than 99 % mortality at a concentration of 10 mg/mL, using the same solvents. For engorged females, the immersion test was performed at concentrations from 2.5 to 10.0 mg/mL. Percentage control greater than 90 % was observed only at the highest concentrations of L. scoparium and O. vulgare EOs diluted in DMSO and ethanol, while L. cubeba EO did not reach 90 % control in any of the treatments. In tests on L. scoparium fractions, larval mortality in the fractions rich in β-triketones (A2 and A3) was above 97 % at a concentration of 2.5 mg/mL, while in the A1 fraction, rich in sesquiterpene hydrocarbons, at the same concentration (2.5 mg/mL), mortality did not reach 22 %. In the adult immersion test, the percentage control was higher than 98 % at the lowest concentration (2.5 mg/mL) of the A1 fraction, while in the treatments with the fractions A2 and A3, the control levels were 16 and 50 %, respectively. Thus, we can conclude that the EOs of L. scoparium, O. vulgare and L. cubeba have acaricidal activity on R. microplus, as also do the fractions derived from L. scoparium EO., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Virus-like particles (VLPs) are a versatile, safe, and highly immunogenic vaccine platform. Recently, there are developmental vaccines targeting SARS-CoV-2, the causative agent of COVID-19. The COVID-19 pandemic affected humanity worldwide, bringing out incomputable human and financial losses. The race for better, more efficacious vaccines is happening almost simultaneously as the virus increasingly produces variants of concern (VOCs). The VOCs Alpha, Beta, Gamma, and Delta share common mutations mainly in the spike receptor-binding domain (RBD), demonstrating convergent evolution, associated with increased transmissibility and immune evasion. Thus, the identification and understanding of these mutations is crucial for the production of new, optimized vaccines. The use of a very flexible vaccine platform in COVID-19 vaccine development is an important feature that cannot be ignored. Incorporating the spike protein and its variations into VLP vaccines is a desirable strategy as the morphology and size of VLPs allows for better presentation of several different antigens. Furthermore, VLPs elicit robust humoral and cellular immune responses, which are safe, and have been studied not only against SARS-CoV-2 but against other coronaviruses as well. Here, we describe the recent advances and improvements in vaccine development using VLP technology.

The COVID-19 pandemic has spread worldwide. However, as soon as the first vaccines-the only scientifically verified and efficient therapeutic option thus far-were released, mutations combined into variants of SARS-CoV-2 that are more transmissible and virulent emerged, raising doubts about their efficiency. This study aims to explain possible molecular mechanisms responsible for the increased transmissibility and the increased rate of hospitalizations related to the new variants. A combination of theoretical methods was employed. Constant-pH Monte Carlo simulations were carried out to quantify the stability of several spike trimeric structures at different conformational states and the free energy of interactions between the receptor-binding domain (RBD) and angiotensin-converting enzyme II (ACE2) for the most worrying variants. Electrostatic epitopes were mapped using the PROCEEDpKa method. These analyses showed that the increased virulence is more likely to be due to the improved stability to the S trimer in the opened state, in which the virus can interact with the cellular receptor, ACE2, rather than due to alterations in the complexation RBD-ACE2, since the difference observed in the free energy values was small (although more attractive in general). Conversely, the South African/Beta variant (B.1.351), compared with the SARS-CoV-2 wild type (wt), is much more stable in the opened state with one or two RBDs in the up position than in the closed state with three RBDs in the down position favoring the infection. Such results contribute to understanding the natural history of disease and indicate possible strategies for developing new therapeutic molecules and adjusting the vaccine doses for higher B-cell antibody production., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Giron, Laaksonen and Barroso da Silva.)

