[ production of hairy roots from [Datura innoxia L.] by Agrobacterium rhizogenes and effects of salicylic acid and methyl jasmonate biological elicitors on atropine and scopolamine content]

Background: Using Agrobacterium rhizogenes due to create hairy roots, is a useful method to increase secondary metabolites many plants

Objective: Purpose of this research is to transgenic hairy roots culture, in order to produce secondary metabolites in Datura innoxia

Methods: Explants leaf and cotyledon of Datura innoxia were inoculated for two months with A7, A4 and 15834 strains of Agrobacterium rhizogenes; Furthermore injection and Immersion methods were used in this scrutiny. The presence of T-DNA in transgenic hairy roots were confirmed by PCR. Transgenic hairy roots in liquid medium of 1/2MS were cultured. In order to induct elicitors, methyl jasmonate in tow densities of 50 micro M and 100micro M, and salicylic acid in three densities of 1mM, 0.1mM and 0.01 mM were used randomly three times. Atropine and scopolamine content of transgenic hairy roots were examined by HPLC

Results: The highest and lowest rate of transgenic hairy roots production was respectively related to the strains of A4 and 15834. Best explants for inoculation, leaf with A4 strain and cotyledon with A7 strain, were reported. With highest production rate of hairy roots, Simple deposit using a syringes method was recognized as the best method of inoculation. The effect of salicylic acid at a density of 0.1 mM increases the content of atropine concentrations. Also the results showed that usage of Methyl jasmonate at higher doses [100 micro M] reduces the content of atropine and scopolamine

Conclusion: A. rhizogenes as an appropriate method to produce hairy roots and elicitors the best treatment for increase alkaloids

Efficacy of tiotropium as add-on therapy for adults with uncontrolled asthma

@#<p style="text-align: justify;"><strong>OBJECTIVE:</strong> The study aimed to evaluate efficacy of tiotropium as add-on therapy on top of standard regimens for uncontrolled asthma, specifically in terms of FEV1, morning and evening PEF, reduction in exacerbations, rescue medication use, and quality of life improvement.<br /><strong>METHODS:</strong> A search was done for eligible trials after which validity screen and data extraction was performed. Results were presented as mean differences, standard errors, and 95% confidence intervals, and graphically as forest plots. Estimates were pooled using the random effects model with I2 and Chi2 tests used to assess heterogeneity. Adverse events were reported as dichotomous variables.<br /><strong>RESULTS:</strong> Four studies were included totaling 1617 participants. The tiotropium group had statistically significant improvement in FEV1 (95% Cl, 0.14 [0.09, 0.19], p<0.00001), morning (95% Cl, 20.03 [11.71, 28.35], p<0.00001) with trend towards benefit in reduction of rescue medications (95% Cl, 0.12 [-0.17,0.4],p=0.42) and quality of life improvements (95% Cl, 0.1 [-0.05,0.25], p=0.20). Homogeneity (I2= 0%, Chi2= 0.47-3.22) was found across studies.<br /><strong>CONCLUSION:</strong> Tiotropium is associated with significant improvement in pulmonary function among patients with uncontrolled asthma, with possible benefit in reduction of rescue medications and quality of life improvement.</p>

