E2F1 overexpression in quiescent fibroblasts leads to induction of cellular DNA synthesis and apoptosis.
Various experiments have demonstrated a role for the E2F transcription factor in the regulation of cell growth during the G0/G1/S phase transition. Indeed, overexpression of the E2F1 product, a component of the cellular E2F activity, induces DNA synthesis in quiescent fibroblasts. To provide an approach to a more detailed biochemical analysis of these events, we have made use of a recombinant adenovirus containing the E2F1 cDNA in order to efficiently express the E2F1 product in an entire population of cells. We demonstrate an induction of DNA synthesis when quiescent cells are infected with the E2F1 recombinant virus. However, we also find that the induction does not lead to a complete replication of the cellular genome, as revealed by flow cytometry. The incomplete nature of cellular DNA replication is due, at least in part, to the fact that E2F1 overexpression leads to massive cell death that is characteristic of apoptosis. This E2F1-mediated induction of apoptosis is largely dependent on endogenous wild-type p53 activity and can be subverted by introducing mutant forms of p53 into these cells or by overexpressing E2F1 in fibroblasts derived from p53-null mouse embryos. We conclude that E2F1 can induce events leading to S phase but that the process is not normal and appears to result from the activation of a cell death pathway.


Immune Globulin Subcutaneous (Human) 20% (Hizentra®): A Review in Chronic Inflammatory Demyelinating Polyneuropathy.
Intravenous immunoglobulin (IVIg) is well-established in the treatment of patients with chronic inflammatory demyelinating polyneuropathy (CIDP). Immune globulin subcutaneous (human) 20% liquid (Hizentra®; referred to as IgPro20 hereafter) has recently been approved in a number of countries, including the USA and those of the EU, as maintenance therapy in patients with CIDP. In the pivotal phase III PATH trial in adults with CIDP who were first stabilized on IVIg therapy, maintenance therapy with IgPro20 for 24 weeks significantly reduced CIDP relapse or study withdrawal rates versus placebo. Efficacy was sustained during ≤ 48 weeks of additional treatment with IgPro20 in the open-label PATH extension study. IgPro20 was generally well tolerated, with low rates of systemic adverse events (AEs); the most common AEs were local reactions (e.g. infusion-site erythema, infusion-site swelling). In PATH, more than one-half of IgPro20 recipients preferred this therapy to their previous IVIg therapy. IgPro20 offers a convenient alternative to IVIg with a better systemic AEs profile and thus extends the options for maintenance therapy in CIDP.


Understanding the influence of disorder on the exciton dynamics and energy transfer in Zn-phthalocyanine H-aggregates.
The photophysics of 9(19),16(17),23(24)-tri-tert-butyl-2-[ethynyl-(4-carboxymethyl)phenyl]phthalocyaninatozinc(ii) and its H-aggregates is studied in different solvents by means of ultrafast non-linear optical spectroscopy and computational modeling. In non-coordinating solvents, both stationary and time-resolved spectroscopies highlight the formation of extended molecular aggregates, whose dimension and spectral properties depends on the concentration. In all the explored experimental conditions, time-resolved transient absorption experiments show multi exponential decay of the signals. Additional insights into the excited state relaxation mechanisms of the system is obtained with 2D electronic spectroscopy, which is employed to compare the deactivation channels in the absence or presence of aggregates. In ethanol and diethylether, where only monomers are present, an ultrafast relaxation process among the two non-degenerate Q-states of the molecule is evidenced by the appearance of a cross peak in the 2D-maps. In chloroform or CCl4, where disordered H-aggregates are formed, an energy transfer channel among aggregates with different composition and size is observed, leading to the non-radiative decay towards the lower energy dark state of the aggregates. Efficient coupling between less and more aggregated species is highlighted in two-dimensional electronic spectra by the appearance of a cross peak. The kinetics and intensity of the latter depend on the concentration of the solution. Finally, the linear spectroscopic properties of the aggregate are reproduced using a simplified structural model of an extended aggregate, based on Frenkel Hamiltonian Calculations and on an estimate of the electronic couplings between each dimer composing the aggregate computed at DFT level.


