In vitro antitumor effects of aqueous extract and protease inhibitors from Sterculia striata st. Hil. et naud and metabolite profiling.

The present study aims to assess the cytotoxic effect of the aqueous and protease inhibitors extracts of Sterculia striata on breast cancer cell lines. The in vitro results showed significant reductions in the highest concentrations from the S. striata seed extract for all cell lines. The aqueous extract reduced the viability by up to 35% in the MCF-7, 25% in the 4T1, and 35% in the MDA-MB-231 cell lines. Regarding the protease inhibitor extract, a 50% reduction in cell viability was observed in the MDA-MB-231 at concentration of 333 µg/mL. The aqueous and the protease inhibitor extracts showed mild reduction in the viability of macrophage cell lines. Chemical characterisation analysis revealed several polyphenols such as flavonoids, tannins, phenolic acids, and other secondary metabolites including terpenes, steroids, fatty acids, and organic acids, which may be related to the promising bioactivity observed. The S. striata showed antitumor activity, emphasising its pharmacological potential.

Protease inhibitors characterisation by SDS-PAGE and MALDI-TOF from Alocasia macrorrhizos and their modulation of macrophage immune-inflammatory properties.

This study describes the extraction and identification by electrophoretic and spectrometric techniques of protease inhibitor from the medicinal plant Alocasia macrorrhizos as well as investigates their immunomodulatory properties and cell viability. The A. macrorrhizos tubers were subjected to protease inhibitor extractions and characterised using SDS-PAGE and MALDI-TOF. The protein extracts were assessed for activities trypsin inhibition stoichiometry, haemagglutinating, cell viability, NO and TNF-α production inhibition. Concerning the protease inhibitors analysis through SDS-PAGE, the results showed two bands with 11 and 24 kDa, and the MS analysis detected the ions more intense of m/z 4276.795 and 8563.361 in the roasted protein extract. The IC50 of trypsin inhibition was 0.119 and 0.302 mg L-1 in the roasted and crude tuber, respectively. The protease inhibitors extract from the roasted tubers showed a reduction in the production of NO and TNF-α at concentrations lower than 100 µg mL-1, without a reduction in cell viability.

Assessing the Effectiveness of Chemical Marker Extraction from Amazonian Plant Cupuassu (Theobroma grandiflorum) by PSI-HRMS/MS and LC-HRMS/MS.

Employing a combination of liquid chromatography electrospray ionization and paper spray ionization high-resolution tandem mass spectrometry, extracts from cupuassu (Theobroma grandiflorum) pulp prepared with either water, methanol, acetonitrile or combinations thereof were subjected to metabolite fingerprinting. Among the tested extractors, 100% methanol extracted preferentially phenols and cinnamic acids derivatives, whereas acetonitrile and acetonitrile/methanol were more effective in extracting terpenoids and flavonoids, respectively. And while liquid chromatography- mass spectrometry detected twice as many metabolites as paper spray ionization tandem mass spectrometry, the latter proved its potential as a screening technique. Comprehensive structural annotation showed a high production of terpenes, mainly oleanane triterpene derivatives. of the mass spectra Further, five major metabolites with known antioxidant activity, namely catechin, citric acid, epigallocatechin-3'-glucuronide, 5,7,8-trihydroxyflavanone, and asiatic acid, were subjected to molecular docking analysis using the antioxidative enzyme peroxiredoxin 5 (PRDX5) as a model receptor. Based on its excellent docking score, a pharmacophore model of 5,7,8-trihydroxyflavanone was generated, which may help the design of new antioxidants.

Metabolic profiling by LC-DAD-MS, FTIR, NMR and CE-UV of polyphenols with potential against skin pigmentation disorder.

In traditional Brazilian medicine, tubers extracts from Alocasia macrorrhizos are widely used in the treatment of skin pigmentation disorder. However, studies that evaluate its benefits in the treatment of this disorder are non-existent. Thus, this work aims to investigate the bioactivity of A. macrorrhizos extracts in cell culture and murine model of Vitiligo and correlating with its phenolic profile. The metabolic profiling from the bioactive extracts was obtained by LC-DAD-MS, FTIR, NMR, and CE-UV. The murine model of Vitiligo was induced with 5% hydroquinone in C57BL/6 male mice, which were treated or not with 100 mg/kg of roasted tuber aqueous extract. In Vitiligo model assay was observed hair follicle repigmentation and reduction of the epidermal layer thickness at the histopathological level, in the animals treated with aqueous extract of roasted tubers. The present study provides new molecular insight and scientific evidence on the potential utility of the extract of A. macrorrhizos against Vitiligo.

