Synthesis, conformation and antiproliferative activity of isothiazoloisoxazole 1,1-dioxides.

Sixteen new isothiazoloisoxazole 1,1-dioxides, one new isothiazolotriazole and one new isothiazolopyrazole have been synthesised by using 1,3-dipolar cycloadditions to isothiazole 1,1-dioxides. One sub-set of these isothiazoloisoxazoles showed low µM activity against a human breast carcinoma cell line, whilst a second sub-set plus the isothiazolotriazole demonstrated an interesting restricted rotation of sterically hindered bridgehead substituents. A thiazete 1,1-dioxide produced from one of the isothiazole 1,1-dioxides underwent conversion into an unknown 1,2,3-oxathiazolin-2-oxide upon treatment with Lewis acids, but was inert towards 1,3-dipoles and cyclopropenones. Six supporting crystal structures are included.

Isolation and structural determination of non-racemic tertiary cathinone derivatives.

The racemic tertiary cathinones N,N-dimethylcathinone (1), N,N-diethylcathinone (2) and 2-(1-pyrrolidinyl)-propiophenone (3) have been prepared in reasonable yield and characterized using NMR and mass spectroscopy. HPLC indicates that these compounds are isolated as the anticipated racemic mixture. These can then be co-crystallized with (+)-O,O'-di-p-toluoyl-D-tartaric, (+)-O,O'-dibenzoyl-D-tartaric and (−)-O,O'-dibenzoyl-L-tartaric acids giving the single enantiomers S and R respectively of 1, 2 and 3, in the presence of sodium hydroxide through a dynamic kinetic resolution. X-ray structural determination confirmed the enantioselectivity. The free amines could be obtained following basification and extraction. In methanol these are reasonably stable for the period of several hours, and their identity was confirmed by HPLC and CD spectroscopy.

Introduction to suspension levels: radiotherapy.

In 2007, the European Commission (EC) commissioned a group of experts to undertake the revision of Report Radiation Protection (RP 91) 'Criteria for acceptability of radiological (including radiotherapy) and nuclear medicine installations' written in 1997. The revised draft report was submitted to the EC in 2010, who issued it for public consultation. The EC has commissioned the same group of experts to consider the comments of the public consultation for further improvement of the revised report. The EC intends to publish the final report under its Radiation Report Series as RP 162. This paper describes the background to the selection of the key performance parameters for radiotherapy equipment and sets out the sources of their criteria of acceptability including suspension levels for a wide range of radiotherapy equipment.

Practical application of suspension criteria scenarios: radiotherapy.

In 2007, the European Commission (EC) commissioned a group of experts to undertake the revision of Report RP91 'Criteria for Acceptability of Radiological (including Radiotherapy) and Nuclear Medicine Installations' written in 1997. The revised draft report was submitted to the EC in 2010, which issued it for public consultation. The EC commissioned the same group of experts to consider the comments of the public consultation for further improvement of the revised report. The EC intends to publish the final report under its Radiation Report Series as RP162. This paper presents a selection of practical applications of suspension criteria scenarios in radiotherapy, mostly in brachytherapy, with special emphasis on the critical roles and responsibilities of qualified radiotherapy staff (radiation oncologists, medical physicists and radiotherapy technicians).

The revision of RP 91 on criteria for acceptability of radiological (including radiotherapy) and nuclear medicine installations.

In 1997 the European Commission published Radiation Protection 91: 'Criteria for acceptability of radiological (including radiotherapy) and nuclear medicine installations'. This document specified the minimum criteria for acceptability. It has been used to this effect in legislation, codes of practice and by individual professionals. In a single document, it defined a level of performance at which remedial action was required. The document specified a series of parameters which characterised equipment performance and acceptable levels of performance. In its time it proved to be a useful document which was applied in member states to various degrees. Since the publication of Report 91 in 1997, a series of weaknesses emerged over time. Development of new radiological systems and technologies, as well as improvements in traditional technologies, has created circumstances where the acceptability criteria were in need of review. These weaknesses were recognised by the European Commission and a tender for its revision was issued. The criteria were developed by a team drawn from a broad range of backgrounds including hospitals, industry, government bodies, regulators and standardisation organisations. Representatives were mainly from Europe, but individuals from the American Association of Physicists in Medicine and International Atomic Energy Agency were included in the drafting process. This study describes the process employed in developing the revised document and the consultation process involved. One of the major difficulties the revision team encountered was related to an understanding of the actual meaning of the EC Directive. The view taken by the revision team was that Article 8, paragraph 3 places responsibilities on both the holders of radiological equipment and competent authorities. The acceptability criteria have been produced consistent with the European Commission's Medical Exposures Directive, which requires that patient exposures are optimised and justified.

