Ilex paraguariensis: the effect of genotypes and growth phase on biomass, secondary metabolism and antioxidant activity of in vitro cultivated calli / Ilex paraguariensis: el efecto de los genotipos y la fase de crecimiento sobre la biomasa, metabolismo secundario y la actividad antioxidante de los callos cultivados in vitro

Yerba mate ( Ilex paraguariensis ) produces several secondary metabolites of interest to the phar maceutical industry, such as chlorogenic acids and methylxanthines. These compounds have been produced in vitro by callus culture from different species. However, for I. paraguariensis , no studies upon the production of these compounds in vitro have been p erformed to date. In this work, we show that the concentration of secondary metabolites from I. paraguariensis callus is possible and highly dependent on the callus growth phase. We observed that the best phase for the production of secondary compounds in calli of yerba mate is the stationary growth phase on both genotypes tested. In this phase, higher levels of phenolic compounds, chlorogenic acid and 3,5 - dicaffeoylquinic acid and greater antioxidant activity were observed. Chlorogenic acid and 3,5 - dicaffe oylquinic acid presented positive correlation with antioxidant activity. For the first time, secondary compounds were reported in yerba mate calli cultivated in vitro .

La yerba mate ( Ilex paraguariensis ) produce varios metabolitos secundarios de interés para la industria farmacéutica, como los ácidos clorogénicos y las metilxantinas. Estos compuestos se han producido in vitro mediante cultivo de ca llos de diferentes especies. Sin embargo, para I. paraguariensis , hasta la fecha no se han realizado estudios sobre la producción de estos compuestos in vitro . En este trabajo, mostramos que la concentración de metabolitos secundarios desde callos de I. pa raguariensis es posible y altamente dependiente de la fase de crecimiento del callo. Observamos que la mejor fase para la producción de compuestos secundarios en callos de yerba mate es la fase de crecimiento estacionario en ambos genotipos probados. En es ta fase se observaron niveles más altos de compuestos fenólicos, ácido clorogénico y ácido 3,5 - dicafeoilquínico y una mayor actividad antioxidante. El ácido clorogénico y el ácido 3,5 - dicafeoilquínico presentaron correlación positiva con la actividad antio xidante. Por primera vez, se reportaron compuestos secundarios en callos de yerba mate cultivados in vitro .

Development of Physiologically Based Pharmacokinetic Model and Assessment of the Impact of Renal Underdevelopment in Preterm Infants on the Pharmacokinetics of Aminophylline

Objective:To develop a physiologically based pharmacokinetic (PBPK) model for individualization of the dosing regimen considering the physiological requirements of these preterm neonates. Methods: The study comprised preterm newborns with fewer than 34 weeks of gestation and six apneic episodes in 24 h. A PBPK model was created using PK-SIM (version 9, update 1, GitHub, San Francisco, CA, USA). A PBPK model is built using a typical loading dosage of 5 mg/kg and a maintenance dose of 1.5 mg/kg. Based on the verified base model, a PBPK model representing renal underdevelopment based on nRIFLE/pRIFLE categorization was developed. Results: The PK parameters of Aminophylline were computed using the PBPK model. As per the model prediction, T1/2 and area under the curve reduced as postnatal age increased, and in the event of renal underdevelopment, even while Cmax for patients under R (RISK), I (injury) was within the therapeutic range; it was greater compared to preterm without any renal complications. Mean Cmax (mol/L) was 59.53 and for R, I, and F (FAILURE) categories the values were 83.04, 99.69, and 126.98, respectively. Conclusion: The model was created using appropriate drug, study subject, and dosage protocol inputs. The established PBPK model could help in individualizing aminophylline dose in preterm babies.