Dereplication of Siparuna brasiliensis extract with in vivo anti-inflammatory activity.

Siparuna brasiliensis is a medicinal plant widely used by indigenous communities of the Amazon rainforest to treat inflammatory diseases and related pathologies. Considering its ethnopharmacological application, it constitutes an important source of biologically active molecules in the development of anti-inflammatory drugs. This study describes a dereplication methodology of the bioactive extract from S. brasiliensis leaves and the evaluation of the anti-inflammatory potential in an in vivo inflammatory model with mice of the BALB/c lineage and in vitro using cell lines, as well as determining the production of an inflammatory mediator. From their charge-to-mass ratios (m/z) and elemental composition obtained through Ultrahigh-resolution mass spectrometry analysis by ESI(-)-Orbitrap MS and chromatographic profile by RP-HPLC-PDA, it was possible to annotate polyphenols with anti-inflammatory properties classified as flavonoids and organic acids. The administration of the extract significantly inhibited carrageenan-induced paw edema and showed effects similar to those of drug dexamethasone without affecting cell viability.

Metabolic Profiling of Inga Species with Antitumor Activity.

This work evaluated the metabolic profiling of Inga species with antitumor potential. In addition, we described the antigenotoxicity of polyphenols isolated from I. laurina and a proteomic approach using HepG2 cells after treatment with these metabolites. The in vitro cytotoxic activity against HepG2, HT-29 and T98G cancer cell lines was investigated. The assessment of genotoxic damage was carried out through the comet assay. The ethanolic extract from I. laurina seeds was subjected to bioassay-guided fractionation and the most active fractions were characterized. One bioactive fraction with high cytotoxicity against HT-29 human colon cancer cells (IC50 = 4.0 µg mL-1) was found, and it was characterized as a mixture of p-hydroxybenzoic acid and 4-vinyl-phenol. The I. edulis fruit peel (IC50 = 18.6 µg mL-1) and I. laurina seed (IC50 = 15.2 µg mL-1) extracts had cytotoxic activity against the cell line T98G, and its chemical composition showed a variety of phenolic acids. The chemical composition of this species indicated a wide variety of aromatic acids, flavonoids, tannins, and carotenoids. The high concentration (ranging from 5% to 30%) of these polyphenols in the bioactive extract may be responsible for the antitumor potential. Regarding the proteomic approach, we detected proteins directly related to the elimination of ROS, DNA repair, expression of tumor proteins, and apoptosis.

Phytochemical, metabolic profiling and antiparasitic potential from Inga semialata leaves (Fabaceae).

Natural Products phytochemical provide a rich source for therapeutic discovery and has led to the development of many drugs. Thus, the aim of this study was to obtaining a metabolic profiling from ethanol extract of Inga semialata leaves (EEIS) selected by bioassay antimalarial and nematostatic and identify metabolites in mixture by co-injection experiments and NMR spectroscopy. The chemical composition of this species indicated a wide variety of aromatic acids (vanillic acid, 3,4,5-trimethoxy benzoic acid, gallic acid, methyl gallate, p-coumaric acid and ferulic acid), flavonoids (quercetin, myricetin-3-O-rhamnoside and myricetin-3-O-(2"-O-galloyl)-α-rhamnopyranoside), triterpenes (lupeol, α-amyrin, friedelin and oleanolic acid) and the 2-hydroxyethyl-dodecanoate. The antimalarial assay showed that the I. semialata n-hexane fraction presented higher inhibition percentage than the Chloroquine standard and may be considered a potential source of compounds with antimalarial activity while the EEIS and its fractions showed nematostatic potential below 17% in the assay of nematostatic evaluation against the parasite Meloidogyne incognita.

Fatty acid profiles of the plasma and follicular fluid mares fed a combination of linseed and salmon oil.

