Bridging the data gaps in the epidemiology of hepatitis C virus infection in Malaysia using multi-parameter evidence synthesis.
Collecting adequate information on key epidemiological indicators is a prerequisite to informing a public health response to reduce the impact of hepatitis C virus (HCV) infection in Malaysia. Our goal was to overcome the acute data shortage typical of low/middle income countries using statistical modelling to estimate the national HCV prevalence and the distribution over transmission pathways as of the end of 2009. Multi-parameter evidence synthesis methods were applied to combine all available relevant data sources - both direct and indirect - that inform the epidemiological parameters of interest. An estimated 454,000 (95% credible interval [CrI]: 392,000 to 535,000) HCV antibody-positive individuals were living in Malaysia in 2009; this represents 2.5% (95% CrI: 2.2-3.0%) of the population aged 15-64 years. Among males of Malay ethnicity, for 77% (95% CrI: 69-85%) the route of probable transmission was active or a previous history of injecting drugs. The corresponding proportions were smaller for male Chinese and Indian/other ethnic groups (40% and 71%, respectively). The estimated prevalence in females of all ethnicities was 1% (95% CrI: 0.6 to 1.4%); 92% (95% CrI: 88 to 95%) of infections were attributable to non-drug injecting routes of transmission. The prevalent number of persons living with HCV infection in Malaysia is estimated to be very high. Low/middle income countries often lack a comprehensive evidence base; however, evidence synthesis methods can assist in filling the data gaps required for the development of effective policy to address the future public health and economic burden due to HCV.


A critical role for CCR2/MCP-1 interactions in the development of idiopathic pneumonia syndrome after allogeneic bone marrow transplantation.
Idiopathic pneumonia syndrome (IPS) is a major complication after allogeneic bone marrow transplantation (allo-BMT) and involves the infiltration of donor leukocytes and the secretion of inflammatory cytokines. We hypothesized that leukocyte recruitment during IPS is dependent in part upon interactions between chemokine receptor 2 (CCR2) and its primary ligand monocyte chemoattractant protein-1 (MCP-1). To test this hypothesis, IPS was induced in a lethally irradiated parent --> F1 mouse BMT model. Compared with syngeneic controls, pulmonary expression of MCP-1 and CCR2 mRNA was significantly increased after allo-BMT. Transplantation of CCR2-deficient (CCR2-/-) donor cells resulted in a significant reduction in IPS severity compared with transplantation of wild-type (CCR2+/+) cells and in reduced bronchoalveolar lavage (BAL) fluid cellularity and BAL fluid levels of tumor necrosis factor-alpha (TNF-alpha) and soluble p55 TNF receptor (sTNFRI). In addition, neutralization of MCP-1 resulted in significantly decreased lung injury compared with control-treated allogeneic recipients. Experimental data correlated with preliminary clinical findings; patients with IPS have elevated levels of MCP-1 in the BAL fluid at the time of diagnosis. Collectively, these data demonstrate that CCR2/MCP-1 interactions significantly contribute to the development of experimental IPS and suggest that interventions blocking these receptor-ligand interactions may represent novel strategies to prevent or treat this lethal complication after allo-BMT.


Differential sex mortality in a rural nineteenth-century population: Escazu, Costa Rica.
Cross-cultural studies of sex-specific mortality indicate that, whereas males experience their greatest mortality in industrialized societies, females experience their greatest mortality in populations with low life expectancy. The higher mortality of females in low-life-expectancy communities has been interpreted as a reflection of nutritional and health-care discrimination against females. Cross-cultural demographic studies also indicate that males have a higher frequency of violent and accidental deaths, possibly because of more frequent risky behaviors. This study focuses on Escazu, a rural nineteenth-century population from Costa Rica with low life expectancy. I investigate whether Escazu males had higher violent and accidental deaths and whether females had higher diarrhea-related deaths, an indication of nutritional discrimination. An analysis of mortality by cause of death indicates that males and females did not experience significantly different diarrhea-related death rates, although males did experience greater violent mortality. This study illustrates that more anthropological community-specific studies of mortality are needed to elucidate variation of death rates within large national or international regions.


Complement activation product C4d in oral and oropharyngeal squamous cell carcinoma.
Complement C4d-containing fragments have been proposed as diagnostic markers for lung cancer. The purpose of this study was to evaluate the presence of C4d in oropharyngeal (OPSCC) and oral (OSCC) squamous cell carcinomas. C4d staining was analyzed by immunohistochemistry in 244 OPSCC surgical specimens. C4d levels were quantified by ELISA in resting saliva samples from 48 patients with oral leukoplakia and 62 with OSCC. Plasma samples from 21 patients with leukoplakia and 30 with oral carcinoma were also studied. C4d staining in OPSCC specimens was associated with nodal invasion (P = 0.001), histopathologic grade (P = 0.014), disease stage (P = 0.040), and focal-adhesion kinase expression (P < 0.001). No association was found between C4d and prognosis. Saliva C4d levels were higher in patients with oral cancer than in subjects with leukoplakia (0.07 ± 0.07 vs 0.04 ± 0.03 μg ml(-1) , P = 0.003). The area under the ROC curve was 0.63 (95%CI: 0.55-0.71). Salivary C4d levels in stage IV patients were higher than in patients with earlier stages (P = 0.028) and correlated with tumor size (P = 0.045). Plasma C4d levels also correlated with salivary C4d levels (P = 0.041), but differences between patients with oral cancer and subjects with leukoplakia were not significant (1.26 ± 0.59 vs 1.09 ± 0.39 μg ml(-1) , P = 0.232). C4d-containing fragments are detected in oral primary tumors and are increased in saliva from patients with OSCC.


Self-Association of Rafoxanide in Aqueous Media and Its Application in Preparing Amorphous Solid Dispersions.
Our primary objective is to characterize the self-association of rafoxanide in alkaline media. The second objective is to illustrate the feasibility of using rafoxanide micellar solution as the feed solution to prepare amorphous solid dispersion via spray drying. Rafoxanide is a poorly water-soluble drug. It is a weak acid, and its poor aqueous solubility is due to its hydrophobicity. The surface-active property of rafoxanide has not been previously reported. It was discovered that the addition of a small percentage of organic solvents is required to elevate the solubility of rafoxanide above the critical micelle concentration to allow for the formation of micelles. Our fluorescence decay study confirms the self-association of rafoxanide in a cosolvent consisting of 70%, v/v, NaOH solution and 30%, v/v, acetone. The position of each functional group in the micellar structures using the 1H NMR technique was identified. The critical micelle concentration of rafoxanide in the cosolvent is determined to be 302 μg/mL using a surface tension method. The solubility of rafoxanide in 0.1 N NaOH solution is less than 11 μg/mL. Interestingly, the apparent solubility increased to 38,400 μg/mL in the presence of 30% acetone as the result of micelle formation. This unique solubility characteristic makes it feasible to prepare rafoxanide amorphous solid dispersions by spray drying a predominantly aqueous (70% 0.1 N NaOH solution and 30% acetone) based feed solution. Povidone and copovidone were both used as polymeric carriers. Based on solid-state characterization, including differential scanning calorimetry, X-ray powder diffraction, and hot-stage polarized light microscopy, our results indicate that rafoxanide solid dispersions prepared using this novel process are amorphous. Approximately 750-fold increase in the concentration of rafoxanide in aqueous media at pH 6.8 was achieved with the amorphous solid dispersions.


Genomics-Driven Discovery of NO-Donating Diazeniumdiolate Siderophores in Diverse Plant-Associated Bacteria.
Siderophores are key players in bacteria-host interactions, with the main function to provide soluble iron for their producers. Gramibactin from rhizosphere bacteria expands siderophore function and diversity as it delivers iron to the host plant and features an unusual diazeniumdiolate moiety for iron chelation. By mutational analysis of the grb gene cluster, we identified genes (grbD and grbE) necessary for diazeniumdiolate formation. Genome mining using a GrbD-based network revealed a broad range of orthologous gene clusters in mainly plant-associated Burkholderia/Paraburkholderia species. Two new types of diazeniumdiolate siderophores, megapolibactins and plantaribactin were fully characterized. In vitro assays and in vivo monitoring experiments revealed that the iron chelators also liberate nitric oxide (NO) in plant roots. This finding is important since NO donors are considered as biofertilizers that maintain iron homeostasis and increase overall plant fitness.


Crystallization and preliminary X-ray diffraction analysis of vaccinia virus H1L phosphatase.
The cysteine-based protein phosphatase H1L was the first reported dual-specificity protein phosphatase. H1L is encapsidated within the vaccinia virus and is required for successful host infection and for the production of viable vaccinia progeny. H1L has therefore been proposed as a target candidate for antiviral compounds. Recombinant H1L has been expressed in a catalytically inactive form using an Escherichia coli host, leading to purification and crystallization by the microbatch method. The crystals diffract to 2.1 A resolution using synchrotron radiation. These crystals belong to space group P422, with unit-cell parameters a = b = 98.31, c = 169.15 A, and are likely to contain four molecules in the asymmetric unit. A sulfur SAD data set was collected to 2.8 A resolution on beamline BM14 at the ESRF to facilitate structure determination. Attempts to derivatize these crystals with xenon gas changed the space group to I422, with unit-cell parameters a = b = 63.28, c = 169.68 A and a single molecule in the asymmetric unit. The relationship between these two crystal forms is discussed.


Population Graphs: the graph theoretic shape of genetic structure.
Patterns of intraspecific genetic variation result from interactions among both historical and contemporary evolutionary processes. Traditionally, population geneticists have used methods such as F-statistics, pairwise isolation by distance models, spatial autocorrelation and coalescent models to analyse this variation and to gain insight about causal evolutionary processes. Here we introduce a novel approach (Population Graphs) that focuses on the analysis of marker-based population genetic data within a graph theoretic framework. This method can be used to estimate traditional population genetic summary statistics, but its primary focus is on characterizing the complex topology resulting from historical and contemporary genetic interactions among populations. We introduce the application of Population Graphs by examining the range-wide population genetic structure of a Sonoran Desert cactus (Lophocereus schottii). With this data set, we evaluate hypotheses regarding historical vicariance, isolation by distance, population-level assignment and the importance of specific populations to species-wide genetic connectivity. We close by discussing the applicability of Population Graphs for addressing a wide range of population genetic and phylogeographical problems.


