Introduction: Patient safety can be defined as the reduction to an acceptable minimum of the risk of unnecessary harm associated with healthcare. Exams are carried out in a complex environment, where there are procedures and equipment, especially human knowledge, in order to guarantee good results that guide diagnostic and therapeutic decisions. The existence of high rates of unacceptable errors which results in wrong, inappropriate and missed diagnoses. Despite all the measures that can be taken to mitigate risks to patient safety, we need to be alert and design a quality system that allows the rapid identification of occurrences and their immediate communication, reducing the impact on patients' health. Objective: Evaluate processes in the pre-analytical, analytical and post-analytical phases, in a clinical analysis laboratory, from the perspective of patient safety. Method: This is an exploratory, observational, descriptive study with a quantitative approach. Data collection took place in the Clinical Analysis Laboratory in a military hospital in the Federal District, in a semi-structured form for each of the phases: Results: The study was carried out based on the analysis of data from 378 patients, equivalent to the assessment of 3,000 thousand laboratory tests. In the pre-analytical phase, 57% of the exams were carried out without identifying the patient, 50% of the time the collection tubes were not placed in the correct order, 88% of the time the venous stasis time was prolonged and in 75% The time between collection and centrifugation respected the blood clotting time. In the analytical phase, equipment failure occurred 21% of the time, controls were not in compliance in 13%, reagent batches were changed in 16%, control batches were changed in 8% and batches were changed in 10%. of calibrator. In the postanalytical phase, there was an interface failure in transcribing the report in 18%, in 5.9% there was instability in the laboratory information system. The result release time is 17% inadequate. Discussion: Patient safety is a global public health priority according to the World Health Organization. In the context of laboratory tests, used for diagnosis, it is essential that the processes are reliable, so that they guide all decision-making regarding health of people. In the study hospital, important flaws were noticed, such as in patient identification, the number 1 goal of patient safety. Furthermore, although the processes in the analytical and post-analytical phases have fewer errors, failures in the pre-analytical phase compromise everything else. The hospital lacks the implementation of quality management processes in order to continually make the necessary improvements. Conclusion: This is a necessary study to expand knowledge about the diagnostic process in laboratory medicine, which proved to be a flawed process in the study hospital. A multicenter study is suggested to understand the reality in Brazil.

The growing number of multidrug-resistant microorganisms is currently one of the biggest global health problems, according to the WHO. The present work proposed to evaluate the effect of postbiotics secreted by Lactobacillus spp. and short-chain fatty acids present in the intestinal microbiota, on the growth and virulence of Staphylococcus aureus and Klebsiella pneumoniae, which, according to data from the Ministry of Health, are the 1st and 3rd pathogens with the highest number of isolates in Brazil, respectively. Lactobacillus spp.. were cultivated for 48h at 37 ± 1°С in LACTOBACILLI MRS BROTH Acumedia® 29 culture medium (Neogen do Brasil, São Paulo, Brazil). S. aureus and K. pneumoniae strains were previously cultivated aerobically for 24h at 37 ± 1°С in Mueller Hinton (MH) BROTH medium. In the preparation of the postbiotic from Lactobacillus spp. pure and fresh cultures were used, centrifuged for 10 minutes at 2000 rpm and subsequently filtered with a 0.25 µm sterile membrane filter. The Minimum Inhibitory Concentration (MIC) assays were standardized following BrCAST (Brazilian Antimicrobial Sensitivity Test Committee) for broth microdilution assay, with adaptations. The media for the MIC assays were MH BROTH plus species-specific (v/v) postbiotics from L. reuteri and L. rhamnnosus, and a 500mM sodium butyrate solution was also prepared, both test compounds were added to the first well following serial dilution in the others. Each column of the 96-well plate represents a group (control or test) and each line represents serial dilution, starting in the first row (more concentrated) and moving on to the last row (less concentrated). The data presented in this study prove a reduction in the growth of S. aureus and K. pneumoniae with postbiotics from L. Reuteri and L. rhamnnosus, wells 1 to 8 at concentrations of 50% to 0.39% (v/v) and butyrate of sodium from well 1 with 50 mM followed by serial dilution in the others. In this way, a new complementary therapeutic approach is suggested for the decolonization of resistant strains of S. aureus and K. pneumoniae, proposing a methodology that allows investigating which microbial interactions can favor the host.