The importance of facets of quality of life to older adults: an international investigation.
To describe the relative importance of 38 facets of quality of life (QOL) to older adults in 22 countries, by gender, age group, and health status. A secondary analysis of data was conducted from the WHOQOL-Old pilot study. Questionnaires were distributed by mail or in person. Independent t-tests for differences between means were calculated between importance scores by genders, age groups, and healthy/unhealthy participants. Hierarchical multiple regression analysis was used to identify which QOL facets explained importance of overall QOL. RESULTS ; Of 7,401 older adults from 22 countries (mean 73.1 years), 57.8% were women and 70.1% were 'healthy'. While there are significant differences in the importance of various facets of QOL on the basis of gender, age, and health status, when these variables were controlled in a stepwise multiple regression analysis, the most important QOL facets were having energy, being happy, having well-functioning senses, and being free from pain. It appears that many of the physical aspects of QOL such as energy, freedom from pain, ability to do activities of daily living and to move around are particularly important to older adults. Practitioners should give greater attention to energy, sensory abilities, and pain.


Enhanced negative feedback sensitivity of the hypothalamus-pituitary-adrenal axis in chronic myogenous facial pain.
Dysregulations of the hypothalamus-pituitary-adrenal (HPA) axis, as a physiological substrate of stress, have been observed in patients with different stress-related and chronic pain disorders. In this study, we investigated possible dysregulations of the HPA axis in patients with masticatory muscles pain. In 20 patients with myogenous facial pain and 20 healthy controls, awakening cortisol responses, i.e.cortisol rise in the first hour after awakening, as well as a short circadian free cortisol profile, i.e. four cortisol samples over 12h during the day, were assessed before and after administration of 0.5mg dexamethasone. In comparison to controls, chronic myogenous facial pain patients showed enhanced and prolonged suppression of cortisol after the administration of 0.5mg dexamethasone. Unstimulated cortisol response (before dexamethasone-intake) to awakening and cortisol levels during the day did not differ between the groups. Dysregulation in terms of enhanced negative feedback suppression exists in chronic myogenous facial pain. These results are in line with a multifactorial etiology of chronic facial pain, shifting the perspective away from a local towards a more central etiology with dysregulations in the stress and pain modulating system.


Lakeside cemeteries in the Sahara: 5000 years of holocene population and environmental change.
Approximately two hundred human burials were discovered on the edge of a paleolake in Niger that provide a uniquely preserved record of human occupation in the Sahara during the Holocene ( approximately 8000 B.C.E. to the present). Called Gobero, this suite of closely spaced sites chronicles the rapid pace of biosocial change in the southern Sahara in response to severe climatic fluctuation. Two main occupational phases are identified that correspond with humid intervals in the early and mid-Holocene, based on 78 direct AMS radiocarbon dates on human remains, fauna and artifacts, as well as 9 OSL dates on paleodune sand. The older occupants have craniofacial dimensions that demonstrate similarities with mid-Holocene occupants of the southern Sahara and Late Pleistocene to early Holocene inhabitants of the Maghreb. Their hyperflexed burials compose the earliest cemetery in the Sahara dating to approximately 7500 B.C.E. These early occupants abandon the area under arid conditions and, when humid conditions return approximately 4600 B.C.E., are replaced by a more gracile people with elaborated grave goods including animal bone and ivory ornaments. The principal significance of Gobero lies in its extraordinary human, faunal, and archaeological record, from which we conclude the following: The early Holocene occupants at Gobero (7700-6200 B.C.E.) were largely sedentary hunter-fisher-gatherers with lakeside funerary sites that include the earliest recorded cemetery in the Sahara.Principal components analysis of craniometric variables closely allies the early Holocene occupants at Gobero with a skeletally robust, trans-Saharan assemblage of Late Pleistocene to mid-Holocene human populations from the Maghreb and southern Sahara.Gobero was abandoned during a period of severe aridification possibly as long as one millennium (6200-5200 B.C.E).More gracile humans arrived in the mid-Holocene (5200-2500 B.C.E.) employing a diversified subsistence economy based on clams, fish, and savanna vertebrates as well as some cattle husbandry.Population replacement after a harsh arid hiatus is the most likely explanation for the occupational sequence at Gobero.We are just beginning to understand the anatomical and cultural diversity that existed within the Sahara during the Holocene.