Justification of diagnostic medical exposures: some practical issues. Report of an International Atomic Energy Agency Consultation.

OBJECTIVES: The Radiation Protection of Patients Unit of the International Atomic Energy Agency (IAEA) is concerned about the effectiveness of justification of diagnostic medical exposures. Recent published work and the report of an initial IAEA consultation in the area gave grounds for such concerns. There is a significant level of inappropriate usage, and, in some cases, a poor level of awareness of dose and risk among some key groups involved. This article aims to address this. METHODS: The IAEA convened a second group of experts in November 2008 to review practical and achievable actions that might lead to more effective justification. RESULTS: This report summarises the matters that this group considered and the outcome of their deliberations. There is a need for improved communication, both within professions and between professionals on one hand, and between professionals and the patients/public on the other. Coupled with this, the issue of consent to imaging procedures was revisited. The need for good evidence-based referral guidelines or criteria of acceptability was emphasised, as was the need for their global adaptation and dissemination. CONCLUSION: Clinical audit was regarded as a key tool in ensuring that justification becomes an effective, transparent and accountable part of normal radiological practice. In summary, justification would be facilitated by the "3 As": awareness, appropriateness and audit.

Impacts of long-term enhanced UV-B radiation on bryophytes in two sub-Arctic heathland sites of contrasting water availability.

BACKGROUND AND AIMS: Anthropogenic depletion of stratospheric ozone in Arctic latitudes has resulted in an increase of ultraviolet-B radiation (UV-B) reaching the biosphere. UV-B exposure is known to reduce above-ground biomass and plant height, to increase DNA damage and cause accumulation of UV-absorbing compounds in polar plants. However, many studies on Arctic mosses tended to be inconclusive. The importance of different water availability in influencing UV-B impacts on lower plants in the Arctic has been poorly explored and might partially explain the observed wide variation of responses, given the importance of water in controlling bryophyte physiology. This study aimed to assess the long-term responses of three common sub-Arctic bryophytes to enhanced UV-B radiation (+UV-B) and to elucidate the influence of water supply on those responses. METHODS: Responses of three sub-Arctic bryophytes (the mosses Hylocomium splendens and Polytrichum commune and the liverwort Barbilophozia lycopodioides) to +UV-B for 15 and 13 years were studied in two field experiments using lamps for UV-B enhancement with identical design and located in neighbouring areas with contrasting water availability (naturally mesic and drier sites). Responses evaluated included bryophyte abundance, growth, sporophyte production and sclerophylly; cellular protection by accumulation of UV-absorbing compounds, ß-carotene, xanthophylls and development of non-photochemical quenching (NPQ); and impacts on photosynthesis performance by maximum quantum yield (F(v) /F(m)) and electron transport rate (ETR) through photosystem II (PSII) and chlorophyll concentrations. RESULTS: Responses were species specific: H. splendens responded most to +UV-B, with reduction in both annual growth (-22 %) and sporophyte production (-44 %), together with increased ß-carotene, violaxanthin, total chlorophyll and NPQ, and decreased zeaxanthin and de-epoxidation of the xanthophyll cycle pool (DES). Barbilophozia lycopodioides responded less to +UV-B, showing increased ß-carotene and sclerophylly and decreased UV-absorbing compounds. Polytrichum commune only showed small morphogenetic changes. No effect of UV-B on bryophyte cover was observed. Water availability had profound effects on bryophyte ecophysiology, and plants showed, in general, lower growth and ETR, together with a higher photoprotection in the drier site. Water availability also influenced bryophyte responses to +UV-B and, in particular, responses were less detectable in the drier site. CONCLUSIONS: Impacts of UV-B exposure on Arctic bryophytes were significant, in contrast to modest or absent UV-B effects measured in previous studies. The impacts were more easily detectable in species with high plasticity such as H. splendens and less obvious, or more subtle, under drier conditions. Species biology and water supply greatly influences the impact of UV-B on at least some Arctic bryophytes and could contribute to the wide variation of responses observed previously.