Financing and investment issues have been part of the research agendas on the Unified Health System (SUS). Although it is recognized that there are resource management problems, it is argued that the system that aims to be “universal” and “integral” operates with underfunding. Financing is also an essential factor in the availability of medicines, a key element in a resolute health system. That said, this study aimed to analyze public investments made by federated entities in medicines and identify potential inequalities in the application of these resources that could have impacts on the supply of medicines through the Unified Health System (SUS). The thesis is presented in 03 chapters. In chapter 01 - Threatened equity: Regional asymmetries in investments in medicines in Brazil, an analysis of investments in health and medicines by the Union, states and municipalities between 2010 and 2019 was carried out, aiming to identify regional differences in the application of these resources and it was observed that although per capita investments in health were higher by municipalities (R$ 926.54), when it comes to medicines, this per capita investment was higher by the Union (R$ 61.89). It was also identified that municipalities in the Northeast and North regions invest less in purchasing medicines when compared to the national average (R$40.41), R$24.20 and R$23.58 respectively. By exploring the percentage of investment in medicines compared to the total investment in health among entities, it was possible to verify that states and municipalities have been decreasing their percentages of investment in medicines, highlighting the greatest reduction by states, where this decrease was 72 .8% during the evaluated period. Chapter 02 presents an analysis of the resources for purchasing Primary Care medicines among the responsibilities of federated entities since 2016, highlighting municipal characteristics. This is a retrospective study, in which the evolution of the values of CBAF counterparts by entities was identified and compared. The amounts spent on medicines by the municipalities were higher than the counterparts from the Ministry of Health (MS) or MS+Estado in the period analyzed. The average percentage of transfers of federal resources and municipal expenditures vary depending on the Brazilian region. The average per capita value invested in medicines by municipalities increased between 2016 and 2020, with the impact being greater for municipalities with lower MHDI. The Popular Pharmacy Program mainly affects municipalities with the largest population sizes and highest MHDI, and is therefore not sufficient to address the inequities highlighted. It is concluded, from this chapter, that there has been an increase in inequalities in the capacity of municipalities to ensure access to medicines, especially among the most vulnerable municipalities, accumulating more risks of injuries and deaths from diseases sensitive to Primary Care. And in chapter 03, with the proposal of the RE indicator, which measures the quality of the price charged in the acquisition of medicines, the prices charged in the acquisition of the most used medicines in Primary Care in Brazilian municipalities were analyzed in the years 2016, 2018 and 2020 using data collected in the National Database of Pharmaceutical Assistance Actions and Services in the SUS (BNAFAR). This study also showed the importance of the practice of combined purchasing in drug prices, in addition to detecting differences in drug prices in relation to analyzes by population strata, by region and by MHDI. It is concluded that there is a regional asymmetry in investments in health and medicines in Brazil, in addition to the different evolution of these investments in the period analyzed in each chapter, with the most vulnerable regions being those that invested least. The increase in inequalities in the capacity of municipalities to ensure access to medicines was shown, especially among the most vulnerable municipalities, accumulating even more risks of injuries and deaths from diseases sensitive to primary care. The relevance of the SUS to promote access to medicines is well-known, however it is suggested that governments pay greater attention to investing in medicines to serve the population, as barriers to access to medicines can have a negative result on adherence to treatment, and achieving universal health coverage for medicines reduces household expenditures on healthcare costs, healthcare litigation, outpatient care, hospitalization, and mortality. It is also suggested that public policies be developed that reduce asymmetries in the evolution of investments in health and medicines by all entities and that contribute to promoting equity in investments in health and medicines throughout the country.”

Curcumin is a substance of natural origin found in the rhizome of Curcuma longa that has been proposed for the treatment of several types of cancer. However, curcumin has some disadvantages that impact its pharmacokinetics, such as low aqueous solubility and sensitivity to physiological pH. Thus, in the case of superficial tumors, such as oral cancer and melanoma, its topical use would be a promising alternative for releasing the drug directly to its site of action. Considering the difficulty that a lipophilic drug must have in penetrating the skin and mucosa, the present study proposes the development of bioadhesive films containing curcumin as an alternative for the topical treatment of skin and oral tumors. First, an analytical method was developed using high-performance liquid chromatography capable of quantifying curcumin recovered from the skin and oral mucosa. The method was able to measure curcumin from the biological matrices and proved to be adequate considering all the validation parameters evaluated. The chemical compatibility between the drug and the formulation excipients was demonstrated using thermal analysis techniques and the results indicated compatibility between them. Then, three films containing chitosan, polyvinyl alcohol, Poloxamer®407 and propylene glycol (FA-5, FC-5, and FD-5) were developed and selected, which were characterized considering aspect, weight, thickness, bending resistance, pH, curcumin content, swelling index, rupture force, mucoadhesiveness and morphology. The films had a smooth appearance, were flexible, free of bubbles, cracks, or precipitates, with uniform weight and thickness. They have a pH compatible with the application sites (from 5.2 to 5.5), were resistant to folding and the dosage was very close to the theoretical concentration (78.6 µg/cm2) of curcumin. They presented tensile strength and swelling necessary to guarantee mucoadhesion and drug release. The morphological analysis reinforced the formation of the structure compatible with the proposed system and elucidated small differences between the formulations. The stability study demonstrated the maintenance of parameters such as appearance, thickness, pH, and content during the experiment period (90 days) at room temperature. Drug release in physiological media was determined in vitro for 24 h. All films controlled the release of curcumin in the first 8 h, with 51.6 ± 4.8%, 65.6 ± 13.0% and 55.2 ± 11.2% of the drug released, for the FA- films. 5, FC-5 and FD-5, respectively. In vitro penetration studies showed that all developed films significantly increased (p<0.05) the penetration of curcumin into the mucosa (static permeation: 14.2 ± 3.1 µg/cm2, 7.5 ± 1.0 µg /cm2 and 8.2 ± 0.8 µg/cm2, dynamic permeation: 1.6 ± 0.1 µg/cm2, 1.6 ± 0.4 µg/cm2 and 1.9 ± 0.1 µg/cm2 for the FA5, FC-5 and FD-5 films, respectively) compared to the control containing curcumin in solution, which did not penetrate the mucosa in quantifiable amounts. In relation to the skin, the films also promoted permeation in quantifiable amounts (stratum corneum: 1.9 ± 0.8 µg/cm2, 1.4 ± 0.1 µg/cm2 and 1.3 ± 0.1 µg/cm2; remaining skin: 2.7 ± 0.2 µg/cm2, 2.3 ± 0.4 µg/cm2 and 1.7 ± 0.1 µg/cm2). Cytotoxicity tests carried out on 3 types of tumor cells (FaDu, SCC-9 and MeWo) and healthy cells (HaCaT) demonstrated cytotoxic activity of curcumin at doses close to those found in the permeation test and the evaluation in association with radiotherapy demonstrated that the treatment with single doses of 4, 8 and 12 Gy radiotherapy would be unnecessary. Finally, the irritation test proved that the films are safe for topical application. Therefore, the films developed should represent a promising alternative for the topical treatment of oral and skin tumors using curcumin.