This study evaluated the presence of polyunsaturated fatty acids in circulating blood and in the ovarian follicular fluid of mares, after supplementation of the diet with linseed oil. Six Mangalarga Marchador mares, weighing 397.00±31.89 kg, were kept on native pasture, and assigned to the current study. In a switch over design, mares were randomly allocated to receive 150 ml of vegetable oil daily, containing polyunsaturated fatty acids n3 (62.23 g ALA, 20.34 g LA, 2.27 g EPA, 2.32 g DHA), (n=3) or no supplementation (n=3) in two replicates. Blood and follicular fluid samples were taken on the first day (D0) and every 30 days until the end of the supplementation period (D60). After 60 days of supplementation, mares were switched across the treatments. Plasma concentrations of linolenic acid in total fatty acids were higher (P=0.006) in the supplemented compared to the control group (1.89±0.13 vs. 1.49±0.13%). There were positive correlations between plasma linoleic acid and follicular fluid arachidonic acid (P=0.0106; r2=0.13) and between plasma alpha linolenic acid and follicular fluid EPA (P=0.0004; r2=0.2544). Data indicated a low to moderate relationship between the dietary linseed-based oil supplementation studied and circulating and follicular fluid polyunsaturated fatty acids contents in mares.

Miconia albicans and Curcuma longa herbal medicines positively modulate joint pain, function and inflammation in patients with osteoarthritis: a clinical study.

This study aims to evaluate the analgesic and modulating effect of Curcuma longa and Miconia albicans herbal medicines in knee's osteoarthritis (OA) treatment. This longitudinal study evaluated 24 patients with OA. The patients were divided into three groups: ibuprofen (1200 mg/day), C. longa (1000 mg/day) and M. albicans (1000 mg/day). The medications were applied orally for 30 days. The synovial fluid of the knee joint was collect at the first (day 0) and the last medical (day 30) consultation. The groups treated with herbal medicines presented the same results when compared to Ibuprofen. The comparison of the means of Total WOMAC for M. albicans before and after treatment presented a statistically significant difference (mean day 0 = 57.19; mean day 30 = 31.02) as well as variation of Total WOMAC for C. longa (mean day 0 = 54.79; mean day 30 = 37.08). The WOMAC Total and the VASP were compared, it was found that there was a significant decrease in the means in the C. longa and M. albicans groups, as well as in the Ibuprofen group after treatment. The study demonstrated that the treatment of knee OA with C. longa or M. albicans positively interferes with patients pain and functionality, decreased WOMAC and VASP scores, leading to functional improvement of these patients. This is the first clinical study demonstrating the analgesic and anti-inflammatory effect on knee osteoarthritis from M. albicans comparable to Ibuprofen drug.

Nasomaxillary effects of miniscrew-assisted rapid palatal expansion and two surgically assisted rapid palatal expansion approaches.

This retrospective study was performed to evaluate nasomaxillary changes in 36 patients at an advanced stage of skeletal maturity who underwent miniscrew-assisted rapid palatal expansion (MARPE) or surgically assisted rapid palatal expansion (SARPE) with/without an alar base cinch. Cone beam computed tomography images taken before and after expansion were analysed. Changes in the width of the dental arch (D66S, D66I), maxillary base (MxMol), and nasal floor and nasal cavity in the molar and canine regions (NaFMol, NaFCan, NaCMol, NaCCan) were compared, as well as changes in the choanal aperture (CA) and nasal soft tissue (NW). The MARPE technique produced smaller dental changes (D66S; P=0.025) and greater nasomaxillary expansion (MxMol, P=0.010; NaCMol, P=0.016; NaCCan, P=0.017; NaFMol, P=0.001; CA, P=0.002) than both SARPE techniques. Changes in NW did not differ significantly between the groups (P=0.200). MARPE uniformly increased the anterior and posterior widths of the nasal cavity. SARPE expanded the nasal cavity in a 'V-shape' pattern. Changes in the nasal cavity and choanal aperture related to the amount of dental arch expansion were greater for MARPE than for SARPE. All three approaches increased the width of the nasal soft tissue, although the cinch in SARPE limited this increase.