Agriculture and the new challenges for photosynthesis research.

A rising human population and changing patterns of land use mean that world food production rates will need to be increased by at least 50% by 2050, a massive rise in harvestable yield per hectare of the major crops such as rice (Oryza sativa) and wheat (Triticum aestivum). Combinations of breeding for improved morphology-related traits such as harvest index and increased inputs of water and fertilizer, which have sustained yield increases since the 1960s, will be neither sufficient nor sustainable. An important limiting factor will be the capacity to produce sufficient biomass during favourable growing periods. Here we analyse this problem in the context of increasing the efficiency of conversion of solar energy into biomass, that is, leaf and canopy photosynthesis. Focussing on crops carrying out C3 photosynthesis, we analyse the evidence for 'losses' in the process of conversion of solar energy into crop biomass and we explore novel mechanisms of improving biomass production rates, which have arisen from recent research into the fundamental primary processes of photosynthesis and carbohydrate metabolism. We show that there are several lines of evidence that these processes are not fully optimized for maximum yield. We put forward the hypothesis that the chloroplast itself should be given greater prominence as a sensor, processor and integrator of highly variable environmental signals to allow a more efficient transduction of energy supply into biomass production.

Attenuation characteristics of MagnaDense high-density concrete at 6, 10 and 15 mv for use in radiotherapy bunker design.

The attenuation characteristics of MagnaDense high-density concrete are measured using the x-ray beams from two Varian linear accelerators at nominal 6, 10, and 15 megavolt energies. The tenth value layers for the primary beam are evaluated under broad- and narrow-beam conditions. The attenuation of secondary radiation is also investigated. Measured data are compared with existing data used in radiotherapy bunker design, derived from the scaling of published tenth value layers for ordinary concrete according to physical density. Instantaneous dose rates around an existing bunker with MagnaDense concrete walls and a conventional concrete roof are predicted using the various different data and compared to actual dose-rate measurements. Primary beam tenth value layer values derived from the broad-beam measurements are found to represent the attenuation properties of the MagnaDense more accurately than those produced by density-based scaling or from narrow-beam measurements.

Coordination chemistry of tetra- and tridentate ferrocenyl polyphosphines: an unprecedented [1,1'-heteroannular and 2,3-homoannular]-phosphorus-bonding framework in a metallocene dinuclear coordination complex.

PalladiumII and nickelII halide complexes of the ferrocenyl polyphosphines 1,1',2,3-tetrakis(diphenylphosphino)ferrocene (1), and 1,1',2-tris(diphenylphosphino)-4-tert-butylferrocene (5) were prepared and characterized by multinuclear NMR. The metallo-ligand 1, the palladium [Pd2Cl4(1)] (3b) and nickel [NiCl2(5)] (6) coordination complexes were additionally characterized by X-ray diffraction crystallography. The behavior of 1 toward coordination to nickel and palladium was surprisingly different because the coordination of a second metal center after the initial 1,2-phosphorus-bonding of nickel was markedly difficult. The preference of nickel for 1,2-P coordination on 1,1'-bonding was confirmed by the exclusive formation of 6 from 5. The changes noted between the solid state structure of the ligand 1 and the structure obtained for the dinuclear palladium complex 3b reveal the rotational flexibility of this tetraphosphine. This flexibility is at the origin of the unique framework for a metallocenic dinuclear metal complex in which both coexist a 1,1'-heteroannular chelating P-bonding and a 2,3-homoannular chelating P-bonding with two palladium centers. Some reported specimens of ferrocenyl polyphosphines of constrained geometry have previously revealed that phosphorus lone pair overlap can lead to very intense "through-space" 31P31P nuclear spin-spin coupling constants (JPP) ( J. Am. Chem. Soc. 2004, 126 (35), 11077-11087] in solution phase. In these cases, an internuclear distance between heteroannular phosphorus atoms below 4.9 A, with an adequate orientation of the lone-pairs in the solid state and in solution, was a necessary parameter. The flexibility of the new polyphosphines 1 and 5 does not allow that spatial proximity (internuclear distances between heteroannular phosphorus above 5.2 A in the solid state); accordingly the expected through-space nuclear spin-spin coupling constants were not detected in any of their coordination complexes nor in 1.

