Nonsurgical Endodontic Treatment of Type II Dens Invaginatus in A Maxillary Lateral Incisor: A Case Report.

Dens invaginatus (DI) is one of the developmental dental anomalies that results in an invagination of the enamel organ into the dental papila during odontogenesis. The purpose of this study is to report a case of nonsurgical endodontic treatment of an Oehlers type II DI in a right maxillary lateral incisor with an extensive periapical damage, along with the two-year clinical and tomographic follow-up. A 30-year-old patient was referred for endodontic treatment of tooth #12. On clinical examination, a change in the shape and color of the crown was observed. The tooth responded negative to pulp sensibility, percussion, palpation and mobility tests. After tomographic evaluation, an Oehlers type II DI was visualized, in addition to an extensive periradicular lesion. The diagnosis was asymptomatic apical periodontitis. The treatment was carried out in two sessions, through intense enhancement of the auxiliary chemical substance with passive ultrasonic irrigation, XP-Endo Finisher and the use of hydroxide-based intracanal medication. Appropriate treatment in cases with anatomic variations requires an accurate and early diagnosis based on clinical examination and radiographic images. A two-year follow-up of the present case showed that the correct diagnosis associated with appropriate instrumentation techniques, supplementary disinfection, and adequate three-dimensional sealing of the canal with filling material, resulted in regression of the periradicular lesion and bone repair.

Effects of acute hypothyroidism on plasma melatonin and Aanat and Asmt expression in the pineal gland and gonads of rats.

Introduction: The reproductive system is tightly regulated by environmental and physiological signals. Melatonin, known as the hormone of darkness, plays a crucial role in regulating both the circadian and reproductive systems in mammals. Hypothyroidism is a key endocrine disorder that harms the reproductive system. Despite many studies on melatonin's effects on the reproductive system, there is conflicting information regarding melatonin synthesis modulation in hypothyroidism. The objective of this study was to investigate the modulation of plasma melatonin levels and gene expression of Aanat and Asmt in the pineal gland and gonads of rats with hypothyroidism at different times of the day. Methods: Female and male Wistar rats were divided into control and hypothyroid groups. Hypothyroidism was induced using propylthiouracil (PTU) for 15 days, rats were euthanized six hours after lights on (ZT6), before lights off (ZT11.5), and six hours after lights off (ZT18). Free thyroxine (FT4) and melatonin were quantified in plasma, and gene expressions of melatonin synthesizing enzymes (Aanat and Asmt) were measured in pineal and sexual organs (testis and ovary). Also, morphological analysis was performed in sexual organs. Results: The results reveal some disparities between the sexes. Hypothyroidism reduced antral and primary follicles in the ovary, and reduced the weight of testis, epididymis, and prostate. In relation to gene expression, we observed a reduction in Aanat expression in the pineal gland during the light phase (ZT6), and in males, this reduction occurred during the dark phase (ZT18). Regarding Asmt expression, there was a decrease in females also during the dark phase (ZT18). In the gonads, there was an increase in expression in both sexes at ZT11.5. Additionally, it was interesting to observe the association between FT4 levels and Asmt expression in the gonads. Conclusions: This study showed that acute hypothyroidism can affect components of the melatonergic system in gonads, particularly gene expression of melatonin synthesis enzymes (Aanat and Asmt) contributing to changes in reproduction organs during disease progression. These findings enhance our understanding of melatonin synthesis in the reproductive system during hypothyroidism, showing distinct responses in male and female rats, and suggest that hypothyroidism affects the circadian rhythmicity of melatonin synthesis in a sex-dependent manner.

QM/MM Study of the Reaction Mechanism of L-Tyrosine Hydroxylation Catalyzed by the Enzyme CYP76AD1.