There is a great demand on the market for rejuvenating cosmetics and the use of injectable collagen biostimulating substances to manage the signs of aging. Calcium Hydroxyapatite (CaHA) is a natural mineral from human teeth and bones, used injectably to treat wrinkles and stimulate collagen production. Its use in regions at risk and incorrectly can lead to complications such as tissue necrosis, blindness and stroke. This study aims to develop a cosmetic formulation for topical application of CaHA and evaluate iontophoresis and microneedling as promoters of its permeation. To this end, CaHA acquired from the supplier with a size of ~10 µm (Commercial CaHA) were dissolved and subjected to drying using two different equipment/processes: (i) Nanospray Dryer and (ii) Spray Dryer. Particles were produced using or not using lactose as an anti-aggregant. The nanoparticles obtained from the Nanospray Dryer (Nano CaHA, Nano CaHA Lac Mix and Nano CaHA Lac) were 829.45 ± 27.08 nm, 504.07 ± 0.25 nm and 566.13 ± 39.79 nm. , and polydispersity index (PdI) 0.6 ± 0.06, 0.3 ± 0.11 and 0.1 ± 0.04, respectively, and the microparticle obtained from the Spray Dryer (Micro CaHA) presented size 2239 .3 ± 309.46 nm. The morphology of all particles was evaluated using scanning electron microscopy (SEM). Thermal analysis of the particles was carried out, demonstrating the interaction of lactose with CaHA during the drying process using both methods. Preliminary assays were carried out to evaluate the effect on cell proliferation, DNA fragmentation and gene expression of Col 1, Col 4 and Col 6, IL-10 and INF-γ in human fibroblast cultures, indicating that the nanometric particles increase division cellular and gene expression of genes related to inflammation and collagen production in human fibroblasts. The particle with the best performance, Nano CaHA Lac, was then selected for further studies. Gene expression of Col 1, Col 3, IL-4, IL-13 and TGF-b was evaluated in co-culture of human fibroblasts and PBMC, in addition to the production of Col 1 in human fibroblasts, in comparison to injectable commercial presentation RadiesseÒ, pointing to a greater action of the particle obtained in this study. Such studies included cells not treated with the obtained particles as controls. The irritant potential of Nano CaHA Lac 1% suspension was evaluated using the HET-CAM method, classifying it as non-irritating. A bioanalytical method using flame atomic absorption spectrometry (FAAS) was developed and validated to quantify calcium (Ca) after in vitro permeation assays and histology of damaged pig ear skin was performed after the use of microneedling. Finally, the permeation of Nano CaHA Lac was evaluated passively, after microneedling and with iontophoresis, demonstrating penetration of 90.5 ± 38.9 µg/cm2, 159.4 ± 39.2 µg/cm2 and 92 .9 ± 25.4 µg/cm2 in the skin layers. From this study it was possible to infer that Nano CaHA Lac suspensions are suitable for topical application of CaHA, as they are safe and capable of significantly stimulating the penetration of CaHA into the deeper layers of the skin, with such penetration being significantly increased in skin pre-treated with microneedling. Thus, the combination of the use of the developed formulation and the microneedling technique can benefit the topical treatment of sagging skin.