Trends in leaf photosynthesis in historical rice varieties developed in the Philippines since 1966.

Crop improvement in terms of yield is rarely linked to leaf photosynthesis. However, in certain crop plants such as rice, it is predicted that an increase in photosynthetic rate will be required to support future grain yield potential. In order to understand the relationships between yield improvement and leaf photosynthesis, controlled environment conditions were used to grow 10 varieties which were released from the International Rice Research Institute (IRRI) between 1966 and 1995 and one newly developed line. Two growth light intensities were used: high light (1500 micromol m(-2) s(-1)) and low light (300 micromol m(-2) s(-1)). Gas exchange, leaf protein, chlorophyll, and leaf morphology were measured in the ninth leaf on the main stem. A high level of variation was observed among high light-grown plants for light-saturated photosynthetic rate per unit leaf area (P(max)), stomatal conductance (g), content of ribulose bisphosphate carboxylase-oxygenase (Rubisco), and total leaf protein content. Notably, between 1966 and 1980 there was a decline in P(max), g, leaf protein, chlorophyll, and Rubisco content. Values recovered in those varieties released after 1980. This striking trend coincides with a previous published observation that grain yield in IRRI varieties released prior to 1980 correlated with harvest index whereas that for those released after 1980 correlated with biomass. P(max) showed significant correlations with both g and Rubisco content. Large differences were observed between high light- and low light-grown plants (photoacclimation). The photoacclimation 'range' for P(max) correlated with P(max) in high light-grown plants. It is concluded that (i) leaf photosynthesis may be systematically affected by breeding strategy; (ii) P(max) is a useful target for yield improvements where yield is limited by biomass production rather than partitioning; and (iii) the capacity for photoacclimation is related to high P(max) values.

Spike sorting based upon machine learning algorithms (SOMA).

We have developed a spike sorting method, using a combination of various machine learning algorithms, to analyse electrophysiological data and automatically determine the number of sampled neurons from an individual electrode, and discriminate their activities. We discuss extensions to a standard unsupervised learning algorithm (Kohonen), as using a simple application of this technique would only identify a known number of clusters. Our extra techniques automatically identify the number of clusters within the dataset, and their sizes, thereby reducing the chance of misclassification. We also discuss a new pre-processing technique, which transforms the data into a higher dimensional feature space revealing separable clusters. Using principal component analysis (PCA) alone may not achieve this. Our new approach appends the features acquired using PCA with features describing the geometric shapes that constitute a spike waveform. To validate our new spike sorting approach, we have applied it to multi-electrode array datasets acquired from the rat olfactory bulb, and from the sheep infero-temporal cortex, and using simulated data. The SOMA sofware is available at http://www.sussex.ac.uk/Users/pmh20/spikes.

The effect of fertility control on the transmission of bovine tuberculosis in wild brushtail possums.

AIM: To determine the effect of fertility control on the rate of transmission of bovine tuberculosis (Tb), caused by Mycobacterium bovis, in brushtail possums (Trichosurus vulpecula). METHODS: At two study sites with a history of Tb infection in the resident possum population, a sample of adult male and female possums (n=50), determined by palpation to be Tb-free, was surgically sterilised by gonadectomy. A sample of untreated Tb-free male and female possums (n=54) served as controls. Each study site was trapped every 2 months over a 3-year period, and the Tb status of the individuals in the trial assessed. At the conclusion of the trial, all remaining experimental individuals were killed, necropsied and examined for characteristic lesions typical of Tb. The rate of transmission of Tb infection was estimated using the incidence of tuberculous cases in the experimental animals and comparing it between sites, sex and sterilisation treatment. RESULTS: Sterilisation by gonadectomy resulted in a reduction in the rate of transmission of Tb in male possums by 53%, but a corresponding increase in sterilised females for reasons that are still unclear. However, this interaction was statistically weak (p=0.10). When the sexes were combined, there was no overall effect of sterilisation on the rate of transmission of Tb (p=0.43). Sterility treatment notwithstanding, there was a highly significant difference in the rate of transmission between the sexes (p=0.01), being almost one order of magnitude higher in untreated males compared with untreated females. CONCLUSIONS: Although lacking strong statistical support, these results suggest that fertility control that targets endocrine control of fertility may provide some additional benefits for disease control over that achieved by reductions in population density, by reducing the rate of transmission of M. bovis in male possums. However, additional studies are needed to confirm the validity of these results. The large difference in the rate of transmission of M. bovis in untreated males compared with untreated females suggests that transmission of Tb among males may be an important driver of the dynamics of Tb infection in possums.

