Aline Rigon Zimmer

Antioxidant and anti-inflammatory properties of Capsicum baccatum: from traditional use to scientific approach.

ETHNOPHARMACOLOGICAL RELEVANCE Peppers from Capsicum species (Solanaceae) are native to Central and South America, and are commonly used as food and also for a broad variety of medicinal applications. AIM OF THE STUDY The red pepper Capsicum baccatum var. pendulum is widely consumed in Brazil, but there are few reports in the literature of studies on its chemical composition and biological properties. In this study the antioxidant and anti-inflammatory activities of Capsicum baccatum were evaluated and the total phenolic compounds and flavonoid contents were determined. MATERIALS AND METHODS The antioxidant property was assayed by scavenging abilities using DPPH and the anti-inflammatory activity was tested through the carrageenan-induced pleurisy model in mice. The total phenolic compounds and flavonoid contents were determined spectrophotometrically. RESULTS The ethanolic and butanol extracts (200mg/kg, p.o.) presented a significant anti-inflammatory activity toward carrageenan-induced pleurisy model in mice in comparison to dexamethasone (0.5mg/kg, s.c.). Among the parameters evaluated, the treatment with these samples inhibited leukocyte migration and reduced the formation of exudate. The contents of flavonoids and total phenolic compounds could be correlated with the antioxidant and anti-inflammatory activities observed for Capsicum baccatum. CONCLUSIONS Our findings suggest that Capsicum baccatum contains potential antioxidant and anti-inflammatory compounds which could be tested as drug candidates against oxidative and inflammation-related pathological processes in medicinal chemistry studies.

Influence of penetration enhancers and molecular weight in antifungals permeation through bovine hoof membranes and prediction of efficacy in human nails.

This work aimed to evaluate the effect of different substances on the permeation of geraniol through bovine hoof membranes. Different penetration enhancers were able to increase the permeability up to 25 times compared to control. It was demonstrated that acetilcysteine in association with ascorbic acid increased the permeation, even in acid formulations. In addition, some antifungal drugs were incorporated into a gel formulation of HPMC containing acetylcysteine 5% and ascorbic acid 0.2% and then the permeation coefficient through bovine hoof membranes was evaluated. The relationship between permeability and molecular weight was established for fluconazole, miconazole, terbinafine, butenafine, geraniol and nerol. Geraniol and nerol, the antifungals with lower molecular weight, had the better permeability results. Permeability coefficients for nail plates were estimated and geraniol demonstrated similar or even better efficacy index values against T. rubrum, T. menthagrophytes and M. canis compared with terbinafine and miconazole.

Nandrolone-induced aggressive behavior is associated with alterations in extracellular glutamate homeostasis in mice.

Nandrolone decanoate (ND), an anabolic androgenic steroid (AAS), induces an aggressive phenotype by mechanisms involving glutamate-induced N-methyl-d-aspartate receptor (NMDAr) hyperexcitability. The astrocytic glutamate transporters remove excessive glutamate surrounding the synapse. However, the impact of supraphysiological doses of ND on glutamate transporters activity remains elusive. We investigated whether ND-induced aggressive behavior is interconnected with GLT-1 activity, glutamate levels and abnormal NMDAr responses. Two-month-old untreated male mice (CF1, n=20) were tested for baseline aggressive behavior in the resident-intruder test. Another group of mice (n=188) was injected with ND (15mg/kg) or vehicle for 4, 11 and 19days (short-, mid- and long-term endpoints, respectively) and was evaluated in the resident-intruder test. Each endpoint was assessed for GLT-1 expression and glutamate uptake activity in the frontoparietal cortex and hippocampal tissues. Only the long-term ND endpoint significantly decreased the latency to first attack and increased the number of attacks, which was associated with decreased GLT-1 expression and glutamate uptake activity in both brain areas. These alterations may affect extracellular glutamate levels and receptor excitability. Resident males were assessed for hippocampal glutamate levels via microdialysis both prior to, and following, the introduction of intruders. Long-term ND mice displayed significant increases in the microdialysate glutamate levels only after exposure to intruders. A single intraperitoneal dose of the NMDAr antagonists, memantine or MK-801, shortly before the intruder test decreased aggressive behavior. In summary, long-term ND-induced aggressive behavior is associated with decreased extracellular glutamate clearance and NMDAr hyperexcitability, emphasizing the role of this receptor in mediating aggression mechanisms.

