Peripheral and central antinociceptive effects of the butanolic fraction of Byrsonima verbascifolia leaves on nociception-induced models in mice.

Byrsonima verbascifolia (Malpighiaceae), commonly known as 'murici', is used in folk medicine, for example, in the treatment of inflammation. The anti-inflammatory activity of the butanolic fraction of B. verbascifolia leaves (BvBF) was previously reported by our group, and the present study was designed to evaluate their antinociceptive effects. BvBF (25, 50, and 100 mg/kg) administered intraperitoneally (i.p.) inhibited acetic acid induced abdominal writhing. In the formalin test, BvBF (10, 30 and 100 mg/kg, i.p.) caused a reduction in licking time in both the neurogenic and inflammatory phases. Moreover, we demonstrated that BvBF (30 and 100 mg/kg, i.p.) caused an increase in the latency to response in the hot-plate test. These results demonstrate that BvBF possesses marked peripheral and central antinociceptive activities. Pre-treatment with the non-selective receptor antagonist naloxone (5 mg/kg, i.p.) abolished the antinociceptive effects of BvBF (100 mg/kg, i.p.) in the neurogenic phase of the formalin and hot-plate tests. The anti-inflammatory activity of BvBF (previously reported) as well as the participation of the opioidergic system seems to be responsible, at least in part, for these antinociceptive effects. Finally, BvBF at the doses investigated (25, 50 and 100 mg/Kg) did not cause any toxicity signals, showing that the antinociceptive activity is devoid of sedative and hypomotility effects.

Effects of Maytenus ilicifolia on reproduction and embryo-fetal development in Wistar rats.

Maytenus ilicifolia (Celastraceae), popularly known as espinheira-santa, is a native plant from the Atlantic forest and is commonly used in popular medicine to treat inflammation and as an abortifacient. To evaluate the effects of M. ilicifolia on pregnant rats during the organogenic period (T1) or throughout the gestational period (T2), an extract obtained using an acetone-water mixture at a 70:30 ratio was administered via gavage at a dose of 15.11 mg·kg(-1)·day(-1) over 2 treatment periods (T1 and T2). No clinical signs of maternal toxicity were observed. Term fetuses did not present malformations or anomalies as the number of implantations, reabsorptions, live, and dead fetuses were similar to the control group. In conclusion, M. ilicifolia hydroacetonic extract is non-toxic to pregnant rats and appears to not interfere with the progress of embryo-fetal development.

Evaluation of mutagenic, teratogenic, and immunomodulatory effects of Annona nutans hydromethanolic fraction on pregnant mice.

Plants such as Annona nutans used in folk medicine have a large number of biologically active compounds with pharmacological and/or toxic potential. Moreover, pregnant women use these plants indiscriminately, mainly in the form of teas, without being aware of the harm that they could cause to the health of the embryo/fetus. Therefore, it is necessary to analyze the potential toxic effects of medicinal plants during gestation. The present study aimed to evaluate the effects of A. nutans hydromethanolic fraction leaves (ANHMF) on mutagenic and immunomodulatory activity, reproductive performance, and embryo-fetal development in pregnant female mice. The animals (N=50 female and 25 male) were divided into 5 groups: Control, Pre-treatment, Organogenesis, Gestational, and Pre+Gestational. The results indicate that ANHMF mainly contains flavonoid and other phenolic derivatives. It was found that it does not exhibit any mutagenic or immunomodulatory activity, and it does not cause embryo-fetal toxicity. Based on the protocols used in the present studies, our analyses confirm that it is safe to use ANHMF during pregnancy.

Pre-treatment with glutamine reduces genetic damage due to cancer treatment with cisplatin.