Synaptic interactions among excitatory afferents to nucleus accumbens neurons: hippocampal gating of prefrontal cortical input.
The interactions among excitatory inputs arising from the prefrontal cortex, amygdala, and hippocampus, and innervating nucleus accumbens neurons were studied using in vivo intracellular recording techniques. Neurons recorded in the accumbens displayed one of three activity states: (1) silent, (2) spontaneously firing at low, constant rates, or (3) a bistable membrane potential, characterized by alternating periods of activity and silence occurring in concert with spontaneous transitions between two steady-state membrane potentials (average, -77.3 +/- 7.1 mV base, -63.0 +/- 7.4 mV plateau). These neurons also exhibited a high degree of convergence of responses elicited by stimulation of each of the three excitatory inputs tested. Activation of hippocampal afferents, but not cortical, amygdaloid, or thalamic afferents, induced bistable cells to switch to the depolarized (active) state. In contrast, no bistable cells were encountered in the nucleus accumbens following an acute transection of the fornix. Furthermore, microinjection of lidocaine in the vicinity of the hippocampal afferents at the level of the fornix caused a reversible elimination of the plateau phase in bistable cells. These data suggest that hippocampal input is necessary for accumbens neurons to enter a depolarized, active state. Furthermore, activation of prefrontal cortical inputs fail to evoke spike firing in accumbens neurons unless they are in this active state. Consequently, the hippocampus appears to be capable of gating prefrontal corticoaccumbens throughput.


Operator training requirements and diagnostic accuracy of Fibroscan in routine clinical practice.
Fibroscan is a quick, non-invasive technique used to measure liver stiffness (kPa), which correlates with fibrosis. To achieve a valid liver stiffness evaluation (LSE) the operator must obtain all the following three criteria: (1) ≥10 successful liver stiffness measurements; (2) IQR/median ratio <0.30 and (3) ≥60% measurement success rate. To assess the operator training requirements and the importance of adhering to the LSE validity criteria in routine clinical practice. We retrospectively analysed the LSE validity rates of 2311 Fibroscans performed (1 August 2008 to 31 July 2011) in our tertiary liver outpatients department at the University Hospital Birmingham, UK. The diagnostic accuracy of Fibroscan was assessed in 153 patients, by comparing LSE (valid and invalid) with the modified Ishak fibrosis stage on liver biopsy. Learning curve analysis highlighted that the greatest improvement in validity of LSE rates occurs in the operator's first 10 Fibroscans, reaching 64.7% validity by the 50th Fibroscan. The correlation between LSE and the fibrosis stage on liver biopsy was superior in patients with a valid LSE (n=97) compared with those with an invalid LSE (n=56) (rs 0.577 vs 0.259; p=0.022). Area under receiving operating characteristics for significant fibrosis was greater when LSE was valid (0.83 vs 0.66; p=0.048). Using an LSE cut-off of 8 kPa, the negative predictive value of valid LSE was superior to invalid LSE for the detection of significant (84% vs 71%) and advanced fibrosis (100% vs 93%). Fibroscan requires minimal operator training (≥10 observed on patients), and when a valid LSE is obtained, it is an accurate tool for excluding advanced liver fibrosis. To ensure the diagnostic accuracy of Fibroscan it is essential that the recommended LSE validity criteria are adhered to in routine clinical practice.


Uncertainty quantification of a DNA origami mechanism using a coarse-grained model and kinematic variance analysis.
Significant advances have been made towards the design, fabrication, and actuation of dynamic DNA nanorobots including the development of DNA origami mechanisms. These DNA origami mechanisms integrate relatively stiff links made of bundles of double-stranded DNA and relatively flexible joints made of single-stranded DNA to mimic the design of macroscopic machines and robots. Despite reproducing the complex configurations of macroscopic machines, these DNA origami mechanisms exhibit significant deviations from their intended motion behavior since nanoscale mechanisms are subject to significant thermal fluctuations that lead to variations in the geometry of the underlying DNA origami components. Understanding these fluctuations is critical to assess and improve the performance of DNA origami mechanisms and to enable precise nanoscale robotic functions. Here, we report a hybrid computational framework combining coarse-grained modeling with kinematic variance analysis to predict uncertainties in the motion pathway of a multi-component DNA origami mechanism. Coarse-grained modeling was used to evaluate the variation in geometry of individual components due to thermal fluctuations. This variation was incorporated in kinematic analyses to predict the motion pathway uncertainty of the entire mechanism, which agreed well with experimental characterization of motion. We further demonstrated the ability to predict the probability density of DNA origami mechanism conformations based on analysis of mechanical properties of individual joints. This integration of computational analysis, modeling tools, and experimental methods establish the foundation to predict and manage motion uncertainties of general DNA origami mechanisms to guide the design of DNA-based nanoscale machines and robots.