Neuroprotective Effect of Compound Anisodine in a Mouse Model with Chronic Ocular Hypertension / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Compound anisodine (CA) is a compound preparation made from hydrobromide anisodine and procaine hydrochloride. The former is an M-choline receptor blocker with the function of regulating the vegetative nervous system, improving microcirculation, and so on. The latter is an antioxidant with the activities of neuroprotection. This study aimed to investigate the potential neuroprotection of CA, which affects the degeneration of the retinal ganglion cells (RGCs) in an animal model with chronic ocular hypertension.</p><p><b>METHODS</b>Female C57BL/6J mice (n = 24) were divided randomly into four groups: normal control group without any treatment (Group A, n = 6); CA control group with feeding the CA solution (Group B, n = 6); microbeads (MBs) control group with injecting MB into the anterior chamber (Group C, n = 6); CA study group with MB injection and with feeding the CA solution (Group D, n = 6). Intraocular pressure (IOP) was measured every 3 days after MB injection. At the 21st day, neurons were retrograde-labeled by Fluoro-Gold (FG). Animals were sacrificed on the 27th day. Retinal flat mounts were stained immunohistologically by α2-III-tubulin. FG-retrograde-labeled RGCs, α2-III-tubulin-positive RGCs, and α2-III-tubulin-positive nerve fibers were quantified.</p><p><b>RESULTS</b>Mice of Groups C and D expressed the incidence of consistent IOP elevation, which is above the IOP level of Group A with the normal one. There is no significant difference in IOP between Groups A and B (P > 0.05). On the 27th day, there were distinct loss in stained RGCs and nerve fibers from Groups C and D compared with Group A (allP < 0.001). The quantity was significantly higher in Group D as compared to Group C (allP < 0.001) but lower than Group A (allP > 0.001). There was no significant difference in the quantity of RGCs and nerve fibers between Groups A and B (allP > 0.05).</p><p><b>CONCLUSIONS</b>These findings suggest that CA plays an importantly neuroprotective role on RGCs in a mouse model with chronic ocular hypertension.</p>

Evaluation of the efficacy and safety of tiotropium bromide (5 µg) inhaled via Respimat in Chinese patients with chronic obstructive pulmonary disease / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>A pharmacokinetic study in an Asian population showed that tiotropium 5 µg via Respimat leads to the same plasma levels compared to 18 µg via HandiHaler. The objective of the trial was to compare the efficacy and safety of longterm treatment (1 year) with tiotropium bromide (5 µg) via Respimat® with placebo in patients with chronic obstructive pulmonary disease (COPD).</p><p><b>METHODS</b>A total of 3991 patients were randomized in this double-blind, placebo controlled, parallel group study, while in China 338 patients (309 males, 29 females) received either tiotropium bromide (n = 167) or placebo (n = 171). Tiotropium bromide solution or matching placebo was delivered via Respimat® at a dosage of 5 µg (2×2.5 µg/puff) once daily for 48 weeks. Co-primary endpoints were trough forced expiratory volume in one second (FEV1) and the time to first exacerbation.</p><p><b>RESULTS</b>Statistically significant improvements in trough FEV1 and trough forced vital capacity (FVC) in the tiotropium group were achieved at weeks 4, 24, and 48 compared with those in the placebo group. A statistically significant difference (P = 0.0027) in favour of tiotropium was also observed for the time to first exacerbation. The total numbers of exacerbations during treatment were 90 and 128 in the tiotropium and placebo groups, respectively, with a rate ratio of 0.69 (P = 0.0164). The difference between the treatment groups in the adjusted mean changes from baseline of St. George Respiratory Questionnaire (SGRQ) total score was -3.9 (95% CI: -7.5, -0.2) and was of statistical significance (P = 0.0367). The incidences of serious adverse events (SAEs) in the tiotropium and placebo groups were 16.2% and 17.0%, respectively. Seven deaths occurred whilst patients were on treatment, four in the tiotropium group and three in the placebo group, all of which were assessed as non-related study drugs by the investigators.</p><p><b>CONCLUSIONS</b>Tiotropium significantly improved lung function and quality of life, delayed the time to first exacerbation, reduced the number of exacerbations. Overall, tiotropium was well tolerated.</p>

Short term effects of tiotropium on COPD patients treated with long acting bronchodilators

The results of bronchodilator therapy in chronic obstructive pulmonary disease [COPD] are not satisfactory and because of this, many drugs are administered for treatment of disease. We examined the short-term additive bronchodilator effects of tiotropium in patients with stable COPD already treated with salmeterol twice daily. In a double-blind, double- dummy randomized study we evaluated the acute bronchodilator efficacy of a single 18 micro g dose of tiotropium in patients with COPD under chronic treatment with a long acting beta-adrenergic [salmeterol 50 micro g twice a day]. Measurements of forced vital capacity [FVC], forced expiratory volume in 1 second [FEV1] and six-minute walking test [6 MWT] as indicator of daily activities were made before and after giving the drug. A total of 129 outpatients with stable COPD [Gold level 2 -3- 4] were enrolled and 92 completed the study. Tiotropium elicited a significantly greater bronchodilation than the placebo. The action of the drug was elicited as increase in FEV1 [p

effect of scopolamine on avoidance memory and hippocampal neurons in male wistar rats