Cisplatin is an effective antineoplastic drug. However, it provokes considerable collateral effects, including genotoxic and clastogenic activity. It has been reported that a diet rich in glutamine can help inhibit such collateral effects. We evaluated this activity in 40 Swiss mice, distributed into eight experimental groups: G1 - Control group (PBS 0.1 mL/10 g body weight); G2 - cisplatin group (cisplatin 6 mg/kg intraperitoneally); G3, G4, G5 - glutamine groups (glutamine at 150, 300, and 600 mg/kg, respectively; orally); G6, G7, G8 - Pre-treatment groups (glutamine at 150, 300, and 600 mg/kg, respectively; orally and cisplatin 6 mg/kg intraperitonially). For the micronucleus assay, samples of blood were collected (before the first use of the drugs at T0, then 24 (T1) and 48 (T2) hours after the first administration). For the comet assay, blood samples were collected only at T2. The damage reduction percentages for the micronucleus assay were 90.0, 47.3, and 37.3% at T1 and 46.0, 38.6, and 34.7% at T2, for G6, G7, and G8 groups, respectively. For the comet assay, the damage reduction percentages were 113.0, 117.4, and 115.0% for G6, G7, and G8, respectively. We conclude that glutamine is able to prevent genotoxic and clastogenic damages caused by cisplatin.

Correlation between the SARC-F Score and Hydration Status in Older Gastrointestinal Cancer Outpatients.

OBJECTIVES: The aim of this study was to assess the association between the extracellular water/total body weight ratio (ECW/TBW) and SARC-F scores among elderly gastrointestinal cancer patients. MEASUREMENTS: A cross-sectional study was performed with 57 older male patients with gastrointestinal cancer. Muscle function was assessed using the SARC-F questionnaire. Total body water (TBW) and extracellular water (ECW) were determined using bioelectrical impedance analysis, and fluid retention was assessed as the ratio of ECW to TBW (ECW/TBW). Pearson´s correlation analysis was used to assess the relationship between the SARC-F score and ECW/TBW, TBW and water intake. Results were considered significant at p < 0.05. RESULTS: Of the 57 older patients evaluated (65 ± 7 y), 13 ± 8% presented severe weight loss in the last 6 months. The median SARC-F score was 1.0 (0-10), and only four patients had SARC-F ≥4, which indicates the risk of sarcopenia. There was a positive correlation between the SARC-F score and ECW/TBW (r = 0.26, p = 0.02). However, no correlation was found between daily water intake or TBW and the SARC-F score. CONCLUSION: In older gastrointestinal cancer outpatients, we found a positive, albeit low, correlation between the SARC-F score and the ECW/TBW ratio. This outcome indicates the likelihood of muscle function loss due to accumulation of extracellular fluid.

Antineoplastic activity of selected constituents of Duguetia glabriuscula.

The cytotoxic effects of seven constituents isolated from Duguetia glabriuscula were evaluated against Hep2 human larynx carcinoma cells. The cytotoxicity exhibited by beta-sitosterol was as strong as that of cis-platin. (+)-Alloaromadendran-10,14beta-diol caused inhibition of cellular growth with IC50 values lower than 25 microg/ml, a feature that was considered as revealing significant activity. Polycarpol showed borderline cytotoxicity, whereas the other compounds were inactive.

Non-clinical studies required for new drug development - Part I: early in silico and in vitro studies, new target discovery and validation, proof of principles and robustness of animal studies.

This review presents a historical overview of drug discovery and the non-clinical stages of the drug development process, from initial target identification and validation, through in silico assays and high throughput screening (HTS), identification of leader molecules and their optimization, the selection of a candidate substance for clinical development, and the use of animal models during the early studies of proof-of-concept (or principle). This report also discusses the relevance of validated and predictive animal models selection, as well as the correct use of animal tests concerning the experimental design, execution and interpretation, which affect the reproducibility, quality and reliability of non-clinical studies necessary to translate to and support clinical studies. Collectively, improving these aspects will certainly contribute to the robustness of both scientific publications and the translation of new substances to clinical development.

Non-clinical studies in the process of new drug development - Part II: Good laboratory practice, metabolism, pharmacokinetics, safety and dose translation to clinical studies.

