Purpose-L-leucine (LEU) is an amino acid with promising benefits for patients with cirrhosis. However, the bitter flavor of this amino acid and the reduced dissolution in water are challenges to the effectiveness of their supplementation. Thus, we evaluated the development and sensory aspects of using chocolate as a vehicle for LEU supplementation for cirrhosis patients. Design/methodology/approach-Seven chocolate bar formulations were developed with 7.5 g of added LEU, using different types of chocolate (semi-sweet, milk, white and blends). The samples were analyzed for total protein (micro-Kjeldahl method), phenolic compounds (Folin-Ciocalteu method) and microbiological quality. The sensory analysis evaluated the acceptance of color, flavor, appearance, aftertaste and overall impression by nine-point hedonic scale and intention to buy used five-point scale. Internal preference mappings were obtained with the results from acceptance and intention to buy tests. Findings-The LEU-chocolate had a higher protein content (MD = 33.67; 33.66-33.90g/100g) than the chocolate without LEU (MD= 9.8; 9.41-9.81 g/100g)[X-2 (5)=12,251; p = 0.031]. The samples of semi-sweet chocolates presented the highest concentration of polyphenolic compounds (F1: LEU-semi-sweet chocolate, 9.69 +/- 0.11; F3: LEU-blend chocolate, 9.49 +/- 0.23; F4: LEU-semi-sweet Cargill, 9.76 +/- 0.06; F5: semi-sweet chocolate without LEU, 9.87 +/- 0.25) when compared with the milk chocolate sample (9.05 +/- 0.65) (p = 0.033). The samples of LEU-chocolate showed similar acceptance (p > 0.05) for all attributes regardless of the kind of chocolate used (semi-sweet, milk or blend). Originality/value-This study is the first to evaluate the acceptability of LEU-chocolate on patients with cirrhosis. The addition of LEU did not significantly alter the sensory aspects of chocolate, suggesting that chocolate could be a beneficial vehicle for LEU supplementation.

Starch is widely used as an excipient in the pharmaceutical industry because it is cheap, nontoxic, and biocompatible. Commercial starches used are extracted from cereals as corn, wheat, and rice, tubers as potato, and roots as cassava. Potato and cassava originated in South America, the latter being the main starch, after corn, used for the manufacture of medicines. Native starches from cereals and starchy root and tuber crops have different characteristics for viscosity, transparency, and water loss through gels (syneresis), but it is possible to adjust these characteristics by modifying the native starch. So, these starches may suffer physical, chemical, enzymatic, and genetic modifications to present novel characteristics that will improve functional properties and their use in pharmaceutical formulations. This chapter presents different types of conventional and modified starches applied for pharmaceutical use.

The recent discovery of hazardous side effects associated with sunscreen ingredients has prompted the search for natural alternatives that can effectively absorb UV rays. This study focuses on evaluating the chemical profile and potential photoprotective and antiphotomutagenic effects of an ethanolic extract from Baccharis dracunculifolia using the Saccharomyces cerevisiae model. An analysis conducted using ESI-QToF-LC-MS confirmed the presence of flavonoids and phenylpropanoids in the B. dracunculifolia extract, including compounds such as apigenin, hydroxycinnamic acid derivatives, caffeoylquinic acids, and chlorogenic acid. In the yeast model, the extract demonstrated the ability to protect S. cerevisiae yno1 strain cells against cytotoxic lesions induced by high Solar Simulated Light (SSL) irradiation exposure. Furthermore, the extract exhibited antioxidant properties, as evidenced by its protection against SSL-induced oxidative damage in the yno1 strain, which is an indicator of oxidative damage. Additionally, the extract showed potential as an antiphotomutagenic agent, as the yno1-treated strain displayed a lower frequency of photomutagenicity in terms of CanR/107 mutant cells, thereby reducing both lethal cytotoxic lesions induced by SSL and protecting the cells against SSL-induced oxidative mutagenic lesions. These findings suggest that the B. dracunculifolia extract holds promise as a natural and effective photoprotective ingredient, offering a green alternative to traditional sunscreens by reducing the need for harmful chemical filters.