Impact of temperature and precipitation on propagation of intestinal schistosomiasis in an irrigated region in Ethiopia: suitability of satellite datasets.
To assess the suitability of satellite temperature and precipitation datasets for investigating the dependence of Schistosoma mansoni disease transmission on meteorological conditions in an irrigated agricultural region in Ethiopia. Data used were monthly number of patients infected with S. mansoni and seeking treatment at the local hospital, monthly maximum air temperature from a local weather station, monthly average land surface temperature from MODIS satellite data, monthly total precipitation from a local rain gauge and precipitation estimates from four widely used satellite products, namely, TMPA 3B42RT, TMPA 3B42, CMORPH and PERSIANN. The number of patients was used as proxy for vector abundance. Temperature and precipitation play a role in the transmission of S. mansoni disease. There is a weak but significant positive correlation between monthly maximum air temperature derived from a meteorological station (or average land surface temperature derived from MODIS satellite product) and the number of patients in the same month. There is a significant negative correlation between monthly precipitation volume (derived from rain gauge or satellite data) and number of patients at lags of 1 and 2 months. Satellite temperature and precipitation products provide useful information to understand and infer the relationship between meteorological conditions and S. mansoni prevalence.


Metals, oxidative stress and neurodegenerative disorders.
The neurodegenerative diseases, Alzheimer's disease (AD) and Parkinson's disease (PD), are age-related disorders characterized by the deposition of abnormal forms of specific proteins in the brain. AD is characterized by the presence of extracellular amyloid plaques and intraneuronal neurofibrillary tangles in the brain. Biochemical analysis of amyloid plaques revealed that the main constituent is fibrillar aggregates of a 39-42 residue peptide referred to as the amyloid-β protein (Aβ). PD is associated with the degeneration of dopaminergic neurons in the substantia nigra pars compacta. One of the pathological hallmarks of PD is the presence of intracellular inclusions called Lewy bodies that consist of aggregates of the presynaptic soluble protein called α-synuclein. There are various factors influencing the pathological depositions, and in general, the cause of neuronal death in neurological disorders appears to be multifactorial. However, it is clear, that the underlying factor in the neurological disorders is increased oxidative stress substantiated by the findings that the protein side-chains are modified either directly by reactive oxygen species (ROS) or reactive nitrogen species (RNS), or indirectly, by the products of lipid peroxidation. The increased level of oxidative stress in AD brain is reflected by the increased brain content of iron (Fe) and copper (Cu) both capable of stimulating free radical formation (e.g. hydroxyl radicals via Fenton reaction), increased protein and DNA oxidation in the AD brain, enhanced lipid peroxidation, decreased level of cytochrome c oxidase and advanced glycation end products (AGEs), carbonyls, malondialdehyde (MDA), peroxynitrite, and heme oxygenase-1 (HO-1). AGEs, mainly through their interaction with receptors for advanced glycation end products (RAGEs), further activate signaling pathways, inducing formation of proinflammatory cytokines such as interleukin-6 (IL-6). The conjugated aromatic ring of tyrosine residues is a target for free-radical attack, and accumulation of dityrosine and 3-nitrotyrosine has also been reported in AD brain. The oxidative stress linked with PD is supported by both postmortem studies and by studies showing the increased level of oxidative stress in the substantia nigra pars compacta, demonstrating thus the capacity of oxidative stress to induce nigral cell degeneration. Markers of lipid peroxidation include 4-hydroxy-trans-2-nonenal (HNE), 4-oxo-trans-2-nonenal (4-ONE), acrolein, and 4-oxo-trans-2-hexenal, all of which are well recognized neurotoxic agents. In addition, other important factors, involving inflammation, toxic action of nitric oxide (NO·), defects in protein clearance, and mitochondrial dysfunction all contribute to the etiology of PD. It has been suggested that several individual antioxidants or their combinations can be neuroprotective and decrease the risk of AD or slow its progression. The aim of this review is to discuss the role of redox metals Fe and Cu and non-redox metal zinc (Zn) in oxidative stress-related etiology of AD and PD. Attention is focused on the metal-induced formation of free radicals and the protective role of antioxidants [glutathione (GSH), vitamin C (ascorbic acid)], vitamin E (α-Tocopherol), lipoic acid, flavonoids [catechins, epigallocatechin gallate (EGCG)], and curcumin. An alternate hypothesis topic in AD is also discussed.


Development of a new classificatory model of malingering.
Inclusion criteria for the classification of malingering are shaped and largely predetermined by our explanatory theories. Current theories have postulated the motivation to malinger is either the product of underlying psychopathology (pathogenic model) or criminal backgrounds (DSM III-R model). I have proposed a third model that malingering is typically an adaptive response to adverse circumstances which may best be understood in the context of decision theory. Based on this approach I have argued that indices of malingering should be empirically derived and focused on clinical presentation. Finally, I have proposed a preliminary model for the classification of malingerers which combines clinical data with corroborative evidence.


Modal Symmetry Controlled Second-Harmonic Generation by Propagating Plasmons.
A new concept for second-harmonic generation (SHG) in an optical nanocircuit is proposed. We demonstrate both theoretically and experimentally that the symmetry of an optical mode alone is sufficient to allow SHG even in centro-symmetric structures made of centro-symmetric material. The concept is realized using a plasmonic two-wire transmission-line (TWTL), which simultaneously supports a symmetric and an antisymmetric mode. We first confirm that emission of second-harmonic light into the symmetric mode of the waveguide is symmetry-allowed when the fundamental excited waveguide modes are either purely symmetric or antisymmetric. We further switch the emission into the antisymmetric mode when a controlled mixture of the fundamental modes is excited simultaneously. Our results open up a new degree of freedom into the designs of nonlinear optical components and should pave a new avenue toward multifunctional nanophotonic circuitry.


Pneumatic transport exacerbates interference of room air contamination in blood gas samples.
To characterize and control the potential interference to P(O2) determinations when blood contaminated with air is sent via a pneumatic tube system (PTS). Both tonometered blood at P(O2)s of 65, 75, 142, and 339 mm Hg and arterial blood gas samples from patients with baseline P(O2)s from 70 to 400 mm Hg were analyzed for P(O2) to determine possible effects of air contamination from PTS transport. A large teaching hospital in which a variety of personnel routinely send samples to the laboratory by PTS transport. Twenty patients under anesthesia for elective surgery and 21 patients in an intensive care unit who had a wide range of P(O2)s. Several additional patients with a preexisting lung pathology likely to cause hypoxemia were selected to provide samples with low P(O2)s. Measurement of bias in P(O2) between samples sent via PTS and samples walked to the laboratory. Interference from air contamination was worse after PTS transport compared with manual transport of the specimen. Over a wide range, the P(O2) in specimens after PTS transport tended toward 160 mm Hg. Samples from hypoxemic patients were prone to errors in P(O2) that could have resulted in clinical misinterpretation; 5 of 10 samples with a baseline P(O2) less than 85 mm Hg had increases of 10 mm Hg or more when contaminated with air. Cooling samples with high P(O2)s minimized changes to P(O2), probably by increasing the solubility of oxygen. Mechanical buffering by various liners used in the carriers did little to alleviate the interference. Decreasing the speed of pneumatic transport by 50% lessened the effect on P(O2). Interference can be minimized by carefully purging samples of all air bubbles using the following protocol: invert syringe to check for air bubbles, then retap and reexpel bubbles if necessary. Personnel that collect and send blood gas samples via PTS should be educated about the problem of interference. Modifications both to pneumatic sample transport systems and to blood gas syringes should be investigated to minimize the effect.


Transcatheter aortic valve implantation with preexisting mechanical mitral prosthesis - use of CT angiography.
A 72-year-old Chinese female with prior mitral valve replacement with a mechanical prosthesis 13 years before was diagnosed with severe symptomatic aortic stenosis. She was considered a high-risk surgical candidate and was considered for transcatheter aortic valve implantation using a balloon-expandable device. The close proximity of the non-compliant mechanical mitral prosthesis to the aortic annulus raised concerns that the deployment of the device may be affected. Gated cardiac computed tomographic angiography was used to assess the distance between the mitral prosthesis and the aortic annulus. Transcatheter aortic valve implantation using the transapical approach was subsequently performed successfully.


Dynamic QTL analysis and candidate gene mapping for waterlogging tolerance at maize seedling stage.
Soil waterlogging is one of the major abiotic stresses adversely affecting maize growth and yield. To identify dynamic expression of genes or quantitative trait loci (QTL), QTL associated with plant height, root length, root dry weight, shoot dry weight and total dry weight were identified via conditional analysis in a mixed linear model and inclusive composite interval mapping method at three respective periods under waterlogging and control conditions. A total of 13, 19 and 23 QTL were detected at stages 3D|0D (the period during 0-3 d of waterlogging), 6D|3D and 9D|6D, respectively. The effects of each QTL were moderate and distributed over nine chromosomes, singly explaining 4.14-18.88% of the phenotypic variation. Six QTL (ph6-1, rl1-2, sdw4-1, sdw7-1, tdw4-1 and tdw7-1) were identified at two consistent stages of seedling development, which could reflect a continuous expression of genes; the remaining QTL were detected at only one stage. Thus, expression of most QTL was influenced by the developmental status. In order to provide additional evidence regarding the role of corresponding genes in waterlogging tolerance, mapping of Expressed Sequence Tags markers and microRNAs were conducted. Seven candidate genes were observed to co-localize with the identified QTL on chromosomes 1, 4, 6, 7 and 9, and may be important candidate genes for waterlogging tolerance. These results are a good starting point for understanding the genetic basis for selectively expressing of QTL in different stress periods and the common genetic control mechanism of the co-localized traits.


Pharmacokinetics and pharmacodynamics of meldonium in exercised thoroughbred horses.
Although developed as a therapeutic medication, meldonium has found widespread use in human sports and was recently added to the World Anti-Doping Agency's list of prohibited substances. Its reported abuse potential in human sports has led to concern by regulatory authorities about the possible misuse of meldonium in equine athletics. The potential abuse in equine athletes along with the limited data available regarding the pharmacokinetics and pharmacodynamics of meldonium in horses necessitates further study. Eight exercised adult thoroughbred horses received a single oral dose of 3.5, 7.1, 14.3 or 21.4 mg/kg of meldonium. Blood and urine samples were collected and analyzed using liquid chromatography tandem mass spectrometry. Pharmacokinetic parameters were determined using non-compartmental analysis. Maximum serum concentrations ranged from 440.2 to 1147 ng/mL and the elimination half-life from 422 to 647.8 h. Serum concentrations were below the limit of quantitation by days 4, 7, 12 and 12 for doses of 3.5, 7.1, 14.3 and 21.4 mg/kg, respectively. Urine concentrations were below the limit of detection by day 44 following administration of 3.5 mg/kg and day 51 for all other dose groups. No adverse effects were observed following meldonium administration. While the group numbers were small, changes in heart rate were observed in the 3.5 mg/kg dose group (n = 1). Glucose concentrations changed significantly in all dose groups studied (n = 2 per dose group). Similar to that reported for humans, the detection time of meldonium in biological samples collected from horses is prolonged, which should allow for satisfactory regulation in performance horses. Copyright © 2017 John Wiley & Sons, Ltd.