“Introduction: The development of clinical skills necessary for the pharmaceutical care of selflimiting health problems in community pharmacy needs to be improved, thus it is important to use tools that structure the training process through simulations and allow a standardized way and validated assessment of these skills. Objective: To develop an instrument to assess clinical skills in the management of self-limiting health problems and to analyze the performance in the pharmaceutical consultation simulation. Methodology: The work was divided into two parts, in the first part, the development and validation, through the Delphi technique, of the clinical competence assessment instrument to be used in clinical simulations (PSAL-BRASIL). In the second part, clinical cases were prepared to be used in realistic simulations on the management of self-limiting health problems and the performance of pharmacists and pharmacy students was analyzed using the validated instrument. Results: Of the 26 experts invited to participate in the validation, 19 (73%) responded to the first round of Delphi. Items (2) Verbal communication and (15) Guidance and monitoring did not reach consensus on clarity (79%) and objectivity (79%), respectively. After the changes suggested by the experts, a second round was performed, in which all items reached consensus, ICV > 0.8. In the second part, 81 pharmacy professionals and students participated in clinical case simulations. The performance of the participants was lower in requesting/performing physical and laboratory examinations and verification of vital signs, followed by the items “guidance and monitoring” and “guidelines for the use of medication”. Discussion: The instrument developed in this research was validated and serves as a reference in the training of pharmaceutical professionals and pharmacy students in the development of their clinical skills in the management of self-limiting health problems. In addition, the use of simulations of clinical cases in order to develop the clinical skills of the participants corroborates the evidence that the use of protocols for the care of these problems in the community pharmacy increases patient safety and is therefore a contribution of the pharmacist to the system health public, as it assists in self-care and selfmedication (AMADOR et al., 2022). Conclusion: The validation of PSAL-BRASIL instrument contributed to the standardization in the assessment of clinical skills necessary for self-limiting health problems care in community pharmacies. ”

“Systemic administration of lipopolysaccharide (LPS) induces several responses controlled by the brain, including fever. Fever is the increase in body temperature, which occurs as a defense response of the body against inflammatory or infectious processes. Despite its beneficial nature, it is known that prolonged fevers or those above 42◦C require high metabolic demand and are associated with brain damage. In a previous study of quantitative proteomic analysis carried out by our group, it was verified that during the fever induced by LPS in rats, there is activation of inflammatory and metabolic pathways, with emphasis on the increase in the abundance of protein diacylglycerol kinase (DGK) in the hypothalamus. DGK belongs to a family of lipid protein kinases responsible for the phosphorylation of diacylglycerol, converting it into phosphatidic acid. Little is known about the participation of DGK in the febrile response and in vivo studies regarding its inhibition are scarce. Thus, this work aimed to investigate the effects of pharmacological inhibition of DGK in a model of fever induced by LPS in rats. Wistar rats received intracerebroventricular administration of the DGK inhibitor, R59949, 30 minutes before the intravenous injection of LPS, and body temperature was monitored for 5 h. Inhibition of DGK led to attenuation of LPS-induced fever, accompanied by decreased hypothalamic PGE2 production. Expression of the proinflammatory genes AKT, NFκB, IL-6 and TNFα in the hypothalamus and circulating levels of reactive species were not altered by treatment with the DGK inhibitor. On the other hand, there was a decrease in the concentration of nitrosylated hemoglobin, which indicates a reduction in the bioavailability of nitric oxide. These results are consistent with the involvement of DGK in the febrile response and, particularly, in hypothalamic PGE2 synthesis."

Antibacterial agents are often used to stop the growth of bacteria such as Staphylococcus aureus. Among such agents, copper oxide nanoparticles (CuO-NP) are a promising candidate. Aim: The dose response in growth inhibition of Staphylococcus aureus treated with colloidal copper oxide nanoparticles (CuO-NP) were evaluated. Methods: An in vitro microbial viability assay was conducted with CuO-NP concentrations spreading over the 0.4–848.0 μg/ml range. The dose–response curve was modeled with a double Hill equation. Visible UV absorption and photoluminescence spectroscopies allowed tracking concentration-dependent modifications in CuO-NP. Results: Two specific phases separated by the critical concentration of 26.5 μg/ml were observed in the dose–response curve, with each exhibiting proper IC50 parameters, Hill coefficients and relative amplitudes. Spectroscopy techniques reveal the concentration-triggered aggregation of CuO-NP starting from this critical concentration. Conclusion: The findings demonstrate a dose-related change in S. aureus sensitivity to CuO-NP, which probably arises from the aggregation of this agent.