The progress of viral diseases such as the new coronavirus (COVID-19) can be influenced not only by social isolation policies, but also by climatic factors. Understanding how these factors affect the progress of the pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) may be essential to know the risks each country is facing because of the disease. In this study, we verified the existence of a relationship between the basic reproduction number (R0) of SARS-CoV-2 with different climate variables, while also considering the Global Health Security Index (GHS). We collected data from confirmed cases of COVID-19 along with their respective GHS notes and climate data, from December 31, 2019 to April 13, 2020, for 52 countries. The generalized additive model (GAM) was applied to explore the effect of temperature, relative humidity, solar radiation index, and GHS score on the spread rate of COVID-19. The countries that showed similarity to each other were grouped into clusters using the Kohonen self-organizing map methodology to investigate the importance of each variable in the dissemination of the disease. The temperature variable presented a linear relationship ( p  < 0.001) with the R0, with an explained variation of 36.2%, while the relative humidity variable did not present a significant relationship with the R0. The response curve of the solar radiation variable presented a significant nonlinear relationship ( p  < 0.001) with an explained variation of 32.3%. The GHS index variable, with a significant nonlinear relationship ( p  < 0.001), presented the largest explanatory response in the control of COVID-19, with an explained variation of 38.4%; further, it was observed that the countries with the largest GHS index scores were less influenced by climate variables., Competing Interests: None., (© 2021 Published by Elsevier B.V.)

Viruses can impact and affect human populations in a severe way. The appropriate differentiation among several species or strains of viruses is one of the biggest challenges for virology and infectiology studies. The detection of measurables-quantified discrepancies allows for more accurate clinical diagnoses and treatments for viral diseases. In the present study, we have used a computational approach to explore the dynamical properties of the nonstructural protein 1 from two strains of Zika virus. Our results show that despite a high sequence similarity, the two viral proteins from different origins can exhibit significant dissimilar structural dynamics, which complement their reported differential virulence. The present study opens up new ways in the understanding of the infectivity for these biological entities.

The emission of mercury (Hg) by chlor-alkali plants can pollute soils and sediments, posing risks to the environment and human health. Mangrove ecosystems are particularly sensitive to Hg contamination. Here, we studied the Hg spatial distribution and associated human and ecologic risks in mangrove soils impacted by a chlor-alkali plant. Sixty-six samples of superficial soils were collected from the mangrove of the Botafogo River, Brazil. Mercury contents were determined and ecological and human health risks were estimated from the soil. The Hg contents exceeded the local Hg background by up to 180 times, indicating the substantial anthropic contribution that occurred in the area. Mercury concentrations followed a gradient as a function of the distance from the chlor-alkali plant, with an apparent contribution from the estuary's hydrodynamic regime. The ecological risk was considered high in all the soils evaluated, while the daily average exposure for humans, considering multiple exposure routes to soil, is below the tolerable dose recommended by the World Health Organization (WHO). However, the risk to human health was unacceptable in the estuary section closest to the plant, mainly for children. Vapor inhalation was the main route for estimating non-carcinogenic risk. The results of this study indicate a severe scenario of Hg pollution with unacceptable risks to the ecosystem and the health of human beings, especially of the communities that live from fishery and shellfish colletion and are exposed daily to soils polluted by mercury. Studies on the organomercurial species in the food chain and Hg levels in individuals living close to the estuary are warranted. This research is an important reference in the world regarding the contamination of mangrove areas by Hg., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

A new betacoronavirus named SARS-CoV-2 has emerged as a new threat to global health and economy. A promising target for both diagnosis and therapeutics treatments of the new disease named COVID-19 is the coronavirus (CoV) spike (S) glycoprotein. By constant-pH Monte Carlo simulations and the PROCEEDpKa method, we have mapped the electrostatic epitopes for four monoclonal antibodies and the angiotensin-converting enzyme 2 (ACE2) on both SARS-CoV-1 and the new SARS-CoV-2 S receptor binding domain (RBD) proteins. We also calculated free energy of interactions and shown that the S RBD proteins from both SARS viruses binds to ACE2 with similar affinities. However, the affinity between the S RBD protein from the new SARS-CoV-2 and ACE2 is higher than for any studied antibody previously found complexed with SARS-CoV-1. Based on physical chemical analysis and free energies estimates, we can shed some light on the involved molecular recognition processes, their clinical aspects, the implications for drug developments, and suggest structural modifications on the CR3022 antibody that would improve its binding affinities for SARS-CoV-2 and contribute to address the ongoing international health crisis., (Copyright © 2020. Published by Elsevier B.V.)

