Molecular library design using multi-objective optimization methods. Advancements in combinatorial chemistry and high-throughput screening technology have enabled the synthesis and screening of large molecular libraries for the purposes of drug discovery. Contrary to initial expectations, the increase in screening library size, typically combined with an emphasis on compound structural diversity, did not result in a comparable increase in the number of promising hits found. In an effort to improve the likelihood of discovering hits with greater optimization potential, more recent approaches attempt to incorporate additional knowledge to the library design process to effectively guide the search. Multi-objective optimization methods capable of taking into account several chemical and biological criteria have been used to design collections of compounds satisfying simultaneously multiple pharmaceutically relevant objectives. In this chapter, we present our efforts to implement a multi-objective optimization method, MEGALib, custom-designed to the library design problem. The method exploits existing knowledge, e.g. from previous biological screening experiments, to identify and profile molecular fragments used subsequently to design compounds compromising the various objectives.


Application of Free-Wilson selectivity analysis for combinatorial library design. In this chapter we present an application of in silico quantitative structure-activity relationship (QSAR) models to establish a new ligand-based computational approach for generating virtual libraries. The Free-Wilson methodology was applied to extract rules from two data sets containing compounds which were screened against either kinase or PDE gene family panels. The rules were used to make predictions for all compounds enumerated from their respective virtual libraries. We also demonstrate the construction of R-group selectivity profiles by deriving activity contributions against each protein target using the QSAR models. Such selectivity profiles were used together with protein structural information from X-ray data to provide a better understanding of the subtle selectivity relationships between kinase and PDE family members.


Application of QSAR and shape pharmacophore modeling approaches for targeted chemical library design. Optimization of chemical library composition affords more efficient identification of hits from biological screening experiments. The optimization could be achieved through rational selection of reagents used in combinatorial library synthesis. However, with a rapid advent of parallel synthesis methods and availability of millions of compounds synthesized by many vendors, it may be more efficient to design targeted libraries by means of virtual screening of commercial compound collections. This chapter reviews the application of advanced cheminformatics approaches such as quantitative structure-activity relationships (QSAR) and pharmacophore modeling (both ligand and structure based) for virtual screening. Both approaches rely on empirical SAR data to build models; thus, the emphasis is placed on achieving models of the highest rigor and external predictive power. We present several examples of successful applications of both approaches for virtual screening to illustrate their utility. We suggest that the expert use of both QSAR and pharmacophore models, either independently or in combination, enables users to achieve targeted libraries enriched with experimentally confirmed hit compounds.


Combinatorial library design from reagent pharmacophore fingerprints. Combinatorial and parallel chemical synthesis technologies are powerful tools in early drug discovery projects. Over the past couple of years an increased emphasis on targeted lead generation libraries and focussed screening libraries in the pharmaceutical industry has driven a surge in computational methods to explore molecular frameworks to establish new chemical equity. In this chapter we describe a complementary technique in the library design process, termed ProSAR, to effectively cover the accessible pharmacophore space around a given scaffold. With this method reagents are selected such that each R-group on the scaffold has an optimal coverage of pharmacophoric features. This is achieved by optimising the Shannon entropy, i.e. the information content, of the topological pharmacophore distribution for the reagents. As this method enumerates compounds with a systematic variation of user-defined pharmacophores to the attachment point on the scaffold, the enumerated compounds may serve as a good starting point for deriving a structure-activity relationship (SAR).


Docking methods for structure-based library design. The drug discovery process mainly relies on the experimental high-throughput screening of huge compound libraries in their pursuit of new active compounds. However, spiraling research and development costs and unimpressive success rates have driven the development of more rational, efficient, and cost-effective methods. With the increasing availability of protein structural information, advancement in computational algorithms, and faster computing resources, in silico docking-based methods are increasingly used to design smaller and focused compound libraries in order to reduce screening efforts and costs and at the same time identify active compounds with a better chance of progressing through the optimization stages. This chapter is a primer on the various docking-based methods developed for the purpose of structure-based library design. Our aim is to elucidate some basic terms related to the docking technique and explain the methodology behind several docking-based library design methods. This chapter also aims to guide the novice computational practitioner by laying out the general steps involved for such an exercise. Selected successful case studies conclude this chapter.


Design of screening collections for successful fragment-based lead discovery. A successful fragment-based lead discovery (FBLD) campaign largely depends on the content of the fragment collection being screened. To design a successful fragment collection, several factors must be considered, including collection size, property filters, hit follow-up considerations, and screening methods. In this chapter, we will discuss each factor and how it was applied to the design and assembly of one or more fragment collections in a major pharmaceutical company setting. We will also present examples and statistics of screening results from such collections and how subsequent collections can be improved. Lastly, we will provide a summary comparison of selected fragment collections from literature.


Structure-based library design in efficient discovery of novel inhibitors. Structure-based library design employs both structure-based drug design (SBDD) and combinatorial library design. Combinatorial library design concepts have evolved over the past decade, and this chapter covers several novel aspects of structure-based library design together with successful case studies in the anti-viral drug design HCV target area. Discussions include reagent selections, diversity library designs, virtual screening, scoring/ranking, and post-docking pose filtering, in addition to the considerations of chemistry synthesis. Validation criteria for a successful design include an X-ray co-crystal complex structure, in vitro biological data, and the number of compounds to be made, and these are addressed in this chapter as well.


Structure-based and property-compliant library design of 11β-HSD1 adamantyl amide inhibitors. Multiproperty lead optimization that satisfies multiple biological endpoints remains a challenge in the pursuit of viable drug candidates. Optimization of a given lead compound to one having a desired set of molecular attributes often involves a lengthy iterative process that utilizes existing information, tests hypotheses, and incorporates new data. Within the context of a data-rich corporate setting, computational tools and predictive models have provided the chemists a means for facilitating and streamlining this iterative design process. This chapter discloses an actual library design scenario for following up a lead compound that inhibits 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) enzyme. The application of computational tools and predictive models in the targeted library design of adamantyl amide 11β-HSD1 inhibitors is described. Specifically, the multiproperty profiling using our proprietary PGVL (Pfizer Global Virtual Library) Hub is discussed in conjunction with the structure-based component of the library design using our in-house docking tool AGDOCK. The docking simulations were based on a piecewise linear potential energy function in combination with an efficient evolutionary programming search engine. The library production protocols and results are also presented.


Fragment-based drug design.

Fragment-based drug design. Fragment-based drug design (FBDD), which is comprised of both fragment screening and the use of fragment hits to design leads, began more than 15 years ago and has been steadily gaining in popularity and utility. Its origin lies on the fact that the coverage of chemical space and the binding efficiency of hits are directly related to the size of the compounds screened. Nevertheless, FBDD still faces challenges, among them developing fragment screening libraries that ensure optimal coverage of chemical space, physical properties and chemical tractability. Fragment screening also requires sensitive assays, often biophysical in nature, to detect weak binders. In this chapter we will introduce the technologies used to address these challenges and outline the experimental advantages that make FBDD one of the most popular new hit-to-lead process.


The design, annotation, and application of a kinase-targeted library. We present here a workflow for designing a kinase-targeted library (KTL) with the goal of capturing known kinase inhibitor chemical space. We validated our design retrospectively using recent, high-throughput screening data and found significant enrichment of kinase inhibitor hits while retaining majority of the active kinase inhibitor series. To further assist kinase projects in triaging KTL screen hits, we also developed a methodology to systematically annotate known kinase inhibitors in the KTL with regard to their binding modes.


LEAP into the Pfizer Global Virtual Library (PGVL) space: creation of readily synthesizable design ideas automatically. Pfizer Global Virtual Library (PGVL) of 10(13) readily synthesizable molecules offers a tremendous opportunity for lead optimization and scaffold hopping in drug discovery projects. However, mining into a chemical space of this size presents a challenge for the concomitant design informatics due to the fact that standard molecular similarity searches against a collection of explicit molecules cannot be utilized, since no chemical information system could create and manage more than 10(8) explicit molecules. Nevertheless, by accepting a tolerable level of false negatives in search results, we were able to bypass the need for full 10(13) enumeration and enabled the efficient similarity search and retrieval into this huge chemical space for practical usage by medicinal chemists. In this report, two search methods (LEAP1 and LEAP2) are presented. The first method uses PGVL reaction knowledge to disassemble the incoming search query molecule into a set of reactants and then uses reactant-level similarities into actual available starting materials to focus on a much smaller sub-region of the full virtual library compound space. This sub-region is then explicitly enumerated and searched via a standard similarity method using the original query molecule. The second method uses a fuzzy mapping onto candidate reactions and does not require exact disassembly of the incoming query molecule. Instead Basis Products (or capped reactants) are mapped into the query molecule and the resultant asymmetric similarity scores are used to prioritize the corresponding reactions and reactant sets. All sets of Basis Products are inherently indexed to specific reactions and specific starting materials. This again allows focusing on a much smaller sub-region for explicit enumeration and subsequent standard product-level similarity search. A set of validation studies were conducted. The results have shown that the level of false negatives for the disassembly-based method is acceptable when the query molecule can be recognized for exact disassembly, and the fuzzy reaction mapping method based on Basis Products has an even better performance in terms of lower false-negative rate because it is not limited by the requirement that the query molecule needs to be recognized by any disassembly algorithm. Both search methods have been implemented and accessed through a powerful desktop molecular design tool (see ref. (33) for details). The chapter will end with a comparison of published search methods against large virtual chemical space.


PGVL Hub: An integrated desktop tool for medicinal chemists to streamline design and synthesis of chemical libraries and singleton compounds. PGVL Hub is an integrated molecular design desktop tool that has been developed and globally deployed throughout Pfizer discovery research units to streamline the design and synthesis of combinatorial libraries and singleton compounds. This tool supports various workflows for design of singletons, combinatorial libraries, and Markush exemplification. It also leverages the proprietary PGVL virtual space (which contains 10(14) molecules spanned by experimentally derived synthesis protocols and suitable reactants) for lead idea generation, lead hopping, and library design. There had been an intense focus on ease of use, good performance and robustness, and synergy with existing desktop tools such as ISIS/Draw and SpotFire. In this chapter we describe the three-tier enterprise software architecture, key data structures that enable a wide variety of design scenarios and workflows, major technical challenges encountered and solved, and lessons learned during its development and deployment throughout its production cycles. In addition, PGVL Hub represents an extendable and enabling platform to support future innovations in library and singleton compound design while being a proven channel to deliver those innovations to medicinal chemists on a global scale.