Cholinergic systems are involved in learning and memory. Scopolamine, a muscarinic acetylcholine receptor antagonist, is used as a standard/ reference drug for inducing cognitive deficits in healthy humans and animals. The purpose of this study was to evaluate the effects of scopolamine on avoidance memory and number of neurons in rat's hippocampus. Thirty five male albino Wistar rats [200 +/- 20 g] were used in this study. The rats were divided randomly into five groups: control group [healthy samples], sham [saline] and 3 experimental groups 0.2, 0.5 and 1 mg/kg [intraperitoneally - single dose of Scopolamine]. Animals were tested by passive avoidance method [shuttle box]. After one week, a memory test was taken from rats. Finally, with dissection of the rats' brains and tissue operations, neurons were stained with cresyl violet. Photographs of the samples in hippocampal areas were prepared, and neurons were counted. Our results showed that the number of neurons in all experimental groups was lower than that in the control group. The highest decrease in number of neurons was shown in response to 1 mg/kg scopolamine compared to the control group in all regions of hippocampus. Also, we found that in comparison to the saline-treated animals, the injection of scopolamine to rats after training, caused memory destruction. We concluded that memory impairment-induced by scopolamine is probably associated with neuronal loss and this decrease was dose dependent

Therapeutic effect of compound anisodine for primary open angle glaucoma / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To evaluate the therapeutic effect of compound anisodine (CA) for patients with primary open angle glaucoma (POAG).</p><p><b>METHODS</b>According to the modified Hodapp-Parrish-Anderson Visual Fields Grading System, 46 patients with moderate stage POAG were randomized to receive compound anisodine injection (CA group) or venoruton tablets (control group). Visual acuity (VA), IOP, fundus, visual fields (VF) and the blood flow of optic nerve were observed.</p><p><b>RESULTS</b>The mean of defect (MD) was decreased in CA group after treatment. The PSV and EDV of ophthalmic artery were remarkably improved in both groups, as well as the PSV, EDV and RI of retinal central artery. Compound anisodine was superior in improving hemodynamics of ophthalmic artery and retinal central artery to venoruton.</p><p><b>CONCLUSION</b>Compound anisodine can protect optic nerve of POAG through improving the visual function and blood supply of optic nerve.</p>

Issues on Safety of Long-Acting Muscarinic Antagonist / 결핵및호흡기질환

The prevention of and the controlling of symptoms, reductions in the frequency of exacerbations, and disease severity are central to the pharmacologic therapy of chronic obstructive pulmonary disease (COPD). COPD patients are inclined to be older, have more comorbidities, and use polypharmacy as a result. Long-acting inhaled muscarinic antagonists (LAMAs) is a preferred treatment modality. However, the cardiovascular (CV) safety of anti-cholinergics, including LAMA, has been an issue. In contrast, the results of the UPLIFT trial and a pooled analysis of data from 30 trials of tiotropium illustrates the association of tiotropium with reductions in the risk of all cause mortality, CV mortality and CV events. And, the UPLIFT trial provides clues regarding the additive advantages of tiotropium in COPD patients who already are using long-acting inhaled beta2 agonists and inhaled corticosteroids. Following the contribution of tiotropium as a first LAMA, new LAMAs such as aclidinium and glycopyrrolate (NVA-237) seem to be emerging.