Optimizing actigraphic estimates of polysomnographic sleep features in insomnia disorder.
Actigraphy is a useful tool for estimating sleep, but less accurately distinguishes sleep and wakefulness in patients with insomnia disorder (ID) than in good sleepers. Specific algorithm parameter settings have been suggested to improve the accuracy of actigraphic estimates of sleep onset or nocturnal sleep and wakefulness in ID. However, a direct comparison of how different algorithm parameter settings affect actigraphic estimates of sleep features has been lacking. This study aimed to define the optimal algorithm parameter settings for actigraphic estimates of polysomnographic sleep features in people suffering from ID and matched good sleepers. We simultaneously recorded actigraphy and polysomnography without sleep diaries during 210 laboratory nights of people with ID (n = 58) and matched controls (CTRL) without sleep complaints (n = 56). We analyzed cross-validation errors using 150 algorithm parameter configurations and Bland-Altman plots of sleep features using the optimal settings. Optimal sleep onset latency and total sleep time (TST) errors were lower in CTRL (8.9 ± 2.1 and 16.5 ± 2.1 min, respectively) than in ID (11.7 ± 0.8 and 29.1 ± 3.4 min). The sleep-wake algorithm, a period duration of 5 min, and a wake sensitivity threshold of 40 achieved optimal results in ID and near-optimal results in CTRL. Bland-Altman plots were nearly identical for ID and controls for all common all-night sleep features except for TST. This systematic evaluation shows that actigraphic sleep feature estimation can be improved by using uncommon parameter settings. One specific parameter setting provides (near-)optimal estimation of sleep onset and nocturnal sleep across ID and controls.


A molecular dynamics study of bond exchange reactions in covalent adaptable networks.
Covalent adaptable networks are polymers that can alter the arrangement of network connections by bond exchange reactions where an active unit attaches to an existing bond then kicks off its pre-existing peer to form a new bond. When the polymer is stretched, bond exchange reactions lead to stress relaxation and plastic deformation, or the so-called reforming. In addition, two pieces of polymers can be rejoined together without introducing additional monomers or chemicals on the interface, enabling welding and reprocessing. Although covalent adaptable networks have been researched extensively in the past, knowledge about the macromolecular level network alternations is limited. In this study, molecular dynamics simulations are used to investigate the macromolecular details of bond exchange reactions in a recently reported epoxy system. An algorithm for bond exchange reactions is first developed and applied to study a crosslinking network formed by epoxy resin DGEBA with the crosslinking agent tricarballylic acid. The trace of the active units is tracked to show the migration of these units within the network. Network properties, such as the distance between two neighboring crosslink sites, the chain angle, and the initial modulus, are examined after each iteration of the bond exchange reactions to provide detailed information about how material behaviors and macromolecular structure evolve. Stress relaxation simulations are also conducted. It is found that even though bond exchange reactions change the macroscopic shape of the network, microscopic network characteristic features, such as the distance between two neighboring crosslink sites and the chain angle, relax back to the unstretched isotropic state. Comparison with a recent scaling theory also shows good agreement.


Development of a swine-specific fecal pollution marker based on host differences in methanogen mcrA genes.
The goal of this study was to evaluate methanogen diversity in animal hosts to develop a swine-specific archaeal molecular marker for fecal source tracking in surface waters. Phylogenetic analysis of swine mcrA sequences compared to mcrA sequences from the feces of five animals (cow, deer, sheep, horse, and chicken) and sewage showed four distinct swine clusters, with three swine-specific clades. From this analysis, six sequences were chosen for molecular marker development and initial testing. Only one mcrA sequence (P23-2) showed specificity for swine and therefore was used for environmental testing. PCR primers for the P23-2 clone mcrA sequence were developed and evaluated for swine specificity. The P23-2 primers amplified products in P23-2 plasmid DNA (100%), pig feces (84%), and swine waste lagoon surface water samples (100%) but did not amplify a product in 47 bacterial and archaeal stock cultures and 477 environmental bacterial isolates and sewage and water samples from a bovine waste lagoon and a polluted creek. Amplification was observed in only one sheep sample out of 260 human and nonswine animal fecal samples. Sequencing of PCR products from pig feces demonstrated 100% similarity to pig mcrA sequence from clone P23-2. The minimal amount of DNA required for the detection was 1 pg for P23-2 plasmid, 1 ng for pig feces, 50 ng for swine waste lagoon surface water, 1 ng for sow waste influent, and 10 ng for lagoon sludge samples. Lower detection limits of 10(-6) g of wet pig feces in 500 ml of phosphate-buffered saline and 10(-4) g of lagoon waste in estuarine water were established for the P23-2 marker. This study was the first to utilize methanogens for the development of a swine-specific fecal contamination marker.


Ancient host shifts followed by host conservatism in a group of ant parasitoids.
While ant colonies serve as host to a diverse array of myrmecophiles, few parasitoids are able to exploit this vast resource. A notable exception is the wasp family Eucharitidae, which is the only family of insects known to exclusively parasitize ants. Worldwide, approximately 700 Eucharitidae species attack five subfamilies across the ant phylogeny. Our goal is to uncover the pattern of eucharitid diversification, including timing of key evolutionary events, biogeographic patterns and potential cophylogeny with ant hosts. We present the most comprehensive molecular phylogeny of Eucharitidae to date, including 44 of the 53 genera and fossil-calibrated estimates of divergence dates. Eucharitidae arose approximately 50 Ma after their hosts, during the time when the major ant lineages were already established and diversifying. We incorporate host association data to test for congruence between eucharitid and ant phylogenies and find that their evolutionary histories are more similar than expected at random. After a series of initial host shifts, clades within Eucharitidae maintained their host affinity. Even after multiple dispersal events to the New World and extensive speciation within biogeographic regions, eucharitids remain parasitic on the same ant subfamilies as their Old World relatives, suggesting host conservatism despite access to a diverse novel ant fauna.


Treatment of sagittal sinus thrombosis associated with cerebral hemorrhage and intracranial hypertension.
Two cases of complete sagittal sinus occlusion with multiple brain hemorrhages, elevated intracranial pressure, and disseminated intravascular coagulation are described. These patients were successfully managed using pentobarbital-induced coma to ameliorate intracranial pressure elevation. This therapy was combined with monitoring of intracranial pressure and intermittent drainage of cerebrospinal fluid to further control intracranial pressure elevations. Thrombus and coagulopathy resolved with pentobarbital alone in one patient and after pentobarbital plus heparin therapy in the second patient. It is suggested that cases of severe distal sagittal sinus thrombosis with brain hemorrhage and intracranial hypertension may benefit from combined pentobarbital coma and intraventricular drainage. This allows for stabilization of bleeding tendencies before instituting heparin therapy when necessary. Management of sagittal sinus thrombosis with barbiturates or ventricular drainage is best performed in an intensive care unit environment with continuous monitoring of intracranial pressure and substantial electrophysiologic and neuroradiologic support.


Percutaneous therapy to maintain dialysis access successfully prolongs functional duration after primary failure.
The role of endovascular therapy for thrombosed dialysis access has grown despite the paucity of data on its viability. The purpose of this study was to characterize the outcomes of a universal endovascular dialysis declot policy at a tertiary medical center. A database of patients undergoing endovascular treatment of thrombosed dialysis access between 1997 and 2003 was maintained. A two-puncture, combined percutaneous mechanical and pharmacologic thrombectomy technique was used. Data were collected on the success rate, complication rate, long-term patency, and presence and location of stenosis. Fistulograms were reviewed in all cases to assess lesion characteristics and pre- and postprocedure results. Results were standardized to current Society of International Radiology and Society for Vascular Surgery criteria. Failure was considered as either an anatomic defect requiring therapy or loss of functionality of the fistula. Life-table analyses were performed to assess time-dependent outcomes. Cox's proportional hazard analyses were performed to identify factors associated with outcomes. Values are the mean +/- standard error of the mean. There were 114 patients (50% male; average age 58 years, range 21-78) who presented with 174 thrombosed grafts. Therapy was performed for 237 thrombotic events (median 2, range 1-5 thrombotic events per hemodialysis access). After successful declot, anastomotic venous stenoses were encountered in 72% and central venous stenoses in 18% of cases; no cause was found in 10%. All stenoses were treated with balloon angioplasty. The technical failure rate was 4.6%. The 30-day all-cause mortality rate was 1.7%, and major morbidity rate was 2.4%. There were 413 interventions (236 percutaneous transluminal angioplasty and/or 183 declot) performed to maintain patency, which amounted to 2.3 interventions per patient. Average primary functional dialysis life span was 6.7 months up to the primary thrombotic event. Aggressive endoluminal therapy added a further average of 12 months of functionality (defined as continued dialysis access). A universal policy of endovascular therapy for occluded dialysis access results in reestablishment of function in the majority of patients and will triple functional longevity. Furthermore, while this approach remains procedure-intensive, it carries low morbidity and mortality and preserves future sites of access.