GLARE: A tool for product-oriented design of combinatorial libraries. Combinatorial chemistry with two or more diversity points often leads to an immense number of theoretical products. It is sensible to select the reagents based on the desired properties of the products in the hope of maximizing the usefulness of the synthesized molecules. The presented tool enables the filtering of reagents such that any further reagent selection will form products matching the desired properties. Virtual combinatorial library leading to thousands of billions of products can be rapidly assessed. The publicly available software ( http://glare.sourceforge.net ) and key algorithmic elements are discussed.


CLEVER: A general design tool for combinatorial libraries. CLEVER is a computational tool designed to support the creation, manipulation, enumeration, and visualization of combinatorial libraries. The system also provides a summary of the diversity, coverage, and distribution of selected compound collections. When deployed in conjunction with large-scale virtual screening campaigns, CLEVER can offer insights into what chemical compounds to synthesize, and, more importantly, what not to synthesize. In this chapter, we describe how CLEVER is used and offer advice in interpreting the results.


Design of targeted libraries against the human Chk1 kinase using PGVL Hub. PGVL Hub is a Pfizer internal desktop tool for chemical library and singleton design. In this chapter, we give a short introduction to PGVL Hub, the core workflow it supports, and the rich design capabilities it provides. By re-creating two legacy targeted libraries against the human checkpoint kinase 1 (Chk1) as a showcase, we illustrate how PGVL Hub could be used to help library designers carry out the steps in library design and realize design objectives such as SAR expansion and improvement in both kinase selectivity and compound aqueous solubility. Finally we share several tips about library design and usage of PGVL Hub.


Male behaviors reveal multiple pherotypes within vine mealybug Planococcus ficus (Signoret) (Hemiptera; Pseudococcidae) populations. The vine mealybug (VM) females collected in Israel produce two sex pheromone compounds: lavandulyl senecioate (LS) and (S)-lavandulyl isovalerate (LI). The males display ambiguous behavior to LI: repulsion in the vineyard and attraction of laboratory-reared males. We addressed the question of individual male behavior, i.e., do males respond to both LS and LI, or might they display a distinct response to each of the two pheromone compounds. We compared male pherotype frequencies between wild-caught and laboratory-reared populations. Then, we examined the relationship between pherotype composition and male capture rates in pheromone traps. Finally, we addressed the heredity of the pherotypes. The Israeli VM populations contain nine different male pherotypes, as defined according to the male behavior to pheromone compounds. The studied Portuguese populations included five of the nine pherotypes; none of the Portuguese males were attracted to LI. It seems that the high frequency of males that were attracted to LI is related to dense VM populations. It is hypothesized that selection for the male pherotypes, I males, those that respond to LI, occur under high-density rearing conditions. This may result from shorter development times of males and females that produce more I male pherotypes. The lower relative frequency of trapping of males in LI-baited traps than expected from the percentage determined in a Petri dish arena suggests that males that respond solely to LS (S males) are better fliers. The results also suggest that the pherotype trait is inherited by both sexes of the VM.


Blood exposure has a negative effect on engineered cartilage. The aim of this study was to investigate the in vitro effect of different concentrations of blood on the morphological and biochemical properties of engineered cartilage. Previous studies have demonstrated a negative effect of blood on native cartilage; however, the effect of the contact of blood on engineered cartilage is unclear. Articular chondrocytes were isolated from swine joints, expanded in monolayer culture, and seeded onto collagen membranes. The seeded membranes were cultured for 3 days in the presence of different concentrations of peripheral blood. Some samples were retrieved at the end of the blood contact, others after 21 additional days of standard culture conditions, in order to investigate the "long-term effect" of the blood contact. All seeded samples showed an increase in the weight and an evident cartilage-like matrix production. A concentration-dependent reduction in the mitochondrial activity due to blood contact was shown at the earlier culture time, followed by a partial recover at the longer culture time. A blood contact of 3 days affected the chondrocytes' activity and determined a delay in the maturation of the engineered cartilage. These findings have clinical relevance, as autologous chondrocytes seeded onto biological scaffolds has become an established surgical method for articular cartilage repair. Therefore, further investigation into material sciences should be encouraged for the development of scaffold protecting the reparative cells from the blood insult.


Study of behaviour on simulated daylight ageing of artists' acrylic and poly(vinyl acetate) paint films. This work proposes a multi-method approach that combines advanced microscopy (SEM/EDX, AFM) and spectroscopy (UV-vis and FTIR) techniques. This approach not only characterises the behaviour of the additives of two commercial poly(vinyl acetate) (PVAc) and acrylic emulsion paints but also simultaneously characterises the changes in chemical composition and morphology observed in the paint films as a result of ageing due to the paints being exposed to an intense source of simulated daylight. In parallel, a series of mechanical tests were performed that correlate the chemical changes in composition and the changes observed in the films' mechanical properties. This work was a comparative study between both types of acrylic and PVAc paints. The results obtained are of great interest for the modern paint conservation field as they provide valuable information on the mid- and long-term behaviours of these synthetic paints.


Muscle biopsies from the supraspinatus in retracted rotator cuff tears respond normally to passive mechanical testing: a pilot study. The aim of the present study was to assess the function of the isolated muscle component in retracted rotator cuff tears. Muscle biopsies were harvested from the supraspinatus and the ipsilateral deltoid in seven patients undergoing surgery for a large, retracted rotator cuff tear. Single fibres and fibre bundles were subjected to passive stretching in vitro with subsequent recordings of tension and sarcomere lengths using the laser diffraction technique. Stress-strain curves were plotted, and the elastic modulus was calculated for all preparations. Morphology was evaluated with regard to collagen fraction, ratio between fast and slow fibres, fibre size and fibre size variability using standard staining techniques. Intra-individual comparisons of the stress-strain curves showed a high degree of conformity in terms of both shape and tangent values, and there were no statistically significant differences in the elastic modulus for single fibres and bundles in the deltoid and supraspinatus muscles, respectively, supported by the analysis of the observed confidence interval of the differences between the paired values of the elastic modulus. There were no differences in collagen content, fibre size and ratio between fast and slow fibres in the deltoid and supraspinatus muscles, respectively. We conclude that muscle biopsies from the supraspinatus in retracted rotator cuff tears respond normally to mechanical testing in vitro.


Wrist muscle activation, interaction torque and mechanical properties in unskilled throws of different speeds. An unexpected property of unskilled overarm throws is that wrist flexion velocity at ball release does not increase in throws of increasing speed. We investigated the nature of the interaction torques and wrist mechanical properties that have been proposed to produce this property. Twelve recreational throwers made seated 2-D throws, which were used as a model for unskilled throwing. Joint motions were computed from recordings made with search coils; joint torques were calculated from inverse dynamics. Wrist flexion velocity at ball release was actually smaller in fast throws than in slow throws. This was associated in fast throws with the decrease in a large wrist flexor muscle torque (i.e., a calculated residual torque) in the last 40 ms before ball release, and its reversal to an extensor torque. Consequently, wrist flexor muscle torque was unable to oppose a small maintained wrist extensor interaction torque that arose from continuing elbow extension acceleration. The decrease in wrist flexor muscle torque was not associated with a decrease in wrist flexor EMG activity, nor with an increase in wrist extensor EMG activity. These findings support the hypothesis that the smaller wrist flexion velocity at ball release in fast 2-D throws results from a wrist extensor interaction torque and from a large wrist extensor viscoelastic torque. We propose that in fast 3-D throws skilled subjects decelerate elbow extension before ball release to help overcome these wrist extensor torques.


Surface texture can bias tactile form perception. The sense of touch is believed to provide a reliable perception of the object's properties; however, our tactile perceptions could be illusory at times. A recently reported tactile illusion shows that a raised form can be perceived as indented when it is surrounded by textured areas. This phenomenon suggests that the form perception can be influenced by the surface textures in its adjacent areas. As perception of texture and that of form have been studied independently of each other, the present study examined whether textures, in addition to the geometric edges, contribute to the tactile form perception. We examined the perception of the flat and raised contact surface (3.0 mm width) with various heights (0.1, 0.2, 0.3 mm), which had either textured or non-textured adjacent areas, under the static, passive and active touch conditions. Our results showed that texture decreased the raised perception of the surface with a small height (0.1 mm) and decreased the flat perception of the physically flat surface under the passive and active touch conditions. We discuss a possible mechanism underlying the effect of the textures on the form perception based on previous neurophysiological findings.


Automatic movement error detection and correction processes in reaching movements. Manual aiming movements can be amended during their execution. Recent evidence suggests that error detection and correction are based on automatic and even reflexive processing of afferent information. In this study, we wanted to determine whether these processes are affected by the occurrence of successive events requiring adjustments of the originally planned movement. To reach our goal, we used a video-aiming task. For a small proportion of the trials, the cursor moved by the participant was translated laterally by 15 mm (cursor jump) soon after movement initiation. For some of the cursor-jump trials, a second cursor jump occurred 100 ms after the first one and canceled or doubled the initial cursor translation. Results showed that participants were able to cancel or double the size of the correction in response to the second cursor jump. More importantly, in double-jump trials, the correction latency for the first and second cursor jumps did not differ from that of single-jump trials. Moreover, the correction for the second cursor jump blended seamlessly with the correction for the first cursor jump. These observations suggest that the processes leading of a correction for a cursor jump do not interfere with incoming visual information.


Temporal evolution of the phase correction response in synchronization of taps with perturbed two-interval rhythms. Human sensorimotor synchronization is flexible but subject to temporal constraints. Previous research has shown that musicians tend to lose synchrony with target tones in an isochronous sequence when the sequence rate exceeds 8-10 Hz, presumably because phase correction ceases to function. The present study investigated directly the time required for an immediate phase correction response (PCR). Musicians tapped in synchrony with cyclic two-interval (short-long) rhythms, using the two hands in alternation. Perturbations were applied to the long interval, and the compensatory shift of the next tap (the PCR) was measured following the short interval, whose duration was varied from 100 to 300 ms. The PCR was found to increase gradually within this range, being nearly absent at 100 ms. Similar results were obtained when participants tapped only with the second tone in each rhythmic group, which confirms that the PCR is based on the preceding tone rather than on the preceding tap-tone asynchrony, and also when the second tone was omitted in the pacing sequence, which indicates that the PCR occurs automatically even when there is no synchronization target for the critical tap. These results extend earlier findings regarding rate limits of synchronization and also provide further support for an event-based phase resetting account of the PCR.


