Insights into nucleoside hydrolase from Leishmania donovani inhibition: A new bioaffinity chromatography-based screening assay and docking studies.

Leishmaniasis, a group of neglected infectious diseases, encompasses a serious health concern, particularly with visceral leishmaniasis exhibiting potentially fatal outcomes. Nucleoside hydrolase (NH) has a fundamental role in the purine salvage pathway, crucial for Leishmania donovani survival, and presents a promising target for developing new drugs for visceral leishmaniasis treatment. In this study, LdNH was immobilized into fused silica capillaries, resulting in immobilized enzyme reactors (IMERs). The LdNH-IMER activity was monitored on-flow in a multidimensional liquid chromatography system, with the IMER in the first dimension. A C18 analytical column in the second dimension furnished the rapid separation of the substrate (inosine) and product (hypoxanthine), enabling direct enzyme activity monitoring through product quantification. LdNH-IMER exhibited high stability and was characterized by determining the Michaelis-Menten constant. A known inhibitor (1-(ß-d-Ribofuranosyl)-4-quinolone derivative) was used as a model to validate the established method in inhibitor recognition. Screening of three additional derivatives of 1-(ß-d-Ribofuranosyl)-4-quinolone led to the discovery of novel inhibitors, with compound 2a exhibiting superior inhibitory activity (Ki = 23.37 ± 3.64 µmol/L) compared to the employed model inhibitor. Docking and Molecular Dynamics studies provided crucial insights into inhibitor interactions at the enzyme active site, offering valuable information for developing new LdNH inhibitors. Therefore, this study presents a novel screening assay and contributes to the development of potent LdNH inhibitors.

Nucleoside hydrolase immobilized on magnetic particles as a tool for onflow screening and characterization of inhibitors.

Nucleoside Hydrolases (NH) are considered a target for the development of new antiprotozoal agents. The development of new and automated screening assays for the identification of NH inhibitors can accelerate the first stages of the drug discovery process. In this work, NH from Leishmania donovani (LdNH) was covalently immobilized onto magnetic particles (LdNH-MPs) and trapped by magnets into a TFE tube to yield an immobilized enzyme reactor (IMER). For an automated assay, the LdNH-MP-IMER was connected in-line to an analytical column in an HPLC-DAD system to monitor the enzyme activity through quantification of the product hypoxanthine. Kinetic studies provided a KM value of 2079 ± 87 µmol.L-1 for the inosine substrate. Validation of the LdNH-MP-IMER for onflow screening purposes was performed with a library containing 12 quinolone ribonucleosides. Among them, three were identified as new competitive LdNH inhibitors, with Ki values between 83.5 and 169.4 µmol.L-1. This novel in-line screening assay has proven to be reliable, fast, low cost, and applicable to large libraries of compounds.

Ligand fishing as a tool to screen natural products with anticancer potential.

Cancer is the second leading cause of death in the world and its incidence is expected to increase with the aging of the world's population and globalization of risk factors. Natural products and their derivatives have provided a significant number of approved anticancer drugs and the development of robust and selective screening assays for the identification of lead anticancer natural products are essential in the challenge of developing personalized targeted therapies tailored to the genetic and molecular characteristics of tumors. To this end, a ligand fishing assay is a remarkable tool to rapidly and rigorously screen complex matrices, such as plant extracts, for the isolation and identification of specific ligands that bind to relevant pharmacological targets. In this paper, we review the application of ligand fishing with cancer-related targets to screen natural product extracts for the isolation and identification of selective ligands. We provide critical analysis of the system configurations, targets, and key phytochemical classes related to the field of anticancer research. Based on the data collected, ligand fishing emerges as a robust and powerful screening system for the rapid discovery of new anticancer drugs from natural resources. It is currently an underexplored strategy according to its considerable potential.

Frequency of germline genetic variants in women with a personal or family history of breast cancer from Brazil.

BACKGROUND: About 5-10% of breast cancer cases are related to genetic and hereditary factors. The application of Next Generation Sequencing (NGS) in oncology has allowed the identification of genetic variants present in several genes related to the increased risk of breast cancer. This study aimed to determine the frequency of germline genetic variants in patients with a family and/or personal history of breast cancer. METHODS: An analysis of positive reports from NGS panels was carried out in female individuals with a personal and/or family history of breast cancer, present in the database of a private laboratory in Brazil. RESULTS: From about 2000 reports, 183 individuals presented 219 different germline genetic variants. The genes with the highest number of variants were BRCA2 (16.0%), ATM (15.0%) and BRCA1 (12.8%). Among the variants found, 78 were either pathogenic or probably pathogenic, accounting for 35% of all variants discovered. The gene with the highest proportion of pathogenic/probably pathogenic variants was TP53 (80%) and the most frequent pathogenic variant was also reported in this gene (c.1010G > A p.(Arg337His)). Furthermore, the study obtained a high proportion of variants of uncertain significance (VUS) (65%) and approximately 32% of the variants found were in genes of moderate penetrance. CONCLUSIONS: Our results could improve the risk estimation and clinical follow-up of Brazilian patients with a history of breast cancer.

Magnetic particles for enzyme immobilization: A versatile support for ligand screening.