Efeito de curto prazo do tiotrópio nos portadores de DPOC em tratamento com B2-agonista / Short-term effect of tiotropium in COPD patients being treated with a B2 agonist

OBJETIVO: Avaliar o impacto de curto prazo do uso de tiotrópio em pacientes com DPOC grave e muito grave com queixas de dispneia apesar do tratamento com outros broncodilatadores. MÉTODOS: Estudo prospectivo incluindo pacientes com DPOC grave ou muito grave, com queixa de dispneia de pequenos esforços ou ao repouso. A cada 15 dias, o tratamento broncodilatador foi modificado: salmeterol, tiotrópio e associação salmeterol+tiotrópio. Ao final de cada regime, foram realizados testes de função pulmonar e teste de caminhada de seis minutos (TC6). Também foram avaliados o grau de dispneia e a capacidade de realização de atividades de vida diária. Para a avaliação das atividades de vida diária, foi utilizada a escala London Chest Activity of Daily Living (LCADL) validado para uso no Brasil. RESULTADOS: Foram avaliados 52 pacientes. Desses, 30 completaram o estudo. A introdução de tiotrópio como monoterapia resultou em uma melhora significativa (p < 0,05) da dispneia basal (média do escore da escala do Medical Research Council de 3,0 para 2,5) e ao final do TC6 (média do escore da escala de Borg de 6,1 para 4,5), e as diferenças foram significativas (p < 0,05 para ambos). O uso da associação salmeterol+tiotrópio resultou em um aumento significativo médio de 81 mL no VEF1 e na melhora de 5,7 pontos no escore da escala LCADL. CONCLUSÕES: A introdução de tiotrópio no tratamento de pacientes com DPOC grave a muito grave em uso de β2-agonistas de longa duração causa melhora na função pulmonar e alivio sintomático perceptível pelos pacientes a curto prazo. Esses resultados, obtidos em regime de atendimento de vida real, dão suporte ao uso da associação salmeterol+tiotrópio em protocolos de assistência específicos a esses pacientes.

OBJECTIVE: To evaluate the short-term impact of tiotropium in patients with severe or very severe COPD who complain of dyspnea despite being currently treated with other bronchodilators. METHODS: A prospective study including patients with severe or very severe COPD and complaining of dyspnea at rest or on minimal exertion. Every 15 days, the bronchodilator treatment regimen was altered, from salmeterol to tiotropium to salmeterol+tiotropium. At the end of each regimen, pulmonary function tests and the six-minute walk test (6MWT) were performed. The degree of dyspnea and the ability to perform activities of daily living were also assessed. To evaluate patient ability to perform activities of daily living, we employed the London Chest Activity of Daily Living (LCADL), validated for use in Brazil. RESULTS: We evaluated 52 patients, 30 of whom completed the study. The use of tiotropium in isolation resulted in significant improvement in dyspnea at baseline (mean Medical Research Council scale score reduced from 3.0 to 2.5) and at the end of 6MWT (mean Borg scale score reduced from 6.1 to 4.5), and the differences were significant (p < 0.05 for both). The use of the salmeterol+tiotropium combination resulted in a significant (81 mL) increase in FEV1 and a 5.7 point improvement in the LCADL score. CONCLUSIONS: The introduction of tiotropium into the treatment of patients with severe or very severe COPD and using long-acting β2 agonists improves pulmonary function and provides symptomatic relief, as perceived by patients in the short term. These results, obtained under real life treatment conditions, support the use of the salmeterol+tiotropium combination in specific treatment protocols for these patients.

Therapeutic effect of tiotropium bromide powder inhalation in patients with stable bronchiectasis / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the therapeutic effect of tiotropium bromide powder inhalation on stable bronchiectasis.</p><p><b>METHODS</b>Twenty-two patients with stable bronchiectasis received inhalation of totropium bromide powder at the daily dose of 18 microg, and on days 1 and 28, the patients were examined for forced expiratory volume in one second (FEVl), predicted value [FEVl(%)], forced expiratory volume (FEV), and FEVl/FVC. The symptom score and BODE index were also recorded.</p><p><b>RESULTS</b>After 1 month of inhalation therapy, the FEV1% of the patients showed a moderate increase but the increment was not statistically significant (t=-1.875, P>0.05); the symptom score and BODE index decreased significantly after the therapy (t=7.091, P<0.001; t=2.982, P<0.05).</p><p><b>CONCLUSION</b>Long-term inhalation of tiotropium bromide powder can improve the clinical symptoms and BODE index and enhance the exercise tolerance and quality of life of the patients with bronchiectasis.</p>