Three generations of hereditary long-QT syndrome with complete penetrance caused by the p.G316E KCNQ1 mutation. This report describes a three-generation family with a severe phenotype of long-QT syndrome-1 (LQTS-1) caused by a single nucleotide mutation in the KQT-like, voltage-gated potassium channel-1 gene (KCNQ1; MIM 607542). Two members of the family died suddenly in their childhood, and all eight surviving members with prolonged QT have a heterozygous missense mutation resulting in a glycine-to-glutamate amino acid substitution at position 316 of the potassium channel. In this family, the newly reported mutation, guanine-to-adenosine at position 947 in the KCNQ1 gene, exhibits a dominant trait of LQTS with complete penetrance, in contrast to the relatively reduced clinical penetrance found in most LQTS cases.


Simultaneous resection of primary colorectal cancer and synchronous liver metastases.


Parathyroid carcinoma encountered after minimally invasive focused parathyroidectomy may not require further radical surgery. Parathyroid carcinoma accounts for <1% of tumors in primary hyperparathyroidism (PHPT). Distinguishing parathyroid malignancy from benign disease is difficult both before and after initial surgery. Despite the improved specificity of a malignant diagnosis with immunohistochemistry for parafibromin and PGP9.5, proven metastatic behavior remains the gold standard of diagnosis. Minimally invasive focused parathyroidectomy (MIP) is widely performed in patients with PHPT and positive localization studies; thus, it is inevitable that some parathyroid carcinomas will be encountered at MIP. We present our experience of this rare entity. The present study represents a surgical case series of patients with parathyroid carcinoma encountered after MIP. The clinicopathological features of benign and malignant parathyroid tumors were compared. Multiple regression analysis was undertaken to compare indicators of malignancy. Between May 1999 and April 2010, a total of 1,292 patients underwent MIP at the University of Sydney Endocrine Surgical Unit, and a histopathological diagnosis of parathyroid carcinoma was made in seven patients (0.5%). Staining for parafibromin and/or PGP9.5 was abnormal in five carcinomas (71%). Despite subsequent unilateral thyroid lobectomy and lymphadenectomy in six patients, no further malignancy was identified in any specimens. Compared to controls, preoperative calcium (p = 0.04) and parathyroid hormone (p = 0.01) were significantly higher in patients with malignancy. The positive predictive value of these parameters for carcinoma was 56 and 75%, respectively. In patients diagnosed with parathyroid carcinoma after MIP where preoperative imaging had already demonstrated localized disease, revision en bloc surgery did not reveal any residual disease. The benefits of further radical surgery for parathyroid carcinoma after MIP remain controversial.


Caldicoprobacter algeriensis sp. nov. a new thermophilic anaerobic, xylanolytic bacterium isolated from an Algerian hot spring. A thermophilic anaerobic bacterium (strain TH7C1(T)) was isolated from the hydrothermal hot spring of Guelma in the northeast of Algeria. Strain TH7C1(T) stained Gram-positive, was a non-motile rod appearing singly, in pairs, or as long chains (0.7-1 × 2-6 μm(2)). Spores were never observed. It grew at temperatures between 55 and 75°C (optimum 65°C) and at pH between 6.2 and 8.3 (optimum 6.9). It did not require NaCl for growth, but tolerated it up to 5 g l(-1). Strain TH7C1(T) is an obligatory heterotroph fermenting sugars including glucose, galactose, lactose, raffinose, fructose, ribose, xylose, arabinose, maltose, mannitol, cellobiose, mannose, melibiose, saccharose, but also xylan, and pyruvate. Fermentation of sugars only occurred in the presence of yeast extract (0.1%). The end-products from glucose fermentation were acetate, lactate, ethanol, CO(2), and H(2). Nitrate, nitrite, thiosulfate, elemental sulfur, sulfate, and sulfite were not used as electron acceptors. The G+C content of the genomic DNA was 44.7 mol% (HPLC techniques). Phylogenetic analysis of the small-subunit ribosomal RNA (rRNA) gene sequence indicated that strain TH7C1(T) was affiliated to Firmicutes, order Clostridiales, family Caldicoprobacteraceae, with Caldicoprobacter oshimai (98.5%) being its closest relative. Based on phenotypic, phylogenetic, and genetic characteristics, strain TH7C1(T) is proposed as a novel species of genus Caldicoprobacter, Caldicoprobacter algeriensis, sp. nov. (strain TH7C1(T) = DSM 22661(T) = JCM 16184(T)).


Temperature affects sole carbon utilization patterns of Campylobacter coli 49941. Campylobacter spp. are small, asaccharolytic bacteria exhibiting unique nutritional and environmental requirements. Campylobacter spp. exist as commensal organisms in some animal species, yet are estimated to be the most common causative agents of foodborne illness in humans. C. jejuni is most often associated with poultry, while C. coli are more frequently associated with swine. Temperature has been suggested to trigger potential colonization or virulence factors in C. jejuni, and recent studies have demonstrated temperature-dependent genes are important to colonization. It is possible that temperature-dependent colonization factors are in part responsible for the species-specific colonization characteristics of C. coli also. We determined utilization of 190 different sole carbon substrates by C. coli ATCC 49941 at 37 and 42°C using phenotype microarray (PM) technology. Temperature did affect amino acid utilization. L-asparagine and L-serine allowed significantly (P = 0.004) more respiration by C. coli ATCC 49941 at the lower temperature of 37°C as compared to 42°C. Conversely, L-glutamine was utilized to a significantly greater extent (P = 0.015) at the higher temperature of 42°C. Other organic substrates exhibited temperature-dependent utilization including succinate, D,L-malate, and propionate which all supported active respiration by C. coli to a significantly greater extent at 42°C. Further investigation is needed to determine the basis for the temperature-dependent utilization of substrates by Campylobacter spp. and their possible role in species-specific colonization.


Infection incidence and relative density of the bacteriophage WO-B in Aedes albopictus mosquitoes from fields in Thailand. We have used real-time quantitative PCR to measure, for the first time, the relative phage WO-B orf7 density and infection incidence in Aedes albopictus mosquitoes from fields in Thailand. Our results showed that the infection incidence of phage WO-B in this mosquito, sampled from geographically different places in Thailand, was 97.9%. Average relative densities of the offspring were different when collected from diverse parts and reared under the same conditions in the laboratory. Our results also revealed that geographical differences within Thailand did not influence the maternal transmission rate of bacteriophage WO-B. In addition, the orf7 loci might not be strictly associated with Wolbachia, because less than 100% of them were maternally inherited. This discovery does not support the hypothesis that bacteriophage WO-B is involved in Aedes albopictus' cytoplasmic incompatibility. Whether this bacteriophage actually is involved in Wolbachia-induced cytoplasmic incompatibility in this mosquito thus needs further investigation, and additional densities of phage WO-B loci should be integrated.


Phase II trial of the ribonucleotide reductase inhibitor 3-aminopyridine-2-carboxaldehydethiosemicarbazone plus gemcitabine in patients with advanced biliary tract cancer. 3-Aminopyridine-2-carboxaldehydethiosemicarbazone (3-AP) is a novel small molecule ribonucleotide reductase (RR) inhibitor which is more potent than hydroxyurea, the prototype of RR inhibitors. 3-AP enhances the cellular uptake and DNA incorporation of gemcitabine in tumor cell lines. We evaluated the combination of 3-AP plus gemcitabine in advanced biliary tract adenocarcinoma. Thirty-three patients with advanced adenocarcinoma of the gall bladder or biliary tract received gemcitabine (1,000 mg/m(2) on days 1, 8, and 15 every 28 days) 1 h after completing a 4-h infusion of 3-AP given at a dose of 105 mg/m(2) in patients with normal liver function (stratum A) or 80 mg/m(2) if abnormal liver function (stratum B). The trial was designed to determine whether the response rate was at least 30% in stratum A and 20% in stratum B. Objective response occurred in 3 of 23 patients (13%, 95% confidence intervals [CI] 3, 34%) with normal liver function, and in 0 of 10 patients with abnormal liver function. The most common grade 3-4 adverse events in all patients included neutropenia (42%), infection (33%), thrombocytopenia (27%), anemia (18%), and fatigue (15%). Fine needle aspiration of tumor samples obtained before and 24 h after 3-AP therapy showed increased R2 mRNA expression by in situ RT-PCR, suggesting RR inhibition. Despite evidence for RR inhibition in vivo, the 3-AP plus gemcitabine combination is not likely to be associated with a response rate exceeding 30% in patients with adenocarcinoma of the biliary tract.


Vasotocin- and mesotocin-induced increases in short-circuit current across tree frog skin. In adult amphibian skin, Na(+) crosses from outside to inside. This Na(+) transport can be measured as the amiloride-blockable short-circuit current (SCC) across the skin. We investigated the effects of arginine vasotocin (AVT) and mesotocin (MT), and those of antagonists of the vasopressin and oxytocin receptors, on the SCC across Hyla japonica skin. (1) Both AVT (100 pmol/L or more) and MT (1 nmol/L or more) increased the SCC. (2) The AVT- and MT-induced increases in SCC recovered with time (downregulation). (3) These AVT/MT-induced effects were blocked by application of OPC-31260 (vasopressin V(2)-receptor antagonist). (4) The OPC-31260 concentration needed to block the AVT-induced response was lower upon post-application (after application of agonist) than upon pre-application (before application of agonist), suggesting the number of receptors may have decreased after AVT application. (5) Upon repeated application of AVT (100 pmol/L), the induced SCC increase did not differ significantly between the 1st and 2nd applications. (6) The time to reach the half-maximum value of the AVT-induced or MT-induced increase in SCC was not significantly different between washout and post-application of OPC-31260, suggesting that post-application of OPC-31260 cleared AVT and MT from their receptors. The effects of AVT, MT, and their antagonists in H. japonica, which is adapted to a terrestrial habitat, are compared with our previously published data on Rana catesbeiana (=Lithobates catesbeianus), which is adapted to a semiaquatic habitat.


