Synthesis and antitumour activity of the Primin (2-methoxy-6-n-pentyl-1,4-benzoquinone) and analogues.

Cancer is a serious worldwide health threat, killing almost seven million people per year. Quinones are an important class of antitumour agents that are activated by tumour hypoxia. Primin (2-methoxy-6-n-pentyl-1,4-benzo-quinone), a naturally-occurring product obtained from Primula obconica (Primulaceae) has shown antimicrobial and antitumour properties. The synthesis of the Primin to obtain 3-, 5- or 6-alkyl substituted derivatives has been previously attempted seeking antitumour activity. The intermediate reaction products, 2-methoxy-hydroquinone-di-(2'-tetrahydro-pyranyl) ether and 2-methoxy-6-n-pentyl-hydroquinone-di-(2'-tetrahydropyranyl) ether were obtained and evaluated against sarcoma 180 (S-180) and Ehrlich carcinoma, as well as toxicity tests were performed. The antitumour activity tests showed that these intermediate compounds were able to inhibit S-180 sarcoma and Ehrlich carcinoma growth in mice. These results indicated that the tetrahydropyranyl protect group conserved the antitumour activity in comparison with quinone group, however, it exhibited a less toxic effect, with no characteristic of quinones. These results can suggest that compound 2-methoxy-6-n-pentyl-hydroquinone-di-(2'-tetrahydropyranyl) ether may act as a prodrug with some advantages in comparison with the Primin.

Toxicological Evaluation of Essential Oil From the Leaves of Croton argyrophyllus (Euphorbiaceae) on Aedes aegypti (Diptera: Culicidae) and Mus musculus (Rodentia: Muridae).

Plant-derived essential oils can be used as insecticides for vector control. However, to establish their safety, it is necessary to perform toxicological studies. Herein, we evaluated the chemical composition and insecticidal activity of the essential oil from the leaves of Croton argyrophyllus on the third- and fourth-instar larvae and adult Aedes aegypti (L., 1762). We also evaluated the acute toxicity of the essential oil in adult female Mus musculus. The lethal concentration 50 (LC50) and 90 (LC90) of C. argyrophyllus essential oil on larvae of Ae. aegypti were 0.31 and 0.70 mg ml-1, respectively, and 5.92 and 8.94 mg ml-1, respectively, on Ae. aegypti adults. The major components of the essential oil were spathulenol (22.80%), (E)-caryophyllene (15.41%), α-pinene (14.07%), and bicyclogermacrene (10.43%). It also displayed acute toxicity in adults of Mus musculus; the intraperitoneal and oral lethal dose 50 (LD50) were 2,000 mg kg-1 and 2,500 mg kg-1, respectively. The results showed that the essential oil from C. argyrophyllus leaves has insecticidal activity on Ae. aegypti larvae and adults at an average lethal concentration below the median lethal dose needed to cause acute toxicity in the common mouse.

The role of nitric oxide in the emotional learning of rats in the plus-maze.

The present study evaluated the role of nitric oxide (NO) in the transfer latency (TL) paradigm in the elevated plus-maze. Male Wistar rats received i.p. injections of either 0.9% Saline, N(omega) Nitro-L-arginine-methyl-ester (L-NAME, an inhibitor of NO synthesis), d-NAME (inert isomer), scopolamine (SCO, antagonist of muscarinic receptors), or MK-801 (antagonist of NMDA receptors) and, after 30 min, were submitted to TL procedure. In an independent experiment, the ability of the same L-NAME treatments in changing the arterial pressure and blood glucose level (BGL) was evaluated in conscious rats. The treatment with SCO (1 mg kg(-1)), MK-801 (0.15 mg kg(-1)) and L-NAME (10 and 50 mg kg(-1)), but not with D-NAME, impaired the TL learning. The L-NAME-induced TL deficit was counteracted by L-ARG (100 and 200 mg kg(-1)), while the co-administration of sub-effective doses of L-NAME and MK-801 failed to impair the TL learning. The L-NAME (50 mg kg(-1)) treatment failed to alter the BGL. All treatments with L-NAME induced hypertension, but the rats treated with L-NAME (5 mg kg(-1)) were still able to learn the TL task. The data indicate that the TL deficit induced by L-NAME (10 and 50 mg kg(-1)) is not due to either hypertension or changes in the BGL. It is also possible to establish that NO production is important for emotional learning underlying the TL procedure in rats.

