Two‐photon excited fluorescence lifetime measurements through a double‐clad photonic crystal fiber for tissue micro‐endoscopy

This paper presents an endoscopic configuration for measurements of tissue autofluorescence using two–photon excitation and time‐correlated single photon counting detection through a double‐clad photonic crystal fiber (DC‐PCF) without pre‐chirping of laser pulses. The instrument performance was evaluated by measurements of fluorescent standard dyes, biological fluorophores (collagen and elastin), and tissue specimens (muscle, cartilage, tendon). Current results demonstrate the ability of this system to accurately retrieve the fluorescence decay profile and lifetime of these samples. This simple setup, which offers larger penetration depth than one‐photon‐based techniques, may be combined with morphology‐yielding techniques such as photoacoustic and ultrasound imaging. (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Age-specific birth rate and lifetime reproductive success of chimpanzees at Bossou,Guinea

Preliminary calculations of the age-specific birth rate for wild female chimpanzees were based on demographic records from a long-term study at Bossou, Republic of Guinea. Primiparous age was 12-14 years. Birth rate for a female reached a peak of 0.333 a year at 20-23 years, and then decreased until menopause at more than 40 years of age. Mean birth rate for a female of more than 12 years was 0.197 a year. Mean interbirth interval was 4.6 years overall, but 5.1 years, if all cases in which the previous infant had died within its first three years were excluded. Infant survival rate for the first four years was 0.727 and that of juveniles (four to seven years) was 0.714; however, that of adolescents (eight to 11 years) was only 0.222. Some adolescents who disappeared from Bossou are thought to have migrated to adjacent habitats. From age-specific birth rate and survival rate, the lifetime reproductive success was calculated as only 0.74 offspring per female. This rose to 2.39 when adolescent survival rate was assumed to be the same as that of juveniles. This must be less than the minimum reproductive success required for continued survival of the population. The recent and continuing degradation of the environment will lead to the extermination of this species from the area in the near future. © 1994 Wiley-Liss, Inc.     

Blending effect between n-decane and toluene in oxidation: a ReaxFF study

We studied dependency of toluene oxidation-blended n-decane on blending ratio and temperature using the reactive molecular dynamics (RMD) simulations with the newly developed reactive force field (ReaxFF). Different initial reaction pathways of toluene were observed between pure and blended toluene, while that of n-decane showed little contrast. The differences in toluene oxidation paths are related to radical pool, which is largely influenced by H/C ratio. We analysed the influence of H/C ratio on the consumption of intermediate species, and found different dependencies of HCHO consumption on H/C ratio for different temperatures. The difference is attributed to the large active energy difference between the two main HCHO consumption reactions by OH and O₂. For the production part, the OH producing pathway was analysed carefully and shows H/C ratio influences OH production via H production and H abstract reactions. Our RMD simulations show that H/C ratio plays an important role in the oxidation of fuel.     

Real‐time fiber‐based fluorescence lifetime imaging with synchronous external illumination: A new path for clinical translation

Time‐correlated single photon counting is the “gold‐standard” method for fluorescence lifetime measurements and has demonstrated potential for clinical deployment. However, the translation of the technology into clinic is hindered by the use of ultrasensitive detectors, which make the fluorescence acquisition impractical with bright lighting conditions such as in clinical settings. We address this limitation by interleaving periodic fluorescence detection with synchronous out‐of‐phase externally modulated light source, thus guaranteeing specimen illumination and a fluorescence signal free from bright background light upon temporal separation. Fluorescence lifetime maps are generated in real‐time from single‐point measurements by tracking a reference beam and using the phasor approach. We demonstrate the feasibility and practicality of this technique in a number of biological specimens, including real‐time mapping of degraded articular cartilage. This method is compatible and can be integrated with existing clinical microscopic, endoscopic and robotic modalities, thus offering a new pathway towards label‐free diagnostics and surgical guidance in a number of clinical applications.     

Structural evidence for visual arrestin priming via complexation of phosphoinositols

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GUCY2D mutations in retinal guanylyl cyclase 1 provide biochemical reasons for dominant cone–rod dystrophy but not for stationary night blindness

Mutations in the GUCY2D gene coding for the dimeric human retinal membrane guanylyl cyclase (RetGC) isozyme RetGC1 cause various forms of blindness, ranging from rod dysfunction to rod and cone degeneration. We tested how the mutations causing recessive congenital stationary night blindness (CSNB), recessive Leber''s congenital amaurosis (LCA1), and dominant cone–rod dystrophy-6 (CORD6) affected RetGC1 activity and regulation by RetGC-activating proteins (GCAPs) and retinal degeneration-3 protein (RD3). CSNB mutations R666W, R761W, and L911F, as well as LCA1 mutations R768W and G982VfsX39, disabled RetGC1 activation by human GCAP1, -2, and -3. The R666W and R761W substitutions compromised binding of GCAP1 with RetGC1 in HEK293 cells. In contrast, G982VfsX39 and L911F RetGC1 retained the ability to bind GCAP1 in cyto but failed to effectively bind RD3. R768W RetGC1 did not bind either GCAP1 or RD3. The co-expression of GUCY2D allelic combinations linked to CSNB did not restore RetGC1 activity in vitro. The CORD6 mutation R838S in the RetGC1 dimerization domain strongly dominated the Ca²⁺ sensitivity of cyclase regulation by GCAP1 in RetGC1 heterodimer produced by co-expression of WT and the R838S subunits. It required higher Ca²⁺ concentrations to decelerate GCAP-activated RetGC1 heterodimer—6-fold higher than WT and 2-fold higher than the Ser⁸³⁸-harboring homodimer. The heterodimer was also more resistant than homodimers to inhibition by RD3. The observed biochemical changes can explain the dominant CORD6 blindness and recessive LCA1 blindness, both of which affect rods and cones, but they cannot explain the selective loss of rod function in recessive CSNB.     

Phytochrome and photoperiodic induction

The photoreceptor phytochrome has been extensively characterized at the chromophore, protein and gene level. It consists of a family of red/far-red reversible molecules and the genes for three members have been sequenced. Phytochromes are chromoproteins, which probably exist as dimers in vivo. Photoperiodism in higher plants involves the interaction of phytochrome with an endogenous timekeeping system. The interaction is complex, and several distinct actions of light can be distinguished. The possible involvement of different phytochromes in different actions of light in both long-day plants and short-day plants is discussed. Potential roles for different members of the phytochrome family and homo-and hetero-dimers of phytochrome are proposed.