Antiproliferative oleanane saponins from Meryta denhamii

Eight new oleanane saponins (1- 8) together with four know saponins (9-12) were isolated from the aerial parts of Meryta denhamii. Their structures were elucidated by 1D and 2D NMR experiments including 1D TOCSY, DQF-COSY, ROESY, HSQC, and HMBC spectroscopy, as well as ESIMS analysis. The antiproliferative activity of all compounds was evaluated using three murine and human cancer cell lines: J774.A1, HEK-293, and WEHI-164.     

Preliminary investigation of the central nervous system effects of 'Tira-capeta' (Removing the Devil), a cigarette used by some Quilombolas living in Pantanal Wetlands of Brazil

During an ethnopharmacological survey carried out among some Quilombolas living in Brazil, 48 plants with possible central nervous system (CNS) action were cited. A mixture of nine plants, known as 'Tira-capeta' (Removing the Devil) cigarette, has been utilized for years as a tonic for the brain. The effects observed after consuming this cigarette are: dizziness, lightness sensation, humor changes, yawns, heavy eyes, hunger, sleep and relaxation. This study aimed to verify the effects of a hydroalcohol extract of 'Tira-capeta' cigarette (ETC), as well as to evaluate the phytochemical profile. The phytochemical screening carried out through characterization reactions, thin layer chromatography and high efficiency liquid chromatography indicated the presence of tannins, phenolic acids, flavonoids, saponins and alkaloids; tannins and phenolic acids being the principal constituents. The pharmacological tests showed that ETC induced a biphasic effect, with intense initial stimulation of the CNS, followed by a general depressor state; decreased the latency for sleeping and increased the total sleeping time (50, 100 and 500 mg/kg), without causing prejudice in motor coordination (doses up to 200 mg/kg); induced catalepsy in mice, verified 10 and 50 min after drug administration (500 mg/kg). Also, no anxiolytic or anxiogenic effects were verified in rats submitted to the elevated plus-maze.     

A nutraceutical extract from Inula viscosa leaves: UHPLC-HR-MS/MS based polyphenol profile,and antioxidant and cytotoxic activities

Nowadays, advanced extraction techniques and highly sensitive metabolic profiling methods are effectively employed to get new information on plant chemical constituents. Among them wild medicinal plants or their parts, with large and ancient use in folk medicine, are investigated for their potential functional use and cultivation. In this context, Inula viscosa leaves engaged our attention. A simple experimental design, based on Soxhlet extraction and chromatographic fractionation, allowed us to obtain the investigated polyphenol fraction (IvE). UHPLC-HRMS analyses revealed shikimoyl depsides of caffeic acid and unusual dihydrobenzofuran lignans as main secondary metabolites. These compounds, together with cinchonain-type phenols, and hydroxycinnamoyl flavonol glycosides, are reported for the first time in inula. Overall, forty-three secondary metabolites were identified. The extract exerted a remarkable antiradical activity towards DPPH^• and ABTS^+•. Furthermore, it was able to inhibit cell viability and mitochondrial redox activity of neuroblastoma, hepatoblastoma and colon carcinoma cells, whereas it did not affect cell density of HaCaT cells immortalized human keratinocytes. As detected by the oxidant-sensing probe 2′,7′-dichlorodihydrofluorescein diacetate, the inhibitory responses seemed to be related to IvE-induced increase of intracellular reactive oxygen species (ROS). The obtained results highlighted that inula leaves, nowadays even undervalued and unexplored, could be considered a renewable source of nutraceutical compounds.     

CO2 laser therapy of the Bartholin's gland cyst: surgical data and functional short- and long-term results

The Bartholin's gland cyst is a condition that occurs in approximately 2% of women, most of whom are of reproductive age. Although benign, it is associated with significant patient discomfort. Definitive treatment involves the surgical removal of the entire cyst. The objective of this study was to describe an alternative conservative surgical technique with CO(2) laser adopted in our institution and to evaluate the feasibility, complication rate, and results obtained with this technique in an office setting. Our study showed that CO(2) laser treatment of the Bartholin's gland cyst can be carried out safely in an outpatient setting, with minimum patient discomfort and high long-term cure rates.     

Hepatic splenosis misinterpreted as hepatocellular carcinoma in cirrhotic patients referred for liver transplantation: report of two cases.

Liver transplantation is one of the main therapeutic options for hepatocellular carcinoma (HCC) occurring in cirrhotic patients; an accurate diagnosis and staging of this cancer is crucial to selecting the candidates for this treatment. Although the best diagnostic strategy is debated, the guidelines proposed by the European Association for the Study of the Liver (EASL) are used by many centers. We report 2 cases of cirrhotic patients with hepatic splenosis, a rare condition that may be misinterpreted as HCC. In conclusion, the application of the EASL guidelines in the first case would have led to an incorrect staging of the cancer and in the second case would have led to a false diagnosis of HCC.     

Cisplatin-induced kidney injury in the rat: l-carnitine modulates the relationship between MMP-9 and TIMP-3

Renal interstitial fibrosis is a major complication of cisplatin treatment, due to the increased accumulation of extracellular matrix (ECM) proteins whose remodeling is important for the development of normal tissues; indeed, its malfunction might play a role in the etiology of various diseases.Biopharmacological evaluations suggest that l-carnitine can prevent cardiac metabolic damage caused by doxorubicin, as well as can inhibit cisplatin-induced injury in the kidney and in the small intestine, without any interference with the drug's antitumoral properties. Since the glomerular basement membrane and the mesangial matrix constitute the ECM of the renal glomerulus, we examined the localization and expression of MMP-9 and TIMP-3 in normal rat kidney and the changes in their expression over a period of time by treatment with cisplatin, with and without l-carnitine. MMP-9 immunoreaction in cisplatin-treated rat kidney tissue suggests an involution of the basal membrane, an alteration of ECM components and low glomerular function, due to the increased thickness of the mesangium. Our results suggest that the matrix remodeling by MMP-9 and TIMP-3, in the later stages, can play an important role in the development of glomerular sclerosis and interstitial fibrosis after cisplatin treatment. It can also be postulated that l-carnitine protects from cisplatin injury, by modulating the relationship between MMP-9 and TIMP-3.     