We have studied the hydroxylation mechanism of l-Tyr by the heme-dependent enzyme CYP76AD1 from the sugar beet (Beta vulgaris). This enzyme has a promising biotechnological application in modified yeast strains to produce medicinal alkaloids, an alternative to the traditional opium poppy harvest. A generative machine learning software based on AlphaFold was used to build the structure of CYP76AD1 since there are no structural data for this specific enzyme. After model validation, l-Tyr was docked in the active site of CYP76AD1 to assemble the reactive complex, whose catalytic distances remained stable throughout the 100 ns of MD simulation. Subsequent QM/MM calculations elucidated that l-Tyr hydroxylation occurs in two steps: hydrogen abstraction from l-Tyr by CpdI, forming an l-Tyr radical, and subsequent radical rebound, corresponding to a rate-limiting step of 16.0 kcal·mol-1. Our calculations suggest that the hydrogen abstraction step should occur in the doublet state, while the radical rebound should happen in the quartet state. The clarification of the reaction mechanism of CYP76AD1 provides insights into the rational optimization of the biosynthesis of alkaloids to eliminate the use of opium poppy.

Binding studies of promethazine and its metabolites with human serum albumin by high-performance affinity chromatography and molecular docking in the presence of codeine.

"Purple Drank", a soft drink containing promethazine (PMZ) and codeine (COD), has gained global popularity for its hallucinogenic effects. Consuming large amounts of this combination can lead to potentially fatal events. The binding of these drugs to plasma proteins can exacerbate the issue by increasing the risk of drug interactions, side effects, and/or toxicity. Herein, the binding affinity to human serum albumin (HSA) of PMZ and its primary metabolites [N-desmethyl promethazine (DMPMZ) and promethazine sulphoxide (PMZSO)], along with COD, was investigated by high-performance affinity chromatography (HPAC) though zonal approach. PMZ and its metabolites exhibited a notable binding affinity for HSA (%b values higher than 80%), while COD exhibited a %b value of 65%. To discern the specific sites of HSA to which these compounds were bound, displacement experiments were performed using warfarin and (S)-ibuprofen as probes for sites I and II, respectively, which revealed that all analytes were bound to both sites. Molecular docking studies corroborated the experimental results, reinforcing the insights gained from the empirical data. The in silico data also suggested that competition between PMZ and its metabolites with COD can occur in both sites of HSA, but mainly in site II. As the target compounds are chiral, the enantioselectivity for HSA binding was also explored, showing that the binding for these compounds was not enantioselective.

Computational analysis to comprehend the structure-function properties of fibrinolytic enzymes from Bacillus spp for their efficient integration into industrial applications.

Background: The fibrinolytic enzymes from Bacillus sp. are proposed as therapeutics in preventing thrombosis. Computational-based analyses of these enzymes' amino acid composition, basic physiological properties, presence of functional domain and motifs, and secondary and tertiary structure analyses can lead to developing a specific enzyme with improved catalytic activity and other properties that may increase their therapeutic potential. Methods: The nucleotide sequences of fibrinolytic enzymes produced by the genus Bacillus and its corresponding protein sequences were retrieved from the NCBI database and aligned using the PRALINE programme. The varied physiochemical parameters and structural and functional analysis of the enzyme sequences were carried out with the ExPASy-ProtParam tool, MEME server, SOPMA, PDBsum tool, CYS-REC tool, SWISS-MODEL, SAVES servers, TMHMM program, GlobPlot, and peptide cutter software. The assessed in-silico data were compared with the published experimental results for validation. Results: The alignment of sixty fibrinolytic serine protease enzymes (molecular mass 12-86 kDa) sequences showed 49 enzymes possess a conserved domain with a catalytic triad of Asp196, His242, and Ser569. The predicted instability and aliphatic indexes were 1.94-37.77, and 68.9-93.41, respectively, indicating high thermostability. The random coil means value suggested the predominance of this secondary structure in these proteases. A set of 50 amino acid residues representing motif 3 signifies the Peptidase S8/S53 domain that was invariably observed in 56 sequences. Additionally, 28 sequences have transmembrane helices, with two having the most disordered areas, and they pose 25 enzyme cleavage sites. A comparative analysis of the experimental work with the results of in-silico study put forward the characteristics of the enzyme sequences JF739176.1 and MF677779.1 to be considered when creating a potential mutant enzyme as these sequences are stable at high pH with thermostability and to exhibit αß-fibrinogenase activity in both experimental and in-silico studies.