Single/multiple-mode-selection optical nanofilters based on end-coupled split-ring resonators.
A plasmonic mode-selection filter based on end-coupled circular split-ring resonators is proposed and demonstrated. In contrast to regular ring resonators, which can support only the integer modes, extra noninteger resonance modes will be excited by placing metallic nanowalls in the proposed circular ring resonators. Since all the modes are highly sensitive to the position of the nanowall, the associated modes are effectively excited or suppressed by manipulating the position. This proposed scheme offers great flexibility to design the transmission spectrum with expected modes. Moreover, each integer or noninteger resonance mode with high transmittance can be selected individually by cascading two proposed split-ring resonators that share an intersection of transmission peaks. The corresponding spectra and the propagation characteristics are numerically investigated by using the finite-difference time-domain method.


Antiseptic skin agents for percutaneous procedures.
Infections associated with percutaneously implanted devices, such as pacemakers, internal cardiac defibrillators, and endovascular prostheses, create difficult and complex clinical scenarios because management can entail complete device removal, antibiotic therapy, and prolonged hospitalization. A source for pathogens is often thought to be the skin surface, making skin preparation at the time of the procedure a critical part of minimizing implantation of infected devices and prostheses. The most common skin preparation agents used today include products containing iodophors or chlorhexidine gluconate. Agents are further classified by whether they are aqueous-based or alcoholbased solutions. Traditional aqueous-based iodophors, such as povidone-iodine, are one of the few products that can be safely used on mucous membrane surfaces. Alcohol-based solutions are quick, sustained, and durable, with broader spectrum antimicrobial activity. These agents seem ideal for percutaneous procedures associated with prosthesis implantation, when it is critical to minimize skin colony counts to prevent hardware infection.


Biodegradable and compostable alternatives to conventional plastics.
Packaging waste forms a significant part of municipal solid waste and has caused increasing environmental concerns, resulting in a strengthening of various regulations aimed at reducing the amounts generated. Among other materials, a wide range of oil-based polymers is currently used in packaging applications. These are virtually all non-biodegradable, and some are difficult to recycle or reuse due to being complex composites having varying levels of contamination. Recently, significant progress has been made in the development of biodegradable plastics, largely from renewable natural resources, to produce biodegradable materials with similar functionality to that of oil-based polymers. The expansion in these bio-based materials has several potential benefits for greenhouse gas balances and other environmental impacts over whole life cycles and in the use of renewable, rather than finite resources. It is intended that use of biodegradable materials will contribute to sustainability and reduction in the environmental impact associated with disposal of oil-based polymers. The diversity of biodegradable materials and their varying properties makes it difficult to make simple, generic assessments such as biodegradable products are all 'good' or petrochemical-based products are all 'bad'. This paper discusses the potential impacts of biodegradable packaging materials and their waste management, particularly via composting. It presents the key issues that inform judgements of the benefits these materials have in relation to conventional, petrochemical-based counterparts. Specific examples are given from new research on biodegradability in simulated 'home' composting systems. It is the view of the authors that biodegradable packaging materials are most suitable for single-use disposable applications where the post-consumer waste can be locally composted.


Red and processed meat consumption and risk of incident cardiovascular disease and mortality: Isfahan cohort study.
To investigate the cardiovascular risks associated with red and/or processed meat intake in a large population-based cohort study in Iran. A total of 5432 participants from the Isfahan Cohort Study (ICS) were enrolled. Diet was assessed using a validated, 48-item food frequency questionnaire. Cox proportional hazards regression models were applied to calculate hazard ratios (HRs) for the CVD risk associated with red and processed meat intake. Median follow-up was 11.2 years. Compared to the first tertile, the highest tertile of red meat intake, either alone or in combination with processed meat was associated with a 50% reduced risk of stroke (95% CI: 0.31-0.77; 95% CI: 0.33-0.82, respectively), but increased risk of CVD mortality with corresponding HRs of 1.58 (95% CI: 1.06-2.35) and 1.47 (95% CI: 1.02-2.08), respectively. Red meat and red plus processed meat intake were inversely associated with stroke risk, but positively linked with CVD mortality.