Intra- and extracellular osmotic regulation in the hololimnetic Caridea and Anomura: a phylogenetic perspective on the conquest of fresh water by the decapod Crustacea. We investigate extra- and intracellular osmoregulatory capability in two species of hololimnetic Caridea and Anomura: Macrobrachium brasiliense, a palaemonid shrimp, and Aegla franca, an aeglid anomuran, both restricted to continental waters. We also appraise the sharing of physiological characteristics by the hololimnetic Decapoda, and their origins and role in the conquest of fresh water. Both species survive salinity exposure well. While overall hyperosmoregulatory capability is weak in A. franca and moderate in M. brasiliense, both species strongly hyporegulate hemolymph [Cl(-)] but not osmolality. Muscle total free amino acids (FAA) increase slowly but markedly in response to the rapid rise in hemolymph osmolality consequent to hyperosmotic challenge: 3.5-fold in A. franca and 1.9-fold in M. brasiliense. Glycine, taurine, arginine, alanine and proline constitute ≈85% of muscle FAA pools in fresh water; taurine, arginine, alanine each contribute ≈22% in A. franca, while glycine predominates (70%) in M. brasiliense. These FAA also show the greatest increases on salinity challenge. Muscle FAA titers correlate strongly (R = 0.82) with hemolymph osmolalities across the main decapod sub/infraorders, revealing that marine species with high hemolymph osmolalities achieve isosmoticity of the intra- and extracellular fluids partly through elevated intracellular FAA concentrations; freshwater species show low hemolymph osmolalities and exhibit reduced intracellular FAA titers, consistent with isosmoticity at a far lower external osmolality. Given the decapod phylogeny adopted here and their multiple, independent invasions of fresh water, particularly by the Caridea and Anomura, our findings suggest that homoplastic strategies underlie osmotic and ionic homeostasis in the extant freshwater Decapoda.


Feeding into old age: long-term effects of dietary fatty acid supplementation on tissue composition and life span in mice. Smaller mammals, such as mice, possess tissues containing more polyunsaturated fatty acids (PUFAs) than larger mammals, while at the same time live shorter lives. These relationships have been combined in the 'membrane pacemaker hypothesis of aging'. It suggests that membrane PUFA content might determine an animal's life span. PUFAs in general and certain long-chain PUFAs in particular, are highly prone to lipid peroxidation which brings about a high rate of reactive oxygen species (ROS) production. We hypothesized that dietary supplementation of either n-3 or n-6 PUFAs might affect (1) membrane phospholipid composition of heart and liver tissues and (2) life span of the animals due to the altered membrane composition, and subsequent effects on lipid peroxidation. Therefore, we kept female laboratory mice from the C57BL/6 strain on three diets (n-3 PUFA rich, n-6 PUFA rich, control) and assessed body weights, life span, heart, and liver phospholipid composition after the animals had died. We found that while membrane phospholipid composition clearly differed between feeding groups, life span was not directly affected. However, we were able to observe a positive correlation between monounsaturated fatty acids in cardiac muscle and life span.


Overwinter body temperature patterns in captive jerboas (Jaculus orientalis): influence of sex and group. The jerboa (Jaculus orientalis) has been described in the past as a hibernator, but no reliable data exist on the daily and seasonal rhythmicity of body temperature (T (b)). In this study, T (b) patterns were determined in different groups of jerboas (isolated males and females, castrated males and grouped animals) maintained in captivity during autumn and winter, and submitted to natural variations of light and ambient temperature (T (a)). T (b) and T (a) variations were recorded with surgically implanted iButton temperature loggers at 30-min intervals for two consecutive years. About half (6/13) of isolated female jerboas hibernated with a T (b) < 33°C, with hibernation bouts interspersed with short periods of normothermy from November to February. Hibernation bout durations were longer (4-5 days) than those of normothermia phases (1-4 days). During hibernation, the minimum T (b) was low (T (b)min ~10.7°C). In contrast, one of the 12 isolated males showed short hibernation bouts of ca. 2 days late in the hibernation season, February-March. The males had T (b)min values of 15.1°C. In contrast to predictions, no castrated males hibernated. When jerboas were grouped, females and males exhibited concomitant torpor bouts. In males, the longest bouts were observed during the late hibernation season. These data suggest complex regulation of hibernation in jerboas.


Diabetes mellitus abrogates erythropoietin-induced cardioprotection against ischemic-reperfusion injury by alteration of the RISK/GSK-3β signaling. Recent studies reported cardioprotective effects of erythropoietin (EPO) against ischemia-reperfusion (I/R) injury through activation of the reperfusion injury salvage kinase (RISK) pathway. As RISK has been reported to be impaired in diabetes and insulin resistance syndrome, we examined whether EPO-induced cardioprotection was maintained in rat models of type 1 diabetes and insulin resistance syndrome. Isolated hearts were obtained from three rat cohorts: healthy controls, streptozotocin (STZ)-induced diabetes, and high-fat diet (HFD)-induced insulin resistance syndrome. All hearts underwent 25 min ischemia and 30 min or 120 min reperfusion. They were assigned to receive either no intervention or a single dose of EPO at the onset of reperfusion. In hearts from healthy controls, EPO decreased infarct size (14.36 ± 0.60 and 36.22 ± 4.20% of left ventricle in EPO-treated and untreated hearts, respectively, p < 0.05) and increased phosphorylated forms of Akt, ERK1/2, and their downstream target GSK-3β. In hearts from STZ-induced diabetic rats, EPO did not decrease infarct size (32.05 ± 2.38 and 31.88 ± 1.87% in EPO-treated and untreated diabetic rat hearts, respectively, NS) nor did it increase phosphorylation of Akt, ERK1/2, and GSK-3β. In contrast, in hearts from HFD-induced insulin resistance rats, EPO decreased infarct size (18.66 ± 1.99 and 34.62 ± 3.41% in EPO-treated and untreated HFD rat hearts, respectively, p < 0.05) and increased phosphorylation of Akt, ERK1/2, and GSK-3β. Administration of GSK-3β inhibitor SB216763 was cardioprotective in healthy and diabetic hearts. STZ-induced diabetes abolished EPO-induced cardioprotection against I/R injury through a disruption of upstream signaling of GSK-3β. In conclusion, direct inhibition of GSK-3β may provide an alternative strategy to protect diabetic hearts against I/R injury.


Mannitol-1-phosphate dehydrogenase activity in Ectocarpus siliculosus, a key role for mannitol synthesis in brown algae. Mannitol represents a major end product of photosynthesis in brown algae (Phaeophyceae), and is, with the β-1,3-glucan laminarin, the main form of carbon storage for these organisms. Despite its importance, little is known about the genes and enzymes responsible for the metabolism of mannitol in these seaweeds. Taking benefit of the sequencing of the Ectocarpus siliculosus genome, we focussed our attention on the first step of the synthesis of mannitol (reduction of the photo-assimilate fructose-6-phosphate), catalysed by the mannitol-1-phosphate dehydrogenase (M1PDH). This activity was measured in algal extracts, and was shown to be regulated by NaCl concentration in the reaction medium. Genomic analysis revealed the presence of three putative M1PDH genes (named EsM1PHD1, EsM1PDH2 and EsM1PDH3). Sequence comparison with orthologs demonstrates the modular architecture of EsM1PHD1 and EsM1PDH2, with an additional N-terminal domain of unknown function. In addition, gene expression experiments carried out on samples harvested through the diurnal cycle, and after several short-term saline and oxidative stress treatments, showed that EsM1PDH1 is the most highly expressed of these genes, whatever the conditions tested. In order to assess the activity of the corresponding protein, this gene was expressed in Escherichia coli. Cell-free extracts prepared from bacteria containing EsM1PDH1 displayed higher M1PDH activity than bacteria transformed with an empty plasmid. Further characterisation of recombinant EsM1PDH1 activity revealed its very narrow substrate specificity, salt regulation, and sensitivity towards an inhibitor of SH-enzymes.


Solvent-free geranyl oleate production by enzymatic esterification. This work reports the optimization of geranyl propionate production by esterification of geraniol and propionic acid in a solvent-free system using a commercial lipase as catalyst. For this purpose, a sequential strategy was performed applying two experimental designs. The operating conditions that optimized geranyl propionate production were determined to be 40 °C, geraniol to propionic acid molar ratio of 3:1, 150 rpm and 10 wt% of enzyme, with a resulting reaction conversion of about 93%. After determining the optimum reaction parameters, a kinetic study was carried out evaluating the influence of substrates molar ratio, enzyme concentration and temperature on reaction conversion. Results obtained in this step allow to conclude that an excess of alcohol (acid to alcohol molar ratio of 1:6), relatively low enzyme concentration (5 wt%), temperature of 40 °C and substrates molar ratio of 1:1 afforded nearly complete reaction conversion after 30 min of reaction. New experimental data on enzymatic esterification of geraniol and propionic acid for geranyl propionate production are reported in this work, showing a promising perspective of the technique to overcome the well-known drawbacks of the chemical-catalyzed route.


Determination of the complete nucleotide sequence of a lupine potyvirus isolate from the Czech Republic reveals that it belongs to a new member of the genus Potyvirus. The complete nucleotide sequence of the ssRNA genome of a lupine potyvirus (LP) isolate was determined. It comprised 10,113 nucleotides excluding the poly(A) tail. Phylogenetic analysis of CP protein sequences identified pepper veinal mottle virus, narcissus yellow stripe virus, and chili veinal mottle virus as the closest relatives, sharing coat protein amino acid sequence identities of only about 64% with the LP isolate. Thus, LP can be regarded as a member of a newly described potyvirus species, for which the name Lupine mosaic virus (LuMV) is proposed.


Molecular mass spectrometry imaging in biomedical and life science research. This review describes the current state of mass spectrometry imaging (MSI) in life sciences. A brief overview of mass spectrometry principles is presented followed by a thorough introduction to the MSI workflows, principles and areas of application. Three major desorption-ionization techniques used in MSI, namely, secondary ion mass spectrometry (SIMS), matrix-assisted laser desorption ionization (MALDI), and desorption electrospray ionization (DESI) are described, and biomedical and life science imaging applications of each ionization technique are reviewed. A separate section is devoted to data handling and current challenges and future perspectives are briefly discussed at the end.