Influence of storage time and temperature on the chemical composition of the essential oil of Hyptis pectinata L. Poit / Influência do tempo de armazenamento e da temperatura na composição química do óleo essencial de Hyptis pectinata L. Poit

ABSTRACT Hyptis pectinata, popularly known as “sambacaitá”, is a native medicinal herb used by local people for treating disease. Its pharmacological, antimicrobial, and medicinal properties have been reported in several studies. The essential oil of the leaves is rich in terpene compounds responsible for its properties. Because there are factors that can influence the chemical composition of essential oils, the objective of this study was to evaluate the influence of storage on Hyptis pectinata essential oil for 360 days at two different temperatures. The essential oil was distilled from dried leaves of seven-month-old plants using the hydrodistillation method, and the chemical composition was determined using GC-MS and GC/FID. The effects of storage on the essential oil Hyptis pectinata were tested over the course of a year (0, 15, 30, 60, 90, 120, 150, 180, 240, 300, and 360 days), using two temperatures: room (±32ºC) and freezer (-20ºC). The predominant compounds in the essential oil are β–caryophyllene, caryophyllene oxide, and germancrene D. The concentration of the chemical constituents of the essential oil varied depending on the storage temperature, and over the 360 days. The storage of the essential oil at room temperature over one year resulted in higher concentrations of β-elemene, α-copaene, germacrene D, caryophyllene oxide, and (E,E)-α-farnesene and lower content of α-humulene and β-caryophyllene, compared to the results obtained from storing the essential oil in a freezer. These results indicate that the essential oil should be stored in a freezer, which provides greater stability to the concentration of the chemical constituents.

RESUMO Hyptis pectinata, conhecida popularmente como Sambacaitá é uma erva medicinal nativa utilizada pela população local para o tratamento de enfermidades, cujas propriedades farmacológicas, antimicrobianas e medicinais foram relatadas em diversos estudos. O óleo essencial das folhas é rico em compostos terpênicos responsáveis pelas suas propriedades. Devido à existência de fatores que podem influenciar a composição química dos óleos essenciais, o objetivo deste trabalho foi avaliar a influência do armazenamento do óleo essencial Hyptis pectinata durante 360 dias em duas temperaturas diferentes. O óleo essencial foi destilado a partir de folhas secas de plantas de sete meses de idade, utilizando o método de hidrodestilação e a composição química foi determinada usando CG-EM e CG-DIC. Os efeitos do armazenamento no óleo essencial Hyptis pectinata foram testados ao longo de um ano (0, 15, 30, 60, 90, 120, 150, 180, 240, 300 e 360 dias), utilizando duas temperaturas: ambiente (±32ºC) e freezer (-20ºC). Os compostos predominantes no óleo essencial foram o β-cariofileno, óxido de cariofileno e germancreno D e houve variação na concentração dos constituintes químicos do óleo essencial, dependendo da temperatura de armazenamento, bem como ao longo dos 360 dias. O armazenamento do óleo essencial, à temperatura ambiente ao longo de um ano resultou em maiores concentrações de β-elemeno, α-copaeno, germacreno D, óxido de cariofileno e (E, E) -α-farneseno e menores teores de α-humuleno e β-cariofileno, quando comparado com os resultados obtidos a partir de armazenamento do óleo essencial no freezer. Estes resultados indicam que o óleo essencial deve ser estocado no freezer, o que propicia maior estabilidade na concentração dos constituintes químicos.

Development and characterization of novel microsatellite markers in Hyptis pectinata (Lamiaceae).

A microsatellite-enriched library was constructed and a set of 19 SSR markers were developed to characterize a germplasm collection of Hyptis pectinata (L.) Poit., maintained at the Universidade Federal de Sergipe (UFS). Fifteen markers of 19 ranged from moderately to highly polymorphic. A total of 113 alleles were identified, with a mean of 7.52 alleles per locus. The mean HO and HE were 0.582 and 0.657, respectively. The primers developed were efficient tools for accessing the genetic diversity of the germplasm collection analyzed and may also be useful for other studies involving this species and other species in the genus Hyptis.

Macroscopic and histological evaluation of tunica vaginalis dorsal grafting in the first stage of Bracka's urethroplasty: an experimental study in rabbits.

OBJECTIVE: To evaluate the use of tunica vaginalis as a dorsal free graft in the first stage of Bracka's urethroplasty in a rabbit animal model using macroscopic and histological analysis. MATERIAL AND METHODS: Surgery was performed on 16 rabbits, divided into four groups of four according to the number of weeks after surgery at which they were killed (2, 4, 8 and 12 weeks). The penile urethra was removed and only the ventral aspect of the urethra was reconstructed using a tunica vaginalis free graft dorsally fixed in the corpora cavernosa. Epithelial, inflammatory and fibrotic changes in the graft were evaluated macroscopically then histologically at the point when each rabbit group was killed. RESULTS: Macroscopic evaluation showed good graft uptake with minimal retraction (P > 0.05). Histological evaluation showed a reduction in the inflammatory process during the first 4 weeks, after which inflammation stabilized. The mesothelium was partially healed at 2 and 8 weeks with a metaplasia defined by a stratified squamous, non-keratinized lining. The grafted area was thinner than the surrounding penile skin and similar to urothelial native epithelium. CONCLUSION: Tunica vaginalis used as a free graft placed dorsally on the corpora cavernosa in the first stage of Bracka's urethroplasty shows good tissue integration and develops into a stratified non- keratinized epithelium (metaplasia) without significant retraction, despite exposure to the external environment.