Ethnopharmacological relevance: Lippia alba (Mill.) N.E.Br. ex Britton & P. Wilson is traditionally used in Brazil as an adjunct in the relief of mild anxiety, as an antispasmodic, and as an antidyspeptic. This medicinal species was included in the Phytotherapeutic Form of the Brazilian Pharmacopeia 2nd edition (2021) and has already been described as the most used medicinal plant in a study with patients from an Anticoagulation Clinic in Brazil. Meanwhile, no studies were found that support the safety of the use of L. alba in patients using anticoagulants, a drug with several safety limitations.Aim of the study: Provide scientific evidence to ensure the safety of the concomitant use of L. alba and warfarin and support the management of these patients by evaluating its in vitro anticoagulant effect and chemical composition. And, as a timely complementation, evaluate the potential of this medicinal species in the devel-opment of new antithrombotics.Methods: The chemical profile of L. alba derivatives was analyzed by chromatographic methods such as Ultra -Performance Liquid Chromatography (UPLC) coupled with electrospray ionization mass spectrometry (ESI-MS), qualitative UPLC using Diode-Array Detection, and Thin Layer Chromatography. The anticoagulant activity was evaluated by the innovative Thrombin Generation Assay by Calibrated Automated Thrombogram method and using traditional coagulometric tests: prothrombin time, activated partial thromboplastin time, and plasma fibrinogen measurement.Results: Extracts and fractions prolonged the coagulation time in all the tests and reduced thrombin formation in thrombin generation assay. Coagulation times with the addition of ethanloic extract (2.26 mg/mL) was 17.78s, 46.43s and 14.25s respectively in prothrombin time, activated partial thromboplastin time and fibrinogren plasma measurement. In thrombin generation test, this same extract showed ETP as 323 nM/min compared to control (815 nM/min) with high tissue factor and 582 nM/min compared to control (1147 nM/min) using low tissue factor. Presence of flavonoids, phenylpropanoids, and triterpenes were confirmed by chromatographic methods and 13 compounds were identified by UPLC-ESI-MS. Based on these results and on the scientific literature, it is possible to propose that phenylpropanoids and flavonoids are related to the anticoagulant activity observed.Conclusion: The results demonstrate the in vitro anticoagulant activity of L. alba, probably due to the activation of intrinsic and extrinsic pathways. It is concluded, then, that there is a potential for interaction, which needs to be further studied, between L. alba and warfarin. Also, this medicinal species shows a great potential for use in the development of new antithrombotics.

Introduction: Lippia alba, popularly known as "cidreira" (lemon balm) is used to treat gastrointestinal disorders, anxiety, and insomnia. However, this species is known for its phenotypic plasticity and genome variation, resulting in variation of the chemical composition that can alter the pharmacological effect. This species was identified as one of the most consumed by patients undergoing warfarin treatment and there are no studies on its safety in concomitant use with anticoagulants.Methods: Ethanolic extract from 18 different accessions of L. alba was studied for its anticoagulant activity using a thrombin generation test and for its chemical composition using ultra-efficient liquid chromatography-diode array detector. Hierarchical Cluster Analysis and Principal Component Analysis were performed to analyze the relationship between anticoagulant activity and chemical composition of L. alba accessions.Results: In the phytochemical analysis it was possible to identify the presence of flavonoids and phenylpropanoids in all L. alba accessions. Sixteen of them (89 %) were able to reduce thrombin formation compared to the control, but there was a large difference in anticoagulant activity between the accessions.Discussion/conclusion: In general, the most active accessions are diploids while tetraploids were less active. All triploid accessions have compound 1, which is rarely found in diploids and tetraploids. Chemometric analyses demonstrate similarity of chemical composition within accessions of the same ploidy (2x, 3x, 4x) and indicate that the anticoagulant activity is due to the synergism between flavonoids and phenylpropanoids. Therefore, it is important that this plant species be used with caution in patients using oral anticoagulants.

