RESUMEN
Procedure for isolation of pyrrolizidine alkaloids (PAs) from Rindera umbellata Bunge plant species was optimised. Different extraction media (methanol, ethanol and sulphuric acid), concentration and volume of sulphuric acid, pH of PA solution for alkaline extraction, extraction time and techniques (maceration, ultrasonic and overhead rotary mixer assisted extraction) were investigated. The yields of six PAs (7-angeloyl heliotridane, 7-angeloyl heliotridine, lindelofine, 7-angeloyl rinderine, punctanecine and heliosupine) were monitored by GC-MS/FID. The best results for the isolation all of six PAs were obtained when the extraction was performed with 1 M sulphuric acid (30 mL per 1.00 g of dried sample) by overhead rotary mixer during three days. Optimal pH value for alkaline extraction of PAs with CH2Cl2 was 9, and the extraction should be performed with four portions of 30 mL of CH2Cl2. This procedure could be also useful for a plant sample preparation for GC and LC analyses of PAs.
Asunto(s)
Boraginaceae/química , Fraccionamiento Químico/métodos , Alcaloides de Pirrolicidina/aislamiento & purificación , SolventesRESUMEN
Preoperative diagnosis of papillary thyroid carcinoma (PTC) comprises numerous diagnostic procedures which are mostly applicable in tertiary institutions. Normal thyroid function depends on the presence of many trace elements and copper (Cu) and zinc (Zn) are some of those. The study is based on retrospective review of 118 patients with preoperatively diagnosed benign thyroid disease (BTD) and 12 with PTC, who underwent thyroid surgery at the Center for Endocrine Surgery Clinical Center of Serbia, Belgrade, between 2010 and 2012. The objective was to evaluate concentrations of Cu and Zn in serum as possible prediction markers for PTC in patients who underwent surgery for preoperatively diagnosed BTD. Concentrations of Cu and Zn ions in serum were measured using atomic absorption spectrophotometer. Data were analyzed using methods of descriptive statistics, Anova and t-test (p < 0.05 was considered statistically significant). Definitive pathohistological findings revealed PTC in 23 (19.5%) and papillary microcarcinoma-mPTC in 13 (11.0%) of BTD patients. The concentrations of Cu ions in serum of PTC patients as well as in serum of patients with mPTC were significantly higher than in serum of BTD patients (p < 0.05). The concentrations of Zn ions and Cu/Zn ratio in serum of PTC and mPTC patients were not significantly higher than in serum of BTD patients. The concentration of Cu ions in serum of patients before thyroid surgery can be useful, easy available, and a low-cost tool in prediction of preoperatively undiagnosed PTC in patients with BTD.
Asunto(s)
Carcinoma Papilar/sangre , Carcinoma Papilar/diagnóstico , Carcinoma/sangre , Carcinoma/diagnóstico , Cobre/sangre , Neoplasias de la Tiroides/sangre , Neoplasias de la Tiroides/diagnóstico , Carcinoma/cirugía , Carcinoma Papilar/cirugía , Femenino , Humanos , Masculino , Cáncer Papilar Tiroideo , Glándula Tiroides/metabolismo , Glándula Tiroides/patología , Neoplasias de la Tiroides/cirugía , Zinc/sangreRESUMEN
The examination of the aerial parts, roots, and seeds of the endemic plant Rindera umbellata is reported in this paper for the first time. Phytochemical investigation of R. umbellata led to the isolation and characterization of ten pyrrolizidine alkaloids and eleven fatty acids in the form of triglycerides. Pyrrolizidine alkaloids 1-9 were found in the aerial parts, 7 and 8 in the roots, and 6-10, together with eleven fatty acids, in the seeds of this plant species. The structures of compounds 1-10 were established based on spectroscopic studies (¹H- and ¹³C-NMR, 2D NMR, IR and CI-MS). After trans-esterification, methyl esters of the fatty acids were analyzed using GC-MS. The effect of lindelofine-N-oxide (7) on tubulin polymerization was determined.