Analysis of Chemical Constituents in Paullinia cupana Dried Seeds by UPLC-Q-TOF-MS / 中国实验方剂学杂志

Objective:The chemical constituents in guarana （<italic>Paullinia cupana</italic> dried seeds） were systematically analyzed to provide a basis for further research， development and utilization of this plant. Method:The contents of crude protein， crude fat， crude polysaccharide and crude fiber in guarana were determined according to national standards and related documents， and the chemical constituents of guarana was qualitatively analyzed by ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF-MS）， ACQUITY UPLC-HSS-T3 column （2.1 mm×100 mm， 1.8 μm） was used with 0.1% formic acid aqueous solution （A）-0.1% formic acid acetonitrile solution （B） as mobile phase for gradient elution （0-5 min， 2%-10%B； 5-6 min， 10%-20%B； 6-9 min， 20%-30%B； 9-9.5 min， 30%-35%B； 9.5-10.5 min， 35%-45%B； 10.5~13 min， 45%-55%B； 13-15 min， 55%-80%B； 15-19 min， 80%-98%B； 19-20 min， 98%B； 20-20.3 min， 98%-2%B； 20.3-23 min， 2%B）， the electrospray ionization （ESI） was used for detection in positive and negative ion modes， the scanning range was <italic>m</italic>/<italic>z</italic> 50-1 500， and the structure was identified according to the relative molecular weight and fragment information combined with database matching and comparison of reference substances. Result:The contents crude protein， crude fat， crude polysaccharide and crude fiber in guarana were （0.63±0.03）%， （2.73±0.09）%， （3.23±0.12）% and （8.89±0.59）%， respectively. A total of 42 chemical constituents in guarana were identified by UPLC-Q-TOF-MS， including 3 methylxanthines， 2 nucleosides， 1 amino acid， 3 organic acids， 33 flavonoids， 3 （<italic>L</italic>-tryptophan， epigallocatechin gallate， daidzein） of which were first discovered in guarana. Conclusion:Guarana is rich in nutrients and has good potential to be developed as a functional food. UPLC-Q-TOF-MS technique provides a simple， rapid and accurate method for the identification of chemical constituents in guarana. Methylxanthines and proanthocyanidins are the main chemical constituents of guarana， which is meaningful for quality evaluation and material basis of guarana.

A comparative study of efficacy and tolerability of doxofylline over other methylxanthenes in mild to moderate persistent bronchial asthma patients in a tertiary care hospital

Background: Studies have shown that doxofylline is endowed with a remarkable bronchodilator activity with less extra respiratory effects than other methylxanthines. To compare the efficacy and tolerability of doxofylline over other methylxanthines in mild to moderate persistent bronchial Asthma patients.Methods: It is a comparative study conducted in patients of mild to moderate persistent bronchial Asthma in the outpatient department of regional chest hospital. The subjects were randomly assigned into 2 groups, one group treated with oral doxofylline and the other with theophylline oral or Deriphylline. Efficacy was measured objectively using spirometric parameters like FEV1 (Forced Expiratory Volume at the end of 1 second), FVC (Forced Vital Capacity), FEV1/FVC and PEFR (Peak Expiratory flow Rate), Subjectively by the Asthma control test questionnaire, subjective rating of Asthma control and the need for use of rescue medications like β2 agonists in the previous four weeks. Tolerability were done at base line and at the end of study.Results: Doxofylline compared to other methylxanthines groups was statistically better with respect to subjective parameters like Asthma control tests questionnaire, subjective rating of Asthma control. Doxofylline had equal efficacy with spirometric parameters compared to other methylxanthines. Doxofylline was significantly better tolerated with adverse drug reactions noted in 10% compared to 28% in other methylxanthins group.Conclusions: Doxofylline with better tolerability profile and equivalent efficacy seemed to be a good alternative to other methylxanthines in the treatment of bronchial Asthma.

Alcaloides y polifenoles del cacao, mecanismos que regulan su biosíntesis y sus implicaciones en el sabor y aroma / Cacao alkaloids and polyphenols: Mechanisms that regulate their biosynthesis and its implications on the taste and aroma

El sabor y aroma de los granos de cacao (Theobroma cacao L.) fueron las principales razones que promovieron su domesticación y uso alimentario por los pueblos precolombinos de Mesoamérica. Incluso hoy día, la calidad sensorial determina la clasificación entre cacaos finos y a granel. Muchos compuestos químicos de las almendras son responsables de la calidad sensorial, pero sobresalen los polifenoles y los alcaloides, compuestos que de manera directa inciden en el sabor y palatabilidad de las almendras y de manera indirecta sobre los precursores de aroma. Los alcaloides están asociados con el amargor. Su concentración está relacionada con la variedad y se modifica con el procesamiento. Los polifenoles son responsables, junto con otras moléculas de la astringencia (poco deseable en chocolates), pero también de propiedades antioxidantes deseables por los consumidores. En esta revisión se abordan aspectos de la biosíntesis de estas importantes moléculas en las almendras de cacao, de las implicaciones en el sabor y aroma, así como los cambios que ocurren durante el procesamiento de las mismas(AU)