Comparison for the Effects of Triple Therapy with Salmeterol/Fluticasone Propionate and Tiotropium Bromide versus Individual Components in Patients of Severe COPD Combined with Bronchial Hyperresponsiveness / 결핵및호흡기질환

BACKGROUND: A combination of salmeterol and fluticasone propionate (SFC) and tiotropium bromide (TIO) is commonly prescribed for COPD patients but there is little data on their effectiveness, particularly in COPD patients with bronchial hyperresponsiveness. This study compared the spirometric improvement based on the change in FEV1, FEV1/FVC, and IC as well as the clinical outcomes of the therapeutic strategies with SFC and TIO versus the individual components in patients with severe COPD and bronchial hyperresponsiveness. METHODS: This study examined the spirometric data and clinical outcomes of 214 patients with COPD and hyperresponsiveness, who were divided into three groups according to the therapeutic regimen (TIO only, SFC only, and a triple therapy regimen). RESULTS: All regimen groups showed early improvement in the FEV1 and IC (at 3- and 6 months after treatment). However, long-term beneficial effects were observed only in the SFC group (at 24 months after treatment). However, these beneficial effects decreased after a 36-month follow up. In all spirometric results, the 12-, 24-, and 36-months data showed a similar degree of improvement in the three groups. The triple therapy group showed higher St. George's Respiratory Questionnaire scores and lower acute exacerbations and hospitalization. CONCLUSION: SFC can be a more important component in the pharmacological treatment of severe COPD patients with hyperresponsiveness than TIO, particularly in the spirometric and clinical outcomes.

The Efficacy of Cimetropium Bromide as a Premedication before Esophagogastroduodenoscopy / 대한소화기내시경학회지

BACKGROUND/AIMS: Cimetropium bromide has been used widely as a premedication for endoscopy; however, there are no subjective data pertaining to the effects of cimetropum bromide as a premedication. Thus, the current study was undertaken to compare the effects of cimetropum bromide with placebo as a premedication for esophagogastroduodenoscopy (EGD). METHODS: Two hundred ninety-nine consecutive outpatients who had undergone EGD were enrolled in this study. Thirty minutes before EGD, the patients were randomly given an intramuscular injection of cimetropium bromide (5 mg) or saline using a placebo-controlled, double-blind, randomized technique. Immediately after EGD, all the patients and endoscopists were requested to fill out the questionnaire form. RESULTS: One-hundred patients were injected with cimetropium bromide and 150 patients were injected with placebo. There was no statistically significant difference in the degree of residual gastric secretions, the peristaltic activity detected by endoscopists, and the comfort experienced by the patients in each study group. CONCLUSIONS: The intramuscular injection of cimetropium bromide (5 mg) as a premedication for EGD was not significantly superior to placebo, at least with respect to subjective parameters, in spite of its broad use.

The Combination of Tiotropium and Budesonide in the Treatment of Chronic Obstructive Pulmonary Disease

Because additive effects of inhaled corticosteroids and long-acting anticholinergics are unclear, we undertook this study to compare the efficacy of tiotropium alone and tiotropium plus budesonide in patients with chronic obstructive pulmonary disease. The study subjects were randomized to receive either tiotropium 18 microgram once daily with or without budesonide 200 microgram twice daily for 6 weeks. The efficacy variables were changes in trough forced expiratory volume in one second (FEV1), St. George's Respiratory Questionnaire (SGRQ), 6-minute walk distance (6MWD), and use of rescue medication. One hundred patients were randomized and 81 completed the study. The mean age was 64.0 yr, and the mean FEV1 was 39.7% predicted. Compared with tiotropium alone (N=40), the tiotropium/budesonide combination (N=41) was related to an improvement in the SGRQ total score (tiotropium -2.8 units and tiotropium/budesonide -5.6 units, p=0.003). 6MWD was improved by 13.5 m in the tiotropium group and by 22.5 m in the tiotropium/budesonide group (p=0.031). Changes in trough FEV1 and the use of rescue medication were similar between two groups. In conclusion, compared with tiotropium alone, the tiotropium/ budesonide combination was related to an improved health-related quality of life. These data support that low-dose budesonide may enhance the efficacy of tiotropium.