Enzyme inhibitors represent a substantial fraction of all small molecules currently in clinical use. Therefore, the early stage of drug-discovery process and development efforts are focused on the identification of new enzyme inhibitors through screening assays. The use of immobilized enzymes on solid supports to probe ligand-enzyme interactions have been employed with success not only to identify and characterize but also to isolate new ligands from complex mixtures. Between the available solid supports, magnetic particles have emerged as a promising support for enzyme immobilization due to the high superficial area, easy separation from the reaction medium and versatility. Particularly, the ligand fishing assay has been employed as a very useful tool to rapidly isolate bioactive compounds from complex mixtures, and hence the use of magnetic particles for enzyme immobilization has been widespread. Thus, this review provides a critical overview of the screening assays using immobilized enzymes on magnetic particles between 2006 and 2021.

Carcinoma basoescamoso na pálpebra inferior: reconstrução com enxerto de cartilagem auricular associado ao retalho de Tripier após cirurgia de Mohs / Basosquamous carcinoma in lower eyelid: reconstruction with auricular cartilage graft associated with the Tripier flap after Mohs surgery

O carcinoma basoescamoso tem alto potencial de recorrência local e metástase, especialmente quando recidivado e na localização periocular. Um paciente masculino de 49 anos foi submetido a cirurgia micrográfica de Mohs e reconstrução subtotal da pálpebra inferior com enxerto de cartilagem da hélice da orelha associado ao retalho de Tripier, evoluindo com ótimo resultado funcional e estético

Basosquamous carcinoma has a high potential for local recurrence and metastasis, especially when it recurs and at the periocular area. A 49-year-old man underwent Mohs micrographic surgery and subtotal reconstruction of the lower eyelid with cartilage graft from the helix of the ear associated with the Tripier flap, evolving with excellent functional and aesthetic results

Aesthetic Rehabilitation in Teeth with Wear from Bruxism and Acid Erosion.

BACKGROUND: Bruxism is defined as a repetitive activity of grinding the teeth through lateral or protrusive movements of the mandible, and it is considered the most complex and destructive functional disorder. In addition, erosion caused by diet or reflux can damage the remaining teeth. OBJECTIVE: In this report, a patient with bruxism and dental erosion was treated with a multidisciplinary approach to restore the function and aesthetic. METHODS: This clinical report describes the management of an adult woman, 33 years old, who was dissatisfied with the aesthetics of her smile and complained of joint pain and headaches. As result of her condition, the patient's dentition exhibited generalized wear on the vestibular and incisal surface of the upper incisors incompatible with her age, moderate darkening of the teeth and excess gingival tissue in the upper incisors. After a detailed anamnesis and clinical examination, a diagnosis of bruxism and acid erosion caused by a diet rich in citrus foods and beverages was obtained. Forthwith, a treatment plan was established, and the patient underwent home bleaching, gingivoplasty and ceramic laminates of lithium disilicate on the anterior teeth. After the rehabilitation was completed, a night guard was made to reduce the symptoms of bruxism and avoid fracture of the ceramic restorations. The patient was followed at different time intervals. CONCLUSION: The improvement in the aesthetics of the teeth was significant and remained stable after periodic controls in which no adverse effects were observed.

Effect of maternal cells transferred with colostrum on the health of neonate calves.

The objective of this research was to evaluate the influence of cells from colostrum on the health of neonate calves. Animals were distributed in 2 groups: COL+ (n=9) which received fresh colostrum from their own damns; and COL- (n=10) which received frozen colostrums from donors. Heifers were assessed before colostrum intake - D0; D2; D7; D14; D21 and D28. Heifers were monitored by clinical examination, hematological profile and serum iron. COL- had a higher diarrhea intensity score (typically 3) on D7. Moreover, a single case each of bronchopneumonia and navel inflammation were observed in COL- calves. COL- had fewer red blood cells (RBC) (6.5±0.8×106/µL) and less hemoglobin (Hgb) (8.3±1.4g/dL) than COL+ (RBC=7.2±0.8×106/µL; Hgb=9.6±1.3g/dL) at D14 (P≤0.05). COL- had more anemia on D21 (P=0.03) and on D28 (P=0.02). Iron was lower in COL- (5.6±2.7µM/L) than COL+ (10.7±6.2µM/L) (P=0.03) on D7. Lymphocytes was lower in COL- than COL+ on D7 (3.8±1.0×103/µL COL+ and 5.4±2.2×103/µL COL-, P=0.02). COL- calves had more anemia and lower serum iron concomitant with diarrhea on D7. The number of leukocytes was relatively consistent in the COL+ calves, while COL- calves showed an increasing number of of lymphocytes starting on D7.

Influence of filling technique and bevel preparation on marginal sealing of conventional and low shrinkage composite restorations

AIM: The aim of this study was to evaluate two composite restorations (conventional methacrylate-based and low shrinkage-based silorane), associated or not to beveling and incremental technique, in relation to the microleakage and marginal gap formation tests. MATERIAL AND METHODS: 30 sound human molars had their mesial and distal surfaces prepared with and without bevel. The teeth were divided into six groups according to the type of resin (conventional nanofilled or silorane resin), and restorative filling technique (incremental technique or builk). Microleakage and marginal gap formation data were statistically analyzed by Kruskal-Wallis and Chi-Square tests, with a significance level set at 5%. RESULTS: Group 1 (Z350 XT, without bevel, incremental technique) presented highest rate of microleakage and marginal gap in most of the restorations, with statistically significant differences. Groups 4 and 6 (P90, beveled, builk and P90, beveled and incremental technique) had lower rates of microleakage and absence of marginal gaps in most of the restorations. CONCLUSION: It's concluded that the silorane showed lower rates of marginal gaps and microleakage, when compared to the conventional methacrylate-based resin. Bevel preparation was effective in reducing microleakage and marginal gaps for both resins used. Incremental technique was not necessary when associated with low shrinkage composite resin.