Burning mouth syndrome and saliva: detection of salivary trace elements and cytokines.
Burning mouth syndrome (BMS) is considered a syndrome with an unknown cause. Roles of various trace elements and cytokines in saliva have been implicated in the development of BMS. The aim of the present study was to compare the levels of salivary trace elements [magnesium (Mg), zinc (Zn), copper (Cu)] and interleukin (IL)-2 and IL-6, and to search for a correlation between depression/anxiety and salivary trace elements and cytokines in BMS patients and controls. Thirty patients with BMS and 30 matched healthy controls participated in the study. Unstimulated saliva was collected from participants and salivary flow rates were determined. Mg, Zn and Cu levels were determined by atomic absorbance spectrophotometry. Cytokine immunoassay kits were used to determine the concentration of IL-2 and IL-6 in the whole saliva samples. Anxiety and depression were analyzed by means of the Speilberger State-Trait Anxiety Inventory (SAI-TAI) and Zung Self-Rating Depression Scale. Although subjects in the control group had significantly higher mean levels for Mg compared with BMS patients (P < 0.01), no statistically significant differences were observed in relation to Zn and Cu levels between the two groups (P < 0.001). There were no statistically significant differences in IL-2 and IL-6 levels of BMS and control groups, but subjects in BMS group had slightly, not significantly, higher mean levels for IL-6 compared with controls. Subjects in BMS group had significantly higher mean values for TAI compared with controls (P < 0.05). There were no statistically significant differences in relation to salivary levels of Mg, Zn, Cu, IL-2, IL-6 and depression/anxiety between BMS and control groups. The results of our study indicate that Mg levels could have an impact on symptoms of BMS and further studies are necessary to determine the importance of cytokines in the pathogenesis of BMS.


Improving coeliac disease risk prediction by testing non-HLA variants additional to HLA variants.
The majority of coeliac disease (CD) patients are not being properly diagnosed and therefore remain untreated, leading to a greater risk of developing CD-associated complications. The major genetic risk heterodimer, HLA-DQ2 and DQ8, is already used clinically to help exclude disease. However, approximately 40% of the population carry these alleles and the majority never develop CD. We explored whether CD risk prediction can be improved by adding non-HLA-susceptible variants to common HLA testing. We developed an average weighted genetic risk score with 10, 26 and 57 single nucleotide polymorphisms (SNP) in 2675 cases and 2815 controls and assessed the improvement in risk prediction provided by the non-HLA SNP. Moreover, we assessed the transferability of the genetic risk model with 26 non-HLA variants to a nested case-control population (n=1709) and a prospective cohort (n=1245) and then tested how well this model predicted CD outcome for 985 independent individuals. Adding 57 non-HLA variants to HLA testing showed a statistically significant improvement compared to scores from models based on HLA only, HLA plus 10 SNP and HLA plus 26 SNP. With 57 non-HLA variants, the area under the receiver operator characteristic curve reached 0.854 compared to 0.823 for HLA only, and 11.1% of individuals were reclassified to a more accurate risk group. We show that the risk model with HLA plus 26 SNP is useful in independent populations. Predicting risk with 57 additional non-HLA variants improved the identification of potential CD patients. This demonstrates a possible role for combined HLA and non-HLA genetic testing in diagnostic work for CD.


[Reduced consciousness levels caused by hyperammonaemia].
Hyperammonaemia is an important cause of lethargy. In this article, we describe a lesser-known but potential fatal cause of hyperammonaemia. A 27-year-old woman presented with lethargy caused by hyperammonaemia. She was treated with the emergency regime that is used to treat hyperammonaemia in urea cycle defects. Although this effectively lowered the ammonia levels, the clinical situation of the patient initially deteriorated and she was transferred to the Intensive Care Unit and intubated. Urine culture identified Proteus mirabilis, a urea-splitting bacterium that caused the hyperammonaemia. Prompt and adequate treatment with antibiotics and adequate drainage of urine was started and she completely recovered. Although every patient can get hyperammonaemia caused by urinary tract infection with urea-splitting bacteria, patients with structural bladder abnormalities are at greater risk. Lethargy can be the only presenting symptom. When recognized early, it is quite treatable and has a good prognosis.


Recent advances in intervention in markovian regulatory networks.
Markovian regulatory networks constitute a class of discrete state-space models used to study gene regulatory dynamics and discover methods that beneficially alter those dynamics. Thereby, this class of models provides a framework to discover effective drug targets and design potent therapeutic strategies. The salient translational goal is to design therapeutic strategies that desirably modify network dynamics via external signals that vary the expressions of a control gene. The objective of an intervention strategy is to reduce the likelihood of the pathological cellular function related to a disease. The task of finding an effective intervention strategy can be formulated as a sequential decision making problem for a pre-defined cost of intervention and a cost-per-stage function that discriminates the gene-activity profiles. An effective intervention strategy prescribes the actions associated with an external signal that result in the minimum expected cost. This strategy in turn can be used as a treatment that reduces the long-run likelihood of gene expressions favorable to the disease. In this tutorial, we briefly summarize the first method proposed to design such therapeutic interventions, and then move on to some of the recent refinements that have been proposed. Each of these recent intervention methods is motivated by practical or analytical considerations. The presentation of the key ideas is facilitated with the help of two case studies.