In recent years, Cannabis sativa L. has transitioned from a globally prohibited plant to one that is gaining cultural and legal acceptance in many countries for medicinal and recreational use. As jurisdictions legalize cannabis-based products, the variety and complexity of these products extend beyond dried plant material. While numerous active compounds are present in the plant, the main cannabinoids of regulatory and safety concern are Δ9 -tetrahydrocannabinol (THC), cannabidiol (CBD), their respective acid forms THCA-A and CBDA, and cannabinol (CBN), which can be a product of THC oxidation. Recently, the Brazilian Health Regulatory Agency (Anvisa) removed CBD from the list of prohibited substances and reclassified it as controlled substances, as well as allowed the manufacturing and commercialization of certain medicinal cannabis-based products. Therefore, appropriate methods for quantifying the biologically active constituents are essential to ensure safety and regulatory compliance in the oversight and monitoring of medicinal use products, as well as in drug seizure and combating drug trafficking. In this study, a simple, sensitive, and selective LC-MS/MS method was developed and validated for the simultaneous analysis of CBD, CBDA, THC, THCA-A, CBN, and CBNA in plant material and oil cannabis-based products. The chromatographic method used a Kinetex C18 column with isocratic elution containing 0.1% formic acid in water and acetonitrile, providing successful separation of the analytes. Samples of dried and ground plant material (50 mg) and oil samples (12 mg) were extracted with acetonitrile and methanol (8:2), followed by vortexing, sonication, centrifugation, filtration, and dilution. Dilutions were performed at ratios of 1:104 or 1:105 , depending on the sample concentration, to adjust it within the quantification range established by the calibration curve (1.56-100 ng/mL). The preparations were then injected into the LC-MS/MS system (API 3200, Sciex). The method was validated using a Humulus lupulus control sample as the blank matrix for plant samples and a mixture of sunflower, coconut, and extra virgin olive oils for oil samples. Both blank matrices were fortified with the analytes and internal standards (THC-d3, CBD-d3, CBN-d3, and THCAd3). The linearity of the mobile phase calibration curves was demonstrated with regression coefficients r² ≥ 0.99. The method's limits of quantification ranged from 0.09 to 0.15%, meeting regulatory requirements for cannabis products in Brazil, which cannot contain more than 0.2% THC, except in special cases. To demonstrate the method's applicability, 25 oil samples and 150 plant material samples were analyzed. The study showed that products within the same category could exhibit large variations in cannabinoid profile and levels. The results of the study emphasized the need for cannabinoid quantification in cannabis-based products given the current diverse and rapidly changing regulatory landscape worldwide.

Metabolic dysfunction-associated steatotic liver disease (MASLD) – one of the most common types of chronic liver disease with a global prevalence of 20% to 30% and strongly correlated with obesity, insulin resistance, metabolic syndrome, and genetic components – has been characterized as a pandemic with relevant socioeconomic impacts. Since there are no pharmacological therapies to prevent or reverse the progression of MASLD, medications from different classes, such as antidiabetics, antiobesity, antioxidants, and cytoprotective agents, have been evaluated. Particularly, PPARα and PPARγ agonists have been associated with improving MASLD through receptor expression in the liver, stimulating the uptake of fatty acids, lipid oxidation, and insulin sensitization. The present work describes the planning, synthesis, and evaluation of new PPAR ligands for saturated anacardic acid. In this sense, using the strategy of structural modifications in the salicylic subunit of saturated anacardic acid, 9 target compounds were synthesized in yields ranging from 55% to 97% and characterized by NMR. Results on murine and human PPAR activation demonstrated that target derivatives containing the acidic subunit act as partial and dual agonists with EC50 activation values in the micromolar range. The study of structure-activity relationships (REA) for the 2-aryloxyethanoic series revealed that concerning the analog LDT16 (hPPARα EC50 1.1 µM; hPPARα EC50 3.7 µM; hPPARβ/δ EC50 10 µM), LDT458 (2-hydroxymethyl subunit) and LDT461 (2-carbomethoxy subunit) modulated the agonist profiles in a non-significant (well-tolerated strategy) and significant (tolerated approach) negative way for PPARα; and in a non-significant negative way (well-tolerated approach) for PPARγ, respectively. For the 2-aryloxymethylpropanoic series, considering the analog LDT409 (hPPARα EC50 0.5 µM; hPPARα EC50 0.9 µM; hPPARβ/δ EC50 37 µM), LDT469 (2-carbomethoxyl subunit) modulated the profile negatively significant for PPARα (tolerated strategy) and very significant negative (poorly tolerated approach) for PPARγ. Finally, the acidic derivatives LDT458, LDT461, and LDT469 modulated the agonist profile in a negative pharmacophoric way (deleterious approach) for PPARβ/δ.

Rosacea is a chronic dermatological disease characterized mainly by recurrent centrofacial flushing. There is currently no specific treatment for rosacea, and its control is established with topical formulations or oral treatments. Resveratrol (RES) is a natural substance with anti-inflammatory and antioxidant properties, which has been studied to control rosacea, although there is still no product containing RES on the market intended directly for the treatment of rosacea. The present study proposed the development of a stable oil-in-water (EM) microemulsion using grape seed oil enriched with RES for the topical treatment of rosacea. The ME was developed with the construction of a pseudoternary phase diagram. The oily phase consisted of grape seed oil, Tween 80® and Span 80® (2:1 w/w) were used as surfactants, and the aqueous phase comprised ultrapure water. Thermal analyzes (DSC and TGA) of binary mixtures of RES and formulation components were used to evaluate pharmaceutical compatibility. The ME was characterized by analyzing the size distribution of oil droplets, pH and conductivity. ME drug permeation was studied in vitro using porcine skin coupled to Franz diffusion cells for 12 and 24 hours and compared with a control solution of free RES. The selected ME composition (15% oil, 55% water, 30% surfactants) consisted of a translucent formulation, presenting a droplet size of 22.16±0.92 nm, pH 6.4, conductivity of 90.4 µS/cm and was physically stable for at least 60 days. Thermal analyzes showed the compatibility between the medicine and the components of the formulation. After topical application of the drug, ME increased the penetration of RES into the deeper layers of the skin (2.9±0.1 µg/cm2) by 4 times (p<0.05) compared to the control, while reducing it by 4.5 times (p<0.05) the penetration of the medication into the stratum corneum. The physical characteristics of ME seem suitable for topical application. Furthermore, ME could significantly increase RES in the deeper layers of the skin, which may benefit topical rosacea treatment.