Asunto(s)
Boraginaceae/química , Ácidos Grasos/química , Extractos Vegetales/química , Alcaloides de Pirrolicidina/química , Semillas/química , Moduladores de Tubulina/química , Tubulina (Proteína)/química , Animales , Antineoplásicos/química , Bovinos , Ácidos Grasos/aislamiento & purificación , Componentes Aéreos de las Plantas/química , Raíces de Plantas/química , Polimerizacion , Multimerización de Proteína/efectos de los fármacos , Alcaloides de Pirrolicidina/aislamiento & purificaciónRESUMEN
The most common histological variants of papillary thyroid carcinoma (PTC), classical (CPTC) and follicular (FPTC), have different diagnostic features, molecular biology, and prognosis. Matrix metalloproteinase-9 (MMP-9) endopeptidase which degrades the components of the extracellular matrix is essential in the invasive growth and metastasizing of malignant tumors. The serum copper (Cu)/zinc (Zn) ratios are sensitive diagnostic and prognostic indicators in oncology since Cu- and Zn-dependent enzymes play important roles in the genesis and the progression of tumors. The aim of this study was to examine the expressions of MMP-9 in tissues of CPTC and FPTC, as well as to determine the Cu/Zn ratios in the same samples. MMP-9 was determined immunohistochemically, and the concentrations of copper and zinc in thyroid tissue were determined by means of flame atomic absorption spectrometry. The results obtained revealed significantly higher expressions of MMP-9 in CPTC in comparison with FPTC, as well as higher Cu/Zn ratios in CPTC than in FPTC. Thus, determining MMP-9 activities and the Cu/Zn ratios could improve the accuracy of the standard histopathological diagnosis of these two types of PTC.
Asunto(s)
Carcinoma Papilar Folicular/diagnóstico , Carcinoma/diagnóstico , Cobre/sangre , Metaloproteinasa 9 de la Matriz/metabolismo , Neoplasias de la Tiroides/diagnóstico , Zinc/sangre , Adolescente , Adulto , Anciano , Carcinoma/sangre , Carcinoma/enzimología , Carcinoma Papilar , Carcinoma Papilar Folicular/sangre , Carcinoma Papilar Folicular/enzimología , Diagnóstico Diferencial , Detección Precoz del Cáncer/métodos , Femenino , Humanos , Inmunohistoquímica/métodos , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias , Pronóstico , Espectrofotometría Atómica , Cáncer Papilar Tiroideo , Glándula Tiroides/enzimología , Neoplasias de la Tiroides/sangre , Neoplasias de la Tiroides/enzimología , Tiroidectomía , Adulto JovenRESUMEN
Natural monocrystalline pyrite as a new indicator electrode for the potentiometric titration of weak acids in acetonitrile, propionitrile and benzonitrile was studied. The investigated electrode showed a linear dynamic response for p-toluenesulfonic acid concentrations in the range from 0.1 to 0.001 M, with a Nernstian slope of 74 mV per decade. Sodium methylate, potassium hydroxide and tetrabutylammonium hydroxide (TBAH) proved to be very suitable titrating agent for this titration. The response time was less than (11 s) and the lifetime of the electrode is long. The advantages of the electrode are log-term stability, fast response, and reproducibility, while the sensor is easy to prepare and of low cost.
Asunto(s)
Acetonitrilos/química , Ácidos/química , Hierro/química , Minerales/química , Nitrilos/química , Sulfuros/química , Bencenosulfonatos/análisis , Electroquímica , Electrodos , Potenciometría , Sensibilidad y Especificidad , Soluciones , Factores de TiempoRESUMEN
Pancuronium bromide (PCBr) inhibition effect on enzyme cholinesterase from pooled human serum (Che, EC 3.1.1.8 acylcholine acylhydrolase) was used for development of a spectrophotometric kinetic method for PCBr determination in human serum and urine. Optimal conditions for the basic and inhibitor reactions were established: pH=7.7 and substrate concentration c(benzoylcholine chloride)=1.33 mmol/L. Kinetic parameters were also determined: Michaelis-Menten's constant K(M)=0.40 mmol/L, maximal reaction rate V(max)=52.2 micromol/L min, inhibition constant K(i)=0,56 micromol/L and IC(50)=1.31 micromol/L. Linear dependence between the reaction rate and inhibitor concentration exists in PCBr concentration range 8.20-68.25 nmol/L, which corresponds to the real sample concentrations from 0.328 to 2.730 micromol/L. The method detection and quantification limits were 2.01 nmol/L and 6.67 nmol/L, respectively. Precision of the method was tested for three pancuronium concentrations (10.70, 29.35 and 51.25 nmol/L). Relative standard deviation (RSD) was in the range 0.15-7.45%. Accuracy was examined by standard addition method. Influence of the substances usually present in serum and urine on the reaction rate was tested. The developed method was applied for PCBr content determination in serum model samples, urine model samples and in urine taken during surgery. The method has good sensitivity, accuracy, precision and it is suitable for clinical practice.