[Radioisotopic mapping of the arrhythmogenic focus in patients with chronic chagasic cardiomyopathy and sustained ventricular tachycardia]. / Mapeamento radioisotópico do foco arritmogênico em portadores de cardiopatia chagásica crônica e taquicardia ventricular sustentada.

PURPOSE: To localize the site of the origin of sustained ventricular tachycardia in chronic chagasic cardiomyopathy patients refractory to antiarrhythmic therapy by radionuclide angiography techniques. METHODS: Five patients underwent radionuclide angiography by intravenous administration of 25mCi 99mTc. The images were obtained in sinus rhythm and during sustained ventricular tachycardia induced in the electrophysiologic laboratory for endocardial mapping. Amplitude and phase images were obtained resulting in a contraction wave synchronic to ventricular dispolarization. RESULTS: All patients had haemodynamic stability during the arrhythmia. One patient had incessant ventricular tachycardia. Mean ejection fraction was 0.38. In 4 patients the site of the origin of ventricular tachycardia was posterior and in one it was localized in the interventricular septum. There was identity in the site of the origin of ventricular tachycardia obtained by endocardial mapping or radionuclide angiography in all patients. The therapy was chemical ablation in 3 patients, surgical aneurysmectomy in one and pharmacologic therapy in the last patient. CONCLUSION: The site of the origin of ventricular tachycardia can be estimated by analyzing the contraction wave obtained by radionuclide angiography techniques in patients with hemodynamic stable sustained ventricular tachycardia.

[Catheter ablation in patients with refractory cardiac arrhythmias with radiofrequency techniques]. / Ablação por cateter em pacientes com arritmias cardíacas refratárias utilizando técnicas de radiofreqüência.

PURPOSE: evaluate the efficacy of radiofrequency catheter ablation in patients with refractory cardiac arrhythmias. METHODS: twenty patients with refractory cardiac arrhythmias were undertaken to electrophysiologic studies for diagnosis and radiofrequency catheter ablation of their reentrant arrhythmias. Ten patients were men and 10 women with ages varying from 13 to 76 years (mean = 42.4 years). Nineteen patients had supraventricular tachyarrhythmias: One patient had atrial tachycardia and 1 atrial fibrillation with rapid ventricular rate, 5 patients had reentrant nodal tachycardia, 12 patients had reentrant atrioventricular tachycardia and 1 patient had right ventricular outflow tract tachycardia. RESULTS: the mean time of the procedure was 4.1 hours. The radiofrequency current energy applied was 40-50 V for 30-40 seconds. Ablation was successful in 18/20 (90%) patients; in 15/18 (83%) of successfully treated patients the same study was done for diagnosis and radiofrequency ablation. One patient had femoral arterial occlusion and was treated with no significant sequelae. During a mean follow-up of 4 months no preexcitation or reentrant tachycardia occurred. CONCLUSIONS: the results of our experience with radiofrequency catheter ablation of cardiac arrhythmias suggest that this technique can benefit an important number of patients with cardiac arrhythmias.

[Catheter ablation of ectopic incessant atrial tachycardia using radiofrequency. Reversion of tachycardiomyopathy]. / Ablação por cateter de foco de taquicardia atrial ectópica incessante utilizando radiofreqüência. Reversão de taquicardiomiopatia.

A 8-year-old female patient with refractory incessant atrial tachycardia, very symptomatic and with left ventricular ejection fraction of 0.25. Electrophysiological study and endocardial mapping localized the site of the origin of atrial tachycardia in the superior right atrium. In this site 2 applications of radiofrequency current (25V, 20 and 50 seconds) resulted in termination of the atrial tachycardia. She was discharged off antiarrhythmic drugs and after 2 months ejection fraction was 0.52. She was completely asymptomatic 6 months after ablation procedure.

[Potential drug interactions in patients with sepsis admitted to the intensive care unit]. / Potenciales interacciones medicamentosas en pacientes con sepsis internados en la unidad de terapia intensiva.