ERCC2 Lys751Gln and Asp312Asn polymorphisms and gastric cancer risk: a meta-analysis. Studies investigating the association between excision repair cross-complimentary group 2 (ERCC2) polymorphisms and gastric cancer (GC) risk have reported conflicting results. We performed a meta-analysis of published epidemiological studies to derive a more precise estimation of the relationship. Published literature from PubMed, EMBASE, and China National Knowledge Infrastructure was retrieved. Ten studies with 2,141 GC cases and 5,343 controls were selected. No association between ERCC2 Lys751Gln polymorphism and GC susceptibility for all genetic models was found. When stratified by race, we found the Gln/Gln genotype carriers might be at high risk of GC among Asians, but not among Caucasians. Also, the pooled results showed there was a significant difference in genotype distribution between non-gastric cardia cancer cases and controls. For ERCC2 Asp312Asn polymorphism, significantly elevated GC risk was associated with Asn/Asn genotype (AA vs. GG + GA: OR = 1.36, 95%CI = 1.04-1.77, P = 0.02). We also found this genotype was associated with GC susceptibility among Asians and subjects without Helicobacter pylori infection. No publication bias was found in the present study. This meta-analysis concluded that both ERCC2 Lys751Gln and Asp312Asn polymorphisms might contribute to increased risk of GC among Asians.


Molecular characterization and comparative analysis of pandemic H1N1/2009 strains with co-circulating seasonal H1N1/2009 strains from eastern India. During the peak outbreak (July-September 2009), a total 1886 patients were screened in eastern India, of which 139 (7.37%) and 52 (2.76%) were positive for pH1N1 and seasonal H1N1, respectively. Full-length HA1, NA, NS1 and PB1-F2 genes of representative strains were sequenced. Phylogenetic analysis of deduced amino acid sequences of pH1N1 strains revealed HA1 and NS1 to be of North American swine lineage, and the NA gene of Eurasian swine lineage. Consistent with previous reports, the PB1-F2 gene of pH1N1 strains was unique due to a mutation resulting in a truncated protein of 11 aa. The HA, NA and NS1 genes of H1N1/2009 strains clustered with H1N1 strains of 2000-2009, whereas a subset of strains contained a pH1N1-like truncated PB1-F2. The truncated PB1-F2 may confer the advantage of lower pathogenicity but higher replication and infectivity to the human H1N1 strains. This is the first report of seasonal H1N1/2009 strains with a pH1N1/2009-like gene segment.


Expressed sequence-tag analysis of tobacco sperm cells reveals a unique transcriptional profile and selective persistence of paternal transcripts after fertilization. Transcript analysis of male gametes of Nicotiana tabacum was conducted to gather gene expression data regarding the specialization of male germ cells and transmission of paternal transcripts during fertilization. We constructed a tobacco sperm cell cDNA library yielding 1,864 expressed sequence tags representing 1,050 clusters; 37.2% of these clusters have no homologs in GenBank, and 42% did not match any functionally classified protein. A comparative analysis of tobacco sperm transcripts with those of Arabidopsis and maize confirms that some genes are conserved in sperm specialization, while some are distinct to tobacco germline cells. Using reverse transcription-PCR (RT-PCR) of selected transcripts, we evaluated expression of sperm-obtained sequences in vegetative tissue, isolated egg cells, zygotes, and two-celled proembryos, identifying sperm cell-specific transcripts as potential markers for fertilization analysis. We further confirmed that two clusters of sperm transcripts were detected in zygotes about 10 h after fertilization, offering new examples of apparently paternally transmitted transcripts that may be involved in egg cell activation and/or early embryogenesis.


Brown tumor of the cervical spine: a case report and review of the literature. Brown tumors (BT) are benign focal bone lesions that may appear in the context of primary and secondary hyperparathyroidism (HPT). Involvement of the spine is exceedingly rare. We present a case of brown tumor involving the cervical spine, the third reported in the literature. In the literature review (until August 2010), we found nine cases of spinal BT in primary HPT and 14 cases in secondary HPT. Fifteen patients (65%) had evidence of spinal cord compression. A 34-year-old woman on long-term hemodialysis, with secondary HPT, presented with a 9-month history of persistent neck pain. Radiographs of the cervical spine revealed an expansive osteolytic lesion in the posterior arch of the second cervical vertebra. MR imaging revealed an expansive mass on C2 affecting the vertebral body, odontoid process, right pedicle, laminas, and spinous process; there were no signs of spinal edema. A CT-guided needle biopsy of the lesion showed destruction of trabecular bone, infiltration of the fibroblastic cells, and abundant osteoclast-like multinucleated giant cells with hemorrhage and hemosiderin pigment, and the diagnosis of brown tumor was made. Cervical pain disappeared within a few days of parathyroidectomy, and rapid remineralization of C2 was evident within a few months. BT must always be considered in the context of hyperparathyroidism and osteolytic lesions. Vertebral BT can be particularly devastating due to medullar compression symptoms. Regression or complete disappearance of these lesions after parathyroidectomy is common, but prompt surgical decompression is necessary in case of medullar compression symptoms.


Comparison of two control measures of weatherstripping in reducing blowing dust during hospital renovations. Hospital renovation projects pose risks of invasive infection by fungi from dust that is blown about during the period in question. Control measures to reduce the amount of dust during hospital renovation are thus necessary. Currently, no study has compared different control measures for effectiveness through more than one period of renovation. In this study, we examined the capacities of two control measures of weatherstripping (0.15 mm poly film and adhesive tape) to reduce the amount of blowing dust during two different hospital renovations (in 2008 and 2009). The amount of dust in the air of the hospital before and during the renovation was measured about once a week in both 2008 and 2009, and the between-year and within-year differences were tested. Our study revealed that the weatherstripping used in 2009 (adhesive tape) was significantly more effective than the measures taken in 2008 (0.15 mm poly film) to reduce the amount of dust during the renovations (p < 0.001), while in both years the amount of dust became significantly higher during the renovations than before the renovations. Differences in the effectiveness of weatherstripping during renovations between floors of the hospital were not significant in both 2008 and 2009. The number of Aspergillus-positive samples did not significantly increase compared with the number observed before the start of the hospital renovations (2006-2007) in 2008 and 2009, respectively. The weatherstripping potentially reduced the associated risk of airborne fungal infection.


Pollution of River Mahaweli and farmlands under irrigation by cadmium from agricultural inputs leading to a chronic renal failure epidemic among farmers in NCP, Sri Lanka. Chronic renal failure (CRF) associated with elevated dietary cadmium (Cd) among farming communities in the irrigated agricultural area under the River Mahaweli diversion scheme has reached a significantly higher level of 9,000 patients. Cadmium, derived from contaminated phosphate fertilizer, in irrigation water finds its way into reservoirs, and finally to food, causing chronic renal failure among consumers. Water samples of River Mahaweli and its tributaries in the upper catchment were analyzed to assess the total cadmium contamination of river water and the possible source of cadmium. Except a single tributary (Ulapane Stream, 3.9 μg Cd/l), all other tested tributaries carried more than 5 μg Cd/l, the maximum concentration level accepted to be safe in drinking water. Seven medium-sized streams carrying surface runoff from tea estates had 5.1-10 μg Cd/l. Twenty larger tributaries (Oya), where the catchment is under vegetable and home garden cultivation, carried 10.1-15 μg Cd/l. Nine other major tributaries had extremely high levels of Cd, reaching 20 μg Cd/l. Using geographic information system (GIS), the area in the catchment of each tributary was studied. The specific cropping system in each watershed was determined. The total cadmium loading from each crop area was estimated using the rates and types of phosphate fertilizer used by the respective farmers and the amount of cadmium contained in each type of fertilizer used. Eppawala rock phosphate (ERP), which is mostly used in tea estates, caused least pollution. The amount of cadmium in tributaries had a significant positive correlation with the cadmium loading of the cropping system. Dimbula Tea Estate Stream had the lowest Cd loading (495.9 g/ha/year), compared with vegetable-growing areas in Uma Oya catchment with 50,852.5 g Cd/ha/year. Kendall's τ rank correlation value of total Cd loading from the catchment by phosphate fertilizer used in all crops in the catchment to the Cd content in the tributaries was +0.48. This indicated a major contribution by the cropping system in the upper catchment area of River Mahaweli to the eventual Cd pollution of river water. Low soil pH (4.5-5.2), higher organic matter content (2-3%), and 18-20 cmol/kg cation exchange capacity (CEC) in upcountry soil have a cumulative effect in the easy release of Cd from soil with the heavy surface runoff in the upcountry wet zone. In view of the existing water conveyance system from upcountry to reservoirs in North Central Province (NCP) through diversion of River Mahaweli, in addition to their own nonpoint pollution by triple superphosphate fertilizer (TSP), this demands a change in overall upper catchment management to minimize Cd pollution through agriculture inputs to prevent CRF due to elevated dietary cadmium among NCP farmers.


Spectrofluorimetric methods for the determination of gemifloxacin in tablets and spiked plasma samples. Two new, sensitive and selective spectrofluorimetric methods have been developed for the determination of gemifloxacin (GFX) in tablets and spiked plasma samples. Gemifloxacin, as a primary amine compound, reacts with 7-chloro-4-nitrobenzofurazon (NBD-Cl) (for method A) and fluorescamine (for method B) which are a highly sensitive fluorogenic reagents used in many investigations. For method A, the reaction product was measured spectrofluorimetrically at 516 nm with excitation at 451 nm. The reaction proceeded quantitatively at pH 8.5, 80 °C in 7 min. For method B, the method was based on the reaction between GFX and fluorescamine in borate buffer solution of pH 8.5 to give highly fluorescent derivatives that were measured at 481 nm using an excitation wavelength of 351 nm. The fluorescence intensity was directly proportional to the concentration over the range 40-200 ng mL(-1) and 100-1,200 ng mL(-1) for method A and B, respectively. Successful applications of the developed methods, for the drug determination in pharmaceutical preparations and spiked plasma samples, were performed.