Modulation by cell trypsinization of Sendai virus expression in African green monkey kidney cells: first infection and establishment of a carrier state.
Two African green monkey kidney (AGMK) cell lines, 37RC (interferon-producing) and Vero (non-interferon-producing), were infected by egg-grown Sendai virus passaged in eggs at high and low m.o.i. The appearance of haem-adsorption, and cytopathic effect (c.p.e.) as well as the presence of haemagglutinating virions in the supernates were much more pronounced with a virus seed obtained with 10(-3) diluted passages than with a seed obtained with undiluted inoculum. They were also independent of interferon production (data obtained in 37RC and Vero cells were almost superimposable). In studies carried out with the virus seed prepared at high dilution the establishment of infection was maximal when monolayers were infected as soon as 5 h after trypsinization and cell seeding, regardless of cell density. Virus in supernates obtained from cultures infected 5, 20 or 50 h after seeding exhibited a greatly reduced infectivity for monkey cells, but not apparently for chick embryos. Trypsin treatment of the virus supernates restored their infectivity for AGMK cells more efficiently for virus released from cells infected 5 h after seeding than for virus released from cells infected later after seeding. In keeping with these observations, virus in supernates from cultures infected 5, 20 or 50 h after seeding contained increasing amounts of auto-interfering virions. The decreased infectivity obtained in cell supernates was not accounted for by major differences in virus RNA synthesis. Similarly, the optimum infection established in cells seeded only for 5 h was not due to increased virus adsorption. Several lines of cells surviving first infection with egg-grown Sendai virus have been obtained and kept in cultures for 3 to 18 months. Virus release and c.p.e. apparently become reduced in the cells surviving the first infection until the newly repopulated monolayers must undergo trypsinization. Weekly protease treatments of the cells reactivate all parameters of virus infection which again will tend to disappear slowly and then reappear following each trypsinization ('intermittent' carrier state). The establishment and the 'intermittent' reactivation of these lines were not prevented by the inclusion in the medium of anti-Sendai virus serum. Temperature-sensitive ts functions do not seem to play an important role in this virus-host relationship.


Clinical implications of monitoring nivolumab immunokinetics in non-small cell lung cancer patients.
The PD-1-blocking antibody nivolumab persists in patients several weeks after the last infusion. However, no study has systematically evaluated the maximum duration that the antibody persists on T cells or the association between this duration and residual therapeutic efficacy or potential adverse events. To define the duration of binding and residual efficacy of nivolumab after discontinuation, we developed a simplified strategy for T cell monitoring and used it to analyze T cells from peripheral blood from 11 non-small cell lung cancer patients previously treated with nivolumab. To determine the suitability of our method for other applications, we compared transcriptome profiles between nivolumab-bound and nivolumab-unbound CD8 T cells. We also applied T cell monitoring in 2 nivolumab-treated patients who developed progressive lung tumors during long-term follow-up. Prolonged nivolumab binding was detected more than 20 weeks after the last infusion, regardless of the total number of nivolumab infusions (2-15 doses) or type of subsequent treatment, in 9 of the 11 cases in which long-term monitoring was possible. Ki-67 positivity, a proliferation marker, in T cells decreased in patients with progressive disease. Transcriptome profiling identified the signals regulating activation of nivolumab-bound T cells, which may contribute to nivolumab resistance. In 2 patients who restarted nivolumab, T cell proliferation markers exhibited the opposite trend and correlated with clinical response. Although only a few samples were analyzed, our strategy of monitoring both nivolumab binding and Ki-67 in T cells might help determine residual efficacy under various types of concurrent or subsequent treatment. University Hospital Medical Information Network Clinical Trials Registry, UMIN000024623. This work was supported by Japan Society for the Promotion of Science KAKENHI (JP17K16045, JP18H05282, and JP15K09220), Japan Agency for Medical Research and Development (JP17cm0106310, JP18cm0106335 and JP18cm059042), and Core Research for Evolutional Science and Technology (JPMJCR16G2).


A series of two-photon absorption pyridinium sulfonate inner salts targeting endoplasmic reticulum (ER), inducing cellular stress and mitochondria-mediated apoptosis in cancer cells.
In this study, a two-photon active DiphenthioER1 was screened from the four pyridinium sulfonate salt derivatives (TriphenER1-2 and DiphenthioER1-2) for imaging endoplasmic reticulum (ER) and tracking the dynamics of the ER morphology. The photophysical properties of TriphenER1-2 and DiphenthioER1-2 were systemically investigated both experimentally and theoretically, revealing that they possess large Stokes shifts, and large two-photon absorption cross-sections from 163 GM to 2023 GM in the near infrared region using a Z-scan method by avoiding spontaneous fluorescence, deep tissue penetration, and low cell damage in living cells. Among them, the DiphenthioER1 compound was found to exhibit the highest cellular uptake ability and two-photon fluorescence signals using confocal microscopy. DiphenthioER1 successfully targeted the ER and induced ER-stress, subsequently nuclear misshaping and mitochondria-mediated apoptosis have been displayed. This study thus provides great insights into designing novel two-photon fluorescent materials with dual functionality and offers tools to understand the ER-stress related mechanism for cell and chemical biologists.


Older people's experience of participating in a reminiscence group with a gerotranscendental perspective: reminiscence group with a gerotranscendental perspective in practice.
Tornstam's theory of gerotranscendence provides the theoretical underpinning for two small reminiscence groups that met for eight weekly sessions in a Swedish daycentre. Tornstams's theoretical perspective provides a developmental model for understanding positive progressive changes in values, attitudes and behaviour in the second half of life. Practical guidelines derived from this theory informed the group-work approach used. Participants' opinions about their reminiscence experience were analysed to determine the relevance of gerotranscendental theory as a basis for understanding older people's use of reminiscence in the ageing process. The study aimed at investigating older peoples' experiences of participating in a reminiscence group with a gerotranscendental perspective. The study used a qualitative approach. Older people were invited to participate in reminiscence group sessions arranged at a daycentre. When the sessions were finished, participants were interviewed about their experience of the reminiscence group. Data were analysed and categorized by using qualitative methods. The participants were satisfied with the arrangement of the group sessions. All participants believed they had reminisced and thought much more about their childhood and also recalled other memories from their lives during the period. Three quite different views emerged of the recall experience and effects of participating in the reminiscence group: 'An activity like any other; an activity that led to thoughts about memories from life or an activity that influenced my thoughts about life.' Findings suggest that it is possible to arrange reminiscence groups from a gerotranscendental perspective that serve as an intervention in gerontological nursing. This paper provides some guidance concerning how this type of group may be arranged and the various categories of participant response that may be expected.


Uterine disorders and pregnancy complications: insights from mouse models.
Much of our knowledge of human uterine physiology and pathology has been extrapolated from the study of diverse animal models, as there is no ideal system for studying human uterine biology in vitro. Although it remains debatable whether mouse models are the most suitable system for investigating human uterine function(s), gene-manipulated mice are considered by many the most useful tool for mechanistic analysis, and numerous studies have identified many similarities in female reproduction between the two species. This Review brings together information from studies using animal models, in particular mouse models, that shed light on normal and pathologic aspects of uterine biology and pregnancy complications.


Plasmodium species mixed infections in two areas of Manhiça district, Mozambique.
We compared the distribution patterns of individual Plasmodium species and mixed-species infections in two geographically close endemic areas, but showing environmental differences. Comparisons concerned circulating Plasmodium infections in both human and mosquito vector populations in the dry and wet seasons, at a micro-epidemiological level (households). Both areas revealed a very high overall prevalence of infection, all year-round and in all age groups. Plasmodium falciparum was the predominant species, being found in the vast majority of infected individuals regardless of the presence of other species. Plasmodium malariae and Plasmodium ovale occurred almost exclusively in mixed infections. Seasonal variation in P. malariae prevalence was observed in one area but not in the other. A decrease in P. malariae prevalence concurred with a marked increase of P. falciparum prevalence. However this was strongly dependent on age and when analysing infections at the individual level, a different pattern between co-infecting species was unveiled. Regarding transmission patterns, in both areas, P. falciparum gametocytes predominated in single infections regardless of age and P. malariae gametocyte carriage increased when its overall prevalence decreased.


The rapid synthesis and dynamic behaviour of an isophthalamide [2]catenane.
A serendipitous [2]catenane has been prepared in three steps from commercially available starting materials. The interlocked topology of the catenane has been confirmed by single crystal X-ray structural determination. The rings of the catenane may rotate relative to one another - a process that may be controlled by varying solvent or temperature.


Effects of extensive practice on bradykinesia in Parkinson's disease: improvement, retention and transfer.
We investigated the effects of extensive practice of rapid aiming on bradykinesia and, more specifically, generation of peak velocity, in discrete rapid aiming and in transfer to reach-to-grasp. Twenty-one participants (seven young adults, seven older adults, and seven adults with mild to moderate Parkinson's disease (PD) while on medication) engaged in eight practice periods per week for three weeks (> 700 trials), with changes in performance measured weekly. Retention was measured weekly for three weeks postpractice. Movement time decreased with one week of practice, primarily due to a decrease in time-to-peak velocity. With practice and after retention, the PD group generated peak velocity as consistently as both neurologically healthy groups, but remained more variable in time-to-peak velocity. Transfer was observed in the neurologically healthy groups, but not in the PD group. We concluded that short-term practice (one week in our paradigm) is sufficient for decreasing movement time, but more extensive practice is needed to improve consistency of rapid aiming performance for people with mild to moderate PD.


When groups are more competitive than individuals: the domain of the discontinuity effect.
The related goals of the research were to delineate the domain of discontinuity, 1st by demonstrating its occurrence in a nonmatrix situation and, 2nd, by establishing the antecedent outcome conditions necessary for producing a discontinuity effect. The 1st goal was met by designing a mixed motive situation involving the production of origami products. Under these conditions, the magnitude of the discontinuity effect did not significantly differ from that in a matrix-only condition. The 2nd goal was met by a separate experiment that used H. H. Kelley and J. W. Thibaut's (1978) analysis of degree of noncorrespondence of outcomes. This experiment demonstrated that as noncorrespondence increased, so did the rate of competitive responding by groups but not by individuals. This pattern was qualified by an interaction with gender such that competitiveness was more markedly affected by noncorrespondence for groups of women than for groups of men.


Arbuscular mycorrhizal fungi enhance the copper tolerance of Tagetes patula through the sorption and barrier mechanisms of intraradical hyphae.
Arbuscular mycorrhizal fungi (AMF) are widespread soil fungi that can form endosymbiotic structures with the root systems of most plants and can improve the tolerance of host plants to heavy metals. In the present study, we investigated the effects of AMF (Glomus coronatum) inoculation on the tolerance of Tagetes patula L. to Cu. Almost all of the non-mycorrhizal plants exposed to 100 μM Cu died after 3 d, whereas phytotoxicity was only observed in mycorrhizal plants that were exposed to Cu concentrations greater than 100 μM. Analysing the dynamic accumulation of Cu indicated that, after 7 d of Cu exposure, less Cu was absorbed or accumulated by mycorrhizal plants than by control plants, and significantly less Cu was translocated to the shoots. Meanwhile, analysing the root morphology, the integrity of the root plasma membranes, the photosynthesis rate, and the content of essential elements of plants growing in cultures with 50 μM Cu revealed that AMF inoculation markedly alleviated the toxic effects of Cu stress on root system activity, photosynthesis rate, and mineral nutrient accumulation. In addition, to understand the Cu allocation, an energy spectrum analysis of Cu content at the transverse section of root tips was conducted and subsequently provided direct evidence that intraradical hyphae at the root endodermis could selectively immobilise large amounts of Cu. Indeed, the sorption and barrier mechanisms of AMF hyphae reduce Cu toxicity in the roots of T. patula and eventually enhance the plants' Cu tolerance.


