Femtosecond fluorescence upconversion investigations on the excited-state photophysics of curcumin

The demonstration of curcumin as a photodynamic therapy agent has generated a high level of interest in understanding the photoinduced chemical and physical properties of this naturally occurring, yellow-orange medicinal compound. Important photophysical processes that may be related to photodynamic therapy effects including excited-state intramolecular hydrogen atom transfer (ESIHT) occur within the femtosecond to picosecond time scales. Femtosecond fluorescence upconversion spectroscopy has sufficient time resolution to resolve and investigate these important photophysical processes. In this review, recent advances in using femtosecond fluorescence upconversion to reveal ultrafast salvation and ESIHT of curcumin are presented. The excited-state photophysics of curcumin has been investigated in alcohols and micellar solutions. The results of curcumin in methanol and ethylene glycol reveal the presence of two decay components in the excited-state kinetics with time scales of 12–20 ps and ,100 ps. Similarly, in a micellar solution, biphasic kinetics are present with the fast decay component having a time constant of 3–8 ps, the slow decay component 50–80 ps. Deuteration of curcumin in bothmedia leads to a pronounced isotope effect in the slow decay component, which suggests that ESIHT is an important photophysical process on this time scale. The results of multiwavelength fluorescence upconversion studies show that the fast component in the excited-state kinetics is due to ultrafast solvation. These advances forma part of the continuing efforts to elucidate the photodynamic therapy properties of curcumin.Tak W. Kee, Ramkrishna Adhikary, Philip J. Carlson, Prasun Mukherjee and Jacob W. Petric

Relationship between nicotine tolerance questionnaire scores and plasma cotinine

The Fagerstrom Tolerance Questionnaire (TQ) is often used in both research and treatment contexts to evaluate nicotine tolerance and physiological dependence in cigarette smokers. Recently, however, questions about its validity and its usefulness in comparison to other easily collected measures have been raised. In the present study, 100 male subjects reporting for experimental sessions (Sample I) and 50 male and female subjects entering a smoking cessation clinic program (Sample II) were administered the TQ and determinations of plasma continine during ad libitum smoking were made. TQ scores were found to be correlated with continine levels in both samples, and several of the individual items proved to have statistically significant discriminatory value. Other schemes for determining degree of dependence were considered and found not to be superior to the TQ. Suggestions for further refining the TQ are reviewed.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/28790/1/0000624.pd

First record of the pantropical blue tick Rhipicephalus microplus in Namibia

The invasive pantropical blue tick, Rhipicephalus microplus, has recently been collected from cattle in Namibia. A cross-sectional study aimed at recording the geographic distribution of Rhipicephalus decoloratus and establishing whether R. microplus is present in Namibia was conducted towards the end of summer (March–April) 2013. Ticks were collected from cattle on 18 privately owned farms across a large geographical scale. Ticks were collected from three to five cattle per farm and species belonging to the genera Hyalomma and Rhipicephalus were recovered. Rhipicephalus decoloratus was present on all farms and R. microplus was recorded on four of the farms. The small numbers of R. microplus compared to R. decoloratus collected in the mixed infestations, suggests that the introduction events were recent.Stellenbosch University and the National Research Foundation (IFR2011032500004)http://link.springer.com/journal/10493hb2013ab201

Natural and Engineered Photoactivated Nucleotidyl Cyclases for Optogenetic Applications*

Cyclic nucleotides, cAMP and cGMP, are ubiquitous second messengers that regulate metabolic and behavioral responses in diverse organisms. We describe purification, engineering, and characterization of photoactivated nucleotidyl cyclases that can be used to manipulate cAMP and cGMP levels in vivo. We identified the blaC gene encoding a putative photoactivated adenylyl cyclase in the Beggiatoa sp. PS genome. BlaC contains a BLUF domain involved in blue-light sensing using FAD and a nucleotidyl cyclase domain. The blaC gene was overexpressed in Escherichia coli, and its product was purified. Irradiation of BlaC in vitro resulted in a small red shift in flavin absorbance, typical of BLUF photoreceptors. BlaC had adenylyl cyclase activity that was negligible in the dark and up-regulated by light by 2 orders of magnitude. To convert BlaC into a guanylyl cyclase, we constructed a model of the nucleotidyl cyclase domain and mutagenized several residues predicted to be involved in substrate binding. One triple mutant, designated BlgC, was found to have photoactivated guanylyl cyclase in vitro. Irradiation with blue light of the E. coli cya mutant expressing BlaC or BlgC resulted in the significant increases in cAMP or cGMP synthesis, respectively. BlaC, but not BlgC, restored cAMP-dependent growth of the mutant in the presence of light. Small protein sizes, negligible activities in the dark, high light-to-dark activation ratios, functionality at broad temperature range and physiological pH, as well as utilization of the naturally occurring flavins as chromophores make BlaC and BlgC attractive for optogenetic applications in various animal and microbial models