Alzheimer's disease remains one of the great public health challenges, whose number of affected patients will increase from 35 million to an astonishing 135 million by 2050. Uncertainty about the etiology and multifactorial nature of Alzheimer's disease (AD) are reasons for the lack of effective drugs while being the basis for developing multi-target ligands (MTDLs). As cases increase in developing countries, there is a need for new drugs that are not only effective but also affordable. With that motivation, we describe the first sustainable MTDLs, derived from cashew nut shell liquid (CNSL) – inexpensive food waste with anti-inflammatory properties. We apply a combination of functionalized CNSL component structures and well-established acetylcholinesterase (AChE)/butyrylcholinesterase (BChE) tacrine templates. MTDLs were selected based on hepatic, neuronal, and microglial cell toxicity. Enzymatic studies revealed potent and selective AChE/BChE inhibitors (43, 44, and 61) with subnanomolar activities. Investigation in BV-2 microglial cells revealed antineuroinflammatory and neuroprotective activities for 43 and 44 (at 0.01 µM), confirming the rational design for this class of compounds.

Epigenetic modifications, including DNA methylation and histone modification, control gene expression and play a crucial role in tumorigenesis. The EHMT family of histone methyltransferasesconsisting of EHMT1 and EHMT2-is dysregulated in various cancers. In this study, we investigated the effects of EHMT1/EHMT2 on cancer cell lines, MeWo and A549. For this purpose, we used the selective inhibitor of EHMT1/EHMT2, UNC0646, and examined the following processes: Cell proliferation, viability, death, mitochondrial function, migration, and gene expression. In addition, we performed in silico analyses and searched public databases of clinical and laboratory data from lung and skin cancer samples. Through these databases, we identified genes that are differentially expressed according to the expression of EHMT1/EHMT2, biological processes modulated by these enzymes, and their impact on the survival of patients with lung and skin cancer. Then, considering the right IC50 and IC75 after MTT assay, we examined the occurrence of cellular apoptosis by flow cytometry (Annexin V/ PI), the production of lactate dehydrogenase (LDH) by colorimetric assay, and the activity of caspase 3/7 and caspase-1 by luminescence. We examined the integrity of mitochondrial membrane potential using the rhodamine 123 probe, migratory ability, cell cycle progression, and proliferation of cell lines after inhibition of EHMT1/EHMT2. Finally, the transcriptional profile of genes related to proliferation, cell death, and migration were examined. Inhibition of EHMT1/EHMT2 enzymes caused a significant decrease in the viability of MeWo and A549 cells in a dose-dependent manner. Subsequently, we identified cell death associated with cell apoptosis by the annexin-V/PI assay. In addition, we observed that inhibition of EHMT1/EHMT2 also promoted an increase in LDH, depolarization of mitochondrial membrane potential, and activation of caspases 3/7. Furthermore, the influence of EHMT1/EHMT2 affected A549 migratory ability and proliferation profile after 48 hours of treatment. Interestingly, EHMT1/EHMT2 modification modulates BCL2, NLRP1, KI67, CDK1 gene expression in A549 and BAX, BAK, BCL2, CDK1 in MeWo. Finally, our in silico analysis highlights the importance of EHMT1/EHMT2 in the cancer models studied, considering that some important biological processes are affected, including apoptosis and necrosis. Our results show that inhibition of EHMT1/EHMT2 promotes cell death in skin and lung cancer. These results are important and suggest that these histone methyltransferases may be epigenetic targets for the treatment of lung adenocarcinoma and malignant melanoma.

Access to medicines is one of the pillars of the right to health provided for in the Federal Constitution and the State develops actions to ensure that this duty is fulfilled. Medicines can also be considered socially and politically sensitive products, which is why almost all countries exercise control over their prices. In Brazil, drug pricing is carried out by the Medicines Market Regulation Chamber (CMED), which aims to define norms that promote pharmaceutical services to the population, through mechanisms that stimulate the supply of medicines and competitiveness in the sector. These norms have been in force in the country for 19 years with minor updates. It is noted that with the advancement of pharmaceutical technological innovation, the process of drug pricing has been an increasingly common challenge in all countries due to high prices. Ensuring sustainable access to lifesaving medicines is no longer just a problem for developing countries. In this sense, this study analyzed aspects of the regulatory policy for drug prices in Brazil to understand its implementation in the current scenario of the pharmaceutical market; and compared the rules of economic regulation in selected countries to try to identify the best practices for promoting access to medicines at fair prices. Therefore, a quantitative and qualitative research, of an exploratory type, was carried out to identify the variation in entry prices and prices practiced for medicines marketed in Brazil and in other countries. Based on the analyzed data, it was possible to list the numerous challenges for drug pricing in Brazil: (1) entry price definition methodologies are similar in most countries compared, however Brazil uses only one criterion, while most countries combines several methodologies in search of prices that are more suitable to the characteristics of the country and the market; (2) the active monitoring of the market may not be carried out effectively, as products were found that do not follow regulatory rules and the price variations characterizes an unregulated market in fact; (3) entry prices for new drugs in Brazil are generally the lowest in the year of registration, but over time they become one of the highest in the world, mainly due to the lack of periodical review; (4) the pricing rules for biosimilars have generated significant distortions in prices and do not help in the development of a market with perfect competition, and (5) the regulatory flow for disruptive drugs in Brazil can be an obstacle to the availability and access to population to innovative products. The information presented in this thesis shows that it is urgent to revise the current policy of economic regulation of the pharmaceutical market in Brazil to ensure that the population has access to medicines with prices that suit the social conditions of the country and that encourages the availability of innovative technologies, in a sustainable and isonomic way for the Unified Health System (SUS).