Soil microbial characteristics at the monitoring plots on windthrow areas of the Tatra National Park (Slovakia): their assessment as environmental indicators. This study focused on the responses of soil microorganisms to different management regimes on disturbed windthrow areas. Microbial parameters potentially serving as indicators of environmental changes within a long-term monitoring of forest development after large-scale disturbance events were assessed. Basal and substrate-induced respiration, N mineralisation, catalase activity, microbial biomass as well as functional diversity based on Biolog assay were determined in soil samples from three disturbed plots and an undisturbed reference plot in the Tatra National Park (Slovakia) since 2006. A relative congruence of inter-annual trends of microbial activity indicators at all plots results from a common response of microbiota to changes of climate at the landscape level after forest stands were destroyed. While catalase activity and functional diversity proved to be useful indicators of temporal trends, microbial biomass seems to reflect different management regimes at the disturbed plots.


Comparative cancer risk assessment of THMs in drinking water from well water sources and surface water sources. This research evaluates the lifetime cancer risks from trihalomethanes in Tehran's drinking water. The Trihalomethanes were measured in seven different water districts. Sixty-three samples were taken from tap water across the city for 7 months. The samples were analyzed for trihalomethanes using US EPA method 524.2. The average concentration of total trihalomethanes in different districts were between 0.81 and 9.0 μg/L, and the highest concentrations were detected in district 2 at 19.5 μg/L. Total lifetime cancer risks assessment from exposure to trihalomethanes in drinking water (ingestion, inhalation, and skin routes) were performed for people living in different districts in Tehran. The lifetime cancer risk was 7.19 × 10(-5) in district 2 (a more affluent neighborhood) where mostly surface water sources is used to supply drinking water and 9.38 × 10(-6) in district 7 (a less affluent neighborhood) which is mainly supplied with well water sources. Based on the population data, the total expected lifetime cancer cases from exposure to trihalomethanes are 104, 108, 81, 81, 41, 27, and three for districts 1 through 7, respectively. The average lifetime cancer risk was 4.33 × 10(-5) which means a total of 606 lifetime cancer cases for the entire province of Tehran. The highest risk from THMs seems to be from the inhalation route followed by ingestion and dermal contacts.


Preservation of positional identity in fetus-derived neural stem (NS) cells from different mouse central nervous system compartments. Neural stem (NS) cells are a self-renewing population of symmetrically dividing multipotent radial glia-like stem cells, characterized by homogeneous expansion in monolayer. Here we report that fetal NS cells isolated from different regions of the developing mouse nervous system behave in a similar manner with respect to self-renewal and neuropotency, but exhibit distinct positional identities. For example, NS cells from the neocortex maintain the expression of anterior transcription factors, including Otx2 and Foxg1, while Hoxb4 and Hoxb9 are uniquely found in spinal cord-derived NS cells. This molecular signature was stable for over 20 passages and was strictly linked to the developmental stage of the donor, because only NS cells derived from E14.5 cortex, and not those derived from E12.5 cortex, carried a consistent transcription factor profile. We also showed that traits of this positional code are maintained during neuronal differentiation, leading to the generation of electrophysiologically active neurons, even if they do not acquire a complete neurochemical identity.


Use of feathers of feral pigeons (Columba livia) as a technique for metal quantification and environmental monitoring. This study quantifies metal concentrations in washed and unwashed feathers of feral pigeons (Columba livia). The material was collected in two different sites: Goiânia (capital of Goiás State), with high anthropic activities, and Jataí (a city in Goiás), with low anthropic activities. Results revealed that metals such as Pb, Cr, and Cd were deposited in the feathers originating from air and soil (exogenous process). Other metals such as Cu, Fe, Mn, and Zn, which are part of the tissue constitution of a living being, were absorbed through the pigeons' diet (endogenous process). There was a statistically significant difference between mean values of Cd, Pb, and Cr in the three collection sites. The order of metal concentrations were Civic Square > Zoo > Jataí, which shows highest levels of metal contamination in anthropic environments. Significant differences between Cd, Pb, and Cr in washed and unwashed feathers suggest that metal concentrations are directly related to exposure time due to exogenous sources. We concluded that it is possible to quantify and monitor metal levels in the environment by analyzing the feathers of urban feral pigeons. Furthermore, unwashed feathers were especially efficient to investigate endogenous and exogenous absorption of the metals in our study.


Use of fish functional traits to associate in-stream suspended sediment transport metrics with biological impairment. Loss of ecological integrity due to excessive suspended sediment in rivers and streams is a major cause of water quality impairment in the USA. Current assessment protocols for development of sediment total maximum daily loads (TMDLs) lack a means to link temporally variable sediment transport rates with specific losses of ecological functions as loads increase. In order to accomplish this linkage assessment, a functional traits-based approach was used to correlate site occurrences of 17 fish species traits in three main groups (preferred rearing habitat, trophic feeding guild, and spawning behavior) with suspended sediment transport metrics. The sediment transport metrics included concentrations, durations, and dosages for a range of exceedance frequencies; and mean annual suspended sediment yields (SSY). In addition, this study in the Northwestern Great Plains Ecoregion examined trait relationships with three environmental gradients: channel stability, drainage area, and elevation. Potential stressor responses due to elevated suspended sediment concentration (SSC) levels were correlated with occurrences of five traits: preferred pool habitat; feeding generalists, omnivores, piscivores, and nest-building spawners; and development of ecologically based TMDL targets were demonstrated for specific SSC exceedance frequencies. In addition, reduced site occurrences for preferred pool habitat and nest-building spawners traits were associated with unstable channels and higher SSY. At an ecoregion scale, a functional traits assessment approach provided a means to quantify relations between biological impairment and episodically elevated levels of suspended sediment, supporting efforts to develop ecologically based sediment TMDLs.


Advancing effects analysis for integrated, large-scale wildfire risk assessment. In this article, we describe the design and development of a quantitative, geospatial risk assessment tool intended to facilitate monitoring trends in wildfire risk over time and to provide information useful in prioritizing fuels treatments and mitigation measures. The research effort is designed to develop, from a strategic view, a first approximation of how both fire likelihood and intensity influence risk to social, economic, and ecological values at regional and national scales. Three main components are required to generate wildfire risk outputs: (1) burn probability maps generated from wildfire simulations, (2) spatially identified highly valued resources (HVRs), and (3) response functions that describe the effects of fire (beneficial or detrimental) on the HVR. Analyzing fire effects has to date presented a major challenge to integrated risk assessments, due to a limited understanding of the type and magnitude of changes wrought by wildfire to ecological and other nonmarket values. This work advances wildfire effects analysis, recognizing knowledge uncertainty and appropriately managing it through the use of an expert systems approach. Specifically, this work entailed consultation with 10 fire and fuels program management officials from federal agencies with fire management responsibilities in order to define quantitative resource response relationships as a function of fire intensity. Here, we demonstrate a proof-of-concept application of the wildland fire risk assessment tool, using the state of Oregon as a case study.


Formulation and stabilisation of the biocontrol yeast Pichia anomala. The yeast Pichia anomala has antifungal activities and its potential in biocontrol and biopreservation has previously been demonstrated. To practically use an organism in such applications on a larger scale the microbe has to be formulated and stabilised. In this review we give an overview of our experience of formulating and stabilising P. anomala strain J121 in a wider perspective. The stabilisation techniques we have evaluated were liquid formulations, fluidised bed drying, lyophilisation (freeze-drying) and vacuum drying. With all methods tested it was possible to obtain yeast cells with shelf lives of at least a few months and in all cases the biocontrol activity was retained. Fluidised bed drying was dependent on the addition of cottonseed flour as a carrier during the drying process. In liquid formulations a sugar, preferentially trehalose, was a required additive. These two kinds of microbial stabilisation are easily performed and relatively inexpensive but in order to keep the cells viable the biomaterial has to be stored at cool temperatures. However, there is room for optimization, such as improving the growth conditions, or include preconditioning steps to enable the cells to produce more compatible solutes necessary to survive formulation, desiccation and storage. In contrast, lyophilisation and vacuum drying require a lot of energy and are thus expensive. On the other hand, the dried cells were mostly intact after one year of storage at 30°C. Inevitably, the choice of formulation and stabilisation techniques will be dependent also on the intended use.


Assessment of left ventricular function: visual or quantitative?


A review of the pharmacological effects of Arctium lappa (burdock). Arctium lappa, commonly known as burdock, is being promoted/recommended as a healthy and nutritive food in Chinese societies. Burdock has been used therapeutically in Europe, North America and Asia for hundreds of years. The roots, seeds and leaves of burdock have been investigated in view of its popular uses in traditional Chinese medicine (TCM). In this review, the reported therapeutic effects of the active compounds present in the different botanical parts of burdock are summarized. In the root, the active ingredients have been found to "detoxify" blood in terms of TCM and promote blood circulation to the skin surface, improving the skin quality/texture and curing skin diseases like eczema. Antioxidants and antidiabetic compounds have also been found in the root. In the seeds, some active compounds possess anti-inflammatory effects and potent inhibitory effects on the growth of tumors such as pancreatic carcinoma. In the leaf extract, the active compounds isolated can inhibit the growth of micro-organisms in the oral cavity. The medicinal uses of burdock in treating chronic diseases such as cancers, diabetes and AIDS have been reported. However, it is also essential to be aware of the side effects of burdock including contact dermatitis and other allergic/inflammatory responses that might be evoked by burdock.


Incorporation of in silico biodegradability screening in early drug development--a feasible approach? The concentration of a pharmaceutical found in the environment is determined by the amount used by the patient, the excretion and metabolism pattern, and eventually by its persistence. Biological degradation or persistence of a pharmaceutical is experimentally tested rather late in the development of a pharmaceutical, often shortly before submission of the dossier to regulatory authorities. To investigate whether the aspect of persistence of a compound could be assessed early during drug development, we investigated whether biodegradation of pharmaceuticals could be predicted with the help of in silico tools. To assess the value of in silico prediction, we collected results for the OECD 301 degradation test ("ready biodegradability") of 42 drugs or drug synthesis intermediates and compared them to the prediction of the in silico tool BIOWIN. Of these compounds, 38 were predictable with BIOWIN, which is a module of the Estimation Programs Interface (EPI) Suite™ provided by the US EPA. The program failed to predict the two drugs which proved to be readily biodegradable in the degradation tests. On the other hand, BIOWIN predicted two compounds to be readily biodegradable which, however, proved to be persistent in the test setting. The comparison of experimental data with the predicted one resulted in a specificity of 94% and a sensitivity of 0%. The results of this study do not indicate that application of the biodegradation prediction tool BIOWIN is a feasible approach to assess the ready biodegradability during early drug development.