Optimisation of derivatisation for the analysis of estrogenic compounds in water by solid-phase extraction gas chromatography-mass spectrometry.
An optimisation of derivatisation methods for the simultaneous determination of endocrine disrupting chemicals (EDCs) in water by solid-phase extraction (SPE) gas chromatography-mass spectrometry (GC-MS) was developed in this study. Seven highly potent EDCs including 17beta-estradiol (E2), estrone (E1), 16alpha-hydroxyestrone, 17alpha-ethynylestradiol (EE2), bisphenol A, 4-nonylphenol and 4-tert-octylphenol were selected as the target compounds. The SPE technique, followed by the derivatisation with N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) was used for the extraction recoveries of compounds from water and effluent samples. The stability of the silylation derivatives under different reaction conditions was investigated. The combined use of BSTFA and pyridine as derivatisation reagents, together with the use of hexane as the final solvent, was preferred in order to generate more stable derivatives of EDCs. The relative response factor (RRF) of all derivatives except that of EE2 was stable 120 h after derivatisation. The addition of pyridine as derivatisation reagent with BSTFA can prevent the conversion of EE2 to other products during the reaction. Several parameters that may affect the recovery of EDCs, such as the SPE flow rate, and water properties including aquatic colloid content and surfactant concentration were tested. The results showed that the flow rate (1-25 mL min(-1)), colloid concentration (0-50 mg L(-1)) and surfactants concentration (0-10 microg L(-1)) did not cause significant decrease in the EDCs recovery.


Characteristics of Rathke's cleft cyst based on cyst location with a primary focus on recurrence after resection.
Rathke's cleft cysts (RCCs) are benign lesions with a location that is entirely intrasellar, intrasellar with suprasellar extension (intrasuprasellar), or purely suprasellar. The recurrence of RCC is relatively uncommon. The present study was conducted to report clinical characteristics, histological features, and outcomes based on location of the cyst with a primary focus on analyzing the predictors of squamous metaplasia and recurrence in these 3 types of RCCs. A retrospective review of the medical records of patients with symptomatic RCCs who had undergone resection at the authors' institution was conducted. Data points, including clinical presentation, preoperative endocrine status, operative details, imaging findings, pathology, and clinical outcomes, were reviewed. A multivariable regression model was used to identify predictors of recurrence. The mean age of the 87 eligible patients, 64 females and 23 males, was 41 ± 14 years (range 10-73 years). Sixteen patients (18%) had an entirely intrasellar RCC, 21 (24%) had a purely suprasellar cyst, and 50 (58%) had an intrasuprasellar RCC. The mean cyst volume was 2.4 ± 0.9 cm(3) (range 0.36-4.9 cm(3)). Headache was the most frequent initial symptom (76%) followed by visual disturbance (45%). The transsphenoidal approach was performed for all intrasellar RCCs (16 cysts) and 33 of 50 intrasuprasellar RCCs. The transcranial route was used for all suprasellar cysts (21 cysts) and 17 of 50 intrasuprasellar RCCs. Squamous metaplasia was present in 27 (31%) of 87 RCCs. The occurrence of squamous metaplasia was associated with cyst location (p = 0.027), T1 signal intensity (p = 0.004) and ring enhancement on Gd-enhanced MRI (p = 0.017), and cyst volume (p = 0.045). A suprasellar location (p = 0.048, OR 3.89, 95% CI 1.010-15.020), ring enhancement on Gd-enhanced MRI (p = 0.028, OR 3.922, 95% CI 1.158-13.288), hypointensity on T1-weighted MRI (p = 0.002, OR 6.86, 95% CI 1.972-23.909), and cyst volume (p = 0.01, OR 0.367, 95% CI 0.170-0.789) were independent predictors of squamous metaplasia. The mean time to reaccumulation (11 [12.6%] of 87 cases) and recurrence (7 [8%] of 87 cases) was 14 ± 6 months. Recurrence-free survival was 84.5% at a mean of 98.2 ± 4.6 months after treatment. A suprasellar cyst location (p = 0.007, OR 7.7, 95% CI 1.75-34.54), the occurrence of squamous metaplasia (p = 0.007, OR 19.3, 95% CI 2.25-165.18), and isointensity on T2-weighted MRI (p = 0.041, OR 10.29, 95% CI 1.094-96.872) were the independent predictors of RCC recurrence. A suprasellar cyst location, the occurrence of squamous metaplasia, and isointensity on T2-weighted MRI were independent predictors of RCC recurrence. The extent of resection and type of surgical approach used were not associated with recurrence. A tailored extent of resection based on cyst location and predictive factors is recommended.


Eleven ancestral gene families lost in mammals and vertebrates while otherwise universally conserved in animals.
Gene losses played a role which may have been as important as gene and genome duplications and rearrangements, in modelling today species' genomes from a common ancestral set of genes. The set and diversity of protein-coding genes in a species has direct output at the functional level. While gene losses have been reported in all the major lineages of the metazoan tree of life, none have proposed a focus on specific losses in the vertebrates and mammals lineages. In contrast, genes lost in protostomes (i.e. arthropods and nematodes) but still present in vertebrates have been reported and extensively detailed. This probable over-anthropocentric way of comparing genomes does not consider as an important phenomena, gene losses in species that are usually described as "higher". However reporting universally conserved genes throughout evolution that have recently been lost in vertebrates and mammals could reveal interesting features about the evolution of our genome, particularly if these losses can be related to losses of capability. We report 11 gene families conserved throughout eukaryotes from yeasts (such as Saccharomyces cerevisiae) to bilaterian animals (such as Drosophila melanogaster or Caenorhabditis elegans). This evolutionarily wide conservation suggests they were present in the last common ancestors of fungi and metazoan animals. None of these 11 gene families are found in human nor mouse genomes, and their absence generally extends to all vertebrates. A total of 8 out of these 11 gene families have orthologs in plants, suggesting they were present in the Last Eukaryotic Common Ancestor (LECA). We investigated known functional information for these 11 gene families. This allowed us to correlate some of the lost gene families to loss of capabilities. Mammalian and vertebrate genomes lost evolutionary conserved ancestral genes that are probably otherwise not dispensable in eukaryotes. Hence, the human genome, which is generally viewed as being the result of increased complexity and gene-content, has also evolved through simplification and gene losses. This acknowledgement confirms, as already suggested, that the genome of our far ancestor was probably more complex than ever considered.


Concurrent multiple outbreaks of varicella, rubeola, german measles in unvaccinated children of co-educational mount carmel senior secondary school, thakurdwara palampur of northern himachal, India.
In April, 2009, in a co-education school, we investigated suspected triple outbreak; varicella first and then with chance detection of rubeola and rubella. The aim was to confirm diagnosis and recommend remedial measures to prevent further outbreaks. We defined a case of varicella with maculopapulovesicular rash without other apparent cause in students or staff of the school and residents of neighboring villages of Khalet and Roady since 23(rd) March to 14(th) October, 2009. We line listed case patients and collected information on age, sex, residence, date of onset, symptoms, signs, traveling, treatment history, and vaccination status. The outbreak was described by time, place, and person characteristics. Diagnosis was confirmed epidemiologically and serologically; first to chickenpox, measles, and german measles viruses. We identified 505 case patients from mixed outbreaks of varicella, measles, and german measles (30/505 clinically, 467/505 epidemiologically linked and 8/505 laboratory confirmed case patients from a study population of 3280. We investigated the suspected outbreak with case definition of varicella but measles 20/3280 (0.60%) and rubella 34/3280 (1.03%) cases were also observed. The overall attack rate (AR) was 15% while in school; it was 22% but highest (56%) in Nursery up to 4(th) standard with index case in first standard. Sex-specific AR was (23%) more in boys. Triple concurrent infection caused 05% complications but no death was reported. Severity of the symptoms was more in 5(th) standard onwards with 49-249 lesions and severer in poor villages Roady and Khalet (P < 0.05). Only 4% were immunized against varicella/german measles privately. Seventeen percent of the cases went for traditional treatment vs modern medicine (P < 0.001). 5/10 samples for IgM antibodies for chickenpox and 2/10 samples were positive for rubella. Triple infection of varicella, measles, and rubella was confirmed epidemiologically and serologically. We recommended local authorities for MMRV in the school and near villages with aggressive IEC activities in affected areas.


The benefit of therapeutic medial branch blocks after cervical operations.
Persistent neck pain is a common problem after surgery of the cervical spine. No therapy recommendation exists for these patients. The objective of this study was to determine if a therapeutic medial branch block is a rational treatment for patients with postoperative neck pain after cervical spine operations. Retrospective practice audit. Review of charts of all patients who underwent cervical spine operations for degenerative reasons during a time period of 3 years. Patients with persistent postsurgical pain were treated with therapeutic medial branch blocks (local anesthetic and steroid). A positive treatment response was defined if at least 80% reduction of pain could be achieved or if the patient was sufficiently satisfied with the relief. All patients with a minimum follow up time of 6 months were included. Of the 312 operations performed, 128 were artificial disc operations, 125 were stand alone cages, and 59 were fusions with cage and plate. Persistent neck pain occurred in 33.3% of the patients. There was no difference between the patients with neck pain and the whole group of patients. More than half of the patients with neck pain--52.9%--were treated successfully with therapeutic medial branch blocks. Since no further treatment was necessary, the initial treatment was considered successful. Nearly a third--32.2%--of the patients were initially treated successfully, but their pain recurred and further diagnostics and treatments were necessary. In this group of patients, significantly more with double level operations were found (P = 0.003). Patients not responding to the medial branch block were 14.9%. This audit is retrospective and observational, and therefore does not represent a high level of evidence. However, to our knowledge, since this information has not been previously reported and no recommendation for the treatment of post-operative zygapophysial joint pain exists, it appears to be the best available research upon which to recommend treatment and to plan higher quality studies. For persistent postsurgical neck pain only limited therapy recommendations exist. This study suggests treating these patients in a first instance with therapeutic medial branch blocks. The success rate is 52.9 %.