The need for non-animal performance tests for screening new ophthalmic formulations is a challenge for pharmaceutical sciences. In this way, we developed a new ex vivo method of ocular penetration of drugs with simulated tear flow and an innovative device that simulates the dynamics and physicalchemical barriers of the eye to evaluate the performance of ophthalmic drugs in vitro, the OphtalMimic. The first model model with simulated tear flow was coupled to a commercial rotary perfusion pump (the system operated with a flow of 48 µL/min during the first 2 minutes followed by a flow of 33 µL/min for 13 minutes of the experiment, after the application of 300 µL of formulation in the donor compartment). The system was challenged by different formulations with different viscosities and mucoadhesive properties (solution; 14, 16 and 20% poloxamer gels (PLX) and 16% PLX gel with chitosan (CS) at 0.5, 1.0 and 1.25%). The “Whole Eye Globe” model with simulated tear flow was able to differentiate formulations that contained a mucoadhesive component (CS) from those that did not (only PLX) in terms of the amount of drug that penetrated the cornea (p<0.05). The static model of conventional excised tissue was not only unable to differentiate these formulations (p>0.05), but also resulted in about 5 times the amount of drug penetrated in samples that lacked the mucoadhesive component, overestimating the performance of the formulation. . The new all-in-vitro device has an exposed area with a three-layer hydrogel-based synthesized cornea accommodated in a support base, upon which resides a simulated eyelid, a simulated cul-de-sac area, and an entrance and exit. for simulated pumping. tear flow. The support base is mounted on a motor that constantly moves from 0° to 50°, which also moves the eyelid. OphtalMimic was challenged by two formulations of Fluconazole, a Poloxamer 16% (PLX16) and a Poloxamer 16% + Chitosan 1.0% (PLX16C10), formulation with higher viscosity and mucoadhesive properties. Each formulation was tested with simulated tear flow of 0.5 mL.min-1 and test time of 10 minutes. All drained liquid was collected, diluted and analyzed by LC-UV. OphtalMimic was able to differentiate both formulations draining 72% ±4 and 65% ±3 of the drug in PLX16 and PLX16C10, respectively. Furthermore, the relative standard deviation was less than 5%, demonstrating the reproducibility of this innovative ophthalmic product performance test device. OphtalMimic is useful for comparing formulations, enabling highthroughput screening during development, research and quality phases. Our future prospect is to accommodate a reconstructed human cornea on the scaffold and provide dynamic results regarding drug permeability along with the dynamic performance of the tested formulations.

Brazil is one of the world’s largest consumers of pesticides. Many of the health hazards related to exposure to these substances are still under investigation. Recent studies show that some of these compounds can act as environmental obesogens, which are substances capable of disrupting the endocrine system, promoting adipogenesis and contributing to the development of obesity. Preliminary and unpublished studies by our research group showed that the pesticides bitertanol and fembutatin oxide promoted lipid accumulation in 3T3-L1 preadipocytes and that fembutatin oxide also activate PPARγ. Considering the intensive use of pesticides in Brazil and the few studies related to their endocrine disrupting effects, the present study aimed to investigate other mechanisms that may be related to the adipogenic effect promoted by the pesticides bitertanol and fembutatin oxide in mammalian cell culture. The evaluation of the effect on the transcriptional activity of the nuclear receptors GRα, PPARα, PPARβ/δ, TRα, TRβ and PXR was observed through a transfection and luciferase reporter assay. For that, HeLa cells were co-transfected with plasmids containing the complementary DNA of the nuclear receptor and with the plasmid containing its responsive element fused to the luciferase gene reporter. After 4 hours the cells were treated with DMSO, known agonists of the nuclear receptors in study or with increasing concentrations of the pesticides bitertanol and fembutatin oxide. This assay was also performed to observe if cysteine 285 is important for the PPARγ activation promoted by fembutatin oxide. In addition, a differentiation assay was performed using a specific PPARγ antagonist, T0070907. Therefore, 3T3-L1 cells were treated for 15 days in the presence or absence of the antagonist and with vehicle (DMSO), rosiglitazone or concentrations of the pesticides that promoted lipid accumulation. In the transfection and gene reporter assay, both bitertanol and fembutatin oxide did not show agonist activity on GRα, PPARα, PPARβ/δ, TRα, TRβ and PXR receptors. When the assay with the mutation in the cysteine 285 was performed, it was observed that the presence of the mutation abolished the transcricional activity promoted by fembutatin oxide suggesting that its essential for its activation and that fembutatin oxide can mimic endogenous ligands. In the differentiation assay the presence of the antagonist T0070907 significantly reduced the lipid accumulation induced by both pesticides suggesting that this effect occurs via PPARγ. The results of this research contribute to a better understanding of the impacts that these compounds can have on human and other animal health. Additionally, these results can support government decisions regarding the use of these compounds, as well as encourage the search for efficient assets with less negative impacts. However, further studies ate needed to better elucidate their mechanisms of action.

