In vitro cultures of Silybum marianum and silymarin accumulation

AbstractIn this study, a protocol for initiation of callus and shoot cultures from leaves and shoot tips explants of different silybium genotypes collected from different locations in Egypt was established. Callus cultures were initiated from leaves explants and exposed to different concentrations of the precursor (coniferyl alcohol). Shoot cultures were initiated from shoot tips explants. Moreover, the produced plants of the different Silybium shoots as well as intact plants were subjected to protein screening using SDS–PAGE analysis.Results obtained revealed that the optimum medium for growth and maintenance of friable callus was MS medium supplemented with 0.25mgL−1 2,4-Dichlorophenoxy acetic acid (2,4-D)+0.25mg L−1 Kinetin (Kin). The best medium for proliferation of high number of shoots was MS-medium with 0.25mgL−1 each of Benzyl Adinine (BA) and Naphthalene Acetic Acid (NAA). Coniferyl alcohol in concentration of 30μM caused an increase in accumulation of silymarin contents in most callus cultures. SDS–PAGE of different Silybium shoots revealed that the protein profiles of 100% of in vitro produced plantlets similar to their control

Denial of Reward in the Neonate Shapes Sociability and Serotonergic Activity in the Adult Rat

BACKGROUND: Manipulations of the early environment are linked to long-lasting alterations of emotionality and social capabilities. Denial of rewarding mother-pup interactions in early life of rats could serve as model for child neglect. Negative consequences for social competence in later life, accompanied by changes in the serotonergic system would be expected. In contrast, rewarding mother-pup contact should promote adequate social abilities. METHODOLOGY/PRINCIPAL FINDINGS: Male Wistar rats trained in a T-maze during postnatal days 10-13 under denial (DER) or permission (RER) of maternal contact were tested for play behavior in adolescence and for coping with defeat in adulthood. We estimated serotonin (5-HT) levels in the brain under basal conditions and following defeat, as well as serotonin receptor 1A (5-HT1A) and serotonin transporter (SERT) expression. DER rats exhibited increased aggressive-like play behavior in adolescence (i.e. increased nape attacks, p<0.0001) and selected a proactive coping style during defeat in adulthood (higher sum of proactive behaviors: number of attacks, flights, rearings and defensive upright posture; p = 0.011, p<0.05 vs RER, non-handled-NH). In adulthood, they had lower 5-HT levels in both the prefrontal cortex (p<0.05 vs RER) and the amygdala (p<0.05 vs NH), increased 5-HT levels following defeat (PFC p<0.0001) and decreased serotonin turnover (amygdala p = 0.008). The number of 5-HT1A immunopositive cells in the CA1 hippocampal area was increased (p<0.05 DER, vs RER, NH); SERT levels in the amygdala were elevated (p<0.05 vs RER, NH), but were lower in the prefrontal cortex (p<0.05 vs NH). CONCLUSIONS/SIGNIFICANCE: Denial of expected maternal reward early in life negatively affects sociability and the serotonergic system in a complex manner. We propose that our animal model could contribute to the identification of the neurobiological correlates of early neglect effects on social behavior and coping with challenges, but also in parallel with the effects of a rewarding early-life environment

Production of carboxyatractyloside and atractyloside by cell suspension cultures of Atractylis gummifera

Cell suspension cultures were grown from explants of Atractylis gummifera in several culture media. The accumulated biomass differed in the various culture media. The presence of carboxyatractyloside and atractyloside was examined in relation to the different culture media, by means of HPLC. Carboxyatractyloside was identified in the biomass produced in only one of the culture media employed, while atractyloside was not present in detectable amounts in any of the biomasses

Isolation and identification of new cannabinoids in cannabis smoke

Three new cannabinoids, furo[1,2-a] 4-n-pentyl-7,7,10-trimethyl-dibenzopyran 1, 2-methylfuro[1, 2-a] 4-n-pentyl-7,7,10 trimethyl-dibenzopyran 2 and 2,3 dimethylfuro(1,2-a)-4-n-pentyl-7,7,10 trimethyl-dibenzopyran 3 were identified in the sublimate of cannabis resin smoke by spectroscopic methods. Spectral data of dehydrocannabifuran 4 are also presented. © 1983

Production of carboxyatractyloside and atractyloside by cell suspension cultures of Atractylis gummifera

Cell suspension cultures were grown from explants of Atractylis gummifera in several culture media. The accumulated biomass differed in the various culture media. The presence of carboxyatractyloside and atractyloside was examined in relation to the different culture media, by means of HPLC. Carboxyatractyloside was identified in the biomass produced in only one of the culture media employed, while atractyloside was not present in detectable amounts in any of the biomasses

Effects of interaction of an early experience of reward through maternal contact or its denial with social stress during adolescence on the serotonergic system and the stress responsiveness of adult female rats

Experiences during critical periods, such as the neonatal and adolescence, play a critical role in determining adult stress-coping behavior. Based on the aforementioned we developed an experimental protocol, which included a neonatal experience and a social stress during adolescence. The serotonergic system is known as an important modulator of coping ability and, in general, emotional balance in both normal and pathological states, such as depression and anxiety, for which females are more vulnerable. Thus in the present work we used female rats and determined 5-HT, 5-hydroxyindoleacetic acid (5-HIAA), and 5-hydroxytryptamine receptor type 1A (5-HT1A) receptor levels in the prefrontal cortex (PFC) and the amygdala (AMY). During postnatal days 10-13 (PND 10-13) rat pups were exposed to a T-maze, one arm of which lead to the mother. One group of animals was allowed contact with the mother (rewarded-receiving expected reward (RER)), whereas the other was denied the expected reward (DER). High performance liquid chromatography (HPLC) analysis revealed that in both the PFC and in AMY, adult RER animals had higher basal 5-HT levels. Furthermore, in the AMY of this group of animals, higher levels of 5-HT1A receptors were detected by Western blot analysis. In adulthood rats were exposed to the Forced Swimming Test/Stress (FST/S). RER animals not exposed to the adolescent stress exhibited longer immobility time during both the first and second day of FST. Corticosterone levels following the FST fell faster in the DER animals. Adolescent stress affected the responses to the adult FSS only in the DER animals, which had decreased 5-HT in the AMY and increased immobility time on both days of the FST, compared with the DER, not stressed in adolescence. The phenotype of the DER animals is in line with the &quot;match-mismatch&quot; hypothesis, which states that if two events during critical periods of life &quot;match&quot; in being mildly stressful, their interaction can be adaptive. © 2012 IBRO