The process of drug development involves non-clinical and clinical studies. Non-clinical studies are conducted using different protocols including animal studies, which mostly follow the Good Laboratory Practice (GLP) regulations. During the early pre-clinical development process, also known as Go/No-Go decision, a drug candidate needs to pass through several steps, such as determination of drug availability (studies on pharmacokinetics), absorption, distribution, metabolism and elimination (ADME) and preliminary studies that aim to investigate the candidate safety including genotoxicity, mutagenicity, safety pharmacology and general toxicology. These preliminary studies generally do not need to comply with GLP regulations. These studies aim at investigating the drug safety to obtain the first information about its tolerability in different systems that are relevant for further decisions. There are, however, other studies that should be performed according to GLP standards and are mandatory for the safe exposure to humans, such as repeated dose toxicity, genotoxicity and safety pharmacology. These studies must be conducted before the Investigational New Drug (IND) application. The package of non-clinical studies should cover all information needed for the safe transposition of drugs from animals to humans, generally based on the non-observed adverse effect level (NOAEL) obtained from general toxicity studies. After IND approval, other GLP experiments for the evaluation of chronic toxicity, reproductive and developmental toxicity, carcinogenicity and genotoxicity, are carried out during the clinical phase of development. However, the necessity of performing such studies depends on the new drug clinical application purpose.

Metal complexes of anhydrotetracycline. 1. A spectrometric study of the Cu(II) and Ni(II) complexes.

Anhydrotetracycline (AHTC), one of the major toxic decomposition products of the antibiotic tetracycline, contains several potential binding sites to metal ions. The acidity constants of the ligand were calculated in aqueous medium (I = 0.1 M) at 25 and 37 degrees C. We found pKa1 = 3.23 +/- 0.08, pKa2 = 5.94 +/- 0.09, and pKa3 = 8.48 +/- 0.02 at 25 degrees C and pKa1 = 3.12 +/- 0.09, pKa2 = 5.86 +/- 0.03, and pKa3 = 8.38 +/- 0.04 at 37 degrees C. The coordination of AHTC to Cu(II) and Ni(II) ions was studied in the solid state as well as in buffered aqueous solution at pH 10.0. At this pH, the formation of the two CuL2 and CuL species was indicated (log beta 1 = 8.41 +/- 0.04 and log beta 2 = 12.55 +/- 0.05), but only the formation of the NiL complex (log beta = 5.74 +/- 0.04) was identified. Spectroscopic data confirm the previous assignment of the C11 and C12 oxygens as the coordination sites, yielding six-membered ring chelates and excluding complexation through any of the potential binding positions on ring A.

Non-clinical studies in the process of new drug development - Part II: Good laboratory practice, metabolism, pharmacokinetics, safety and dose translation to clinical studies

The process of drug development involves non-clinical and clinical studies. Non-clinical studies are conducted using different protocols including animal studies, which mostly follow the Good Laboratory Practice (GLP) regulations. During the early pre-clinical development process, also known as Go/No-Go decision, a drug candidate needs to pass through several steps, such as determination of drug availability (studies on pharmacokinetics), absorption, distribution, metabolism and elimination (ADME) and preliminary studies that aim to investigate the candidate safety including genotoxicity, mutagenicity, safety pharmacology and general toxicology. These preliminary studies generally do not need to comply with GLP regulations. These studies aim at investigating the drug safety to obtain the first information about its tolerability in different systems that are relevant for further decisions. There are, however, other studies that should be performed according to GLP standards and are mandatory for the safe exposure to humans, such as repeated dose toxicity, genotoxicity and safety pharmacology. These studies must be conducted before the Investigational New Drug (IND) application. The package of non-clinical studies should cover all information needed for the safe transposition of drugs from animals to humans, generally based on the non-observed adverse effect level (NOAEL) obtained from general toxicity studies. After IND approval, other GLP experiments for the evaluation of chronic toxicity, reproductive and developmental toxicity, carcinogenicity and genotoxicity, are carried out during the clinical phase of development. However, the necessity of performing such studies depends on the new drug clinical application purpose.

