Characterizing the multi-dimensional reaction dynamics of dihalomethanes using XUV-induced Coulomb explosion imaging.

Site-selective probing of iodine 4d orbitals at 13.1 nm was used to characterize the photolysis of CH2I2 and CH2BrI initiated at 202.5 nm. Time-dependent fragment ion momenta were recorded using Coulomb explosion imaging mass spectrometry and used to determine the structural dynamics of the dissociating molecules. Correlations between these fragment momenta, as well as the onset times of electron transfer reactions between them, indicate that each molecule can undergo neutral three-body photolysis. For CH2I2, the structural evolution of the neutral molecule was simultaneously characterized along the C-I and I-C-I coordinates, demonstrating the sensitivity of these measurements to nuclear motion along multiple degrees of freedom.

Photon Acceleration in a Flying Focus.

A high-intensity laser pulse propagating through a medium triggers an ionization front that can accelerate and frequency upshift the photons of a second pulse. The maximum upshift is ultimately limited by the accelerated photons outpacing the ionization front or the ionizing pulse refracting from the plasma. Here, we apply the flying focus-a moving focal point resulting from a chirped laser pulse focused by a chromatic lens-to overcome these limitations. Theory and simulations demonstrate that the ionization front produced by a flying focus can frequency upshift an ultrashort optical pulse to the extreme ultraviolet over a centimeter of propagation. An analytic model of the upshift predicts that this scheme could be scaled to a novel tabletop source of spatially coherent x rays.

The current and future role of the medical oncologist in the professional care for cancer patients: a position paper by the European Society for Medical Oncology (ESMO).

The number of cancer patients in Europe is rising and significant advances in basic and applied cancer research are making the provision of optimal care more challenging. The concept of cancer as a systemic, highly heterogeneous and complex disease has increased the awareness that quality cancer care should be provided by a multidisciplinary team (MDT) of highly qualified healthcare professionals. Cancer patients also have the right to benefit from medical progress by receiving optimal treatment from adequately trained and highly skilled medical professionals. Built on the highest standards of professional training and continuing medical education, medical oncology is recognised as an independent medical specialty in many European countries. Medical oncology is a core member of the MDT and offers cancer patients a comprehensive and systemic approach to treatment and care, while ensuring evidence-based, safe and cost-effective use of cancer drugs and preserving the quality of life of cancer patients through the entire 'cancer journey'. Medical oncologists are also engaged in clinical and translational research to promote innovation and new therapies and they contribute to cancer diagnosis, prevention and research, making a difference for patients in a dynamic, stimulating professional environment. Medical oncologists play an important role in shaping the future of healthcare through innovation and are also actively involved at the political level to ensure a maximum contribution of the profession to Society and to tackle future challenges. This position paper summarises the multifarious and vital contributions of medical oncology and medical oncologists to today's and tomorrow's professional cancer care.

Involvement in research activities and factors influencing research capacity among dietitians.

BACKGROUND: A healthcare professional's aptitude to develop research skills and actively engage in research is necessary to optimise healthcare efficacy. The present study investigated the factors that contribute to research capacity within the Australian dietetic workforce. METHODS: Queensland-based dietitians scored their department and individual skill or success in research on a 10-point scale using an anonymous online survey that incorporated the validated Research Capacity in Context tool. Descriptive statistics were assessed against geographical setting, dietetic experience and the proportion of role (Full Time Equivalent; FTE) designated to research. Research activities were defined by the number of items currently involved in or completed in the past 6 months (n = 11). Factors associated with research activities were assessed by multivariable linear regression. RESULTS: Dietitians (n = 130) identified having a moderate skill or success in 14 research items [mean (SD) 5.1 (1.7)] and perceived that their departments provided a moderate level of research support in 19 research items [mean (SD) 6.1 (2.5)]. Geographical setting, the proportion of role designated to research (FTE) and participation in research activities were associated with individual and department ratings of research skill or success. Research involvement was predicted by the proportion of role (FTE) designated to research (ß = 0.34, t = 4.16, P < 0.001) and years of experience in dietetics (ß = 0.32, t = 2.67, P < 0.009). CONCLUSIONS: A dietitian's capacity for research is related to professional experience and the designation of research in the role description. The findings of the present study will provide a baseline of research capacity and expertise among dietitians, and also inform the strategic development of building research capacity.

High-resolution crystal structure of cytochrome P450cam.