Alport syndrome, mental retardation, midface hypoplasia, and elliptocytosis: a new X linked contiguous gene deletion syndrome?
We describe a family with four members, a mother, two sons, and a daughter, who show clinical features consistent with X linked Alport syndrome. The two males presented with additional features including mental retardation, dysmorphic facies with marked midface hypoplasia, and elliptocytosis. The elliptocytosis was not associated with any detectable abnormalities in red cell membrane proteins; red cell membrane stability and rigidity was normal on ektacytometry. Molecular characterisation suggests a submicroscopic X chromosome deletion encompassing the entire COL4A5 gene. We propose that the additional abnormalities found in the affected males of this family are attributable to deletion or disruption of X linked recessive genes adjacent to the COL4A5 gene and that this constellation of findings may represent a new X linked contiguous gene deletion syndrome.


Reproductive status in adult male long-term survivors of childhood cancer.
This study assessed the long-term effects of cancer therapies on reproductive status in adult male childhood cancer survivors, evaluated the treatment-related risk factors for hypergonadotropic hypogonadism and assessed the association between the FSH levels and the later need for assisted reproductive techniques (ART). The study cohort included adult male 5-year survivors of childhood cancer who were treated in our institution between 1966 and 2003. Data concerning patient and treatment characteristics, FSH, LH and testosterone levels and pregnancy outcome were collected. Multivariate regression analyses were performed to evaluate the treatment-related risk factors for disturbances in reproductive endocrine status. The diagnostic and predictive values of FSH and later need for ART were evaluated. Data on reproductive endocrine status were available for 488 survivors (86.4%) of the 565 male survivors who visited the outpatient clinic in adulthood. The median follow-up time from initiation of treatment to first visit to the outpatient clinic in adulthood was 15 years. The prevalence rates of elevated FSH levels and decreased testosterone levels were 33 and 12%, respectively. The use of procarbazine, cyclophosphamide, vinca-alkaloids, other alkylating agents, pelvic/abdominal irradiation, total body irradiation and testicular surgery were identified as treatment-related risk factors for elevated FSH levels. During the follow-up period, 73 men reported 120 conceptions, which resulted in 103 live births. Of these men, 56 (77%) were able to achieve conception naturally. All men whose partners conceived by assisted reproductive techniques (n = 13) had elevated FSH levels at their first visit after their 18th birthday (sensitivity: 100%; 95% CI: 71-100%) and all male survivors with a normal FSH level did not need assisted reproductive techniques (negative predictive value: 100%; 95% CI: 89-100%). One-third of young adult male survivors of childhood cancer has elevated FSH levels. FSH appears to be a very sensitive marker for the need of assisted reproductive techniques in male childhood cancer survivors.


Impaired structural correlates of memory in Alzheimer's disease mice.
The healthy adult brain demonstrates robust learning-induced neuroanatomical plasticity. While altered neuroanatomical plasticity is suspected to be a factor mitigating the progressive cognitive decline in Alzheimer's disease (AD), it is not known to what extent this plasticity is affected by AD. We evaluated whether spatial learning and memory-induced neuroanatomical plasticity are diminished in an adult mouse model of AD (APP mice) featuring amyloid beta-driven cognitive and cerebrovascular dysfunction. We also evaluated the effect of early, long-term pioglitazone-treatment on functional hyperemia, spatial learning and memory, and associated neuroanatomical plasticity. Using high-resolution post-mortem MRI and deformation-based morphometry, we demonstrate spatial learning and memory-induced focal volume increase in the hippocampus of wild-type mice, an effect that was severely attenuated in APP mice, consistent with their unsuccessful performance in the spatial Morris water maze. These findings implicate impaired neuroanatomical plasticity as an important contributing factor to cognitive deficits in the APP mouse model of AD. Pioglitazone-treatment in APP mice completely rescued functional hyperemia and exerted beneficial effects on spatial learning and memory-recall, but it did not improve hippocampal plasticity.


African-American race and mortality in interstitial lung disease: a multicentre propensity-matched analysis.
We studied whether African-American race is associated with younger age and decreased survival time at diagnosis of interstitial lung disease (ILD).We performed a multicentre, propensity score-matched analysis of patients with an ILD diagnosis followed at five US hospitals between 2006 and 2016. African-Americans were matched with patients of other races based on a time-dependent propensity score calculated from multiple patient, physiological, diagnostic and hospital characteristics. Multivariable logistic regression models were used. All-cause mortality and hospitalisations were compared between race-stratified patient cohorts with ILD, and sensitivity analyses were performed.The study included 1640 patients with ILD, 13% of whom were African-American, followed over 5041 person-years. When compared with patients of other races, African-Americans with ILD were younger at diagnosis (56 years versus 67 years), but in the propensity-matched analyses had greater survival (hazard ratio 0.46, 95% CI 0.28-0.77; p=0.003) despite similar risk of respiratory hospitalisations (relative risk 1.04, 95% CI 0.83-1.31; p=0.709), and similar GAP-ILD (gender-age-physiology-ILD) scores at study entry. Sensitivity analyses in a separate cohort of 9503 patients with code-based ILD diagnosis demonstrated a similar association of baseline demographic characteristics with all-cause mortality.We conclude that African-Americans demonstrate a unique phenotype associated with younger age at ILD diagnosis and perhaps longer survival time.


Direct and indirect control of late sporulation genes by GerR of Bacillus subtilis.
GerR is a sporulation-specific transcriptional factor of Bacillus subtilis that has been identified as a negative regulator of genes transcribed by sigma(E)-containing RNA polymerase and as a positive effector of the expression of three late sporulation genes. Here we confirmed that gerR transcription is dependent on sigma(E)-containing RNA polymerase but also observed that it requires the transcriptional regulator SpoIIID. The study of the role of GerR in regulating the expression of several late sporulation genes allowed us to observe that its effect is strongly positive on spoVIF, cotC, and cotG, weakly positive on cotB, and negative on cotU. The results of chromatin immunoprecipitation (ChIP) experiments indicated that GerR binds to the promoter regions of some, but not all, of the GerR-controlled genes, leading us to propose that GerR controls late sporulation genes in two ways: (i) directly, by acting on the transcription of cotB, cotU and spoVIF; and (ii) indirectly, through the activation of SpoVIF, which stabilizes the transcriptional activator GerE and consequently induces the expression of the GerE-dependent genes cotC and cotG.


Characterization of three low-molecular-weight Glu-D3 subunit genes in common wheat.
Low-molecular-weight glutenins (LMW-GS) in common wheat (Triticum aestivum L.) are of great importance for processing quality of pan bread and noodles. The objectives of this study are to identify LMW-GS coding genes at GluD3 locus on chromosome 1D and to establish relationships between these genes and GluD3 alleles (a, b, c, d, and e) defined by protein electrophoretic mobility. Specific primer sets were designed to amplify each of the three LMW-GS chromosome 1D gene regions including upstream, coding and downstream regions of eight wheat cultivars containing GluD3 a, b, c, d and e alleles. Three LMW-GS genes, designated as GluD3-1, GluD3-2 and GluD3-3, were amplified from the eight wheat cultivars. The allelic variants of these three genes were analysed at the DNA and protein level. GluD3-1 showed two allelic variants or haplotypes, one common to cultivars containing protein alleles a, d and e (designated GluD3-11) and the other was present in cultivars with alleles b and c (designated GluD3-12). Comparing with GluD3-12, a 3-bp deletion was found in the coding region of the N-terminal repetitive domain of GluD3-11, leading to a glutamine deletion at the 116th position. GluD3-2 had three variants at the DNA level in the eight cultivars, which were designated as GluD3-21, GluD3-22 and GluD3-23. In comparison to GluD3-21, a single nucleotide polymorphism (SNP) was detected for GluD3-22 in the signal peptide region, resulting in an amino acid change from alanine to threonine at the 11th position; and 11 mutations were found at GluD3-23, with five in upstream region, four in coding region and two in downstream region, respectively. GluD3-3 had two haplotypes, designated as GluD3-31 and GluD3-32, both belonging to LMW-s glutenin subunits though their first amino acids in N-terminal region are different. Compared with the GenBank GluD3 genes, nucleotide sequences of GluD3-21 and GluD3-23 were the same as X13306 and AB062875, respectively. GluD3-22 and GluD3-11 had only one-base difference from U86027 and AB062865. GluD3-12 was not found in the GenBank database, indicating a newly identified GluD3 gene variation. GluD3-3 was a new gene different from any other known GluD3 genes. Analyses of the relationship between Glu-D3 alleles defined by protein electrophoretic mobility and different GluD3 gene variations at the DNA or protein level provided molecular basis for DNA based identification of glutenin alleles.


Increased use of laparoscopy in acute right iliac fossa pain - is it good for patients?
The use of laparoscopy, with or without appendicectomy, is becoming more common in the management of acute right iliac fossa (RIF) pain, but little is known of the 'unintended' consequences of this change. This study aimed to evaluate the impact of increased use of laparoscopy on the number and type of patients treated surgically and on the rate of negative appendicectomy. A prospective audit was carried out of admissions to a teaching hospital over two, 3-month periods during 2007 and 2008. The management, investigations and outcome of patients presenting with RIF pain were studied. Admissions were stable over the two time-periods. There was a significant increase in the number of laparoscopic operations performed, from 22.5% (14/62) in 2007 to 85.7% (72/84) in 2008 (P < 0.0001), and the percentage of patients undergoing surgery rose from 55.4% (n = 62) in 2007 to 71.2% (n = 84) in 2008 (P < 0.01). In 2008, female patients were more likely to have surgery, an increase from 37.1% to 66.2% (P < 0.001), and were more likely to have a laparoscopic procedure, an increase from 50% to 98% (P < 0.0001). The rate of histologically confirmed appendicitis did not increase significantly (50/122 vs 57/118; P = 0.25), but the number of patients with a normal appendix either left in situ because it was macroscopically normal or found to be histologically normal following excision, increased significantly, from 9.01% in 2007 to 21.2% in 2008 (P < 0.01). The diagnostic value of pelvic ultrasound decreased from 75.6% of examinations in 2007 to 54.5% in 2008 (P = 0.039). An increase in laparoscopic procedures has resulted in more operations in women, an associated higher negative appendicectomy rate and decreased usefulness of pelvic ultrasound. Increased use of laparoscopy needs to be balanced against the diagnostic benefits of 'negative' laparoscopy.