Synthesis and transport of different sphingomyelin species in rat Sertoli cells.

Cellular phospholipids of Sertoli cells from immature rats were labeled with [14C]-choline. Two sphingomyelin bands (SM1 and SM2) were identified by TLC. The incorporation of [14C]-choline over a 45 h period of incubation demonstrated that there are differences in labeling kinetics between SM1 and SM2. The subcellular location of SM1 and SM2 was investigated by accessibility to bacterial sphingomyelinase. The results showed the existence of two SM pools in Sertoli cells, but an equal cellular distribution of SM1 and SM2. SM2 is characterized by a relatively high content of unsaturated fatty acids. The inhibition of vesicular flow by monensin determines a decrease of about 60–70% in incorporation into SM1 and SM2, suggesting the existence of at least two sites of sphingomyelin synthesis. Pulse-chase and time-course experiments indicated a phosphatidylcholine → SM precursor product relationship and differences in kinetic properties between SM1 and SM2. Resynthesis experiments showed that monensin had only a partial inhibitory effect on SM1 resynthesis, and a second sphingomyelinase treatment demonstrated that the resynthesized fraction reached the outer leaflet of the plasma membrane. The 60–70% inhibition of SM synthesis by monensin showed that the trans-Golgi cisternae and the trans-Golgi network are the most likely sites of bulk SM synthesis, and that about 15% of SM was synthesized in the cis/medial Golgi apparatus. Additionally the results indicated that plasma membrane SM synthase activity could be the site of about 15% of SM synthesis in Sertoli cells.

Vaccinium virgatum fruit extract as an important adjuvant in biochemical and behavioral alterations observed in animal model of metabolic syndrome

The aim of this study was to investigate the effect of blueberry (Vaccinium virgatum) fruit extract on metabolic, behavioral and oxidative stress parameters in the hippocampus and cerebral cortex of mice submitted to an experimental model of metabolic syndrome induced by a highly palatable diet (HPD). Mice C57BL/6 were divided into 4 experimental groups: (1) received standard chow and saline orally, (2) received standard chow and blueberry hydroalcoholic extract, (3) received HPD and saline orally, (4) received HPD and blueberry hydroalcoholic extract. The animals were treated for 150days. Our results showed that the animals fed with HPD presented insulin resistance, increased body weight, visceral fat, glucose, triglycerides, and total cholesterol when compared to the control group. The blueberry extract prevented the increase of these metabolic parameters. Also, the extract was able to reduce the levels of thiobarbituric acid reactive substances in the cerebral cortex and hippocampus of animals submitted to HPD. In contrast, no differences were observed in the total thiol content, activity of the antioxidant enzymes catalase and superoxide dismutase. In addition, the HPD fed animals showed a significant increase in immobility time in the forced swimming test and blueberry prevented this alteration, although no changes were observed in the ambulatory behavior, as well as in the anxiolytic profile of these animals. Overall, our findings suggest that chronic consumption of blueberry extract exhibits hypoglycemic, hypolipidemic, antidepressant-like and antiperoxidative effects in an animal model of metabolic syndrome.

HPLC-DAD for the determination of three different classes of antifungals: method characterization, statistical approach, and application to a permeation study.

This study describes and characterizes methods for high-performance liquid chromatography diode array detection (HPLC-DAD) analysis of formulations containing molecules with antifungal activity of three different classes: terbinafine and butenafine (allylamines), miconazole and fluconazole (azoles), and geraniol, neral and geranial (monoterpenes). All methods used the same chromatographic column (RP18 ), enabling the analysis to be performed in a single batch. The specificity was extensively discussed through the establishment of purity peak methods. The analytical parameters (linearity, precision and accuracy) were calculated and discussed in detail using specific statistical approaches. All substances showed satisfactory results for chromatographic and analytical parameters. Limits of 1.3% to mean repeatability and 2.0% for intermediate precision are suggested as acceptance criteria in validation of methods by HPLC-DAD, in situations where there is no extensive pretreatment of the samples. The methods proved to be robust and significant factors were discussed regarding their influence on chromatographic parameters (retention time, resolution, tailing factor and column efficiency). Finally, the application of the developed methods was demonstrated by the results of a permeation study of the antifungal agents through bovine hoof membranes.