Analysis of anisodine and its metabolites in rat plasma by liquid chromatography-tandem mass spectrometry / 药学学报

<p><b>AIM</b>To identify anisodine and its metabolites in rat plasma after ingestion of anisodine by combining liquid chromatography and tandem mass spectrometry (LC-MS(n)).</p><p><b>METHODS</b>Plasma samples from rats after a single orally administration of 20 mg anisodine were added with methanol to precipitate protein. Then, it was analyzed by LC-MS(n). Identification and structural elucidation of the metabolites were performed by comparing their changes in molecular masses, retention-times and full scan MS(n) spectra with those of the parent drug and blank plasma.</p><p><b>RESULTS</b>The results revealed that the parent drug and its four metabolites (norscopine, scopine, hydroxyanisodine, N-oxide anisodine) existed in rat plasma.</p><p><b>CONCLUSION</b>This method is sensitive, rapid, simple, and it is suitable for the rapid identification of drug and its metabolits.</p>

Perfil epidemiológico de la intoxicación con burundanga en la clínica Uribe Cualla S.A. de Bogotá, D.C / Epidemic profile of the intoxication with burundanga in the Clínica Uribe Cualla of Bogotá, D. C

Introducción: Las intoxicaciones con intención delictiva constituyen un motivo frecuente de consulta en urgencias. Objetivo: Presentar el perfil epidemiológico de la intoxicación con burundanga y nueva burundanga en un centro de referencia de Bogotá. Material y métodos: Se hizo una revisión de las historias clínicas de los pacientes que consultaron a la clínica Uribe Cualla SA. entre Enero de 1998 y Julio de 2004. Resultados: Se evaluaron las historias de 860 pacientes, se encontró una mayor frecuencia de pacientes en edad reproductiva (20-50 años), del sexo masculino 79,1 por ciento, en quienes el móvil más frecuente fue el robo 67,44 por ciento. Conclusión: Los datos obtenidos en el estudio concuerdan con los de otras instituciones colombianas que reciben esta clase de pacientes. La Clínica Guillermo Uribe Cualla es un centro de referencia para patologías toxicológicas. Conocer las características de los pacientes atendidos en esta Institución permite una buena aproximación al perfil epidemiológico de estas intoxicaciones en nuestro país y con base en estos datos diseñar estudios que permitan estudiar y mejorar los problemas que traen consigo la intoxicación con estas sustancias y sus consecuencias reales sobre la salud de los Colombianos

Pre and post-synaptic muscarinic receptors of the rat vas deferens: an update

Los receptores muscarínicos pre- y post sinápticos del vas deferens de la rata no son M1 ya que el antagonista muscarínico M1-selectivo pirenzepina (PZ) posee baja afinidad por ambos. Basándonos en este hecho las dos acciones de ACh, pre-0y post-sinápticas, en esta preparación parecen ser mediadas por receptores muscarínicos parecidos al subtipo M2. La siguiente serie de observaciones experimentales revelan que ambas respuestas son mediadas por receptores muscarínicos farmacológicamente distintos. El rango de orden de potencia desplegado por 3 antagonistas muscarínicos (Atropina, N-metil-escopolamina [NMS] y PZ) en cada uno de estos lugares son diferentes. Atropina y PZ son bloqueadores selectivos del receptor muscarínico presente en músculo liso. NMS es un antagonista selectivo del receptor pre-sináptico muscarínico que facilita la liberación de norepinefrina. Por último, PZ y NMS despliegan un antagonismo diferencial, siendo competitivos y no-competitivos pre- y post-sinápticamente, respectivamente. Los resultados sugieren que el receptor muscarínico post-sináptico presente en el músculo liso pertenece a los subtipos M2B (oM3). El receptor pre-sináptico pertenece a los subtipos M2A (o M2) o a una subclase de los receptores muscarínicos M2B (o M3)