Deployment and validation of a smart bed architecture for untethered patients with wireless biomonitoring stickers.

Conventional patient monitoring in healthcare has limitations such as delayed identification of deteriorating conditions, disruptions to patient routines, and discomfort due to extensive wiring for bed-bound patients. To address these, we have recently developed an innovative IoT-based healthcare system for real-time wireless patient monitoring. This system includes a flexible epidermal patch that collects vital signs using low power electronics and transmits the data to IoT nodes in hospital beds. The nodes connect to a smart gateway that aggregates the information and interfaces with the hospital information system (HIS), facilitating the exchange of electronic health records (EHR) and enhancing access to patient vital signs for healthcare professionals. Our study validates the proposed smart bed architecture in a clinical setting, assessing its ability to meet healthcare personnel needs, patient comfort, and data transmission reliability. Technical performance assessment involves analyzing key performance indicators for communication across various interfaces, including the wearable device and the smart box, and the link between the gateway and the HIS. Also, a comparative analysis is conducted on data from our architecture and traditional hospital equipment. Usability evaluation involves questionnaires completed by patients and healthcare professionals. Results demonstrate the robustness of the architecture proposed, exhibiting reliable and efficient information flow, while offering significant improvements in patient monitoring over conventional wired methods, including unrestricted mobility and improved comfort to enhance healthcare delivery.

Interaction of major saffron constituent safranal with trypsin: An experimental and computational investigation.

Trypsin is a serine protease, an important digestive enzyme that digests the proteins in the small intestine. In the present study, we have investigated the interaction of safranal, a major saffron metabolite, with trypsin using spectroscopic and molecular docking analyses. Fluorescence emission spectra of trypsin were largely affected by the inner filter effect from safranal; that's why these were corrected using the standard procedure. The corrected fluorescence spectra have shown that the safranal quenched the intrinsic fluorescence of trypsin with a blue shift in the wavelength of emission maximum, which revealed that the microenvironment of the fluorophore became more hydrophobic. There was approximately 1: 1 fair binding between them, which increased with a rise in temperature. The interaction was favored, principally, by hydrophobic forces, and there was an efficient energy transfer from the fluorophore to the safranal. Synchronous fluorescence spectra suggested that the tryptophan residues were the major ones taking part in the fluorescence quenching of trypsin. Safranal also influenced the secondary structure of trypsin and caused partial unfolding. Molecular Docking and the Molecular Dynamics simulation of the free and complexed trypsin was also carried out. Safranal formed a stable, non-covalent complex within the S2'-S5' subsite. Moreover, two nearby tyrosine residues (Tyr39 and Tyr151) stabilized safranal through π-π interactions. Additionally, the presence of safranal led to changes in the protein flexibility and compactness, which could indicate changes in the surrounding of tryptophan residues, impacting their fluorescence. Furthermore, a loss in compactness is in line with the partial unfolding observed experimentally. Thus, both experimental and computational studies were in good agreement with each other.

Revisiting the reaction pathways for phospholipid hydrolysis catalyzed by phospholipase A2 with QM/MM methods.