A new extra-abdominal channel alternative to the Mitrofanoff principle: experimental and preliminary clinical experience.

INTRODUCTION: The appendix is the gold-standard channel for the Mitrofanoff principle in pediatric urology, but the search for alternatives is justified considering it may not be available or preferably used for colonic stomas (Malone antegrade continence enema). The aim of this study is to report on technical feasibility of a new approach for creating catheterizable channels in a rabbit model and to present our preliminary clinical experience. MATERIAL AND METHODS: We configured a tube from two rectangular skin flaps 1x4 cm opposite each other in the middle line of the lower inferior abdomen. The channel was anastomosed to the bladder dome with embedding sutures to create a valvular mechanism. The experimental study consisted of 12 rabbits, divided in 4 groups according to the sacrifice schedule at 2, 4, 8 and 12 weeks. At 30th postoperative day, an urodynamic evaluation was performed to record continence of the stoma. A histological analysis of the specimens stained with hematoxylin-eosin, Masson trichrome and Picrosirius red was also done in group 2 (sacrifice at 4 weeks postoperatively). We used this method in 3 patients with congenital non-neurogenic bladder disease presenting with massive residual volumes without compliance deficits. RESULT: The technique proved feasible in all animals, 9 of 12 could be easily catheterized and underwent urodynamic study. No stoma leakage was observed in 7 animals at high bladder pressures (> 50 cm H20) and only 2 animals had some leakage at 40 cm H20. Urodynamics performed through the stoma showed urethral leakage at 20 cm H20, therefore demonstrating the efficacy of the valvular mechanism. Histological analysis confirmed good integration between the tube and the bladder. Mean follow-up of the clinical series (3 patients) was 7.2 months. Two patients remained continent up to 4 hours, whereas 1 patient had some leakage after 2 hours. CONCLUSION: We were able to confirm feasibility of a new extra-abdominal channel based on the Mitrofanoff principle and successfully reproduced the method in a clinical setting. Follow-up was short and long term results are required before any conclusive judgment can be made.

A new extra-abdominal channel alternative to the mitrofanoff principle: experimental and preliminary clinical experience

INTRODUCTION: The appendix is the gold-standard channel for the Mitrofanoff principle in pediatric urology, but the search for alternatives is justified considering it may not be available or preferably used for colonic stomas (Malone antegrade continence enema). The aim of this study is to report on technical feasibility of a new approach for creating catheterizable channels in a rabbit model and to present our preliminary clinical experience. MATERIAL AND METHODS: We configured a tube from two rectangular skin flaps 1x4 cm opposite each other in the middle line of the lower inferior abdomen. The channel was anastomosed to the bladder dome with embedding sutures to create a valvular mechanism. The experimental study consisted of 12 rabbits, divided in 4 groups according to the sacrifice schedule at 2, 4, 8 and 12 weeks. At 30th postoperative day, an urodynamic evaluation was performed to record continence of the stoma. A histological analysis of the specimens stained with hematoxylin-eosin, Masson trichrome and Picrosirius red was also done in group 2 (sacrifice at 4 weeks postoperatively). We used this method in 3 patients with congenital non-neurogenic bladder disease presenting with massive residual volumes without compliance deficits. RESULT: The technique proved feasible in all animals, 9 of 12 could be easily catheterized and underwent urodynamic study. No stoma leakage was observed in 7 animals at high bladder pressures (> 50 cm H20) and only 2 animals had some leakage at 40 cm H20. Urodynamics performed through the stoma showed urethral leakage at 20 cm H20, therefore demonstrating the efficacy of the valvular mechanism. Histological analysis confirmed good integration between the tube and the bladder. Mean follow-up of the clinical series (3 patients) was 7.2 months. Two patients remained continent up to 4 hours, whereas 1 patient had some leakage after 2 hours. CONCLUSION: We were able to confirm feasibility of a new extra-abdominal...