Deposition-based passive monitors for assigning radon, thoron inhalation doses for epidemiological studies.

The International Commission on Radiological Protection dose limits for radiation protection have been based on linearly extrapolating the high-dose risk coefficients obtained from the Japanese A bomb survivor data to low doses. The validity of these extrapolations has been questioned from time to time. To overcome this, epidemiological studies have been undertaken across the world on populations chronically exposed to low-radiation levels. In the past decade, the results of these studies have yielded widely differing, and sometimes, contradictory, conclusions. While recent residential radon studies have shown statistically significant radon risks at low doses, high-level natural radiation (HLNR) studies in China and India have not shown any low-dose risks. Similar is the case of a congenital malformation study conducted among the HLNR area populations in Kerala, India. It is thus necessary to make efforts at overcoming the uncertainties in epidemiological studies. In the context of HLNR studies, assigning radon and thoron doses has largely been an area of considerable uncertainty. Conventionally, dosimetry is carried out using radon concentration measurements, and doses have been assigned using assumed equilibrium factors for the progeny species. Gas-based dose assignment is somewhat inadequate due to variations in equilibrium factors and possibly due to significant thoron. In this context, passive, deposition-based progeny dosimetry appears to be a promising alternative method to assess inhalation doses directly. It has been deployed in various parts of India, including HBRAs and countries in Europe. This presentation discusses the method, the results obtained and their relevance to dose assignment in Indian epidemiological studies.

On the preparation, characterization, and enzymatic activity of fungal protease-gold colloid bioconjugates.

We present herein details pertaining to the preparation of bioconjugates of colloidal gold with aspartic protease from the fungus Aspergillus saitoi (F-prot) and their characterization and enzymatic activity. Simple mixing of the colloidal gold and protein solutions under protein-friendly conditions (pH = 3) followed by centrifugation (to remove uncomplexed gold nanoparticles and protein molecules) results in the formation of the fungal protease-gold nanoparticle conjugates. The protein-gold nanoparticle bioconjugate was redispersed in buffer solution and indicated the formation of efficient bioconjugates with intact native protein structures. The bioconjugates in solution were characterized by UV-vis spectroscopy, fluorescence spectroscopy, and biocatalytic activity measurements while drop-dried bioconjugate films on Si (111) substrates were characterized by scanning electron microscopy (SEM), energy dispersive analysis of X-rays (EDAX), and X-ray diffraction (XRD) measurements. Microscopy images do show some aggregate formation, but the intactness of the native structure of the enzyme in the bioconjugate material was verified by fluorescence and biocatalytic activity measurements. The enzyme retains substantial biocatalytic activity in the bioconjugate material and was comparable to that of free enzyme in solution.

A new method for the generation of patterned protein films by encapsulation in arrays of thermally evaporated lipids.

In this article we demonstrate a versatile method for the generation of patterned protein films by encapsulation in arrays of the lipids, octadecylamine (ODA, cationic), and arachidic acid (AA, anionic). A simple 2 x 2 array of ODA and AA was vacuum deposited on different substrates using appropriate masks. Thereafter, the enzymes pepsin and fungal protease as well as the heme-proteins cytochrome c and hemoglobin were encapsulated in the different elements of the array by sequential immersion (combined with judicious masking) of the array elements in the different protein solutions. The proteins are incorporated into the lipid elements by electrostatic interaction between charged amino acid residues on the protein surface and charged functional groups in the lipid matrix. This procedure leads to spatially distinct regions of the different proteins on one substrate and shows promise for single-chip multianalyte immunoassay/multiplex, high-throughput biosensor and catalysis applications. Fourier transform infrared spectroscopy (FTIR) was used to monitor the incorporation of the proteins in the different elements of the array as well as to ascertain whether intermixing of the proteins in a particular array element had occurred. The heme-protein composite regions were further characterized using UV-VIS spectroscopy.

Fabrication, characterization, and enzymatic activity of encapsulated fungal protease--fatty lipid biocomposite films.

Encapsulation of an aspartic protease from the fungus Aspergillus saitoi (F-prot) in thermally evaporated fatty acid films by a simple beaker-based immersion technique under enzyme-friendly conditions is described. The approach is based on diffusion of the enzyme from aqueous solution, driven primarily by attractive electrostatic interaction between charged groups on the enzyme surface and ionized lipid molecules in the film. The encapsulated enzyme molecules could be "pumped out" of the biocomposite film into solution by modulating the electrostatic interaction between the enzyme and fatty acid molecules via solution pH variation. The kinetics of F-prot diffusion into the acid films was followed using quartz crystal microgravimetry measurements while the secondary and tertiary structure of the enzyme in the lipid matrix was studied using Fourier transform infrared (FT-IR) and fluorescence spectroscopies. FT-IR and fluorescence measurements indicated little perturbation to the native structure of the enzyme. A chemical analysis of the F-prot-fatty acid biocomposite film was also performed using X-ray photoelectron spectroscopy. The encapsulated F-prot molecules showed catalytic activity (as estimated by reaction with hemoglobin) comparable to free enzyme molecules in solution, indicating facile access of biological analytes/reactants in solution to the enzyme molecules. The advantages/disadvantages of this approach vis-à-vis methods currently used for encapsulation of biomolecules are briefly discussed.