Adubação fosfatada associada à cama de frango e sua influência na produtividade e no teor de flavonoides da Marcela (Achyrocline satureioides (Lam. ) DC. ) em duas épocas de colheita / Total biomass and flavonoid contents of Achyrocline satureioides in response to phosphate fertilizer and broiler litter at two successive harvests

RESUMO: Objetivou-se com este trabalho avaliar a produção de massa e o teor de flavonoides de plantas de marcela cultivadas com fósforo e cama de frango e colhidas em duas épocas. Estudou-se a combinação de doses de P2O5 (0, 100, 200 e 300 kg ha-1) na forma de superfosfato triplo, com e sem cama de frango (10 t ha-1), arranjados como fatorial 4x2, no delineamento blocos casualizados, com quatro repetições. Foram feitas duas colheitas das plantas, consideradas como parcelas subdivididas no tempo. A propagação foi indireta, com semeio em bandejas e transplante ao campo. As plantas foram colhidas aos 195 e 223 dias após o transplantio - DAT. A altura máxima das plantas foi de 1,14 m com uso da cama de frango e de 0,97 m sem cama, aos 195 dias após o transplantio. A produção máxima de massa fresca de parte aérea das plantas foi de 30,31 t ha-1 na colheita aos 223 DAT, enquanto a de massa seca foi de 11,38 t ha-1, ambas com o uso de 300 kg ha-1 de P2O5.A massa fresca de inflorescências foi maior (4,08 t ha-1) com adição de cama de frango ao solo e menor (3,49 t ha-1) sem cama de frango; em resposta às doses de fósforo, cresceram linearmente, sendo a máxima de 4,65 t ha-1 com uso de 300 kg ha-1 de P2O5, independente da época de colheita. A massa seca de inflorescências foi máxima de 2,38 t ha-1 com o uso de 300 kg ha-1 P2O5 na colheita aos 223 DAT. A maior produção de massa seca e o maior teor de flavonoides das inflorescências foram obtidos com o cultivo das plantas de marcela em solo com cama de frango (10 t ha-1) e com P2O5 (300 kg ha-1) na colheita aos 223 dias após o transplantio.

ABSTRACT: The aim of this experiment was to evaluate the biomass yield and flavonoid contents of Achyrocline satureioides grown under different levels of phosphate fertilizer in association with broiler litter, during two successive harvests. Four levels of P2O5 (0, 100, 200 and 300 kg ha-1), as triple superphosphate, and two levels of broiler litter (0 and 10 t ha-1) were arranged as a 4x2 factorial design in randomized blocks with four replications. Two harvests of plants, considered as split-plot, were made. Seeds were sown in polystyrene trays then transplanted to the experimental field and harvested at 195 and 223 days after transplanting (DAT). The maximum heights of the plants were 1.14 m under the addition of broiler litter and 0.97 m without it, at 195 days after transplanting. The highest yields of total fresh weight (30.31 ton ha-1) and dry weight (11.38 ton ha-1) were obtained at 223 DAT, both with the concentration of 300 kg ha-1 P2O5. The fresh weight of the inflorescences was greater under the addition of broiler litter (4.08 ton ha-1) than without it (3.49 ton ha-1). The plants grew linearly in response to the use of phosphate, and the maximum yield was 4.65 ton ha-1 under the concentration of 300 kg ha-1 P2O5, regardless of the harvest season. The dry weight of the inflorescences reached maximum yield (2.38 ton ha-1) at 223 DAT, in response to the use of 300 kg ha-1 P2O5. The highest yield of dry weight and the highest contents of flavonoids from the inflorescences were obtained when A. satureioides was grown under the concentration of P2O5 (300 kg ha-1) with the addition of broiler litter (10 ton ha-1) and harvested at 223 DAT.