Host genetic influences on highly active antiretroviral therapy efficacy and AIDS-free survival.
We studied the influence of AIDS restriction genes (ARGs) CCR5-Delta32, CCR2-64I, SDF1-3'A, IL10-5'A, CX3CR1-V249I, CX3CR1-T280M, and MDR1-C3435T and haplotypes of the CCR5 P1 promoter and RANTES variants -403A, In1.1C, 3'222C, and -28G among HIV-1 infected patients on highly active antiretroviral therapy (HAART) in the Multicenter AIDS Cohort Study (MACS) and the Multicenter Hemophilia Cohort Study (MHCS). Our results indicate that several ARGs also influence therapy efficacy (ie, the success in viral suppression) and subsequent progression to AIDS while on HAART. CCR5-Delta32 decreased time to viral suppression (<200 HIV RNA copies/mL, relative hazard [RH]=1.40; P=0.008) and was protective against AIDS (RH=0.11; P=or<0.0001), whereas the CCR5 P1 haplotype was associated with delayed viral suppression (RNA<50 copies/mL, odds ratio [OR]=0.65; P=0.03) and accelerated time to AIDS (RH=2.68; P=0.02). SDF1-3'A reduced viral suppression (OR=0.61; P=0.02) and accelerated AIDS (RH=3.18; P=0.009). Accelerated AIDS progression was also observed with the RANTES haplotype carrying RANTES-IN1.1C and RANTES-3'222C (P=0.005 to 0.007). In contrast, the RANTES haplotype H1, which lacks suspected deleterious single-nucleotide polymorphisms, was protective against AIDS. CX3CR1-V249I seemed to accelerate viral suppression (RNA<50 copies/mL, OR=1.27; P=0.01). ARG influence after HAART suggests residual HIV-1 replication, and spread continues even in patients successfully suppressing detectable viral RNA.


Update on beta-lactam allergy diagnosis.
Allergic reactions to penicillin and cephalosporin antibiotics are common. The mechanisms involved are heterogeneous and not yet completely understood. However, clinical and immunologic studies suggest that type I (IgE-mediated) and type IV (cell-mediated) pathogenic mechanisms are involved in most immediate and non-immediate reactions, respectively. The diagnostic procedure has recently been standardized under the aegis of the European Network for Drug Allergy and the European Academy of Allergy and Clinical Immunology interest group on drug hypersensitivity. In this review, we describe the general guidelines for evaluating subjects in whom there is suspicion of allergic reactions to beta-lactams.


Argonaute Utilization for miRNA Silencing Is Determined by Phosphorylation-Dependent Recruitment of LIM-Domain-Containing Proteins.
As core components of the microRNA-induced silencing complex (miRISC), Argonaute (AGO) proteins interact with TNRC6 proteins, recruiting other effectors of translational repression/mRNA destabilization. Here, we show that LIMD1 coordinates the assembly of an AGO-TNRC6 containing miRISC complex by binding both proteins simultaneously at distinct interfaces. Phosphorylation of AGO2 at Ser 387 by Akt3 induces LIMD1 binding, which in turn enables AGO2 to interact with TNRC6A and downstream effector DDX6. Conservation of this serine in AGO1 and 4 indicates this mechanism may be a fundamental requirement for AGO function and miRISC assembly. Upon CRISPR-Cas9-mediated knockout of LIMD1, AGO2 miRNA-silencing function is lost and miRNA silencing becomes dependent on a complex formed by AGO3 and the LIMD1 family member WTIP. The switch to AGO3 utilization occurs due to the presence of a glutamic acid residue (E390) on the interaction interface, which allows AGO3 to bind to LIMD1, AJUBA, and WTIP irrespective of Akt signaling.


Visualizing evolutionary dynamics of self-replicators: a graph-based approach.
We present a general approach for evaluating and visualizing evolutionary dynamics of self-replicators using a graph-based representation for genealogy. Through a transformation from the space of species and mutations to the space of nodes and links, evolutionary dynamics are understood as a flow in graph space. A formalism is introduced to quantify such genealogical flows in terms of the complete history of localized evolutionary events recorded at the finest level of detail. Represented in a multidimensional viewing space, collective dynamical properties of an evolving genealogy are characterized in the form of aggregate flows. We demonstrate the effectiveness of this approach by using it to compare the evolutionary exploration behavior of self-replicating loops under two different environmental settings.


Endometriosis does not confer improved prognosis in ovarian carcinoma of uniform cell type.
The role of endometriosis in ovarian cancer, disease progression, and survival is a subject of active investigation. A series of 144 ovarian cancers with clear cell or endometrioid histology or associated endometriosis, all classified on the basis of strict histologic criteria, was evaluated to further explore the relationship between endometriosis-associated ovarian cancer and age at presentation, FIGO stage, histology, presence of synchronous primary disease elsewhere in the mullerian tract, and survival. Patients with endometrioid carcinomas were significantly younger (mean, 52 y) in comparison with patients with either clear cell carcinoma (mean, 55 y) or mixed tumors (mean, 59 y; P=0.002). Clear cell carcinoma presented as low-stage disease (FIGO I) in 33% of cases compared with endometrioid carcinomas in 97% of cases and mixed carcinomas in 27% of cases. Endometriosis was associated with 53% of clear cell carcinomas, 33% of endometrioid carcinomas, and 45% of mixed tumors (P<0.001). Synchronous primary tumors, observed in 31% of endometrioid tumors, 5% of mixed tumors, and in 2% of clear cell tumors (P<0.001), were unlikely to be associated with endometriosis (P=0.04). Univariate analysis of the aggregate cohort demonstrated that the single best overall predictor of disease-free survival was FIGO stage at presentation (P<0.001), followed by histologic subtype (P=0.003). Endometriosis did not have a significant relationship with disease-free survival (P=0.7). We conclude that the link between endometriosis and ovarian cancer is much stronger for clear cell carcinoma than for other histologic subtypes (P<0.001). Furthermore, when uniform histologic criteria are applied, true mixed endometrioid and clear cell carcinomas are uncommon; most endometriosis-associated mixed tumors are heterogenous mixtures of endometrioid, mucinous, and serous histology with areas of clear cell cytoplasm. Endometriosis per se does not appear to predict prognosis in clear cell and endometrioid tumors, with the possible exception of tumors with mixed histology. Until more data are collected, pathologists should classify ovarian tumors with mixed histology as a separate and potentially unique biological and prognostic group.


Cerebellar Nuclear Neurons Use Time and Rate Coding to Transmit Purkinje Neuron Pauses.
Neurons of the cerebellar nuclei convey the final output of the cerebellum to their targets in various parts of the brain. Within the cerebellum their direct upstream connections originate from inhibitory Purkinje neurons. Purkinje neurons have a complex firing pattern of regular spikes interrupted by intermittent pauses of variable length. How can the cerebellar nucleus process this complex input pattern? In this modeling study, we investigate different forms of Purkinje neuron simple spike pause synchrony and its influence on candidate coding strategies in the cerebellar nuclei. That is, we investigate how different alignments of synchronous pauses in synthetic Purkinje neuron spike trains affect either time-locking or rate-changes in the downstream nuclei. We find that Purkinje neuron synchrony is mainly represented by changes in the firing rate of cerebellar nuclei neurons. Pause beginning synchronization produced a unique effect on nuclei neuron firing, while the effect of pause ending and pause overlapping synchronization could not be distinguished from each other. Pause beginning synchronization produced better time-locking of nuclear neurons for short length pauses. We also characterize the effect of pause length and spike jitter on the nuclear neuron firing. Additionally, we find that the rate of rebound responses in nuclear neurons after a synchronous pause is controlled by the firing rate of Purkinje neurons preceding it.


Bulky side chains and non-native salt bridges slow down the folding of a cross-linked helical peptide: a combined molecular dynamics and time-resolved infrared spectroscopy study.
Multiple 4-micros molecular dynamics (MD) simulations are used to study the folding process of the cross-linked alpha-helical peptide Ac-EACAR(5)EAAAR(10)EAACR(15)Q-NH(2) (EAAAR peptide). The folding kinetics are single exponential at 330 K, while they are complex at 281 K with a clear deviation from single-exponential behavior, in agreement with time-resolved infrared (IR) spectroscopy measurements. Network analysis of the conformation space sampled by the MD simulations reveals four main folding channels which start from conformations with partially formed helical structure and non-native salt-bridges in a kinetically partitioned unfolded state. The independent folding pathways explain the comparable quality of models based on stretched exponential and multiexponential fitting of the kinetic traces at low temperature. The rearrangement of bulky side chains, and in particular their reorientation with respect to the cross-linker, makes the EAAAR peptide a slower folder at 281 K than a similar peptide devoid of the three glutamate side chains. On the basis of this simulation result, extracted from a total MD sampling of 1.0 ms, a mutant with additional bulky side chains (three methionines replacing alanines at positions 2, 7, and 12) is suggested to fold slower than the EAAAR peptide. This prediction is confirmed by time-resolved IR spectroscopy.


Flood prevention dams for arid regions at a micro-scale sub-catchment, case study: Tabuk, Saudi Arabia.
Unexpected flash flooding is one of the periodic hydrological problems affecting the city of Tabuk in Saudi Arabia. The region has high potential for floods as it suffers high rainfall intensity in a short time and also has high urbanization rates and topographic complexity. Constructing flood prevention dams is one option to solve this problem. A cost-effective design requires a detailed feasibility study and analysis for the selection of suitable sites. The aim of this study was to develop a method for selecting a suitable site for flood protection dams in the Abu Saba'a district, the most affected part of the city of Tabuk during the flash flood in January 2013. Spatial analysis was applied using Landsat Thematic Mapper images and Shuttle Radar Topography Mission digital elevation model to select a site in the Abu Saba'a area. A simple model using ArcGIS was built including all suggested parameters. The results showed the best site for a dam was 2 km distance backfrom the area, where all parameter values matched. The results showed that the dynamic properties of land cover can affect site selection. It is therefore suggested that more field and hydrological data should be gathered for greater accuracy.