OBJECTIVE: To analyze, detect and classify potential drug interactions in patients with sepsis admitted to the intensive care unit. METHODS: This prospective observational cohort study of septic patients was conducted between April 2010 and August 2011. Data collection included demographic characteristics, clinical features, and analysis of pharmacotherapy. RESULTS: Of 86 patients, 80% had potential drug interactions, with a mean of 1.84 ± 1.09 interaction per patient. Regarding the classification of drug interactions, 64.2% had a pharmacodynamic profile, 60% were of major severity, 53.3% had a rapid onset of action, and 53.8% had good documentation. The prevalence of interactions was associated with age, number of drugs prescribed, and length of stay in the intensive care unit (p < 0.001). CONCLUSION: This study showed that drug interactions represent an important clinical concern in septic patients admitted to intensive care units.

Objetivo: Analizar, detectar y clasificar las potenciales interacciones medicamentosas (IMs) en pacientes con cuadro de sepsis internados en la Unidad de Terapia Intensiva. Método: El diseño de este estudio fue una cohorte prospectiva y observacional en pacientes sépticos, durante el periodo de abril de 2010 a agosto de 2011. La recolección de datos incluyó datos demográficos, clínicos y análisis de la terapéutica farmacológica. Resultados: De los 86 pacientes, 80% presentaron potenciales IMs, con una media de 1,84 ± 1,09 interacciones por paciente. En cuanto a la clasificación de las IMs, 64,2% presentaron perfil farmacodinámico, 60% gravedad mayor, 53% inicio rápido y 53,8% buena documentación científica. La prevalencia de las interacciones se asociaron con la edad, el número de medicamentos prescriptos y la duración de la internación. Conclusión: El estudio demostró que las interacciones medicamentosas representan una importante preocupación clínica en pacientes sépticos internados en unidades de terapia intensiva.

Expression of a recombinant Phoneutria toxin active in calcium channels.

PnTx3-4 is a toxin isolated from the venom of the spider Phoneutria nigriventer that blocks N-, P/Q-, and R-type voltage-gated calcium channels and has great potential for clinical applications. In this report we used the SUMO system to express large amounts of recombinant PnTx3-4 peptide, which was found in both soluble and insoluble fractions of bacterial extracts. We purified the recombinant toxin from both fractions and showed that the recombinant peptide showed biological activity similar to the native PnTx3-4. In silico analysis of the primary sequence of PnTx3-4 indicated that the peptide conforms to all the criteria of a knottin scaffold. Additionally, circular dichroism spectrum analysis of the recombinant PnTx3-4 predicted that the toxin structure is composed of approximately 53% turns/unordered, 31% α-helix and 16% ß-strand, which is consistent with predicted model of the PnTx3-4 knottin scaffold available at the knottin database (http://knottin.cbs.cnrs.fr). These studies provide the basis for future large scale production and structure-function investigation of PnTx3-4.

Efeito da fração não-oleosa doRicinus communisna biodistribuição comTecnécio-99m em camundongos / Effect of non-oily fraction ofRicinus communis on the bio distribution with Technetium-99m in mice

RESUMO Ricinus communisé uma planta pertence à família Euforbiaceae. A partir de seus frutos é extraído o óleo de rícino e outra fração altamente tóxica (fração não-oleosa). Neste estudo, foi avaliado o efeito da fração não oleosa de R. communis(FNORC) na biodistribuição do Tc-99m em camundongos sadios e em animais transplantados com Sarcoma-180 (S180). Para avaliação da biodistribuição do Tc-99m, o percentual de atividade radioativa total injetada (ATI%) foi mensurada utilizando um contador gama. Os resultados mostraram que, em animais sadios, houve aumento da captação do Tc-99m após o tratamento com FNORC pelos rins, cérebro e estômago. Apenas houve diminuição da absorção deste radioisótopo nos músculos. Em animais com tumor, houve redução significativa da captação do Tc-99m no coração, pulmões e tumor, e foi observado o aumento da captação apenas nos rins. Foi observada também a diminuição da concentração das proteínas plasmáticas totais em animais com tumor após tratamento com FNORC e atividade antitumoral significativa. Diante desses dados, conclui-se que fração não oleosa de R. communis possui atividade antitumoral significativa, porém altera a biodistribuição do Tc-99m, fato este que pode resultar em falhas nos processos de diagnóstico por imagem que utilizem este radioisótopo como marcador.