The crystal structure of Pseudomonas putida cytochrome P450cam with its substrate, camphor, bound has been refined to R = 0.19 at a normal resolution of 1.63 A. While the 1.63 A model confirms our initial analysis based on the 2.6 A model, the higher resolution structure has revealed important new details. These include a more precise assignment of sequence to secondary structure, the identification of three cis-proline residues, and a more detailed picture of substrate-protein interactions. In addition, 204 ordered solvent molecules have been found, one of which appears to be a cation. The cation stabilizes an unfavorable polypeptide conformation involved in forming part of the active site pocket, suggesting that the cation may be the metal ion binding site associated with the well-known ability of metal ions to enhance formation of the enzyme-substrate complex. Another unusual polypeptide conformation forms the proposed oxygen-binding pocket. A localized distortion and widening of the distal helix provides a pocket for molecular oxygen. An intricate system of side-chain to backbone hydrogen bonds aids in stabilizing the required local disruption in helical geometry. Sequence homologies strongly suggest a common oxygen-binding pocket in all P450 species. Further sequence comparisons between P450 species indicate common three-dimensional structures with changes focused in a region of the molecule postulated to be associated with the control of substrate specificity.

Crystallization of single-chain Fv proteins.

Single-chain Fv (sFv) proteins consist of the variable heavy chain (VH) and variable light chain (VL) domains of an antibody, covalently joined by an engineered polypeptide linker. We report the crystallization of single-chain Fv's with specificities for fluorescein (4-4-20 sFv) and the TAG-72 pan-carcinoma glycoprotein antigen (CC49 sFv). Concentration of these proteins, preliminary to crystallization, results in a monomer-multimer equilibrium, causing aggregation which interferes with crystallization. Aggregation has been observed to depend primarily on an intact linker between VL and VH domains, although other factors are likely to modulate this phenomenon as well, including the specific identity of Fv and ligand, presence or absence of the ligand, linker length and possibly sequence. We have found two methods to overcome sFv aggregation, both of which yield X-ray diffraction quality crystals. The first, discovered serendipitously, is by introducing a proteolytic clip into the linker region (effectively yielding an Fv fragment). The second is the purification of the sFv dimer form, with linker regions intact, from an equilibrium mixture of aggregates. The sFv molecular association in a dimer is believed to be unusual in that each VL/VH interface may not be formed by the two linker-connected VL and VH domains, but rather by interaction of VL and VH domains from two distinct sFv monomers. Structure determination of the CC49 sFv dimer, with the 14-residue linker designated 212, is underway to test this model. Increasing linker length, to relieve steric strain on the monomer, and inclusion of the appropriate antigen, to slow transitions between monomeric and multimeric forms, may prove valuable strategies with sFv proteins less amenable to crystallization.

The 1.2 A resolution structure of the Con A-dimannose complex.

The complex between concanavalin A (Con A) and alpha1-2 mannobiose (mannose alpha1-2 mannose) has been refined to 1.2 A resolution. This is the highest resolution structure reported for any sugar-lectin complex. As the native structure of Con A to 0.94 A resolution is already in the database, this gives us a unique opportunity to examine sugar-protein binding at high resolution. These data have allowed us to model a number of hydrogen atoms involved in the binding of the sugar to Con A, using the difference density map to place the hydrogen atoms. This map reveals the presence of the protonated form of Asp208 involved in binding. Asp208 is not protonated in the 0.94 A native structure. Our results clearly show that this residue is protonated and hydrogen bonds to the sugar. The structure accounts for the higher affinity of the alpha1-2 linked sugar when compared to other disaccharides. This structure identifies different interactions to those predicted by previous modelling studies. We believe that the additional data presented here will enable significant improvements to be made to the sugar-protein modelling algorithms.

Biological function made crystal clear - annotation of hypothetical proteins via structural genomics.

Many of the gene products of completely sequenced organisms are 'hypothetical' - they cannot be related to any previously characterized proteins - and so are of completely unknown function. Structural studies provide one means of obtaining functional information in these cases. A 'structural genomics' project has been initiated aimed at determining the structures of 50 hypothetical proteins from Haemophilus influenzae to gain an understanding of their function. Each stage of the project - target selection, protein production, crystallization, structure determination, and structure analysis - makes use of recent advances to streamline procedures. Early results from this and similar projects are encouraging in that some level of functional understanding can be deduced from experimentally solved structures.

Cloning and characterization of the rat glutathione peroxidase gene.