Does patient rurality predict quality colon cancer care?: A population-based study.
More than 50 million people reside in rural America. However, the impact of patient rurality on colon cancer care has been incompletely characterized, despite its known impact on screening. Our study sought to examine the impact of patient rurality on quality and comprehensive colon cancer care. We constructed a retrospective cohort of 123,129 patients with stage 0 to IV colon cancer. Rural residence was established based on the patient medical service study area designated by the registry. The study was conducted using the 1996-2008 California Cancer Registry. All of the patients diagnosed between 1996 and 2008 with tumors located in the colon were eligible for inclusion in this study. Baseline characteristics were compared by rurality status. Multivariate regression models then were used to examine the impact of rurality on stage in the entire cohort, adequate lymphadenectomy in stage I to III disease, and receipt of chemotherapy for stage III disease. Proportional-hazards regression was used to examine the impact of rurality on cancer-specific survival. Of all of the patients diagnosed with colon cancer, 18,735 (15%) resided in rural areas. Our multivariate models demonstrate that rurality was associated with later stage of diagnosis, inadequate lymphadenectomy in stage I to III disease, and lower likelihood of receiving chemotherapy for stage III disease. In addition, rurality was associated with worse cancer-specific survival. We could not account for socioeconomic status directly, although we used insurance status as a surrogate. Furthermore, we did not have access to treatment location or distance traveled. We also could not account for provider or hospital case volume, patient comorbidities, or complications. A significant portion of patients treated for colon cancer live in rural areas. Yet, rural residence is associated with modest differences in stage, adherence to quality measures, and survival. Future endeavors should help improve care to this vulnerable population (see video, Supplemental Digital Content 1, http://links.lww.com/DCR/A143).


Subcutaneous absorption of biotherapeutics: knowns and unknowns.
Subcutaneous administration of biotherapeutics offers several potential advantages compared with intravenous administration. Many biotherapeutics, both marketed or in development, are administered via the subcutaneous route. This minireview provides an overview of the presystemic absorption processes following subcutaneous administration, the resulting pharmacokinetics after subcutaneous administration, and provides recent case examples of the development of subcutaneous administered drugs with a focus on monoclonal antibodies. Subcutaneous absorption of biotherapeutics is relatively slow and mostly incomplete. Knowledge of the subcutaneous tissue is important to understand the absorption kinetics after subcutaneous administration. Transport in the subcutis to the absorbing blood or lymph capillaries appears to be a major contributor to the slow subcutaneous absorption. Larger proteins (>20 kDa) are mostly absorbed via the lymphatic system, although potential species differences are not fully understood yet. Also, the presystemic catabolism leading to incomplete bioavailability is little understood, both the involved enzymes and its translation across species. For IgGs, binding to the neonatal Fc receptor is important to obtain a high bioavailability. Overall, several aspects of subcutaneous absorption are still poorly understood, which hampers, e.g., translation across species. Further research in this area is warranted.


The Demand-Induced Strain Compensation Questionnaire: A Cross-national Validation Study.
The demand-induced strain compensation model is a theoretical job stress model that has been tested in different kinds of empirical study in several countries. To measure key concepts in the model (job demands and job resources), the demand-induced strain compensation questionnaire (DISQ) was developed and has been used in many empirical studies. However, most studies neither focused on the psychometric properties of the DISQ nor tested it cross-nationally. In this study, we examined the psychometric properties of the DISQ tested on Italian (n = 422) and Dutch (n = 1629) employees. Results indicated that the six-factor model, consisting of cognitive, emotional and physical job demands as well as cognitive, emotional and physical job resources, provided a better fit to the data than an alternative two-factor model (demands and resources only). Findings were invariant across Italian and Dutch samples. Internal consistencies were satisfactory. Additionally, the six dimensions of DISQ were found to be meaningfully related to employee active learning behaviour, emotional exhaustion and musculoskeletal disorders. Results suggest the DISQ questionnaire is a valid and reliable instrument to measure the proposed dimensions of job demands and job resources in both Italian and Dutch work contexts.


Striking changes in tea metabolites due to elevational effects.
Climate effects on crop quality at the molecular level are not well-understood. Gas and liquid chromatography-mass spectrometry were used to measure changes of hundreds of compounds in tea at different elevations in Yunnan Province, China. Some increased in concentration while others decreased by 100's of percent. Orthogonal projection to latent structures-discriminant analysis revealed compounds exhibiting analgesic, antianxiety, antibacterial, anticancer, antidepressant, antifungal, anti-inflammatory, antioxidant, anti-stress, and cardioprotective properties statistically (p = 0.003) differentiated high from low elevation tea. Also, sweet, floral, honey-like notes were higher in concentration in the former while the latter displayed grassy, hay-like aroma. In addition, multivariate analysis of variance showed low elevation tea had statistically (p = 0.0062) higher concentrations of caffeine, epicatechin gallate, gallocatechin, and catechin; all bitter compounds. Although volatiles represent a small fraction of the total mass, this is the first comprehensive report illustrating how normal variations in temperature, 5 °C, due to elevational effects impact tea quality.


Discovery of high-level behavior from observation of human performance in a strategic game.
This paper explores the issues faced in creating a system that can learn tactical human behavior merely by observing a human perform the behavior in a simulation. More specifically, this paper describes a technique based on fuzzy ARTMAP (FAM) neural networks to discover the criteria that cause a transition between contexts during a strategic game simulation. The approach depends on existing context templates that can identify the high-level action of the human, given a description of the situation along with his action. The learning task then becomes the identification and representation of the context sequence executed by the human. In this paper, we present the FAM/Template-based Interpretation Learning Engine (FAMTILE). This system seeks to achieve this learning task by constructing rules that govern the context transitions made by the human. To evaluate FAMTILE, six test scenarios were developed to achieve three distinct evaluation goals: 1) to assess the learning capabilities of FAM; 2) to evaluate the ability of FAMTILE to correctly predict human and context selections, given an observation; and 3) more fundamentally, to create a model of the human's behavior that can perform the high-level task at a comparable level of proficiency.


Gemcitabine-associated thrombotic thrombocytopenic purpura and hemolytic uremic syndrome.
A patient being treated for metastatic adenocarcinoma of the pancreas presents to the clinic for a routine appointment. A complete blood count reveals hemoglobin of 6.5 g/dl and a platelet count of 30,000 K/mm3 thought to be from the last of many doses of gemcitabine. On assessment, the only complaint was fatigue with no evidence of bleeding or other abnormal physical findings other than pallor. Past medical history includes hypertension managed with three antihypertensive agents. Additional laboratory tests reveal elevated blood urea nitrogen (69 mg/dl), creatinine (2.76 mg/dl), and lactic dehydrogenase (LDH), was well as indirect bilirubin (2.1 mg/dl). The patient is admitted and transfused with packed red blood cells (pRBCs). The next day, the platelet count drops to 9,000 K/mm3 and the hemoglobin increases, appropriately, to 8.9 g/dl. Urinalysis is positive for hemoglobin (+ 3). The peripheral blood smear is positive for schistocytes (fragmented RBCs). A pheresis catheter is placed after the patient was evaluated by a hematologist and a nephrologist. A presumptive diagnosis of thrombotic thrombocytopenic purpura (TTP) with hemolytic uremic syndrome (HUS) was made.


What drives the distribution of the bloom-forming cyanobacteria Planktothrix agardhii and Cylindrospermopsis raciborskii?
The cyanobacteria Planktothrix agardhii and Cylindrospermopsis raciborskii are bloom-forming species common in eutrophic freshwaters. These filamentous species share certain physiological traits which imply that they might flourish under similar environmental conditions. We compared the distribution of the two species in a large database (940 samples) covering different climatic regions and the Northern and Southern hemispheres, and carried out laboratory experiments to compare their morphological and physiological responses. The environmental ranges of the two species overlapped with respect to temperature, light and total phosphorus (TP); however, they responded differently to environmental gradients; C. raciborskii biovolume changed gradually while P. agardhii shifted sharply from being highly dominated to a rare component of the phytoplankton. As expected, P. agardhii dominates the phytoplankton with high TP and low light availability conditions. Contrary to predictions, C. raciborskii succeeded in all climates and at temperatures as low as 11 °C. Cylindrospermopsis raciborskii had higher phenotypic plasticity than P. agardhii in terms of pigments, individual size and growth rates. We conclude that the phenotypic plasticity of C. raciborskii could explain its ongoing expansion to temperate latitudes and suggest its future predominance under predicted climate-change scenarios.


Pollen tube guidance by the female gametophyte.
In flowering plants, pollen grains germinate on the pistil and send pollen tubes down the transmitting tract toward ovules. Previous genetic studies suggested that the ovule is responsible for long-range pollen tube guidance during the last phase of a pollen tube's journey to the female gametes. It was not possible, however, to unambiguously identify the signaling cells within an ovule: the haploid female gametophyte or the diploid sporophytic cells. In an effort to distinguish genetically between these two possibilities, we have used a reciprocal chromosomal translocation to generate flowers wherein approximately half the ovules do not contain a functional female gametophyte but all ovules contain genotypically normal sporophytic cells. In these flowers, pollen tubes are guided to the normal but not to the abnormal female gametophytes. These results strongly suggest that the female gametophyte is responsible for pollen tube guidance, but leave open the possibility that the gametophyte may accomplish this indirectly through its influence on some sporophytic cells.


Rewiring cellular morphology pathways with synthetic guanine nucleotide exchange factors.
Eukaryotic cells mobilize the actin cytoskeleton to generate a remarkable diversity of morphological behaviours, including motility, phagocytosis and cytokinesis. Much of this diversity is mediated by guanine nucleotide exchange factors (GEFs) that activate Rho family GTPases-the master regulators of the actin cytoskeleton. There are over 80 Rho GEFs in the human genome (compared to only 22 genes for the Rho GTPases themselves), and the evolution of new and diverse GEFs is thought to provide a mechanism for linking the core cytoskeletal machinery to a wide range of new control inputs. Here we test this hypothesis and ask if we can systematically reprogramme cellular morphology by engineering synthetic GEF proteins. We focused on Dbl family Rho GEFs, which have a highly modular structure common to many signalling proteins: they contain a catalytic Dbl homology (DH) domain linked to diverse regulatory domains, many of which autoinhibit GEF activity. Here we show that by recombining catalytic GEF domains with new regulatory modules, we can generate synthetic GEFs that are activated by non-native inputs. We have used these synthetic GEFs to reprogramme cellular behaviour in diverse ways. The GEFs can be used to link specific cytoskeletal responses to normally unrelated upstream signalling pathways. In addition, multiple synthetic GEFs can be linked as components in series to form an artificial cascade with improved signal processing behaviour. These results show the high degree of evolutionary plasticity of this important family of modular signalling proteins, and indicate that it may be possible to use synthetic biology approaches to manipulate the complex spatio-temporal control of cell morphology.