ABSTRACT Ricinus communis is a plant belonging to the Euforbiaceae family. From its fruits, it is extracted the castor oil and another highly toxic fraction (non-oily fraction). In this study, we evaluated the effect of the non-oily fraction of R. communis (NOFRC) on the bio distribution of Tc-99m in healthy mice and in animals transplanted with Sarcoma-180 (S180). To evaluate the bio distribution of theTc-99m, the percentage of the total injected radioactivity (% TIR) was measured through a gamma counter. There was an increase in the capitation of Tc-99m after the treatment with NOFRC in the kidneys, brain, and stomach of healthy animals. A decrease in the reception of this radionuclide was only found in the muscles. In animals with tumour, there was a significant reduction in the uptake of Tc-99m in the heart, lungs and tumour. An increase in the capitation was only observed in the kidneys. It was also observed a reduction in the total plasma protein concentration in animals with tumours after the treatment with NOFRC, besides significant antitumor activity. We concluded that there is antitumor activity in the NOFRC, but that it alters the biodistribution of Tc-99m, a fact which may result in failures in the diagnostic imaging process using this radioisotope as a marker.

Avaliação da toxicidade aguda do extrato das cascas de Pithecellobium cochliocarpum (Gomez) Macbr / Evaluation of acute toxicity of the Pithecellobium cochliocarpum (Gomez) Macbr bark extract

O uso popular, e mesmo o tradicional, não são suficientes para validar as plantas medicinais como medicamentos eficazes e seguros. Para melhor entendimento, é necessário avaliar a relação risco/benefício de seu uso, por meio de estudos toxicológicos. O objetivo desta pesquisa foi estimar a toxicidade aguda do extrato etanólico das cascas secas de Pithecellobium cochliocarpum (Gomez) Macbr através da obtenção da dose letal (DL50) em roedores, e da Concentração letal (CL50) frente à Artemia salina Leach. Foram realizados experimentos por via oral e intraperitoneal utilizando camundongos fêmeas albinos Swiss (Mus musculus) (n=6). Por via oral foram administradas 3 doses (1.000, 3.000 e 5.000 mg Kg-1) e por via entraperitoneal, 5 doses (155, 160, 176, 345,6 e 414,72 Kg-1). Os sinais comportamentais foram avaliados durante uma hora após a administração do extrato, ficando em observação até 48 horas. O número de óbitos foi quantificado para posterior cálculo da DL50. A administração por via intraperitoneal foi realizada em intervalo de 5 minutos para cada animal. Nos ensaios de toxicidade por via oral a solução foi introduzida por via intragástrica através de cânula metálica acoplada a seringa (gavagem) no mesmo intervalo de tempo utilizado pela via intraperitoneal. Os animais do grupo de administração oral apresentaram algumas reações, porém não letais até a dose de 5.000 mg Kg-1. A DL50 para a via intraperitoneal foi 257, 49 mg Kg-1 (muito tóxico, grau 4) (Schuartsman, 1980). A CL50 (543,5 µg Kg-1) demonstrou ser tóxica frente à A. salina. Conclui-se que sob condições agudas de exposição, o extrato do Pithecellobium cochliocarpum é um agente tóxico, devendo ser considerado como tal, dependendo da dose administrada ou absorvida, do etempo e frequência de exposição e das vias de administração.

