Protonation states of buried histidine residues in human deoxyhemoglobin revealed by neutron crystallography

The protonation states of buried histidine residues in human deoxyhemoglobin were unambiguously identified by using a neutron crystallographic technique. Unexpectedly, the neutron structure reveals that both the alpha- and beta-distal histidines (Hisalpha58 and Hisbeta63) adopt a positively charged, fully (doubly) protonated form, suggesting their contribution to the Bohr effect. In addition, the neutron data provide an accurate picture of the alpha1beta1 hydrogen-bonding network and allow us to observe unambiguously the nature of the intradimeric interactions at an atomic level.     

Synchrotron X-ray techniques for the investigation of structures and dynamics in interfacial systems

This review focuses on recent results obtained by synchrotron X-ray techniques applied to the characterization of interfacial systems, with main emphasis on flat interfaces and on colloidal systems. The techniques covered are, for structural determinations: X-ray reflectivity (XRR), grazing incidence X-ray diffraction (GIXRD) and grazing incidence X-ray excited fluorescence (GIXF), while dynamics are investigated by X-ray photon correlation spectroscopy (XPCS) mainly in the grazing-incidence geometry (GIXPCS).The systems reviewed are, in order of growing complexity, floating Langmuir monolayers, supported films of lipids and proteins, polymeric films, buried interfaces, colloidal systems and gels formed by colloids either in 3D or in the form of 2D interfacial layers. Recent results are critically discussed, and some interesting directions of development are outlined, having also in mind new technical developments such as X-ray free electron laser sources and micro-focused synchrotron beamlines.     

Extended automated separation techniques in destructive neutron activation analysis; application to various biological materials,including human tissues and blood

An automated post-irradiation chemical separation scheme for the analysis of 14 trace elements in biological materials is described. The procedure consists of a destruction with sulfuric acid and hydrogen peroxide, a distillation of the volatile elements with hydrobromic acid and chromatography of both distillate and residue over Dowex 2X8 anion exchanger columns. Accuracy, precision and sensitivity are tested with reference materials (BOWEN’s kale, NBS bovine liver, IAEA materials dried animal whole blood, wheat flour, dried potatoes, powdered milk, oyster homogenate) and on a sample of pooled human blood. Blank values due to trace elements in the quartz irradiation vials are also discussed.     

The highly efficient protein crystallography beamline TLS 13B1 at the National Synchrotron Radiation Research Center

Synchrotron protein crystallography (PX) is one of the most powerful and dominant tools for determining the three-dimensional structures of biological macromolecules. At the National Synchrotron Radiation Research Center, TLS 13B1 is the first dedicated energy-tunable and highly automated PX beamline for efficiently solving de novo protein structures in Taiwan and has been open for users since September 2005. With remote access, user support services, and annual PX training courses over 17 years, the number of user groups at TLS 13B1 has grown from 11 to 139. In total, 1,429 projects were supported, and 2090 experiments were hosted during 67,846 hr of beamtime. This paper describes the photon source, beamline components, experimental station equipment, beamline software, beamline application process, and results from our users.     

A conformational study of bafilomycin A1 by x-ray crystallography and nmr techniques

We report the structure determination of bafilomycin A₁ by X-ray crystallography and a reassignment of the ¹H and ¹³C NMR spectra. By the measurement of ¹³C NT₁ and n.O.e. values the solution conformation was shown to be indistinguishable from that in the crystal.     

Adsorption of supercritical CO2 in aerogels as studied by small-angle neutron scattering and neutron transmission techniques

Small-angle neutron scattering (SANS) has been used to study the adsorption behavior of supercritical carbon dioxide (CO2) in porous Vycor glass and silica aerogels. Measurements were performed along two isotherms (T=35 and 80 degrees C) as a function of pressure (P) ranging from atmospheric up to 25 MPa, which corresponds to the bulk fluid densities ranging from rho(CO2) approximately 0 to 0.9 gcm3. The intensity of scattering from CO2-saturated Vycor porous glass can be described by a two-phase model which suggests that CO2 does not adsorb on the pore walls and fills the pore space uniformly. In CO2-saturated aerogels an adsorbed phase is formed with a density substantially higher that of the bulk fluid, and neutron transmission data were used to monitor the excess adsorption at different pressures. The results indicate that adsorption of CO2 is significantly stronger in aerogels than in activated carbons, zeolites, and xerogels due to the extremely high porosity and optimum pore size of these materials. SANS data revealed the existence of a compressed adsorbed phase with the average density approximately 1.07 gcm3, close to the density corresponding to closely packed van der Waals volume of CO2. A three-phase model [W. L. Wu, Polymer 23, 1907 (1982)] was used to estimate the volume fraction phi3 of the adsorbed phase as a function of the fluid density, and gave phi3 approximately 0.78 in the maximum adsorption regime around rho(CO2) approximately 0.374 gcm3. The results presented in this work demonstrate the utility of SANS combined with the transmission measurements to study the adsorption of supercritical fluids in porous materials.     

Recent advances in neutron reflectivity studies of biological membranes

Biological membranes are critical living interfaces which not only compartmentalise cells but also provide specific surfaces on which biochemical reactions take place. Their study is essential for our understanding of life and disease and also for the development of new pharmaceuticals. Because they are dynamic structures reliant on the surrounding aqueous phase for their activity, techniques which can probe them at a molecular scale under physiological conditions are crucial to understand their function. Here, I describe some of the recent developments in neutron reflectivity in this research area and suggest where future developments in the technology might be usefully directed.     