The increased activity of glutathione peroxidase (GSHPx) in rat lungs is associated with the development of tolerance of the animals to hyperoxia. To understand further the regulation of expression of this enzyme, the molecular structure of the corresponding rat gene was characterized. The rat GSHPx gene consists of two exons interrupted by a single intron of 217 base pairs. The same initiation sites for transcription were found to be utilized in both lung and liver. The promoter of the GSHPx gene contains neither a 'TATA' box nor a 'CAAT' box. Instead, it comprises two copies of Sp1 binding motif and one copy of AP-2 binding motif. These features of the promoter may offer a clue to the mechanisms by which the expression of this gene is controlled.

X-Ray crystal structures of Candida albicans dihydrofolate reductase: high resolution ternary complexes in which the dihydronicotinamide moiety of NADPH is displaced by an inhibitor.

X-ray crystallographic analysis of 5-(4'-substituted phenyl)sulfanyl-2,4-diaminoquinazoline inhibitors in ternary complex with Candida albicans dihydrofolate reductase (DHFR) and NADPH revealed two distinct modes of binding. The two compounds with small 4'-substituents (H and CH3) were found to bind with the phenyl group oriented in the plane of the quinazoline ring system and positioned adjacent to the C-helix. In contrast, the more selective inhibitors with larger 4'-substituents (tert-butyl and N-morpholino) were bound to the enzyme with the phenyl group perpendicular to the quinazoline ring and positioned in the region of the active site that typically binds the dihydronicotinamide moiety of NADPH. The cofactor appeared bound to DHFR but with the disordered dihydronicotinamide swung away from the protein surface and into solution. This unusual inhibitor binding mode may play an important role in the high DHFR selectivity of these compounds and also may provide new ideas for inhibitor design.

Binding of nitropyrenes and benzo[a]pyrene to mouse lung deoxyribonucleic acid after pretreatment with inducing agents.

In assessing the biological effects of exposure to a complex chemical mixture, it is important to determine how the behavior of one compound may be influenced by the presence of other compounds in the mixture. In this study the effect of pre-exposure to an organic extract of diesel exhaust or to selected compounds in diesel exhaust on the binding of diesel exhaust compounds to DNA was determined. The amount of radiolabel covalently bound to mouse lung DNA following intratracheal administration of radiolabeled benzo[a]pyrene (BaP), 1-nitropyrene, 1,3,6-trinitropyrene, or a mixture of dinitropyrene was determined following pretreatment with benzo[a]pyrene, 1-nitropyrene, and diesel exhaust extract. Male CD-1 mice, 15-18 weeks of age, received 10 mg/kg of putative inducing agents by intratracheal instillation and, after 24 hr, 0.03 to 1.2 mg/kg radiolabeled putative DNA binding agents. Lung DNA was extracted, and covalent binding was quantitated by liquid scintillation spectroscopy. 1-Nitropyrene was a potent lung DNA binding agent in the absence of inducing agents [Covalent Binding Index (CBI) = 970] and was extremely potent after benzo[a]pyrene pretreatment (CBI = 21,540, comparable to the CBI for aflatoxin B1). Similar results were obtained for DNA binding of dinitropyrene and trinitropyrene with and without BaP pretreatment. DNA binding of BaP was lower (CBI = 40) and less inducible (BaP-pretreatment CBI = 230). Pretreatment with diesel extract caused an elevation in the binding of benzo[a]pyrene but little or no elevation in the binding of the nitropyrenes. Pretreatment with 1-nitropyrene did not increase significantly DNA binding of any of the agents tested. These results indicate that nitropyrenes bind readily to lung DNA and this binding may be increased in the presence of respirable mixtures, especially those containing inducing agents such as BaP.

Functional significance of sympathohippocampal sprouting: changes in single cell spontaneous activity.

The electrophysiological consequences of noradrenergic sympathohippocampal ingrowth following medial septal lesions were investigated. Spontaneous activity of single dentate neurons was recorded in anesthetized rats with and without sympathohippocampal ingrowth. In animals with medial septal lesions (ingrowth) spontaneous firing rates were not different from rates in normal animals. Medial septal lesions combined with bilateral superior cervical ganglionectomy (no ingrowth) resulted in a significant 82% increase in spontaneous activity.

Infection control organization in hospitals in England and Wales, 1986. Report of a survey undertaken by a Hospital Infection Society working party.

Questionnaires were distributed to hospitals in order to obtain information regarding arrangements for infection control. Returns were obtained from 180 of 200 (90%) health districts in England and Wales covering 95% of 'acute' and 85% of 'other' hospital beds listed in the Hospitals Year Book. The results demonstrated that some form of infection control organization was present in all districts, 98% had control of infection officers and 92% control of infection committees. The proportion of health districts with an infection control nurse had risen from 64% in 1979 to 89%, and the regional variation seen in 1979 was no longer marked. The major commitment of medical microbiologists to infection control was underlined.