Gold nanorods (GNRs) are increasingly being studied for diagnostic and therapeutic purposes. Green synthesis based methods with natural compounds as additives stand out as a hope in terms of better synthesis methodology, with advantages of producing potentially less toxic and, perhaps, biologically active GNRs due to influence of natural additives used during synthesis. Exploring green chemistry using different natural phenolic compounds, the present work reveals different in vitro activity of GNRs evaluated against different parasites that causes skin infectious diseases compared to GNRs produced by convencional seed mediated method. This approach brings advantages in producing active GNRs, with ease calling, less cytotoxic and with a better selectivity index (SI) than GNRs synthesized by conventional seed mediated synthesis, opening new possibilities for therapies. Natural compounds used in green syntheses were gallic acid (GA), resveratrol (RSV) and a purified fraction of the hydroalcoholic extract of Stryphnodendron obovatum. GNRs exhibited great activity against Leishmania braziliensis, and the dermatophytes Tricophyton rubrum, T. interdigitale and Microsporum gypseum. The anti-Leishmania and antidermatophytic activity of GNRs reinforce the applicability of GNRs in biomedical field and the influence of synthesis method in biological activity, showing benefits related to the seedless synthesis with natural compounds. In addition, these preliminary results indicate the possibility of exploring at maximum the physical and chemical properties of GNRs in addition to the biological activity itself, such as the development of topical antiparasitic formulations for association with phototherapy.

The efficiency and the voltage dependence of the AC iontophoresis were studied in vitro. Two cylindrical glass cells separated by a cellophane film were used, where the donor cell was filled with the solution of target electrolytes and the receptor cell with distilled water. The sinusoidal AC voltage with a frequency of 1 kHz was applied between the two platinum plates located at the opposite ends of two cells. The time variation of the ion concentration was evaluated by measuring the impedance of the solution in the receptor cell. The transportation velocity of the ions increased with the amplitude of the voltage applied between two platinum plate electrodes apart 20 mm up to ∼15 V, and leveled off above ∼15 V. A theoretical model is proposed on the AC iontophoresis, where each ion moves together with the surrounding water molecules when it is hydrated. The effective Stokes radius of an ion is assumed to be half of the whole size of the ion with hydrating water molecules. When the external alternating electric field strongly vibrates the ion, the ion–dipole interactions between the ion and water molecules are broken, resulting in the reduced effective Stokes radius, which leads to the increase of the diffusion efficiency.

Citrus sinensis and Lippia alba are herbal medicines widely used in the form of tea (infusion, decoction), which ethanolic extracts have already shown great anticoagulant activity in vitro . For this reason, they seem to be excellent candidates for the development of new antithrombotics and also have the potential to interact with them. The aim of this study was to evaluate the activity of aqueous extracts in blood coagulation and platelet aggregation, in addition to analysing the micromolecular composition of these species. Thrombin generation test (TGT) by the Calibrated Automated Thrombogram method and Platelet Aggregation Test by turbidimetry were performed to evaluate the biological activities, while the chemical composition was qualitatively evaluated using high-performance liquid chromatography. Aqueous extracts were elaborated according to the folk use. All extracts were effective in reducing thrombin formation in TGT. Infusion of L. alba and infusion and decoction of C. sinensis at a concentration of 0.6 mg/ml significantly reduced platelet aggregation induced by ADP, and only the decoction of L. alba at the same concentration was able to significantly reduce collagen-induced platelet aggregation. The presence of phenylpropanoids and flavonoids in C. sinensis and L. alba extracts was verified. Furthermore, hesperidin was identified in C. sinensis through coinjection. C. sinensis and L. alba are rich in phenolics and demonstrated an in-vitro effect on important processes of haemostasis (blood coagulation, platelet agreggation), corroborating the potential of C. sinensis and L. alba for the development of antithrombotics and interact with them.

•Seddless synthesis of gold nanorods using natural polyphenolic compounds with pharmacological uses are described.•Natural polyphenolic compounds allows the production of more stable gold nanorods in large volumes.•Natural polyphenolic compounds can stay on gold nanorods surface after the synthesis.•The results reinforce the potencial biomedical aplication of gold nanorods produced by the described method.

Background: Cryptococcosis affects more than 220,000 patients/year, with high mortality even when the standard treatment [amphotericin B (AMB), 5-flucytosin (5-FC) and fluconazole] is used. AMB presents high toxicity and 5-FC is not currently available in Brazil. In a pre-clinical study, pioglitazone (PIO - an antidiabetic drug) decreased AMB toxicity and lead to an increased mice survival, reduced morbidity and fungal burden in brain and lungs. The aim of this trial is to evaluate the efficacy and safety of PIO combined with standard antifungal treatment for human cryptococcosis. Methods: A phase 1/2, randomized, double blind, placebo-controlled trial will be performed with patients from Belo Horizonte, Brazil. They will be divided into three groups (placebo, PIO 15 mg/day or PIO 45 mg/day) and will receive an additional pill during the induction phase of cryptococcosis' treatment. Our hypothesis is that treated patients will have increased survival, so the primary outcome will be the mortality rate. Patients will be monitored for survival, side effects, fungal burden and inflammatory mediators in blood and cerebrospinal fluid. The follow up will occur for up 60 days. Conclusions: We expect that PIO will be an adequate adjuvant to the standard cryptococcosis' treatment.