Endogenous hydrogen sulfide is an anti-inflammatory molecule in dextran sodium sulfate-induced colitis in mice. Endogenous hydrogen sulfide (H(2)S) is increasingly being recognized as an important gaseous physiological mediator. Accumulating evidence shows the functions of H(2)S in various models of disease, but rarely in colitis. In this study, we investigated the role of endogenous H(2)S in a dextran sodium sulfate (DSS)-induced colitis model. Acute colitis was induced using 8% DSS in male BALB/c mice. The mRNA expression of cystathionine γ-lyase (CSE), the primary synthetase of H(2)S in the gastrointestinal tract, and cystathionine-β-synthetase (CBS) was measured by real-time RT-PCR. The amount of H(2)S in the colonic mucosa was measured by gas chromatography. Colitis severity was evaluated clinically, histologically, and biochemically under the condition of co-treatment with DL-propargylglycine (PAG), an irreversible CSE inhibitor, and sodium sulfide (Na(2)S), an H(2)S donor. The mRNA expression levels of CSE and CBS, and the H(2)S content in the colonic mucosa were increased with time after DSS administration. The disease activity index, which was determined by weight loss, stool consistency, and intestinal bleeding, increased after DSS administration. PAG significantly enhanced the increase in the disease activity index scores. PAG also significantly increased tissue-associated myeloperoxidase activity and thiobarbituric acid-reactive substances in the inflamed mucosa. Moreover, Na(2)S counteracted these effects of PAG. Taken together, the results indicated that the inhibition of endogenous H(2)S generation caused the deterioration of DSS-induced colitis. We conclude that physiological H(2)S might act as an anti-inflammatory molecule in colitis.


Anti-inflammatory and immunomodulatory effects of RDV-8 [C18H22N2O2S (ethyl 1-butyl-6-methyl-2-phenyl-4-thioxo-1,4-dihydropyrimidine-5-carboxylate)] in a rat model of induced pleurisy and in vitro lymphoproliferation. RDV-8 [C(18)H(22)N(2)O(2)S (ethyl 1-butyl-6-methyl-2-phenyl-4-thioxo-1,4-dihydropyrimidine-5-carboxylate)] is derived from the 4-thioxopyrimidine, and presents important clinical effects. The present study explored the RDV-8 effects in the proliferation of human peripheral blood mononuclear cells (PBMCs), as well as in a pleurisy-induced rat model. PBMCs were directly plated in four different RDV-8 concentrations (0.0125, 0.025, 0.05 and 0.1 mg/mL). RDV-8 decreased cell proliferation and monocyte chemotactic protein 1 synthesis. The interleukin 1 levels and the cytotoxic effect were not significantly affected by RDV-8 treatment. In the carrageenan-induced pleurisy model, the RDV-8 (3 mg/kg) treatment induced a significant reduction in the exudate volume, in the polymorphonuclear leukocyte migration and in the pleural exudate NO levels. The results indicate that RDV-8 may have an immunomodulatory effect, as well as anti-inflammatory actions suggesting that it could represent a new strategy in the inflammatory response modulation.


Is looking older than one's actual age a sign of poor health? Physicians often begin the physical examination with an assessment of whether a patient looks older than his or her actual age. This practice suggests an implicit assumption that patients who appear older than their actual age are more likely to be in poor health. To determine the sensitivity and specificity of apparent age for the detection of poor health status. Cross-sectional. A total of 126 outpatients (ages 30-70) from four primary care clinics and one general internal medicine clinic at an academic medical institution. With the patient's actual age provided, physicians (n = 58 internal medicine residents and general internal medicine faculty) viewed patient photographs and assessed how old each patient looked. For each physician, we examined the sensitivity and specificity of the difference between how old the patient looked and the patient's actual age for the detection of poor health, defined using SF-12 physical health and mental health scores. Using the threshold of looking ≥ 5 years older than actual age and with poor health defined as an SF-12 score ≥ 2.0 SD below age group norms, median sensitivity was 29% (IQR, 19% to 35%), median specificity 82% (IQR, 77% to 88%), median positive likelihood ratio 1.7 (IQR, 1.3 to 2.2), and median negative likelihood ratio 0.9 (IQR, 0.8 to 0.9). Using the threshold of looking ≥ 10 years older than actual age, median sensitivity was 5% (IQR, 2% to 9%) and median specificity was 99% (IQR, 96% to 100%). The diagnostic value of apparent age depends on how many years older than his or her actual age a patient looks. A physician's assessment that a patient looks ≥ 10 years older than his or her actual age has very high specificity for the detection of poor health.


Rosuvastatin and thapsigargin modulate γ-secretase gene expression and APP processing in a human neuroglioma model. Alzheimer's disease (AD) is a progressive neurodegenerative disorder leading to slow neuronal loss in several brain regions. It is characterised by the presence of cerebral senile plaques comprised of aggregated amyloid-β peptides. Transcriptional regulation of the γ-secretase complex, which cleaves the β-amyloid precursor protein to produce Aβ-peptides, could modulate the pathological phenotype of AD patients. This study investigates whether rosuvastatin, an HMG-CoA reductase inhibitor, modulates the expression of genes involved in the function of the γ-secretase complex, in a human cellular model for Aβ peptide accumulation. In particular, we analysed the effect of the statin combined with apoptotic induction. Experimental apoptosis was induced by thapsigargin treatment, a drug that depletes intracellular calcium stores via inhibition of the calcium ATPase pump. Notably, systemic calcium dysregulation accompanies almost all of the brain pathology processes observed in AD. We found differential transcriptional regulation of some γ-secretase cofactors relative to rosuvastatin treatment, in cells expressing Swedish mutant APP. Interestingly, this statin down-regulated the transcription of some enzyme cofactors, similar to treatment with thapsigargin. However, rosuvastatin neither affected the basal Aβ levels nor counteracted APP processing or Aβ over-production triggered by the thapsigargin. Our results provide evidence that rosuvastatin alters gene expression of the γ-secretase complex without affecting enzyme activity.


Application of modeling to scale-up dissolution in pharmaceutical manufacturing. Liquid mixing scale-up in pharmaceutical industry has often been based on empirical approach in spite of tremendous understanding of liquid mixing scale-up in engineering fields. In this work, we attempt to provide a model-based approach to scale-up dissolution process from a 2 l lab-scale vessel to a 4,000 l scale vessel used in manufacturing. Propylparaben was used as a model compound to verify the model predictions for operating conditions at commercial scale that would result in similar dissolution profile as observed in lab scale. Geometric similarity was maintained between both of the scales to ensure similar mixing characteristics. We utilized computational fluid dynamics (CFD) to ensure that the operating conditions at laboratory and commercial scale will result in similar power per unit volume (P/V). Utilizing this simple scale-up criterion of similar P/V across different scales, results obtained indicate fairly good reproducibility of the dissolution profiles between the two scales. Utilization of concepts of design of experiments enabled summarizing scale-up results in statistically meaningful parameters, for example -90% dissolution in lab scale at a given time under certain operating conditions will result in 75-88% at commercial scale with 95% confidence interval when P/V is maintained constant across the two scales. In this work, we have successfully demonstrated that scale-up of solid dissolution can be done using a systematic process of lab-scale experiments followed by simple CFD modeling to predict commercial-scale experimental conditions.


Potential protective role of IL15Rα during inflammation. We have shown that TNFα specifically activates the interleukin-15 (IL15) system in cerebral endothelial cells composing the blood-brain barrier. To determine the functions of cerebral IL15 signaling in inflammation, we first treated mice with lipopolysaccharide (LPS) and determined the expression of the three receptor subtypes of IL15. Robust time-dependent upregulation occurred in all subunits. We then tested whether IL15Rα knockout (KO) affected the maintenance of body temperature and activity level after a single dose of LPS. Circadian telemetry data were analyzed by the cosinor method. Both wild-type and KO mice had clear 24-h rhythms of basal temperature and activity. KO mice had a significantly higher midline estimating statistic of rhythm (MESOR; approximating 24 h mean) of temperature and delayed 24-h acrophase (peak) of activity than the wild-type mice. LPS disrupted the circadian rhythm of activity more severely in the KO group. Besides a decrease in MESOR and 24-h amplitude of activity after LPS, the KO mice showed a significant reduction of MESOR, amplitude, and changed acrophase for temperature on the second of 2 days. The disrupted circadian rhythm of temperature and activity in the KO mice after LPS suggests that upregulated IL15 receptors may serve a beneficial role to counteract the consequences of neuroinflammation.


Post-traumatic stress disorder in medical settings: focus on the critically ill. The relevance of the post-traumatic stress disorder (PTSD) concept to medically ill patients is becoming increasingly clear. Some medical conditions (e.g., rheumatologic diseases) are likely related to PTSD indirectly. Others, such as myocardial infarction and critical illness/intensive care unit (ICU) treatment, are likely traumatic stressors. Importantly, PTSD seems to be a potent risk factor for fatal and nonfatal cardiac events. Risk factors for medical illness-related PTSD appear similar to risk factors for PTSD in general. PTSD is particularly common among survivors of critical illness and ICU treatment. Further research is needed to determine how specific ICU interventions (e.g., administration of benzodiazepines, corticosteroids, and catecholamines) affect PTSD risk. In the meantime, given the negative impact of PTSD on patients' quality of life, it is important for psychiatrists and other clinicians to be aware of the syndrome in survivors of critical illness so as to ensure that such patients receive the care that they deserve.


Evaluation of the oral ¹³C-bicarbonate tracer technique for the estimation of CO₂ production and energy expenditure in dogs during rest and physical activity. For feeding of working dogs during their daily life, illness, routine jobs or sporting activities, an accurate determination of their nutritional requirements is essential to ensure their optimal health and performance. To predict the appropriate guidelines about how to feed dogs, it appears essential to determine the energy expenditure (EE) in a reliable and feasible way. In the present experiment, the non-invasive oral ¹³C-bicarbonate tracer technique (o¹³CT), i.e. collection of breath samples after oral administration of NaH¹³CO₃, was used for the estimation of CO₂ production and EE in dogs. Measurements were conducted during two days of rest, and during three days with 3 h of exercise per day. Average EE was 483 and 876 kJ kg⁻⁰·⁷⁵ d⁻¹ during rest and exercise, respectively. The o¹³CT seems appropriate to use as a minimal restrictive and non-invasive method to obtain reliable estimates of EE in dogs at different activity levels under near natural conditions.