The flavor and aroma of cacao (Theobroma cacao) beans were the main reasons that promoted its domestication and food-use by pre-Columbian peoples of Mesoamerica. Polyphenols and alkaloids are compounds that directly affect the flavor of the cocoa beans and indirectly on the flavor precursors. The alkaloids are associated with bitterness; its concentration is related to the cultivar and its modifying through the processing. Polyphenols molecules are responsible together with other molecules of the astringency (not desirable in chocolate), but also are responsible for antioxidant properties, very desirable by consumers. This review focuses on aspects of the biosynthesis of these important molecules in cocoa beans as well as implications in taste and flavor. The changes of these molecules that occur during processing are also approached(AU)

The influence of leaf age on methylxanthines, total phenolic content, and free radical scavenging capacity of Ilex paraguariensis aqueous extracts

Yerba-mate (Ilex paraguariensis A. St. Hil., Aquifoliaceae) is a South American native species that is widely used for its industrial potential in the preparation of drinks, teas and cosmetics. Its properties are directly related to the presence of its chemical constituents, such as saponins, methylxanthines and phenolic compounds. This study aimed to investigate the influence of leaf age on methylxanthine and total phenolic contents by High Performance Liquid Chromatography and Ultraviolet Spectroscopy, as well as on free radical scavenging capacity, of aqueous extracts of I. paraguariensis leaves. The results showed great variability in all the metabolites measured. Leaf ageing significantly increased the methylxanthine content and total phenolic content of the extracts. Free radical scavenging capacity was also significantly affected (p < 0.05) by leaf age. A positive correlation was observed, between the antioxidant activity and total phenolic content.

Efecto de la fermentación sobre la actividad antioxidante de diferentes clones de cacao colombiano / Effect of fermentation on the antioxidant activity of different Colombian cocoa clones

Introducción: el cacao es una de las principales fuentes de polifenoles. Estos compuestos se encuentran relacionados con la actividad antioxidante y las características organolépticas del cacao y sus subproductos. Durante el procesamiento, los granos de cacao son sometidos a diferentes etapas que pueden afectar el contenido de esos metabolitos, pero que son cruciales para el desarrollo de la calidad organoléptica. Una de estas etapas es la fermentación. Objetivo: evaluar el efecto de la fermentación sobre el contenido de metabolitos secundarios y la actividad antioxidante en 5 clones de cacao cultivados en Colombia. Métodos: en los clones estudiados se determinó el contenido de fenoles totales, antocianinas totales y taninos condensados, mediante métodos espectrofotométricos; así como catequina, epicatequina, teobromina y cafeína por cromatografía líquida de alta resolución. La capacidad antioxidante se evaluó mediante las metodologías del DPPH. (1,1-diphenyl-2-picrylhydrazyl), FRAP (ferric reducing antioxidant power), ORAC (oxygen radical antioxidant capacity) y la capacidad atrapadora de radicales superóxido. Resultados: el efecto de la fermentación sobre los clones de cacao no fue uniforme, observándose tanto cambios positivos como negativos en los contenidos de los diversos metabolitos secundarios y la actividad antioxidante, en dependencia de la variedad. Sin embargo, los cambios en la actividad antioxidante expresada como TEAC (trolox equivalent antioxidant capacity) DPPH están correlacionados con los diversos cambios en el contenido de fenoles totales durante el proceso de fermentación y descritos por la expresión siguiente: DDPPH= 6,36099*Dfenoles+5,10923, con r²= 0,982. Conclusiones: la fermentación afecta el potencial antioxidante de los clones de cacao, lo cual es importante para el desarrollo de las propiedades organolépticas de los productos finales.