Abstract Objectives Warfarin is the most widely used anticoagulant in the world, but it has several limitations including its narrow therapeutic range, need for dose adjustment and high potential for interactions. The simultaneous use of other drugs or even medicinal plants and certain foods could interfere with its therapeutic activity. In this context, this study aims to investigate the in vitro anticoagulant potential and phytochemical constitution of 17 plants selected from a previous clinical cross-sectional study (2014), that investigated the habits of plant utilization among patients taking warfarin. Methods Ethanol extracts and essential oils were evaluated, in vitro, as to their effect in the prothrombin time (PT) and activated partial thromboplastin time (aPTT) tests. Four species that presented aPTT >50 s were selected for phytochemical evaluation. Results Thirteen of the 17 plants selected demonstrated a significant anticoagulant effect in at least one of the evaluated parameters. Citrus sinensis (PT=14.75 and aPTT=53.15), Mentha crispa (aPTT=51.25), Mikania laevigata (PT=14.90 and aPTT=52.10), and Nasturtium officinale (aPTT=50.55) showed greater anticoagulant potential compared to normal plasma pool (PT=12.25 and aPTT=37.73). Chemical profiles of these four species were obtained, and certain compounds were identified: rosmarinic acid from M. crispa and isoorientin from N. officinale. Conclusions Thus, the results of this study could be a useful indicator for clinical practice towards the possibility of interaction between these plants and anticoagulants, although further clinical research is needed taking into consideration the limitations of in vitro studies. These findings also suggest that further research into the action of these plants could be of real clinical value in identifying potential alternative anticoagulant therapies.

Introduction: The most common treatment for Primary Open-Angle Glaucoma (POAG) is the daily use of eye drops. Sustained-release drug delivery systems have been developed to im-prove patient adherence by achieving prolonged therapeutic drug concentrations in ocular target tis-sues while limiting systemic exposure. The purpose of this study is to compare the efficacy and safety of bimatoprost inserts with bimatoprost eye drops in patients with POAG and Ocular Hyper-tension (OH). Methods: We include OH and POAG patients aged between 40 and 75 years-old. Both OH and POAG patients had intraocular pressure (IOP) greater than 21 and <30 mmHg at 9:00 am without glaucoma medication and normal biomicroscopy. Five normal patients with IOP<14 mmHg consti-tute the control group. A chitosan-based insert of bimatoprost was placed at the upper conjunctival fornix of the right eye. In the left eye, patients used one drop of LumiganTM daily at 10:00 pm. For statistical analysis, a two-way analysis of variance (ANOVA), Student t-test, and paired t-test is used. Results: Sixteen POAG and 13 OH patients with a mean age of 61 years were assessed. In both eyes, IOP reduction was similar during three weeks of follow-up (19.5 +/- 2.2 mmHg and 16.9 +/- 3.1 mmHg), insert, and eye drop, respectively; P=0.165). The percentage of IOP reduction in the third week was 30% for insert and 35% for eye drops (P=0.165). No intolerance or discomfort with the insert was reported. Among the research participants, 58% preferred the use of the insert while 25% preferred eye drops, and 17% reported no preference. Conclusion: Bimatoprost-loaded inserts showed similar efficacy to daily bimatoprost eye drops during three weeks of follow up, without major side effects. This might suggest a possible change in the daily therapeutic regimen for the treatment of POAG and OH.