The objective of this work was to develop an optimized formulation without solvents for delivery of minoxidil (MXS) to the eyebrows with reduced run-off. Currently, there is growing interest in innovative products specializing in the treatment of hair loss in the eyebrows with fewer adverse effects and no sagging to improve treatment adherence. Poloxamer 407 (PLX) and hydroxypropylmethylcellulose (HPMC) were combined at different concentrations (15, 16 and 17% for PLX and 0.4, 0.8 and 1.2% for HPMC) together with MXS. The characterization was evaluated by the tests of sol/gel transition point, viscosity at 25°C, swine ear skin run-off distance and drop weight. Through these data, an optimized formula (FO) and a comparison formulation (FC) were selected by surface response analysis (ASR). The release profile and skin permeation of MXS was evaluated by the Franz static diffusion cell model for 12 hours. Vertical skin permeation in modified Saarbruecken cells evaluated the total skin permeation (divided into three areas: upper, middle (area of application of the gel) and lower for 12 hours. HPMC added to PLX improved the characteristics of the formulation. By SRA One FO (PLX 16% and HPMC 0.4%) was selected. The FO release profile showed a release similar to the MXS solution together with FC (PLX 4% and HPMC 0.7%) and no difference between the layers of the skin in the static permeation between the formulas (p<0.05). However, in the vertical penetration, the FO demonstrated localized delivery of MXS at the contact point, showing a difference between the average area and the other two areas (p<0, 05) resulting from radial penetration, while FC flowed and penetrated beyond the area of interest with a large amount in the lower area. The optimized formulation showed excellent efficiency in delivering MXS to the site of interest (middle third). Therefore, the obtained formulation increased its fixation when in contact with the skin, allowing for easier, safer, and no-slip treatment.

Tuberculosis is an infectious disease caused by the bacteria Mycobacterium tuberculosis and affects 10 million people a year, leading to more than 1 million deaths in the same period. The drugs used for the first-line treatment of tuberculosis are Rifampicin, Isoniazid, Pyrazinamide, and Ethambutol, which can lose their effectiveness due to resistance associated with the various mutations that the bacteria can present. Considering the difficulty in launching innovative drugs, one alternative is the repositioning of drugs that are on the market and have not yet been tested for this specific pathology. One way to reduce costs and time invested in the choice of a drug is the in silico methodology, which consists in performing computational tests to predict whether there is interaction between the molecule of interest and the molecular target. In the present study, the search for potential ligands to molecular targets of the first-line drugs isoniazid (enoyl-ACPreductase) and rifampicin (RNA-polymerase) was performed. The ligand obtained with the best result (for both targets) in silico was Coenzyme Q10, which was then evaluated in vitro to determine its Minimum Inhibitory Concentration (MIC) in order to verify the effect of this molecule directly on tuberculosis bacteria. The result obtained did not show high activity within the concentration range tested, which denotes the importance of biological tests even with relevant computational results. Other ligands should still be tested to confirm if the computational approach achieved any degree of effectiveness.

“Paracetamol is an effective oral analgesic, with few adverse effects, and is the most widely used over-the-counter drug of its class in the world. In addition, it is the drug most involved in cases of poisoning in several countries, since, when administered beyond the therapeutic dose, it can lead to liver damage. It is known that this damage can be potentiated by some conditions such as: eating disorders, previous administration of alcohol, isoniazid, phenobarbital and other drugs that stimulate the metabolism of paracetamol via cytochrome P450-mediated oxidation, generating a highly reactive metabolite, N-acetyl- p-benzoquinoneimine (NAPQI). Acetaminophen poisoning is common and potentially fatal. It is the most prevalent cause of acute liver failure in the western world. The management of intoxications involves decontamination and use of a specific antidote (Nacetylcysteine), which acts in the replacement of cysteine to support glutathione consumption, in addition to providing additional thiol groups that bind directly to NAPQI. In Brazil, paracetamol is one of the most prevalent agents in cases of drug intoxication. At the Centro de Informação e Assistência Toxicológica do Distrito Federal (CIATox-DF), it was the main analgesic involved in cases of poisoning registered between 2011 and 2016. The data obtained by CIATox-DF, however, are spontaneously reported, and do not necessarily reflect the totality cases of intoxication that occurred at the site. Thus, the present study sought data referring to exogenous intoxications that led to liver damage through different sources such as the Notifiable Diseases Notification System (SINAN) and the Mortality Information System (SIM). The results obtained denote the need for a system that better manages intoxication data, since the databases consulted are inconsistent. The number of cases of paracetamol intoxication may be underestimated within the available systems”