Long-Term Oral Administration of Capsicum baccatum Extracts Does Not Alter Behavioral, Hematological, and Metabolic Parameters in CF1 Mice.

Our group showed that crude ethanol (CE) and butanol (BUT) extracts of Capsicum baccatum presented anti-inflammatory and antioxidant properties. Furthermore, the flavonoid and total phenolic contents were positively correlated with both of these properties observed for C. baccatum extracts. The present study demonstrated that 60 days of oral administration of CE and BUT (200 mg/kg) in mice did not cause significant differences in the following parameters evaluated: hematological profile, body weight and relative weight of visceral organs, systemic lipid profile, glucose homeostasis (GTT), kidney and hepatic biochemical markers, and spontaneous locomotion and anxiety-like behavior. Altogether, these results indicate for the first time that the long-term oral administration of C. baccatum extracts does not affect specific aspects of CF1 mice physiology, suggesting their safety, building up the venue to test their efficacy in animal models underlying persistent activation of oxidative and inflammatory pathways.

Hypnotic effect of ecdysterone isolated from Pfaffia glomerata (Spreng.) Pedersen

Neste trabalho foi avaliado, em roedores, o efeito depressor das fracoes cloroformio (CHCl3), acetato de etila (EtOAc) e n-butanol, obtidas das partes subterrâneas de Pfaffia glomerata, empregando-se o teste de tempo de sono barbiturico como referencia. Somente a fracao lipofilica (CHCl3:EtOAc, 1:1, m/m) (i.p. 500 mg/kg; v.o. 1000 mg/kg) potenciou o tempo de sono induzido por pentobarbital. A ecdisterona foi isolada e identificada como constituinte majoritario (1,4% m/m) desta fracao, atraves de cromatografia liquida de alta eficiencia e metodos espectroscopicos, respectivamente. Este composto potenciou o tempo de sono barbiturico (100 mg/kg, i.p.; 400 mg/kg, v.o), sem causar hipotermia. Nestas mesmas doses, a ecdisterona nao alterou a performance dos animais no rota-rod, esquiva inibitoria e labirinto em cruz-elevado, alem de nao alterar o padrao de convulsoes induzidas por pentilenotetrazol. Este perfil indica que esta substância, nestas doses, nao apresenta perfil ansiolitico ou neurotoxico. Estes resultados indicam que a ecdisterona e o componente responsavel pela acao hipnotica apresentada pela fracao lipofilica obtida das partes subterrâneas de P. glomerata.

OPTIMIZATION AND VALIDATION OF A SE-HPLC METHOD FOR ANALYZING AMYLOID BETA OLIGOMERS

AD coupled with the unsustainable growing costs of the disease dictates the necessity for creating innovative approaches to drug discovery. Technological advances have been made over the last decade, particularly with respect to data science tools owing to the exponential increase of data to be processed and analyzed. These tools allow for new insights to be leveraged that were hitherto not visible. This paper discusses a new technological advancement in the search for novel therapeutic approaches using data science and epigenetics to bring new hope to Alzheimer’s. Methods:We developed a software suite to aid in the full-cycle process of ingesting raw data, providing research tools for identification of epigenetically dysregulated genes and pathways and analysis of all known chemical interactions affecting these pathways in Alzheimer’s disease. Using graph theory, natural language processing, artificial intelligence, and machine learning, we applied these tools to identify chemical and supplement interactions to counteract dysregulated pathways in AD. The knowledge that is constructed allows us to optimize our output based on factors including gene-gene, gene-chemical, and chemical-protein interactions. Results: We constructed a proprietary data science platform which generates optimal treatment plans for Alzheimer’s disease based on dysregulated epigenetic pathways and individualized genetic risk factors. The output generated is optimized against an entire pathway, allowing for higher levels of downstream success in the targeted gene. Conclusions:With the continued failure of clinical trials to bring an effective drug to market, a novel interdisciplinary approach grounded in data science will enhance the discovery phase of clinical trials. In addition to drug targets, the software suite can identify scientifically proven over the counter products, offering those living with Alzheimer’s and their caregivers’ additional options, and decreasing the time needed for patients to access effective therapies.