Secreted phospholipase A2 (sPLA2) is a Ca2+-dependent, widely distributed enzyme superfamily in almost all mammalian tissues and bacteria. It is also a critical component of the venom of nearly all snakes, as well as many invertebrate species. In non-venomous contexts, sPLA2 assumes significance in cellular signaling pathways by binding cell membranes and catalyzing ester bond hydrolysis at the sn-2 position of phospholipids. Elevated levels of GIIA sPLA2 have been detected in the synovial fluid of arthritis patients, where it exhibits a pro-inflammatory function. Consequently, identifying sPLA2 inhibitors holds promise for creating highly effective pharmaceutical treatments. Beyond arthritis, the similarities among GIIA sPLA2s offer an opportunity for developing treatments against snakebite envenoming, the deadliest neglected tropical disease. Despite decades of study, the details of PLA2 membrane-binding, substrate-binding, and reaction mechanism remain elusive, demanding a comprehensive understanding of the sPLA2 catalytic mechanism. This study explores two reaction mechanism hypotheses, involving one or two water molecules, and distinct roles for the Ca2+ cofactor. Our research focuses on the human synovial sPLA2 enzyme bound to lipid bilayers of varying phospholipid compositions, and employing adiabatic QM/MM and QM/MM MD umbrella sampling methods to energetically and geometrically characterize the structures found along both reaction pathways. Our studies demonstrate the higher frequency of productive conformations within the single-water pathway, also revealing a lower free energy barrier for hydrolyzing POPC. Furthermore, we observe that the TS of this concerted one-step reaction closely resembles transition state geometries observed in X-ray crystallography complexes featuring high-affinity transition state analogue inhibitors.

Correction: de Oliveira et al. Viper Venom Phospholipase A2 Database: The Structural and Functional Anatomy of a Primary Toxin in Envenomation. Toxins 2024, 16, 71.

In the published publication [...].

Linezolid and vancomycin for nosocomial infections in pediatric patients: a systematic review

Abstract Objective: To investigate the effectiveness of linezolid and vancomycin for the treatment of nosocomial infections in children under 12 years old. Data sources: This is a systematic review in which five randomized clinical trials about the effectiveness of linezolid and vancomycin, involving a total of 429 children with nosocomial infections, were evaluated. They were searched in scientific databases: PubMed, Bvs, and SciELO. Summary of findings: The main nosocomial infections that affected children were bacteremia, skin, and soft tissue infections followed by nosocomial pneumonia. Most infections were caused by Gram-positive bacteria, which all studies showed infections caused by Staphylococcus aureus, with methicillin-resistant S. aureus (MRSA) and methicillin-resistant coagulase-negative staphylococci strains being isolated. Both linezolid and vancomycin showed high therapeutic efficacy against different types of nosocomial infections, ranging from 84.4% to 94% for linezolid and 76.9% to 90% for vancomycin. Patients receiving linezolid had lower rates of rash and red man syndrome compared to those receiving vancomycin. However, despite the adverse reactions, antimicrobials can be safely administered to children to treat nosocomial infections caused by resistant Gram-positive bacteria. Conclusion: Both linezolid and vancomycin showed good efficacy in the treatment of bacterial infections caused by resistant Gram-positive bacteria in hospitalized children. However, linezolid stands out regarding its pharmacological safety. Importantly, to strengthen this conclusion, further clinical trials are needed to provide additional evidence.

DszA Catalyzes C-S Bond Cleavage through N5-Hydroperoxyl Formation.

Due to its detrimental impact on human health and the environment, regulations demand ultralow sulfur levels on fossil fuels, in particular in diesel. However, current desulfurization techniques are expensive and cannot efficiently remove heteroaromatic sulfur compounds, which are abundant in crude oil and concentrate in the diesel fraction after distillation. Biodesulfurization via the four enzymes of the metabolic 4S pathway of the bacterium Rhodococcus erythropolis (DszA-D) is a possible solution. However, the 4S pathway needs to operate at least 500 times faster for industrial applicability, a goal currently pursued through enzyme engineering. In this work, we unveil the catalytic mechanism of the flavin monooxygenase DszA. Surprisingly, we found that this enzyme follows a recently proposed atypical mechanism that passes through the formation of an N5OOH intermediate at the re side of the cofactor, aided by a well-defined, predominantly hydrophobic O2 pocket. Besides clarifying the unusual chemical mechanism of the complex DszA enzyme, with obvious implications for understanding the puzzling chemistry of flavin-mediated catalysis, the result is crucial for the rational engineering of DszA, contributing to making biodesulfurization attractive for the oil refining industry.

Successful embolization of a clinically significant pulmonary arteriovenous malformation.