The popular use, and even the traditional one, is not enough to validate medicinal plants as effective and safe medicines. For a better understanding, it is necessary to assess the risk / benefit ratio of their use through toxicological studies. The aim of this work was to evaluate the acute toxicity of Pithecellobium cochliocarpum (Gomez) Macbr dried bark ethanolic extract through its lethal dose (LD50), in mice, and lethal concentration (LC50) in relation to Artemia salina Leach. Experiments were performed by oral and intraperitoneal route using female Swiss albino mice (Mus musculus) (n = 6). The first three doses were given orally (1,000, 3,000 and 5,000 mg kg-1) and the last five doses were given intraperitoneally (155, 160, 176, 345.6 and 414.72 Kg-1). The behavioral signs were evaluated one hour after administration of the extract, being observed up to 48 hours. The number of deaths was quantified for subsequent calculation of LD50. The intraperitoneal administration was carried out at an interval of 5 minutes for each animal. For the oral toxicity test, the solution was introduced in the digestive system of the animals through a metal cannula coupled to a syringe (gavage) at the same time interval used for the intraperitoneal route. The animals from the oral group presented some reactions, but they were not lethal up to the dose of 5.000 mg kg-1. The LD50 for the intraperitoneal group was 257.49 mg kg-1 (very toxic, grade 4) (Schuartsman, 1980). The LC50 (543.5 mg kg-1) was toxic to A. salina. We can conclude that, under acute conditions of exposure, the Pithecellobium cochliocarpum extract is a poisonous agent and should be considered as such depending on the administered or absorbed dose, the time and frequency of exposure, and the administration routes.

Avaliação biológica de Foeniculum vulgare (Mill. ) (Umbelliferae/Apiaceae) / Biological evaluation of Foeniculum vulgare (Mill. ) (Umbelliferae/Apiaceae)

Foeniculum vulgare Mill. é espécie herbácea da família Umbelliferae, nativa da região do Mediterrâneo e da Ásia Menor, amplamente distribuída em todo o território brasileiro. É conhecida popularmente como funcho ou erva-doce e usada na medicina como analgésico, digestivo, carminativo, diurético, expectorante, lactígeno, anti-inflamatório, e antiespasmódico. O extrato bruto etanólico para a verificação das atividades biológicas foi preparado a partir de sementes compradas no comércio. Para a realização do perfil fitoquímico foi utilizada a cromatografia em camada delgada analítica; a atividade antimicrobiana foi determinada pelo teste de difusão em disco de papel e da concentração inibitória mínima (CIM); a atividade antinociceptiva foi realizada pelo método de contorções abdominais em cobaias. Os micro-organismos testados foram isolados clínicos multirresistentes obtidos do Setor de Bacteriologia do Hospital das Clínicas de Pernambuco. O estudo fitoquímico identificou a maioria dos compostos secundários presentes na fração metanólica das sementes, sendo eles: triterpenos, glicosídeos de flavanóides, terpenos menores (monoterpenóides, sesquiterpenóide e diterpenóides), e açúcares redutores. Os resultados obtidos mostraram que o extrato etanólico apresentou maior atividade frente à Micrococcus spp. (CIM=250µg/mL). Os resultados da avaliação da atividade antinociceptiva demonstraram que apenas a dosagem de 298 mg/Kg quando comparado com o padrão indometacina conseguiu uma redução significativa no número de contorções abdominais dos animais. Estudos posteriores deverão ser realizados para a identificação e isolamento de alguns compostos secundários, bem como a realização de outros protocolos de analgesia.

Foeniculum vulgare Mill. (Umbelliferae) is a herbaceous plant of the family Umbelliferae, native to the Mediterranean and Asia Minor region and widely distributed all over the Brazilian territory. It is commonly known as fennel or "erva-doce" and has been used in medicine as analgesic, digestive, carminative, diuretic, expectorant, lactigenous, anti-inflammatory, and antispasmodic agent. Crude ethanolic extract for the verification of biological activities was prepared from seeds bought in the market. For the phytochemical profile, analytical thin-layer chromatography was used; the antimicrobial activity was determined by the paper disc diffusion test and minimum inhibitory concentration (MIC); the antinociceptive activity was verified by the method of abdominal writhings in the animals. The tested microorganisms were multiresistant clinical isolates obtained from the Bacteriology Sector of the Clinical Hospital of Pernambuco. The phytochemical study identified the majority of secondary compounds present in the methanolic fraction of seeds, as follows: triterpenes, flavanoid glycosides, smaller terpenes (monoterpenoids, sesquiterpenoids and diterpenoids) and reducing sugars. The obtained results showed that the ethanolic extract had greater activity against Micrococcus spp. (MIC=250µg/mL). Results of the evaluation of the antinociceptive activity demonstrated that only the dosage of 298 mg/Kg, compared to the indomethacin pattern, led to a significant reduction in the number of abdominal writhings in the animals. Further studies will be carried out for the identification and isolation of some secondary compounds, as well as other analgesic protocols.