This paper describes an integrated setup for fluorescence recovery after photobleaching (FRAP) for determining translational and rotational Brownian diffusion simultaneously, ensuring that these two quantities are measured under exactly the same conditions and at the same time in dynamic experiments. The setup is based on translational-FRAP with a fringe pattern of light for both the bleaching ...

The original approach to calculating diffusion coefficients of a fluorescent probe from Fluorescence Recovery After Photobleaching (FRAP) measurements assumes bleaching with a circular laser beam of a Gaussian intensity profile. This method was used without imaging the bleached cell. An empirical equation for calculating diffusion coefficients from a rectangular bleaching geometry, created in a...

The recruitment of TATA binding protein (TBP) to gene promoters is a critical rate-limiting step in transcriptional regulation for all three eukaryotic RNA polymerases. However, little is known regarding the dynamics of TBP in live mammalian cells. In this report, we examined the distribution and dynamic behavior of green fluorescence protein (GFP)-tagged TBP in live HeLa cells using fluorescen...

The determination of diffusion coefficients from fluorescence recovery data is often complicated by geometric constraints imposed by the complex shapes of intracellular compartments. To address this issue, diffusion of proteins in the lumen of the endoplasmic reticulum (ER) is studied using cell biological and computational methods. Fluorescence recovery after photobleaching (FRAP) experiments ...

The objective of this study was to evaluate total phenolic content and antioxidant activity of leaf and different parts of fruit (hull, shell, and kernel) of Bene in Ilam province. Total phenolic content was determined with the FolinCiocalteu Reagent (FCR); antioxidant activity of methanolic extracts were examined by DPPH (2,2diphenyl1picrylhydrazyl), FRAP (ferric reducing antioxidant power) an...

Medicinal plants have a lot of type antioxidants, mostly polyphenols, flavonoids which exhibit high antioxidant activity (Rice-Evans et al. 1995). The intake of antioxidants present in food is an important health-protecting factor. Herbal compounds known by ancient medicine are of growing interest in the domain of prevention of diseases. The FRAP assay (ferric reducing ability of plasma), a sim...

To examine endothelial nitric-oxide synthase (eNOS) trafficking in living endothelial cells, the eNOS-deficient endothelial cell line ECV304 was stably transfected with an eNOS-green fluorescent protein (GFP) fusion construct and characterized by functional, biochemical, and microscopic analysis. eNOS-GFP was colocalized with Golgi and plasma membrane markers and produced NO in response to agon...

Background: Chronic obstructive pulmonary disease (COPD) is a major public health problem that needs greater attention. Variability in the susceptibility to develop COPD is related to both genetic and environmental factors. Oxidative stress and inflammation are the major hallmarks of COPD and antioxidant status can be used as a biomarker to assess the risk of chronic diseases. Materials and Met...

The natural product rapamycin has been used to provide temporal and quantitative control of gene expression in animals through its ability to interact with two proteins simultaneously. A shortcoming of this approach is that rapamycin is an inhibitor of cell proliferation, the result of binding to FKBP12-rapamycin-associated protein (FRAP). To overcome this limitation, nontoxic derivatives of ra...

Antioxidant activity was evaluated in six Indian cultivars of lentil (Lens culanaris L.) viz. L-4076, L-4594, L-4147, DPL 15, DPL 62, and IPL 406. Three solvents, 70% ethanol, 100% methanol and 80% acetone, were used for preparation of seed extract. The seed extract was tested for total phenolic content, total flavonoid content and antioxidant activity by FRAP assay. The results showed that val...