We investigate the prevalence, and so the role in the amyloidogenesis, of biased conformations in large ensembles of monomeric forms for Aβ42 and Aβ40 that can trigger the formation and growth of fibrils described by a dock-lock mechanism. We model such biased conformations as the structural monomeric units that constitute the Protein Data Bank fibrils 2beg, 2mxu, and 2lmn. These units were employed as templates to search for similar structures in statistical conformational ensembles of Aβ peptides generated by molecular dynamics with an accurate force field in explicit solvation, whose high quality is revealed by comparison with residual dipolar coupling (RDC) experiments. The conformational ensembles generated by these intrinsically disordered peptides do not contain conformations highly similar to the amyloidogenic templates. This is a consequence of the low thermodynamic stability exhibited by the template-like conformations. A further constant-pH Monte Carlo study has revealed that this stability can be increased by suitable pH conditions, which helps to trigger the fibril elongation. Moreover, our analyses on the free energy landscapes, hydrogen bond prevalences, and principal component analysis distributions emphasize the relevance of many-body long-range cooperative interactions, likely acting over the infrequent preexisting structurally biased conformations, to explain the fibrils' emergence.

Two new species of Polypedilum (Asheum) (Diptera: Chironomidae: Chironominae) are described and figured as adult males, P. (A.) sofiae sp. n. collected in the states of Mato Grosso and Rondônia and P. (A.) mayrahu sp. n. collected in the state of Bahia, Brazil. In addition, the adult male of Polypedilum (Asheum) curticaudatum (Rempel) is redescribed based on material from some localities in the Brazilian Atlantic Forest and Pantanal, and its larva and pupa are described for the first time.

The structure and interactions of proteins play a critical role in determining the quality attributes of many foods, beverages, and pharmaceutical products. Incorporating a multiscale understanding of the structure-function relationships of proteins can provide greater insight into, and control of, the relevant processes at play. Combining data from experimental measurements, human sensory panels, and computer simulations through machine learning allows the construction of statistical models relating nanoscale properties of proteins to the physicochemical properties, physiological outcomes, and tastes of foods. This review highlights several examples of advanced computer simulations at molecular, mesoscale, and multiscale levels that shed light on the mechanisms at play in foods, thereby facilitating their control. It includes a practical simulation toolbox for those new to in silico modeling.

Viruses are enthusiastically studied due to the great impact that these organisms can have on human health. Computational approaches can contribute offering tools that can shed light on important molecular mechanisms that help to design new diagnostic procedures. Several cellular processes between the immune-host system and the pathogenic organism are dependent on specific intermolecular interactions. In this study, we evaluated theoretical approaches to understand some properties of the antigen-antibody interactions considering the titratable properties of all ionizable residues of the nonstructural viral protein 1 (NS1) of the West Nile virus (WNV) and the Zika virus (ZIKV). Constant-pH Monte Carlo simulations were performed to estimate electrostatic properties such as the p K a shifts (Δp K a ). We proposed an alternative criterion for the discrimination of antigenic residues based on Δp K a s. Our outcomes were analyzed by an evaluation of the sensitivity and specificity through a receiver operating characteristic (ROC). As a starting point, we used the known crystallographic structure for the complex of NS1 WNV(176-352) and the specific antibody 22NS1 (PDB ID 4OII ) to differentiate the residues belonging to that interface. With an optimal threshold for the absolute value of the p K a shifts, we found that is possible to predict antigenic epitopes reproducing the interfaces as defined by the X-ray structure. After this validation, we evaluated theoretical predictions based on protein-protein (PP) complexation simulations. From them, we observe amino acids with an antigenic potential and defined the optimum threshold that was applied for two strains of ZIKV (i.e., Uganda and Brazil). Several ionizable residues with antigenic capacity were identified. This is favorably related to some studies that show the high immunogenicity of secreted NS1. This approach opens up an important discussion about what are termed here "electrostatic epitopes" and how they work as an important reference in the paratope-epitope interaction for viral systems.