Study of photodegradation kinetics of melatonin by multivariate curve resolution (MCR) with estimation of feasible band boundaries

Multivariate curve resolution—alternating least squares (MCR‐ALS) has been applied to data collected from UV spectrophotometric analysis of melatonin samples exposed to light with varying irradiance power. MCR‐ALS was able to explain the degradation kinetics of this drug, deducing the pure spectra and concentration changes of the different species present throughout the entire process. Possible rotational ambiguities associated with MCR solutions were investigated and their extent was evaluated. The extent of the rotation ambiguity was estimated from the band boundaries of feasible solutions calculated using the MCR‐BANDS procedure. The use of a non‐linear fitting routine allowed improving kinetic information and provided a method of evaluation of the rate constants of the degradation process. The degradation pathway was found to follow a first‐order reaction model, in which melatonin underwent photo‐oxidation of the indole ring to give a formylamine group. Kinetics of the reaction was shown to be dependent on irradiation conditions, with an increase of the rate constants when light power also increased. Copyright © 2011 John Wiley & Sons, Ltd.     

Biochemical alterations from normal mucosa to gastric cancer by ex vivo magnetic resonance spectroscopy

BACKGROUND AND AIMS: The metabolic profile and morphologic aspects of normal and pathologic human gastric mucosa were studied. The aim of the present research was the application of ex vivo high-resolution magic angle spinning magnetic resonance spectroscopy (HR-MAS MRS) to the human gastric tissue to get information on the molecular steps involved in gastric carcinogenesis and the identification of biochemical markers useful for the development of in vivo MRS methodologies to diagnose gastric pathologies in clinical situations. METHODS: Twelve normal subjects, five with autoimmune atrophic gastritis, five with Helicobacter pylori infection, and five with adenocarcinoma were examined. Ten biopsies were taken during endoscopy from each patient. Specimens from carcinoma were also obtained during gastrectomy. Of the 10 biopsies, 4 were used for histologic evaluation, 4 were fixed in glutaraldehyde and processed for transmission and scanning electron microscopy, and 2 were immersed in liquid nitrogen and stored at -85 degrees C for monodimensional and bidimensional ex vivo HR-MAS MRS analysis. RESULTS: Ex vivo HR-MAS MRS identified glycine, alanine, free choline, and triglycerides as possible molecular markers related to the human gastric mucosa differentiation toward preneoplastic and neoplastic conditions. Ultrastructural studies of autoimmune atrophic gastritis and gastric adenocarcinoma revealed lipid accumulations intracellularly and extracellularly associated with a severe prenecrotic hypoxia and mitochondria degeneration. CONCLUSIONS: This is the first report of synergic applications of ex vivo HR-MAS MRS and electron microscopy in studying the human gastric mucosa differentiation. This research provides useful information about some molecular steps involved in gastric carcinogenesis. The biochemical data obtained on gastric pathologic tissue could represent the basis for clinical applications of in vivo MRS.     

Nanoparticles based on N-trimethylchitosan: evaluation of absorption properties using in vitro (Caco-2 cells) and ex vivo (excised rat jejunum) models.

Among the chitosan derivatives, trimethylchitosan (TMC) has been shown to have penetration enhancement properties also in intestinal environment. In addition, the use of nanoparticulate systems has the advantage of protecting peptidic drugs from intestinal degradations, due to internalisation behaviour. Therefore, the aim of this paper was to evaluate nanoparticulate systems based on TMC. In particular the mucoadhesive and absorption enhancement properties of nanoparticles based on TMC with different quaternization degree (QD) intended for the intestinal administration of macromolecules (peptides) have been evaluated. Comparison with chitosan (CS.HCl) nanoparticles was made. The nanoparticles were loaded with fluorescein isothiocyanate dextran (FD4, MW 4400 Da), used as the model macromolecule. The intestinal penetration enhancement properties of nanoparticles were investigated in an in vitro Caco-2 cell model and an ex vivo rat jejunum model. The mucoadhesion of the nanosystems was evaluated using excised rat jejunum. All of the nanoparticulate systems interacted with the Caco-2 cells decreasing the transepithelial electric resistance (TEER) and increasing Lucifer Yellow (LY) Papp (paracellular pathway marker). All the nanosystems improved FD4 Papp, with the exception of the nanoparticles based on TMC with the highest QD. In this case an entrapment of nanoparticles into Caco-2 cells was supposed. Analogous results were obtained using the excised rat jejunum model. The increase in QD of TMC was seen to favour the mucoadhesion, resulting in a prolonged residence time on intestinal mucosa. The nanoparticle penetration into excised rat jejunum tissue, observed by means of CLSM, suggested that the mucoadhesive properties delayed the absorption of nanoparticles, however they produced an increase in the contact time with intestinal epithelium, offering a better chance for internalisation. The improvement of mucoadhesion and of nanoparticle internalisation with respect to chitosan nanosystems makes the TMCs nanosystems suitable carriers for the intestinal absorption of peptides.