Instrumental neutron activation analysis of Na in biological materials

Sodium contents in seven mixtures of biological materials from varying aliquants of wheat flour and oyster tissue were analyzed using the INAA (instrumental neutron activation analysis) method. The samples were activated in the thermal column (TC) of the 1 MW TRIGA reactor at our laboratory. To analyze and compare the measured results, the ratioa of the epithermal neutron flux to the thermal neutron flux was determined in the TC, the rotating rack (RR) and the pneumatic terminal (PT). Due to the negligible interferences of²⁴Mg(n, p)²⁴Na and²⁷Al(n, )²⁴Na to²³Na(n, )²⁴Na, the Na results were obtained with 97% accuracy and 99% precision. It is shown that the method is suited for accurately determining Na contents in 12 various biological materials, especially for the samples that are low in Na content. From the cadmium ratio measurements of Au (4.9 eV resonance of¹⁹⁸Au) and Sm (8 eV resonance of¹⁵³Sm), ratios ofa_e/₀ were determined at 0.0011, 0.052 and 0.053 in the TC, RR and PT activation positions, respectively.     

Determination of molybdenum in biological materials by neutron activation

A method of the determination of molybdenum in biological materials by neutron activation is described. The method is based on the γ-spectrometric measurement of^99mTc after radiochemical separation of the latter by substoichiometric extraction with tetraphenylarsonium chloride using rhenium as inactive carrier. The method has been tested with reference materials.     

Characterization of biological macromolecules by electrophoresis and neutron activation

As analytical techniques have become more sensitive, the role of trace elements associated with biological macromolecules has become the subject of many studies in the past twenty years. Many biologically significant macromolecules, such as nucleic acids and proteins, have trace elements essential to their function and structure. Instrumental neutron activation analysis (INAA) contributes useful information in such studies. A procedure combining polyacrylamide gel electrophoresis (PAGE) with INAA and autoradiography has been developed to study biological macromolecules and their associated trace elements. Results from the application of this method to several metalloproteins are presented.     

Analysis of selenium in environmental and biological samples by neutron activation

This paper presents the results of the application of a fast rabbit system and a high thermal neutron flux to neutron activation analysis of selenium in environmental samples. The short-lived radionuclide^77mSe, (17.5s) is used for analysis. Results are presented for selenium in sea-water and rain-water, in biological reference materials, in food-stuffs and milk powder, in human hair and human blood-serum.     

Updating analytical techniques for fast neutron activation analysis

Two improvements in the analytical techniques used for fast neutron activation analysis have been developed and put into routine use. The first is neutron shielding of a NaI (TL) counting system, which reduces the counting noise encountered during neutron generation. The second is a modification of batch type irradiation for long lived nuclides so they can be done simultaneously with other test.     

Preconcentration techniques for trace analysis via neutron activation

A review is given of the methods that have been proposed for enrichment of trace elements in samples that are to be analysed by neutron-activation methods. The emphasis is on classification of methods, with full illustrations by means of practical examples.     

Determination of iodine in biological materials by neutron activation analysis

A method of determination of iodine (total and PBI) in serum, urine and other biological materials has been developed. The method consists in a gamma-spectrometric measurement of¹²⁸I activity after its radiochemical separation. The radiochemical separation procedure includes wet decomposition of the samples in a nitric acid medium followed by a few separation steps, the essential step being the substoichiometric extraction of iodide with a chloroform solution of tetraphenylarsonium chloride. Owing to the application of the substoichiometric separation, a high radiochemical purity of the separated iodine is achieved and the determination of the yield of radiochemical separation is not necessary.     

Uncertainty assessment in instrumental neutron activation analysis of biological materials

Summary The expression of measurement uncertainties in a standardized form is a requirement for result reliability as it imposes implications to the interpretation of analytical results. In this work, sample mass, elemental standard mass, element decay constant and sample and elemental standard activities were identified as the most important uncertainty sources for the relative method of instrumental neutron activation analysis. The contribution of these sources to the expanded standard uncertainty in the concentration of As, Co, Cr, Fe, K, Na, Se and Zn in biological materials of marine origin was assessed and sample activity was identified as the major contribution.     

Determination of iodine in biological samples by resonance neutron activation

A method of iodine determination in biological samples by means of resonance neutron activation is described. This method has made it possible to determine the iodine content of such samples as children's thyroid gland, grass frog's skin, etc.     

Thallium determination in biological materials by radiochemical neutron activation analysis

Summary The determination of thallium in biological materials sometimes cause problems because of the low concentrations of this toxic element. In the present work a method is described which optimizes the parameters affecting the specificity and sensitivity of the radiochemical NAA of thallium in biological samples. High thermal neutron flux, complete decomposition of the organic matter by pressurized digestion, TlI precipitations, liquid extraction of HTlBr₄ and La(OH)₃ scavenging purification are the steps leading to the final homogeneous preparation of Tl₂CrO₄ for -activity measurement. The method was applied to various materials as bovine liver, bone and nails. Good agreement was found between certified and determined thallium concentrations of the reference material CRM 176. The chemical yield comes to about 80%, with low deviations. The sensitivity of the method is about 10^–3 g/g, the standard deviations being in the range of 3.6% (CRM 176), 14% (bovine liver), and 17% (bone). Detailed working instructions are given.     

Determination of platinum and gold in biological materials by neutron activation analysis

A procedure for platinum and gold determination utilizing, neutron activation combined with radiochemical separation, has been developed. The reaction¹⁹⁸Pt (n, )¹⁹⁹ is used for Pt determination. Four procedures for gold separation are examined: (1) adsorption on untreated polyurethane foam (UPF). (2) extraction with dibutyl sulphide. (3) reduction of gold to elementary state in conc. H₂SO₄, and extraction of gold as diethyldithiocarbamate complex. The extraction with Cu(DDC)₂ is chosen as the most suitable process and applied to platinum and gold determinations in Bowen's Kale and mice organs, previously treated with Biocisplatinum^® specimens.