ABSTRACT Purpose: The present study aimed at testing a new formulation of mesalazine linked to chondroitin sulfate and its components alone in the treatment of actinic proctitis in rats. Methods: Forty-seven female Wistar rats were submitted to pelvic radiation and divided into eight groups: control A, mesalazine A, chondroitin A, and conjugate A, gavage of the according substance two weeks after irradiation and sacrifice three weeks after oral treatment; control C, mesalazine C, chondroitin C, and conjugate C, sacrifice six weeks after oral treatment. The rectum was submitted to histological characterization for each of the findings: inflammatory infiltrate, epithelial degeneration, mucosal necrosis, and fibrosis. Results: The inflammatory infiltrate was more intense in chondroitin A, mesalazine A, and conjugate C. The collagen deposition was less intense in chondroitin A, and mesalazine A, and more intense in control C. Conclusions: Mesalazine and chondroitin alone were efficacious in inducing a delayed inflammatory response, hence reducing the late fibrosis. The conjugate was able to induce an ever more delayed inflammatory response.

O presente estudo teve como objetivo avaliar a eficácia de um novo produto elaborado a base de óleo essencial de alecrim pimenta sob a redução da contagem de microrganismos presentes na pele de tetos de vacas leiteiras. Para o teste in vivo, avaliou-se a eficácia do novo produto sob a antissepsia de negativos para mastite clínica (192 tetos), em comparação ao tratamento que continha produtos comerciais (192 tetos), durante um período de 42 dias. Os tetos foram diariamente acompanhados clinicamente, havendo semanalmente avaliação da microbiota presente na pele dos mesmos quanto à redução na contagem de Staphylococcus aureus, Estafilococos coagulase negativo (ECN), Echerichia coli e aeróbios mesófilos. Os dois produtos mantiveram a integridade dos tetos, sendo observado ausência de mastite clínica durante o período experimental. Não observou-se diferença significativa na ocorrência de mastite subclínica e de microrganismos nos tetos dos dois grupos em estudo pelo teste de Fisher em nível de significância de 5%. Ao comparar a redução Log da contagem em UFC.mL-1 dos microrganismos para o grupo tratado com produto convencional e com produto experimental para aeróbios mesófilos, E. coli, S. aureus e Estafilococos coagulase negativa, obtidos com o processo de higienização dos tetos foi observada diferença estatística(p>0,05), na contagem de microrganismos, evidenciando que ambos apresentaram efeito semelhante ao controle de microrganismos presentes na pele dos tetos. Sendo assim, o novo produto contendo óleo essencial de alecrim pimenta apresentou eficácia semelhante ao produto convencional clorexidine e iodo para antissepsia dos tetos de vacas leiteiras sem promover reações locais na pele, podendo este vir a ser utilizado como produto alternativo no manejo de ordenha.

Paullinia cupana Kunth., commonly named Guaran?a, is a plant from Brazil used as stimulant. The aim of this study was to evaluate the potential of extracts and tannins-rich and methylxanthines-free fraction from guarana? in the anti-inflammatory and antioxidant effect in vitro. Extract 1 obtained good yields of tannins and methylxanthines and was used to identify a type-A procyanidin trimer by LC-ESI-MS. Fraction 4 was rich in tannins and absent of methylxanthines. The extracts and fraction exhibited strong capacity for scavenging DPPH radical with IC50 between 5.88 and 42.75-?g/mL and inhibited TNF-? release by LPS-activated THP-1 cells when compared with control cells and did not present toxicity to THP-1 cells. The fraction 4, rich in tannins, was highly active, with IC50 5.88 ?g/mL by DPPH method and inhibited TNF-? release in 83.50% at 90 ?g/mL. These results reinforced potential anti-inflammatory of guaran?a and data for new therapeutic approaches.

Doxorubicin (DOX), a chemotherapy drug successfully used in the therapy of various types of cancer, is currently associated with the mucositis development, an inflammation that can cause ulcerative lesions in the mucosa of the gastrointestinal tract, abdominal pain and secondary infections. To increase the safety of the chemotherapy, we loaded DOX into nanostructured lipid carriers (NLCs). The NLC–DOX was characterized by HPLC, DLS, NTA, Zeta potential, FTIR, DSC, TEM and cryogenic-TEM. The ability of NLC–DOX to control the DOX release was evaluated through in vitro release studies. Moreover, the effect of NLC–DOX on intestinal mucosa was compared to a free DOX solution in C57BL/6 mice. The NLC–DOX showed spherical shape, high drug encapsulation efficiency (84.8 ± 4.6%), high drug loading (55.2 ± 3.4 mg/g) and low average diameter (66.0–78.8 nm). The DSC and FTIR analyses showed high interaction between the NLC components, resulting in controlled drug release. Treatment with NLC–DOX attenuated DOX-induced mucositis in mice, improving shortening on villus height and crypt depth, decreased inflammatory parameters, preserved intestinal permeability and increased expression of tight junctions (ZO-1 and Ocludin). These results indicated that encapsulation of DOX in NLCs is viable and reduces the drug toxicity to mucosal structures.