Diffusiophoresis: from dilute to concentrated electrolytes.
Electrolytic diffusiophoresis is the movement of colloidal particles in response to a concentration gradient of an electrolyte. The diffusiophoretic velocity vDP is typically predicted through the relation vDP = DDP ∇log cs, where DDP is the diffusiophoretic mobility and cs is the concentration of the electrolyte. The logarithmic dependence of vDP on cs may suggest that the strength of diffusiophoretic motion is insensitive to the magnitude of the electrolyte concentration. In this article, we emphasize that DDP is intimately coupled with cs for all electrolyte concentrations. For dilute electrolytes, the finite double layer thickness effects are significant such that DDP decreases with a decrease in cs. In contrast, for concentrated electrolytes, charge screening could result in a decrease in DDP with an increase in cs. Therefore, we predict a maximum in DDP with cs for moderate electrolyte concentrations. We also show that for typical colloids and electrolytes , where Ds is the solute ambipolar diffusivity. To validate our model, we conduct microfluidic experiments with a wide range of electrolyte concentrations. The experimental data also reveals a maximum in DDP with cs, in agreement with our predictions. Our results have important implications in the broad areas of electrokinetics, lab-on-a-chip, active colloidal transport and biophysics.


Seasonality of Respiratory Viral Infections.
The seasonal cycle of respiratory viral diseases has been widely recognized for thousands of years, as annual epidemics of the common cold and influenza disease hit the human population like clockwork in the winter season in temperate regions. Moreover, epidemics caused by viruses such as severe acute respiratory syndrome coronavirus (SARS-CoV) and the newly emerging SARS-CoV-2 occur during the winter months. The mechanisms underlying the seasonal nature of respiratory viral infections have been examined and debated for many years. The two major contributing factors are the changes in environmental parameters and human behavior. Studies have revealed the effect of temperature and humidity on respiratory virus stability and transmission rates. More recent research highlights the importance of the environmental factors, especially temperature and humidity, in modulating host intrinsic, innate, and adaptive immune responses to viral infections in the respiratory tract. Here we review evidence of how outdoor and indoor climates are linked to the seasonality of viral respiratory infections. We further discuss determinants of host response in the seasonality of respiratory viruses by highlighting recent studies in the field.


Use of oligodeoxynucleotides containing O6-alkylguanine for the assay of O6-alkylguanine-DNA-alkyltransferase activity.
A sensitive assay procedure was developed for the measurement of the activity of mammalian O6-alkylguanine-DNA-alkyltransferase. The procedure utilized oligodeoxynucleotides containing O6-methylguanine as substrates for the reaction. The oligodeoxynucleotides were end labeled with 32P by the reaction with polynucleotide kinase and [gamma-32P]ATP and allowed to react with organ or cell extracts containing the alkyltransferase. The unmethylated product which was formed was separated from the substrate by reverse-phase high-pressure liquid chromatography. Since the repair by the alkyltransferase is bimolecular, the second order rate constants for the reaction between the labeled oligomer and repair protein from several different sources were determined. The amount of alkyltransferase present was then calculated from the amount of product formed and the appropriate second order rate constant for the reaction. Excellent agreement was obtained between the alkyltransferase levels determined in this procedure and those measured by conventional assay procedures in a variety of cell lines having both high and low activity. The method also gave results in good agreement with other assay procedures for a number of rat tissues, although a few tissues gave anomalous results owing to a high level of nuclease activity which degraded the substrate. This method should prove useful for the measurement of alkyltransferase activity in samples in which the activity is very low or the amount of material available is limited.


Stirring and mixing of liquids using acoustic radiation force.
The possibility of using acoustic radiation force in standing waves for stirring and mixing small volumes of liquids is theoretically analyzed. The principle of stirring considered in this paper is based on moving the microparticles suspended in a standing acoustic wave by changing the frequency so that one standing wave mode is replaced by the other, with differently positioned minima of potential energy. The period-average transient dynamics of solid microparticles and gas microbubbles is considered, and simple analytical solutions are obtained for the case of standing waves of variable amplitude. It is shown that bubbles can be moved from one equilibrium position to another two to three orders of magnitude faster than solid particles. For example, radiation force in a standing acoustic wave field may induce movement of microbubbles with a speed of the order of a few m/s at a frequency of 1 MHz and ultrasound pressure amplitude of 100 kPa, whereas the speed of rigid particles does not exceed 1 cms under the same conditions. The stirring effect can be additionally enhanced due to the fact that the bubbles that are larger and smaller than the resonant bubbles move in opposite directions. Possible applications of the analyzed stirring mechanism, such as in microarrays, are discussed.