The authors present the clinical case of a 59-year-old female patient with a history of peripheral desaturation, which was detected in the perioperative period 4 years earlier. She reported exertional dyspnea, quantified as grade 2 on the Modified Medical Research Council (mMRC) Dyspnea Scale (walks slower than people of the same age because of dyspnea or has to stop for breath when walking at her own pace), and morning cough with mucoid sputum and denied platypnea, epistaxis, telangiectasias and hemoptysis. A computed chest tomography scan revealed a contrast-enhanced lesion on the right upper lobe with an afferent and two efferent vessels compatible with pulmonary arteriovenous malformation. The transesophageal echocardiogram revealed an important right-left shunt compatible with arteriovenous fistula in the pulmonary circulation. An angiography confirmed the diagnosis and a selective embolization of the afferent artery was performed with resolution of symptoms.

Acesso vascular intraósseo: atitudes e práticas dos enfermeiros do serviço de urgência / Intraosseous vascular access: attitudes and practices of emergency department nurses

O Estágio de Natureza Profissional é um período fundamental do segundo ciclo de estudos pois visa completar a formação académica da componente de especialização, onde o estudante, integrado num contexto profissional, imerge em ambiente e situações clínicas complexas, desenvolvendo atividades que lhe permitam adquirir e aprimorar competências comuns e específicas do Enfermeiro Especialista em Enfermagem Médico-Cirúrgica. Pretende-se com este relatório evidenciar as oportunidades de aprendizagem e atividades realizadas no âmbito deste espaço e tempo formativo, no serviço de urgência de um hospital do norte do país, compreendendo a importância e apropriando a intervenção do Enfermeiro Especialista em Enfermagem Médico-Cirúrgica na área da Pessoa em Situação Crítica e as suas competências para gerir cuidados de enfermagem, intervir na formação de equipas de saúde e prestar cuidados altamente qualificados à pessoa doente e família. A investigação que integra este relatório assenta num estudo descritivo-correlacional com o objetivo de analisar as atitudes e práticas dos enfermeiros do serviço de urgência, de um hospital do norte de Portugal, na utilização do acesso vascular intraósseo. Utilizou-se o questionário como instrumento de recolha de dados a uma amostra de 76 enfermeiros, distribuído e preenchido via on-line, entre 4 e 15 de maio de 2023. Os resultados evidenciaram que 97,4% dos participantes reconhecem o acesso intraósseo como importante no contexto do serviço de urgência, embora 85,5% nunca o utilizassem, sendo o acesso por veia central privilegiado por 90,8%, como alternativa à falha da inserção da veia periférica. A insatisfação com os conhecimentos sobre cateterização intraóssea é de 89,5%. Fatores como a falta de treino/formação na realização do procedimento são considerados pela maioria, como os mais condicionantes à realização do procedimento. A principal conclusão é que os enfermeiros do serviço de urgência reconhecem a importância do acesso intraósseo, no entanto, afirmam que é necessário mais formação e treino para que o número de cateterizações intraósseas possa aumentar nas situações que se justifiquem. Deste percurso formativo, destaca-se a importância da intervenção diferenciada do Enfermeiro Especialista na melhoria contínua da qualidade de cuidados, num contexto complexo como é o serviço de urgência. Salienta-se, a nível pessoal, o desenvolvimento de competências especializadas comuns e específicas, na interação com o ambiente clínico envolvente.