Bacteriology of otitis media with effusion.

A study was undertaken to evaluate the prevalence and antibiotic susceptibility of bacteria present in the middle ear of patients with otitis media with effusion. Middle ear effusions (MEE), nasopharyngeal and throat swabs were obtained at operation and cultured for aerobic and anaerobic bacteria. Two hundred and fifty-nine effusions were obtained from 152 subjects examined. Haemophilus influenzae was isolated from 32 (12.3 per cent) effusions, Streptococcus pneumoniae from seven (2.7 per cent), Staphylococcus aureus from seven (2.7 per cent), Branhamella catarrhalis from one (0.4 per cent)--Group A beta haemolytic streptococci from one (0.4 per cent) and Staphylococcus epidermidis from three (1.9 per cent). The occurrence of respiratory pathogens in MEE reflected their prevalence in the upper respiratory tract. Significantly fewer children who had received antibiotics prior to surgery had organisms present in the MEE. Eight and a half per cent of H. influenzae and 64 per cent of B. catarrhalis were resistant to ampicillin. The present study confirms that bacteria are present in the middle ear in a significant number of patients with otitis media with effusion.

Measurements of 220Rn emanation from rocks.

Alpha-particle and gamma-ray measurements are employed to determine the individual emanation and production rates associated with the thorium radioactive series for thirty-seven rock specimens of approximately 200 cm3 individual volume representing igneous, sedimentary, and metamorphic types. These results are combined to establish 220Rn emanating powers for each of these specimens, and a wide range of values is observed. The generally large 220Rn emanation observed for these specimens strongly indicates that non-tortuous diffusion paths are commonly present in the associated structures, which gives more evidence for the existence of well-connected pore networks at the nanometer level in many of the specimens studied. While the results are qualitatively explainable, they are not predictable in terms of the current macroscopic observables. Since 220Rn and 222Rn emanation rates from rock specimens have previously been found to be comparable in magnitude, the relatively fast determination of 220Rn emanation rates described herein (measurements involving 1-h duration) is in reasonable probability indicative of a comparable 222Rn emanation rate: The employment of 220Rn as a convenient screening tool for potentially high 222Rn emanation sources is indicated.

Epidemiology of Haemophilus influenzae type b invasive disease in Wales.

OBJECTIVE: To investigate the epidemiology of invasive disease due to Haemophilus influenzae type b, the clones responsible, and the antibiotic resistance of the isolates. DESIGN: Prospective population based analysis of clinical and epidemiological data collected for Gwynedd during 1980-90 and in the whole of Wales during 1988-90. SETTING: 19 hospitals in Wales; all medical microbiology laboratories in Wales participated. PATIENTS: 82 patients with confirmed invasive infections caused by H influenzae type b in Gwynedd during 1980-90 and 207 in Wales during 1988-90. MAIN OUTCOME MEASURES: Clinical and epidemiological measures; analysis of the clonal types of the isolates based on the electrophoretic mobilities of 17 metabolic enzymes; and antibiotic resistance. RESULTS: The annual incidence of H influenzae type b infections in Gwynedd was 3.2 cases/100,000 and in Wales was 2.5 cases/100,000. Most cases occurred in children aged under 5 years, the highest annual incidence being in those aged under 1 (84.6/100,000 and 56.9/100,000 in Wales). The cumulative risk of acquiring H influenzae type b disease by the fifth birthday was one in 456 in Gwynedd and one in 578 in Wales. Fifteen per cent of cases in Gwynedd and 7% of those in Wales occurred in adults. Predominant clinical conditions were meningitis in children and pneumonia in adults. In Gwynedd 2/70 (3%) children and 5/12 (42%) adults died. Long term neurological sequelae occurred in 8% (4/48) of children who survived haemophilus meningitis. Children presenting with infection were usually the youngest members of their family. No secondary household cases were identified. 100 of 128 (78%) strains were of a single clone, electrophoretic type 12.5, and 4/207 (1.9%) isolates from Wales were resistant to both ampicillin and chloramphenicol. CONCLUSIONS: The annual rate of infection in children aged under 5 in four Welsh counties was 12-44% higher than that previously published for the United Kingdom. The study emphasises the potential value of a vaccine effective in early infancy and provides baseline data to assess its efficacy after its introduction. Alternatives to ampicillin and chloramphenicol should be used as first line, empirical treatment for severe infections that might be caused by H influenzae type b in Wales.