Neospora caninum and coxiella burnetii seropositivity are related to endocrine pattern changes during gestation in lactating dairy cows.
Q fever is a zoonotic infection caused by Coxiella burnetii that is endemic worldwide. Domestic ruminants are a source of infection for humans. Given the suggestion that the bacterium recrudesces during pregnancy in cattle, this study was designed to determine whether C. burnetii infection affects hormonal patterns, such as progesterone, cortisol, pregnancy associated glycoproteins (PAG), and prolactin during gestation in lactating cows. Possible interactions with Neospora caninum were also explored. The study was performed on 58 gestating non-aborting cows. Blood samples for hormone determinations were collected on Days 40, 90, 120, 150, 180, and 210 of gestation. For antibody determinations, blood was collected at day 40 postinsemination and postpartum. By GLM repeated measures analysis of variance, we established the effects of production and reproductive variables as well as Coxiella and Neospora seropositivity related to changes on cortisol, PAG, progesterone, and prolactin levels. Coxiella antibody levels were significantly related to cortisol, PAG, and plasma progesterone concentrations, whereas Neospora seropositivity was linked to plasma progesterone concentrations. The interaction between Coxiella and Neospora seropositivity was correlated with cortisol and plasma progesterone levels, whereas the interaction seropositivity against C. burnetii-plasma cortisol concentration was related to plasma PAG levels. Finally, an effect of lactation number only was observed on plasma prolactin. Our findings suggest that both the N. caninum and C. burnetii infection or the presence of both modify endocrine patterns throughout gestation. Cows seropositive to both, Neospora and Coxiella, showed higher plasma progesterone levels than the remaining animals examined. Seropositivity to C. burnetii was associated with placental damage and diminishing PAG levels throughout the second half of gestation, along with increased plasma cortisol levels on Day 180 of gestation.


Preparing for an institutional self review using the WFME standards - an international medical school case study.
Curriculum reform poses significant challenges for medical schools across the globe. This paper describes the reforms that took place at the medical school of the Royal College of Surgeons in Ireland (RCSI) between 2005 and 2008 and the institutional self review process that accompanied these reforms. Although fully accredited with the Irish Medical Council the RCSI sought additional detailed review of all aspects of its undergraduate medical program. Five medical educationalists were invited to visit the College in 2005 and again in 2008 to act as 'critical friends' and guide the self review using the World Federation for Medical Education (WFME) standards which had recently been adopted in Ireland. The process of institutional self review (as opposed to more high stakes accreditation) can bring about significant reform, especially when supported by a panel of 'critical friends' working alongside faculty to help guide and support sustained curriculum reform. The WFME standards continue to provide a useful framework to consider all medical education activities within a medical school engaged in continuous renewal. Adequate preparation for such reviews is critical to the success of such an undertaking and should be supported by a comprehensive communication strategy and project plan.


Ancient Mammalian and Plant DNA from Late Quaternary Stalagmite Layers at Solkota Cave, Georgia.
Metagenomic analysis is a highly promising technique in paleogenetic research that allows analysis of the complete genomic make-up of a sample. This technique has successfully been employed to archaeological sediments, but possible leaching of DNA through the sequence limits interpretation. We applied this technique to the analysis of ancient DNA (aDNA) from Late Quaternary stalagmites from two caves in Western Georgia, Melouri Cave and Solkota. Stalagmites form closed systems, limiting the effect of leaching, and can be securely dated with U-series. The analyses of the sequence data from the Melouri Cave stalagmite revealed potential contamination and low preservation of DNA. However, the two Solkota stalagmites preserved ancient DNA molecules of mammals (bear, roe deer, bats) and plants (chestnut, hazelnut, flax). The aDNA bearing layers from one of the two Solkota stalagmites were dated to between ~84 ka and ~56 ka BP by U-series. The second Solkota stalagmite contained excessive detrital clay obstructing U-series dating, but it also contained bear bones with a minimum age of ~50 BP uncalibrated years and ancient DNA molecules. The preservation of authentic ancient DNA molecules in Late Quaternary speleothems opens up a new paleogenetic archive for archaeological, paleontological and paleoenvironmental research.