The Professional Internship is a fundamental period of the second cycle of studies because it aims to complete the academic training of the specialisation component, where the student, integrated in a professional context, immerses himself in complex clinical environment and diferent situations, developing activities that allow him to acquire and improve common and specific skills of the Specialist Nurse in Medical-Surgical Nursing. This report aims to highlight the learning opportunities and the activities carried out within this space and time training, in the emergency service of a hospital in the north of the country, understanding the importance and appropriate intervention of the Nurse Specialist in Medical-Surgical Nursing in the area of the Person in a Critical Situation and their skills to manage nursing care, intervene in the training of health care teams and provide highly qualified care to the sick person and their families. The research that integrates this report is based on a descriptive-correlational study with the objective of analysing the attitudes and practises of nurses in the emergency service, of a hospital in the north of Portugal, in the use of intraosseous vascular access. The questionnaire was used as a data collection tool for a sample of 76 nurses, distributed and completed online, between May 4 and 15, 2023. The results showed that 97.4% of the participants recognise intraosseous access as important in the context of the emergency service, although 85.5% never used it, and access by central vein was privileged by 90.8%, as an alternative to the failure of the insertion of the peripheral vein. Dissatisfaction with the knowledge of intraosseous catheterisation is 89.5%. Factors such as the lack of training in the realisation of the procedure are considered by most, as the most conditioning to the realisation of the procedure. The main conclusion is that the nurses of the emergency service recognise the importance of intraosseous access, however, they say that more training is necessary so that the number of intraosseous catheterisations can increase in situations that are justified. From this training path, the importance of the differentiated intervention of the Specialist Nurse in the continuous improvement of the quality of care is highlighted, in a complex context such as the emergency service. It is noteworthy, at a personal level, the development of common and specific specialised skills in the interaction with the surrounding clinical environment.

SCCmec transformation requires living donor cells in mixed biofilms.

Methicillin-resistant Staphylococcus aureus (MRSA) is an important human pathogen that has emerged through the horizontal acquisition of the staphylococcal cassette chromosome mec (SCCmec). Previously, we showed that SCCmec from heat-killed donors can be transferred via natural transformation in biofilms at frequencies of 10-8-10-7. Here, we show an improved transformation assay of SCCmec with frequencies up to 10-2 using co-cultured biofilms with living donor cells. The Ccr-attB system played an important role in SCCmec transfer, and the deletion of ccrAB recombinase genes reduced the frequency ∼30-fold. SCCmec could be transferred from either MRSA or methicillin-resistant coagulase-negative staphylococci to some methicillin-sensitive S. aureus recipients. In addition, the transformation of other plasmid or chromosomal genes is enhanced by using living donor cells. This study emphasizes the role of natural transformation as an evolutionary ability of S. aureus and in MRSA emergence.

Evolution of the Quinoline Scaffold for the Treatment of Leishmaniasis: A Structural Perspective.

Since the beginning of the XXI century, Leishmaniasis has been integrated into the World Health Organization's list of the 20 neglected tropical diseases, being considered a public health issue in more than 88 countries, especially in the tropics, subtropics, and the Mediterranean area. Statistically, this disease presents a world prevalence of 12 million cases worldwide, with this number being expected to increase shortly due to the 350 million people considered at risk and the 2-2.5 million new cases appearing every year. The lack of an appropriate and effective treatment against this disease has intensified the interest of many research groups to pursue the discovery and development of novel treatments in close collaboration with the WHO, which hopes to eradicate it shortly. This paper intends to highlight the quinoline scaffold's potential for developing novel antileishmanial agents and provide a set of structural guidelines to help the research groups in the medicinal chemistry field perform more direct drug discovery and development programs. Thus, this review paper presents a thorough compilation of the most recent advances in the development of new quinoline-based antileishmanial agents, with a particular focus on structure-activity relationship studies that should be considerably useful for the future of the field.

Identification of Inflammatory Mediators in Saliva Samples From Hospitalized Newborns: Potential Biomarkers?

