Micropropagation of Salvia wagneriana Polak and hairy root cultures with rosmarinic acid production

Salvia wagneriana Polak is a tropical species native to Central America, well adapted to grow in the Mediterranean basin for garden decoration. Micropropagation has been assessed from axillary shoots of adult plants using a Murashige and Skoog basal medium, with the addition of 1.33-μM 6-benzylaminopurine for shoot proliferation; the subsequent rooting phase occurred in plant growth regulator-free medium. The plants were successfully acclimatised with high survival frequency. Hairy roots were induced after co-cultivation of leaf lamina and petiole fragments with Agrobacterium rhizogenes and confirmed by PCR. The establishment and proliferation of the selected HRD3 line were obtained in hormone-free liquid medium and the production of rosmarinic acid (RA) was evaluated after elicitation. The analysis of RA was performed by LC-ESI-DAD-MS in the hydroalcoholic extracts. The addition of casein hydrolysate increased the RA production, whereas no enrichment was observed after the elicitation with jasmonic acid

Production of Curcuminoids in different in vitro organs of Curcuma longa.

Curcuma longa L. (turmeric) is one of the most important spice and safe food additives. Its main constituents, curcuminoids, showed anti-inflammatory, antitumor and antioxidant effects. In the present work, an in vitro propagation method was developed to achieve selected plant organs with quantified curcuminoid content. I n vitro plants were obtained from sprouting buds as primary explants. The major curcuminoid constituents, such as curcumin (CUR), demethoxycurcumin (DEM), and bis-demethoxycurcumin (bis-DEM) were examined in different organs by LC-DAD-ESI-MS. A significant production of curcumin (more than 260 μg g−1fresh weight) was obtained from in vitro microrhizomes, especially grown in a Murashige and Skoog medium (MS) supplemented with kinetin (0.1 mg L−1), α-naphthaleneacetic acid (NAA, 1 mg L−1), sucrose (6%), agar (5%) and activated charcoal (0.1%). The analyzed microrhizomes showed reduced amounts of DEM and bis-DEM in comparison with CUR levels. In addition a shoot culture line was suitable to biosynthesize curcuminoids, in a ratio very similar to that identified in the fresh rhizomes of parent plants. This study represents the first direct quantification of curcuminoids in turmeric in vitro shoots and microrhizomes to be used in dietary supplements

Daidzein Production and HeLa Cytotoxicity of Bituminaria bituminosa hairy root Cultures

Bituminaria bituminosa (L.) C.H. Stirt is a perennial species widely distributed in the Mediterranean basin and the Canary Islands. This species is used in folk medicine and currently has considerable pharmaceutical interest for its content in phenylpropanoids, furanocoumarins and pterocarpans. In vitro cultures (shoots and hairy roots) have been performed to obtain plant material useful for the production of these metabolites. Hairy root cultures were successfully established after inoculation of hypocotyls with the LBA 9402 A. rhizogenes strain. The HRPB3 line was selected for further analysis and elicited with chitosan and salicylic acid. All the HRPB3 cultures showed higher polyphenol content and greater DPPH-antioxidant activity than shoots cultured in vitro. The presence of isoflavone daidzein was detected in the hairy root extracts. The cytotoxic effect of HR extracts has been further tested on HeLa cells: the salicylic acid elicited HR exhibited good antiproliferative effects

Physiological and Biochemical Adaptive Traits in Leaves of Four Citrus Species Grown in an Italian Charterhouse

Citrus trees are a very important crops that are cultivated worldwide, but not much knowledge is known about the ecophysiological responses to climatic changes in trees under natural conditions. The aim of this study was to investigate their adaptive capacity in response to seasonal phenological and environmental changes. The trial included Citrus trees (sweet orange, bitter orange, lemon, mandarin) growing under non-regular cropping conditions in a Monumental Charterhouse in Tuscany, in a subtropical Mediterranean climate with hot summer conditions. During a 1-year field trial, we determined the variations in chlorophyll fluorescence parameters and leaf biochemical traits (content of chlorophylls and carotenoids, total phenolic content (TPC), total antioxidant capacity (TAC), and total non-structural carbohydrates). In all Citrus spp., interspecific mean values of photochemical efficiency peaked during the summer, while a marked photoinhibition occurred in the winter in concomitance with higher interspecific mean values of leaf TPC, TAC, and non-structural carbohydrates. The trees showed the pivotal role played by photosynthetic acclimation as a survival strategy to tolerate abiotic stress in the climate change hotspot of Mediterranean environment. This study is included in a wider project aimed at a new valorization of Citrus trees as genetic resource and its by-products with added-value applications for innovative functional foods