Spoilt for choice? Cross-sectional study of care-seeking for health problems during pregnancy in Mumbai slums. This study considers care-seeking patterns for maternal morbidity in Mumbai's slums. Our objectives were to document women's self-reported symptoms and care-seeking, and to quantify their choice of health provider, care-seeking delays and referrals between providers. The hypothesis that care-seeking sites for maternal morbidity mirror those used for antenatal care was also tested. We analysed data for 10,754 births in 48 slum areas and interviewed mothers about their illnesses and care-seeking during pregnancy. Institutional care-seeking was high across the board (>80%), and higher for 'trigger' symptoms suggestive of complications (>88%). Private-sector care was preferred, and increased with socio-economic status, although public providers also played an important role. Most women sought treatment at the same site they received their antenatal care, most were treated within 2 days, and less than 2% were referred to other providers. Our findings suggest that poor women in Mumbai recognise symptoms of obstetric complications and the need for health care. However, that more than 80% also sought care for minor conditions implies that the tendency to seek institutional care for serious conditions reflects a broader picture of care-seeking for all illnesses. The role of private health-care providers needs greater recognition, and further research is required on provider motivations and behaviour.


Dynamic modelling and simulation of ¹⁵N separation by chemical exchange in NO, NO₂-HNO₃ system. The dynamic behaviour of a ¹⁵N separation process by chemical exchange in a NO, NO₂-HNO₃ system has been analysed based on an accurate mathematical model. A nonlinear system of first-order partial differential equations was determined by considering the multiple exchange reactions between the components of the gaseous mixture and the liquid phase constituents. The mathematical model of the process describes the space-time variation of the ¹⁵N mole fraction in gas and liquid phases and provides a better understanding of operating limits and decision support in process design and optimisation.


Biodegradation kinetics of 2,6-dihydroxybenzoic acid and peptone mixture by acclimated microbial culture at low sludge age. This study evaluated the kinetics of 2,6-dihydroxybenozic acid and peptone biodegradation at low sludge age by acclimated culture under aerobic conditions. A laboratory-scale sequencing batch reactor was set and fed with peptone mixture. The system was operated at steady-state at a sludge age of 2 days. In order to assess biodegradation kinetics of 2,6-dihydroxybenozic acid and its impact on peptone utilization, a mixture of 2,6-dihydroxybenzoic acid and peptone was fed to mixed culture. After a period of four days, the system became acclimated to simultaneously remove both 2,6-dihydroxybenzoic acid and peptone mixture. A mechanistic model was developed involving model components and kinetic parameters for both substrates. This model was calibrated with related experimental data such as oxygen uptake rate and COD. Biodegradation characteristics and kinetics of peptone and 2,6-dihydroxybenzoic acid was estimated. The evaluation of calibrated model indicated that a group of microorganisms adjusted their enzymatic tools for the utilization of 2,6-dihydroxybenzoic acid resulting in dual microbial community development at low sludge age.


Natural radionuclide emission from coal-fired power plants in the southwestern of Turkey and the population exposure to external radiation in their vicinity. To evaluate the effect of radionuclide emission on the environment from Yatagan, Yenikoy and Kemerkoy coal-fired power plants which are located in southwestern Anatolia of Turkey, the concentrations of natural radionuclides such as (226)Ra, (232)Th and (40)K in coal, bottom ash and fly ash samples, have been measured, as well as the concentration of the same radionuclides in surface soils. The dose rate arises from the total radioactivity content of soil that the people living by the power plants are exposed to be assessed additionally. The average activity concentrations of (226)Ra for Yatagan CPP is 80 ± 22 Bq kg(-1) ranging from 56 to 131 Bq kg(-1), for Yenikoy CPP is 138 ± 20 Bq kg(-1) ranging from 115 to 189 Bq kg(-1), for Kemerkoy CPP is 238 ± 80 Bq kg(-1) ranging from 134 to 356 Bq kg(-1) in coal; average activity concentrations of (226)Ra in fly ash and in bottom ash for above-mentioned power plants are 334 ± 60 Bq kg(-1) ranging from 291 to 481 Bq kg(-1), 461 ± 33 Bq kg(-1) ranging from 398 to 511 Bq kg(-1), 815 ± 254 Bq kg(-1) ranging from 316 to 1260 Bq kg(-1), 276 ± 51 Bq kg(-1) ranging from 222 to 349 Bq kg(-1), 285 ± 69 Bq kg(-1) ranging from 213 to 409 Bq kg(-1), 743 ± 234 Bq kg(-1) ranging from 366 to 1098 Bq kg(-1), respectively. The radionuclides activity concentrations of surface soil in the vicinity of coal-fired power plants are 32 ± 9 Bq kg(-1) (18-53 Bq kg(-1)) for (226)Ra, 37 ± 16 Bq kg(-1) (17-89 Bq kg(-1)) for (232)Th, 455 ± 165 Bq kg(-1) (203-794 Bq kg(-1)) for (40)K relevant to Yatagan CPP; 42 ± 30 Bq kg(-1) (9-168 Bq kg(-1)) for (226)Ra, 32 ± 14 Bq kg(-1) (6-74 Bq kg(-1)) for (232)Th, 365 ± 151 Bq kg(-1) (117-937 Bq kg(-1)) for (40)K relevant to Yenikoy and Kemerkoy CPP. As a result, average dose rates in the vicinity of coal-fired power plants have been calculated to be 56 ± 16 nGy h(-1) ranging from 30 to 100 nGy h(-1) for Yatagan CPP, 54 ± 22 nGy h(-1) ranging from 15 to 126 nGy h(-1) for Yenikoy and Kemerkoy CPP. To sum up, the natural radionuclide activity concentrations of burnt coal and ashes thrown out from these three power plants are quite high relative to the world average UNSCEAR[1] data. In addition, the average (226)Ra, (232)Th and (40)K activity concentration values of surface soil samples and the calculated gamma dose rates in the vicinity of power plants were located within the worldwide intervals reported by UNSCEAR,[30] with some local differences.


Agrochemical spray drift; assessment and mitigation--a review. During application of agrochemicals spray droplets can drift beyond the intended target to non-target receptors, including water, plants and animals. Factors affecting this spray drift include mode of application, droplet size, which can be modified by the nozzle types, formulation adjuvants, wind direction, wind speed, air stability, relative humidity, temperature and height of released spray relative to the crop canopy. The rate of fall of spray droplets depends upon the size of the droplets but is modified by entrainment in a mobile air mass and is also influenced by the rate of evaporation of the liquid constituting the aerosol. The longer the aerosol remains in the air before falling to the ground (or alternatively striking an object above ground) the greater the opportunity for it to be carried away from its intended target. In general, all size classes of droplets are capable of movement off target, but the smallest are likely to move the farthest before depositing on the ground or a non-target receptor. It is not possible to avoid spray drift completely but it can be minimized by using best-management practices. These include using appropriate nozzle types, shields, spray pressure, volumes per area sprayed, tractor speed and only spraying when climatic conditions are suitable. Field layout can also influence spray drift, whilst crop-free and spray-free buffer zones and windbreak crops can also have a mitigating effect. Various models are available to estimate the environmental exposure from spray drift at the time of application.


Spatiotemporal trends of heavy metal concentrations in fish of the River Morava (Danube basin). The aim of this study was to assess mercury (Hg), cadmium (Cd) and lead (Pb) contamination in muscle tissue of fishes over the longitudinal profile of the River Morava (Czech Republic, Danube basin) and to detect any temporal trends over the past 18 years. Fish samples were collected in 1992, 1998, 2000, 2003 and 2009 at 6 study sites situated just downstream of important pollution sources. Chub (Leuciscus cephalus L.) were selected as indicator species at 5 sites, and brown trout (Salmo trutta m. fario L.) at the uppermost site where chub do not occur. In total, muscle tissue of 175 specimens of chub and 19 specimens of brown trout were analysed. Concentrations of heavy metals ranged as follows: mercury 0.015-0.369 mg/kg; cadmium 0.001-0.254 mg/kg and lead 0.006-1.505 mg/kg. Mercury levels did not exceed the maximum allowed concentration in the Czech Republic (0.5 mg/kg). Content of cadmium and lead in fish muscle exceeded the maximum allowed levels (0.05 and 0.3 mg/kg respectively) in 11 and 4 samples, respectively. On average, the order of metal concentration in fish muscle was: Hg>Pb>Cd. No significant differences were found between sites along the longitudinal profile of the river. Significant differences were found, however, for the interannual comparison of cadmium and lead (but not mercury) at different sites (P < 0.05). A catastrophic flood in 1997 resulted in an increase in metal concentrations, especially cadmium and lead, in the following 1998 season. Our results indicate that the Morava river basin does not represent a threatening source of mercury, cadmium or lead for the River Danube downstream.


Occupational exposure to hazardous airborne pollutants: effects of air mixing and source location. The presence of airborne pollutants in indoor environments has been associated with occupants' discomfort and/or adverse health effects. This study investigates occupational exposure in relation to indoor air mixing and source location relative to a human body. Experimental and computational methods were used to provide information about the pollutant distribution in the vicinity of the human body for different levels of room air mixing. Study results show that the often used assumption of uniform pollutant distribution in an occupied space is not always appropriate for estimation of inhalation exposure. Results also indicate that an occupant may experience very high acute exposure to airborne pollutants when little air mixing exists in a space and the pollutant source is in the vicinity of the occupant. The buoyancy-driven flow induced by the convective heat transfer from an occupant's body can transport pollutants in the occupant's vicinity to the breathing zone. Specific study results reveal that a source located in the occupant's front chest region makes a relatively large contribution to the breathing zone concentration compared with the other sources in the vicinity of the human body. With the source position in this region, exposure can be nine times greater than that calculated with the uniform mixing assumption. The buoyancy-driven convective plume around a body seems to have a significant influence on pollutant transport and human exposure, especially in the absence of room air mixing.