Introduction: cocoa is one of the main sources of polyphenols. These compounds are related with the antioxidant activity and sensory characteristics of cocoa and its sub-products. During processing, cocoa beans are subjected to different stages, which may affect the content of these metabolites. However, these processes are crucial in the development of organoleptic quality of cocoa beans. One of these stages is the fermentation. Objective: to evaluate the effect of fermentation on the content of secondary metabolites and the antioxidant activity in five Colombian cocoa clones. Methods: the total phenol content, total anthocyanins and condensate tannins were determined for the studied clones by spectrophotometric methods. Additionally, the content of catechin, epicatechin, theobromine and caffeine were determined by high-performance liquid chromatography (HPLC). The antioxidant capacity of cocoa clones was determined by the methods DPPH. (1,1-diphenyl-2-picrylhydrazyl), FRAP (ferric reducing antioxidant power), ORAC (oxygen radical absorbance capacity) and superoxide radical scavenging capacity. Results: the effect of fermentation on cocoa clones was not uniform. Both positive and negative changes in the contents of various secondary metabolites and antioxidant activity were observed depending on the variety. However, changes in antioxidant activity expressed as TEAC (trolox equivalent antioxidant capacity) DPPH were correlated with the changes in the total phenol content during fermentation and described by the following expression: DDPPH= 6.36099* Dphenols + 5.10923, with r² = 0.982. Conclusions: fermentation affects the antioxidant potential of cocoa clones, which is important for the development of organoleptic properties of final products.

Comparative evaluation of doxofylline and theophylline in patients of mild bronchial asthma.

Background: Asthma is common chronic disease worldwide. Methylxanthines has been used in the treatment of asthma. The study was undertaken to compare two Methylxanthines theophylline and doxofylline at doses recommended and commonly used in clinical practice in Mild Bronchial Asthma Patients. Methods: Study was conducted in patients of Mild Bronchial Asthma in TB and chest disease department of a medical college hospital. It was randomized, prospective and open label. A total of 107 patients were divided in two group .Group I was administered 400 mg theophylline SR once daily and group II was administered doxofylline 400 mg twice a day orally. Spirometric variables symptom score, and adverse effects were recorded on day 0, 7 and 21 of therapy. Data were compared and analysed using SPSS version 16. Results: Results of the study showed that there was significant improvement in spirometric variables and clinical symptom score compared to pretreatment values after medication in both groups on 7th and 21st days of treatment. But there was no statistically significant difference between improvement in theophylline and doxofylline groups with respect to spirometric variables and symptom score. There was no significant difference in two groups with respect to side effects (p>0.05). Conclusions: It is concluded in Patients of mild Bronchial Asthma Theophylline and doxofylline improve the spirometric and clinical symptoms and doxofylline has no advantage over theophylline in terms of either efficacy or safety on the doses commonly used in current clinical practice.

Characterization of rat heart alkaline phosphatase isoenzymes and modulation of activity

Alkaline phosphatase (ALP) is important in calcification and its expression seems to be associated with the inflammatory process. We investigated the in vitro acute effects of compounds used for the prevention or treatment of cardiovascular diseases on total ALP activity from male Wistar rat heart homogenate. ALP activity was determined by quantifying, at 410 nm, the p-nitrophenol released from p-nitrophenylphosphate (substrate in Tris buffer, pH 10.4). Using specific inhibitors of ALP activity and the reverse transcription-polymerase chain reaction, we showed that the rat heart had high ALP activity (31.73 ± 3.43 nmol p-nitrophenol·mg protein-1·min-1): mainly tissue-nonspecific ALP but also tissue-specific intestinal ALP type II. Both ALP isoenzymes presented myocardial localization (striated pattern) by immunofluorescence. ALP was inhibited a) strongly by 0.5 mM levamisole, 2 mM theophylline and 2 mM aspirin (91, 77 and 84 percent, respectively) and b) less strongly by 2 mM L-phenylalanine, 100 mL polyphenol-rich beverages and 0.5 mM progesterone (24, 21 to 29 and 11 percent, respectively). â-estradiol and caffeine (0.5 and 2 mM) had no effect; 0.5 mM simvastatin and 2 mM atenolol activated ALP (32 and 36 percent, respectively). Propranolol (2 mM) tended to activate ALP activity and corticosterone activated (18 percent) and inhibited (13 percent) (0.5 and 2 mM, respectively). We report, for the first time, that the rat heart expresses intestinal ALP type II and has high total ALP activity. ALP activity was inhibited by compounds used in the prevention of cardiovascular pathology. ALP manipulation in vivo may constitute an additional target for intervention in cardiovascular diseases.