Fructan metabolism in tall fescue calli under different environmental condition

Several grasses usually accumulate high levels of fructans at low temperature or in the presence of high sugar supply. Fructans enhance plant resistance in adverse environmental conditions thanks to their vacuolar  localization which allow osmotic adjustments. Tall fescue calli do not show mature vacuoles due to its  anatomic characteristics, and these in vitro cultures can provide more knowledge about the fructan rule in  unspecialized tissues especially during stress conditions. The aim of this study was to evaluate the sugar  metabolism in embryogenic calli of tall fescue fed with different sucrose concentrations (0, 50 or 100 mM)  during growth culture, kept at “control” or “cold” temperatures (23 or 4°C, respectively). In the presence of  sucrose (50 or 100 mM), embryogenic calli accumulated fructans under control conditions. On the other hand,  the accumulation is inhibited by the simultaneously sugar supply and cold treatment while invertase activity  increased. From our data, cold temperature and high sucrose concentration seemed to be negatively  correlated in the fructans biosynthetic pathway in tall fescue in vitro cultures.Key words: Cold, Festuca arundinacea, invertase, sucrose:sucrose fructosyltransferase (SST), sugars

Nicotine dependence and psychological distress: outcomes and clinical implications in smoking cessation

Nicotine dependence is characteristically a chronic and relapsing disease. Although 75%–85% of smokers would like to quit, and one-third make at least three serious lifetime attempts, less than 50% of smokers succeed in stopping before the age of 60. Relevant and complex factors contributing to sustained cigarette consumption, and strongly implicated in the clinical management of smokers, are the level of nicotine dependence and psychological distress. In this review of the literature, these two factors will be examined in detail to show how they may affect smoking cessation outcome and to encourage clinicians to assess patients so they can offer tailored support in quitting smoking

Depressive, anxious, withdrawal symptoms, and craving as possible predictors of abstinence maintenance in smokers attending a 12-week quitting program

BACKGROUND INFORMATION: Depressive, anxious, withdrawal symptoms, and craving might affect differently the probability to maintain abstinence after quitting smoking. OBJECTIVE: The objective of this study was to assess depressive, anxious, nicotine withdrawal symptoms, and craving in a sample of smokers attending a smoking cessation program over a period of 12 weeks. METHODS: A naturalistic study was conducted in which 78 smokers were consecutively recruited for a 12 week evaluation program. Socio-demographic data and clinical information were collected, rating scales were used to assess anxious and depressive symptoms, nicotine dependence, withdrawal symptoms, and craving. RESULTS: Of the 78 recruited smokers, 17 remained abstinent and 61 reverted to smoking during the period of 12 weeks. The probability of maintaining abstinence was increased when low depressive symptoms or low craving occurred during the cessation program. CONCLUSION: The present results strengthen the importance of assessing depressive symptoms and craving over the follow-up of a physician-assisted smoking cessation program to detect abstaining smokers at risk to relapse

Epidemiology of chronic obstructive pulmonary disease: Health effects of air pollution

Abstract:  COPD is one of the leading causes of morbidity and mortality in the industrialized and the developing countries. According to the prediction of the World Health Organization, COPD will become the third leading cause of mortality and the fifth cause of disability in 2020 worldwide. In epidemiology, distinct phenotypic entities converge on the term COPD, so that prevalence and mortality data may be inclusive of chronic bronchitis, emphysema and asthma; moreover, the assessment of prevalence rates may change considerably according to the diagnostic tools used. Thus, a considerable problem is to estimate the real prevalence of COPD in the general population. COPD is determined by the action of a number of various risk factors, among which, the most important is cigarette smoking. However, during the last few decades, evidence from epidemiological studies finding consistent associations between air pollution and various outcomes (respiratory symptoms, reduced lung function, chronic bronchitis and mortality), has suggested that outdoor air pollution is a contributing cause of morbidity and mortality. In conclusion, epidemiological studies suggest that air pollution plays a remarkable role in the exacerbation and in the pathogenesis of chronic respiratory diseases. Thus, respiratory physicians, as well as public health professionals, should advocate for a cleaner environment

Establishment of highly efficient Agrobacterium rhizogenes-mediated transformation for Stevia Rebaudiana Bertoni explants

Leaves and internodes from Stevia rebaudiana Bertoni plants growing in different conditions were used for transformation with two strains of Agrobacterium rhizogenes: ATCC 15384 and LBA 9402. Hairy roots formation was observed and the percentage of the transformed explants depended on the type of explant, time of inoculation and inoculum concentration.Inoculation of explants from ex vitro and in vitro plants with LBA9402 strain led to higher efficiency of transformation than inoculation with ATCC 15384 strain. Growth rate of hairy roots in liquid culture was assessed under light and dark conditions. It was found that the growth of hairy roots decreased significantly under light conditions. Transformation of hairy roots growing in different culture conditions was confirmed at the molecular level using PCR method with primers constructed against rolB and rolC genes from A rhizogenes. Growth rate of hairy roots in liquid culture was assessed under light and dark conditions. It was found that the growth of hairy roots decre ased significantly under light conditions . Transformation of hairy roots growing in different culture conditions was confirmed at the molecular level using PCR method with primers constructed against rolB and rolC genes from A . rhizogene