The antitumoral, trypanocidal and antileishmanial activities of extract and alkaloids isolated from Duguetia furfuracea.

The alkaloid extract and five alkaloids isolated from subterranean stem bark of Duguetia furfuracea (Annonaceae) were investigated for the following activities: antitumoral, trypanocidal and leishmanicidal. Dicentrinone showed weak cytotoxicity, but it had the strongest leishmanicidal activity (IC(50) 0.01 microM). Duguetine and duguetine beta-N-oxide caused considerable antitumoral activity in every cell lines evaluated, although duguetine was more active against trypomastigote forms (IC(50) 9.32 microM) than other alkaloids tested.

A method for bulbospinal trigeminal nucleotomy in the treatment of facial deafferentation pain.

Many types of facial pain are difficult to treat, such as postherpetic, posttraumatic, or pain following denervation procedures used in the treatment of trigeminal neuralgia (anesthesia dolorosa), all of which involve deafferentation of the spinal trigeminal nucleus.

Acute and subacute toxicity of Cochlospermum regium (Mart. & Schr.) pilger.

The toxicity of a hydroethanol extract of the subterranean part of Cochlospermum regium was evaluated in mice and rats. The extract had moderate acute toxicity when administered intraperitoneally and low toxicity upon oral administration. A subacute toxicity test revealed that the extract is well tolerated by these animals.

Plantas medicinais comercializadas por raizeiros no Centro de Campo Grande, Mato Grosso do Sul / Medicinal plants from herb sellers operating in downtown Campo Grande, Mato Grosso do Sul, Brazil

Foi realizado um levantamento das plantas medicinais mais solicitadas a raizeiros do centro da cidade de Campo Grande, Mato Grosso do Sul, ou por eles indicadas. Esse levantamento foi efetuado em duas épocas distintas: 1992 e 2002. A compilação dos dados revelou que 27 espécies foram citadas em 1992 e 32 em 2002. As seis espécies mais freqüentemente consumidas (Maytenus cf. macrodonta, "cancorosa"; Baccharis trimera, "carqueja"; Achyrocline alata, "jateí-kaá"; Stryphnodendron adstringens, "barbatimão"; Cochlospermum regium, "algodãozinho", e Echinodorus macrophyllus, "chapéu-de-couro") foram adquiridas para identificação e avaliação preliminar da sua qualidade. Nas duas épocas, a qualidade das plantas medicinais apresentava itens que reprovavam seu consumo. Em ambos os levantamentos, observou-se uma constância das espécies mais utilizadas e uma coerência satisfatória com os dados disponíveis sobre os hábitos da automedicação da população do município. Os motivos e sintomas mais freqüentes para a aquisição de plantas medicinais foram os relacionados ao uso analgésico, sistemas genitourinário, respiratório e osteomuscular, dispepsia/má digestão e tecido conjuntivo.

A survey of the medicinal plants requested from and/or indicated by herb sellers operating in the central area of Campo Grande, Mato Grosso do Sul, Brazil, was performed at two different times - 1992 and 2002. The first survey revealed 27 species being used; the second one, 32 species. The species most often consumed (Maytenus cf. macrodonta, ‘cancorosa’; Baccharis trimera, ‘carqueja’; Achyrocline alata, ‘jateí-kaá’; Stryphnodendron adstringens, ‘barbatimão’; Cochlospermum regium, ‘algodãozinho’, and Echinodorus macrophyllus, ‘chapéu-de-couro’) were acquired for identification and evaluation of their quality. On both occasions, the quality of the samples was negatively affected by features that made them inappropriate for human consumption. The results of both surveys agreed as to the species most used. Also, a satisfactory degree of coherence was found regarding the data available on the self-medication behavior of the population of Campo Grande. These medicinal plants were more often sought for analgesic use and for symptoms related to the genitourinary, respiratory and skeletomuscular systems, connective tissue and dyspepsia/maldigestion.