The investigation of the effects of three essential oils (EOs) from Taxandria fragrans (FRA), Melaleuca alternifolia (TTO) and Boswellia serrata (IF), alone and combined with ketoconazole (KTZ), and their functionalised gold nanoparticles (AuNP) against Trichophyton interdigitale both in vitro and in vivo indicated that EOs presented activity against T. interdigitale. The combination of EOs and KTZ was antagonistic. FRA, TTO, gold nanoparticles capped with T. fragrans (AuNPFRA) and gold nanoparticles capped with M. alternifolia (AuNPTTO) presented antidermatophytic activity in vivo, with the capacity to reduce fungal burden and to preserve tissue architecture; however, combination treatment with KTZ increased fungal burden and caused tissue damage. The combination of EO with KTZ exhibited antagonistic activity and was histologically harmful. In contrast, FRA, TTO, AuNPFRA and AuNPTTO are promising treatments for dermatophytosis.

Evaluate the in-vitro effect of Mentha crispa extract on blood coagulation, compare the conventional coagulometric tests with thrombin generation test (TGT), and study the qualitative micromolecular composition of M. crispa. Extract of M. crispa was incubated with plasma and used in the coagulometric tests: prothrombin and activated partial thromboplastin times, fibrinogen, and TGT. A phytochemical prospection was performed to evaluate the chemical composition of this extract. The extract was efficient in prolonging prothrombin time and activated partial thromboplastin time, and reducing fibrinogen levels and TGT parameters, indicating that the extract of M. crispa inhibited the intrinsic and extrinsic pathways of blood coagulation. The results obtained in TGT are in agreement with the results of conventional coagulometric tests and the in-vitro anticoagulant activity of M. crispa suggests that its use by patients using oral anticoagulants deserves caution.

PURPOSE:To evaluate different concentrations of ciprofloxacin to prevent infection after open fracture contaminated with S. aureus in rats using absorbable local delivery system. METHODS:Fifty-two Wistar rats were assigned to six groups. After 4 weeks, all animals underwent 99mTc-ceftizoxima scintigraphy evaluation, callus formation measurement and histological analysis. ANOVA, t-Student and Kruskal Wallis were used for quantitative variables statistical analysis, whereas qui square and exact Fisher were used for qualitative variables. RESULTS:Treatment using 25% and 50% of ciprofloxacin incorporated at the fracture fixation device were effective in preventing bone infection compared to control group (p<0.05). Chitosan were not effective in preventing bone infection when used alone compared to control group (p>0.05). Histological findings demonstrated bone-healing delay with 50% of ciprofloxacin. No difference in callus formation were observed (p>0.05). CONCLUSION:Local delivery treatment for contaminated open fracture using chitosan with ciprofloxacin is effective above 25%.

The use of chitosan as a pharmaceutical excipient in the ocular field is already established. Nevertheless, some aspects related to its ocular administration, such as sterilization and excipient's pharmacokinetics, remain unclear. So, in this study, we evaluated those two relevant aspects, related to chitosan administration in eye. We used chitosan-based ocular inserts (CI) as formulation model. CI were produced by solvent/casting method and sterilized by saturated steam. Sterilization was confirmed by direct inoculation of inserts in suitable microbiological growth media. Physicochemical characterization of inserts before and after sterilization was performed. Results suggested that, although steam sterilization changed the arrangement of the matrix, the heat and the humidity did not modify the structure of the main polymeric chain. Pharmacokinetics of CI radiolabeled with technetium-99m (Tc-99m) was assessed by scintigraphic images and ex vivo biodistribution study, after ocular administration in male Wistar rats. Scintigraphic and images analysis and ex vivo biodistribution study showed that the insert remained mainly in the eye until 6 hr after administration and its degradation products began to migrate to the abdominal cavity after 18 hr. Together, these data represent an important step forward the manufacturing and the clinical application of CI in the ophthalmic field.