Applications of multi-variate analysis of variance (MANOVA) to multi-electrode array electrophysiology data.

We have developed an adaptation of multi-variate analysis of variance (MANOVA) to analyze statistically both local and global patterns of multi-electrode array (MEA) electrophysiology data where the activities of many (typically >100) neurons have been recorded simultaneously. Whereas simple application of standard MANOVA techniques prohibits extraction of useful information in this kind of data, our new approach, MEANOVA (=MEA+MANOVA), allows a more useful and powerful approach to analyze such complex neurophysiological data. The MEANOVA test enables the detection of the "hot-spots" in the MEA data and has been validated using recordings from the rat olfactory bulb. To further validate the power of this approach, we have also applied the MEANOVA test to data obtained from a simple computational network model. This MEANOVA software and other useful statistical methods for MEA data can be downloaded from http://www.sussex.ac.uk/Users/pmh20

Acclimation of photosynthesis to high irradiance in rice: gene expression and interactions with leaf development.

Rice (Oryza sativa L.) has been used to study the long-term responses of photosynthesis to high irradiance focusing on the composition of the photosynthetic apparatus and leaf morphology. Typical sun/shade differences in chloroplast composition are seen in the fifth leaf following growth in high irradiance compared with low irradiance (1000 and 200 micromol m(-2) s(-1), respectively): higher light-saturated rates of photosynthesis (P(max)), higher amounts of Rubisco protein, and a lower chlorophyll a:b ratio. In addition, leaves were thicker under high light compared with low light. However, responses appear more complex when leaf developmental stage is considered. Using a system of transferring plants from low to high light in the laboratory responses that occur before and after full leaf extension have been studied. Acclimation of photosynthesis is limited by leaf age: the transfer to high light, post-leaf extension, is characterized by alterations in chlorophyll a:b but not in Rubisco protein, which may be limited by leaf morphology. Microarray analysis of gene expression was carried out on plants that were transferred to high light post-leaf extension. A down-regulation of light-harvesting genes was seen. No change in the expression level of Rubisco genes was observed. Up-regulation of genes involved in photoprotection was observed. It was also shown that high-light leaf morphology is established prior to formation of the zone of cellular elongation and division. The endogenous and environmental factors which establish the characteristics of high light acclimation may be important for attaining high rates of assimilation in leaves and crop canopies, and the fifth leaf in rice provides a convenient model system for the determination of the mechanisms involved.

Physiological interpretation of electrical impedance epigastrography measurements.

Measurement of the electrical impedance of the gastric region is carried out with the epigastrograph. This generates and applies alternating current around the abdominal area and measures the potential difference in order to determine the impedance externally, via electrodes. The change of epigastric impedance for a subject, given a meal after fasting, depends on the conductivity of the meal compared to the stomach and surrounding tissues. Typically a conductive meal has conductivity >7 mS cm(-1), non-conductive <2 mS cm(-1) and neutral about 4.5 mS cm(-1). Half-emptying times (T50s) from gastric emptying studies in volunteers using three test meals of 450 ml volume were obtained and found to be shorter than expected from the literature. The meals were a 10% glucose solution and two milk shakes of energy 1,300 kJ and 2,850 kJ, respectively. These electrical impedance epigastrography (EIE) measurements were carried out with scintigraphy. The T50 values of the latter were significantly longer. The direct comparison of the normalized experimental data obtained by both methods led to the concept that EIE measurements are mainly influenced by gastric secretion. Thus the EIE trace of a 'neutral' meal suggests the hypothesis that the volume of the meal is not the significant factor but is influenced by gastric acid secretions. Physiology of the gastric mucosa during the digestion of a meal and intragastric pH values also suggests this. Gastric function studies using EIE measurements may therefore reflect gastric ionic concentration rather than the volume of the contents of the stomach. In turn this could lead to the development of a non-invasive method for the continuous recording of gastric acid secretions.

Quantitative assessment of the radiation hazards and risks in sentinel node procedures.