Saliva measurements serve as a noninvasive tool for clinically monitoring newborns (NB) and children, a vulnerable population with promising potential for both research and clinical practice. Saliva acts as a repository for various inflammatory biomarkers involved in diverse biological functions. Particularly for children, it offers numerous advantages when compared to plasma and urine sampling. Nevertheless, there is a significant knowledge gap regarding detectable levels of cytokines in the saliva of newborns and children, as well as studies aiming to assess the relationship of this content with physiological and pathological processes. OBJECTIVES: To characterize the levels of 11 inflammatory mediators (IFNg, IL1b, IL2, IL4, IL6, IL8, IL10, IL12, IL17, TNF, and VEGF) in saliva samples from NB on the first and second day of hospitalization in the Neonatal Intensive Care Unit (NICU). METHOD: Exploratory study, descriptive, nested within a primary clinical, observational, and prospective study, conducted in the NICU of a public hospital in São Paulo, Brazil. Demographic data and vital signs were recorded in the clinical records of 90 NB, and five saliva samples from 5 NB were collected between the first and second day of life (D1-D2) at approximately 8-hr intervals (8-9 am, 4-5 pm, and 11-12 pm). Saliva samples were used for the measurement of 11 cytokines (IFNg, IL1b, IL2, IL4, IL6, IL8, IL10, IL12, IL17, TNF, and VEGF). RESULTS: Five NBs participated in this exploratory study, and the vital signs showed variability from the first (D1) to the second day (D2) of hospitalization, variability similar to that of the total population of the primary study. The presence and levels of the 11 cytokines were detected in the saliva samples, as well as a statistical correlation between 10 cytokines (IFNg, IL1b, IL2, IL4, IL6, IL10, IL12, IL17, TNF, and VEGF) and vital signs. CONCLUSIONS: The novelty of measuring inflammatory mediators in saliva samples from hospitalized NBs in the NICU is highlighted, providing support and new perspectives for the development of clinical and experimental research and an opportunity for developing and implementing new salivary biomarkers in different population segments.

Viper Venom Phospholipase A2 Database: The Structural and Functional Anatomy of a Primary Toxin in Envenomation.

Viper venom phospholipase A2 enzymes (vvPLA2s) and phospholipase A2-like (PLA2-like) proteins are two of the principal toxins in viper venom that are responsible for the severe myotoxic and neurotoxic effects caused by snakebite envenoming, among other pathologies. As snakebite envenoming is the deadliest neglected tropical disease, a complete understanding of these proteins' properties and their mechanisms of action is urgently needed. Therefore, we created a database comprising information on the holo-form, cofactor-bound 3D structure of 217 vvPLA2 and PLA2-like proteins in their physiologic environment, as well as 79 membrane-bound viper species from 24 genera, which we have made available to the scientific community to accelerate the development of new anti-snakebite drugs. In addition, the analysis of the sequenced, 3D structure of the database proteins reveals essential aspects of the anatomy of the proteins, their toxicity mechanisms, and the conserved binding site areas that may anchor universal interspecific inhibitors. Moreover, it pinpoints hypotheses for the molecular origin of the myotoxicity of the PLA2-like proteins. Altogether, this study provides an understanding of the diversity of these toxins and how they are conserved, and it indicates how to develop broad, interspecies, efficient small-molecule inhibitors to target the toxin's many mechanisms of action.

Beneficial Effect of Exogenously Applied Calcium Pyruvate in Alleviating Water Deficit in Sugarcane as Assessed by Chlorophyll a Fluorescence Technique.

The growing demand for food production has led to an increase in agricultural areas, including many with low and irregular rainfall, stressing the importance of studies aimed at mitigating the harmful effects of water stress. From this perspective, the objective of this study was to evaluate calcium pyruvate as an attenuator of water deficit on chlorophyll a fluorescence of five sugarcane genotypes. The experiment was conducted in a plant nursery where three management strategies (E1-full irrigation, E2-water deficit with the application of 30 mM calcium pyruvate, and E3-water deficit without the application of calcium pyruvate) and five sugarcane genotypes (RB863129, RB92579, RB962962, RB021754, and RB041443) were tested, distributed in randomized blocks, in a 3 × 5 factorial design with three replications. There is dissimilarity in the fluorescence parameters and photosynthetic pigments of the RB863129 genotype in relation to those of the RB041443, RB96262, RB021754, and RB92579 genotypes. Foliar application of calcium pyruvate alleviates the effects of water deficit on the fluorescence parameters of chlorophyll a and photosynthetic pigments in sugarcane, without interaction with the genotypes. However, subsequent validation tests will be necessary to test and validate the adoption of this technology under field conditions.