Sentinel node localization using an injected radiopharmaceutical and a gamma probe is performed in many hospitals. Employers have a duty to give appropriate training to staff who may not have been previously formally trained to work with unsealed radioactive sources. A study was performed to assess hazards and risks at all stages of the localization procedure. Whole body doses and finger doses of imaging, surgery and pathology staff were determined. The activity remaining in the tumour specimen, excised nodes and disposable waste from the operating theatre was measured. Any radioactive contamination of the operating theatre and equipment was also ascertained. All results were then assessed in light of current UK radiation protection legislation for the protection of staff and members of the public. Results showed that radiation doses are low and no additional procedures are required for protection of staff, provided the usual procedures for biohazards are in place. However, an information sheet has been prepared for the reassurance of staff, and theatre swabs may need to be stored temporarily before disposal. Injecting and imaging on the day before surgery is preferred, compared with injecting and imaging before surgery on the same day, since this gives lower radiation doses to staff, lower activity in excised specimens and waste, and provides a higher count rate giving better image quality.

Plants lacking the main light-harvesting complex retain photosystem II macro-organization.

Photosystem II (PSII) is a key component of photosynthesis, the process of converting sunlight into the chemical energy of life. In plant cells, it forms a unique oligomeric macrostructure in membranes of the chloroplasts. Several light-harvesting antenna complexes are organized precisely in the PSII macrostructure-the major trimeric complexes (LHCII) that bind 70% of PSII chlorophyll and three minor monomeric complexes-which together form PSII supercomplexes. The antenna complexes are essential for collecting sunlight and regulating photosynthesis, but the relationship between these functions and their molecular architecture is unresolved. Here we report that antisense Arabidopsis plants lacking the proteins that form LHCII trimers have PSII supercomplexes with almost identical abundance and structure to those found in wild-type plants. The place of LHCII is taken by a normally minor and monomeric complex, CP26, which is synthesized in large amounts and organized into trimers. Trimerization is clearly not a specific attribute of LHCII. Our results highlight the importance of the PSII macrostructure: in the absence of one of its main components, another protein is recruited to allow it to assemble and function.

Overexpression of beta-carotene hydroxylase enhances stress tolerance in Arabidopsis.

Plant stress caused by extreme environmental conditions is already a principal reason for yield reduction in crops. The threat of global environment change makes it increasingly important to generate crop plants that will withstand such conditions. Stress, particularly stress caused by increased sunlight, leads to the production of reactive oxygen species that cause photo-oxidative cell damage. Carotenoids, which are present in the membranes of all photosynthetic organisms, help protect against such light-dependent oxidative damage. In plants, the xanthophyll cycle (the reversible interconversion of two carotenoids, violaxanthin and zeaxanthin) has a key photoprotective role and is therefore a promising target for genetic engineering to enhance stress tolerance. Here we show that in Arabidopsis thaliana overexpression of the chyB gene that encodes beta-carotene hydroxylase--an enzyme in the zeaxanthin biosynthetic pathway--causes a specific twofold increase in the size of the xanthophyll cycle pool. The plants are more tolerant to conditions of high light and high temperature, as shown by reduced leaf necrosis, reduced production of the stress indicator anthocyanin and reduced lipid peroxidation. Stress protection is probably due to the function of zeaxanthin in preventing oxidative damage of membranes.

Acclimation of Arabidopsis thaliana to the light environment: the existence of separate low light and high light responses.

The capacity for photosynthetic acclimation in Arabidopsis thaliana (L.) Heynh. cv. Landsberg erecta was assessed during growth over a broad range of irradiance. Discontinuities in the response to growth irradiance were revealed for the light- and CO2-saturated rate of photosynthesis (Pmax) and the ratio of chlorophyll a to chlorophyll b (Chl a/b). Three separate phases in the response of Pmax and Chl a/b to growth light were evident, with increases at low and high irradiance ranges and a plateau at intermediate irradiance. By measuring all chlorophyll-containing components of the thylakoid membrane that contribute to Chl a/b we reveal that distinct strategies for growth at low and high irradiance underlie the discontinuous response. These strategies include, in addition to changes in the major light-harvesting complexes of photosystem II (LHCII), large shifts in the amounts of both reaction centres as well as significant changes in the levels of minor LHCII and LHCI components.