Modifying effect of dual antiplatelet therapy on incidence of stent thrombosis according to implanted drug-eluting stent type.
To investigate the putative modifying effect of dual antiplatelet therapy (DAPT) use on the incidence of stent thrombosis at 3 years in patients randomized to Endeavor zotarolimus-eluting stent (E-ZES) or Cypher sirolimus-eluting stent (C-SES). Of 8709 patients in PROTECT, 4357 were randomized to E-ZES and 4352 to C-SES. Aspirin was to be given indefinitely, and clopidogrel/ticlopidine for ≥ 3 months or up to 12 months after implantation. Main outcome measures were definite or probable stent thrombosis at 3 years. Multivariable Cox regression analysis was applied, with stent type, DAPT, and their interaction as the main outcome determinants. Dual antiplatelet therapy adherence remained the same in the E-ZES and C-SES groups (79.6% at 1 year, 32.8% at 2 years, and 21.6% at 3 years). We observed a statistically significant (P = 0.0052) heterogeneity in treatment effect of stent type in relation to DAPT. In the absence of DAPT, stent thrombosis was lower with E-ZES vs. C-SES (adjusted hazard ratio 0.38, 95% confidence interval 0.19, 0.75; P = 0.0056). In the presence of DAPT, no difference was found (1.18; 0.79, 1.77; P = 0.43). A strong interaction was observed between drug-eluting stent type and DAPT use, most likely prompted by the vascular healing response induced by the implanted DES system. These results suggest that the incidence of stent thrombosis in DES trials should not be evaluated independently of DAPT use, and the optimal duration of DAPT will likely depend upon stent type (Clinicaltrials.gov number NCT00476957).


Gradient forests: calculating importance gradients on physical predictors.
In ecological analyses of species and community distributions there is interest in the nature of their responses to environmental gradients and in identifying the most important environmental variables, which may be used for predicting patterns of biodiversity. Methods such as random forests already exist to assess predictor importance for individual species and to indicate where along gradients abundance changes. However, there is a need to extend these methods to whole assemblages, to establish where along the range of these gradients the important compositional changes occur, and to identify any important thresholds or change points. We develop such a method, called "gradient forest," which is an extension of the random forest approach. By synthesizing the cross-validated R2 and accuracy importance measures from univariate random forest analyses across multiple species, sampling devices, and surveys, gradient forest obtains a monotonic function of each predictor that represents the compositional turnover along the gradient of the predictor. When applied to a synthetic data set, the method correctly identified the important predictors and delineated where the compositional change points occurred along these gradients. Application of gradient forest to a real data set from part of the Great Barrier Reef identified mud fraction of the sediment as the most important predictor, with highest compositional turnover occurring at mud fraction values around 25%, and provided similar information for other predictors. Such refined information allows for more accurate capturing of biodiversity patterns for the purposes of bioregionalization, delineation of protected areas, or designing of biodiversity surveys.


An examination of concussion education programmes: a scoping review methodology.
The primary purpose was to review the literature on concussion education programmes. The secondary purpose was to inform knowledge translation strategies for concussion researchers and practitioners. Research on concussion education programmes is relatively new. As a result, the current study implemented a scoping review methodology, which is a type of literary search used to provide a preliminary assessment of the size and scope of a body of literature, as well as identify strengths, weaknesses and gaps in the research. A five-stage process for conducting a scoping review was followed for this study: (a) identifying the research questions, (b) identifying relevant studies, (c) identifying the study selection criteria, (d) charting the data and (e) reporting the results. Concussion education programmes have been developed and implemented with populations ranging in age from 9 to 49 years and have used interactive oral presentations, educational videos and computer-based learning programmes. Although the content of these programmes varied, the topics generally addressed salient aspects of concussion injury and recovery. Quantitative instruments have been the preferred methods for assessment. Education programmes aimed at improving participants' long-term concussion knowledge, behaviours and attitudes of concussions are needed. Researchers must consider using a knowledge translation framework to enhance concussion education programmes. The application of such a framework can lead to novel and interesting ways of disseminating information about concussive injury and recovery.


Hyperglycemia-induced changes in Na+/myo-inositol transport, Na(+)-K(+)-ATPase, and protein kinase C activity in proximal tubule cells.
In many tissues, hyperglycemia alters the activities of the Na(+)-dependent myo-inositol (Na/MI) transporter, Na(+)-K(+)-ATPase, and protein kinase C (PKC). However, little is known concerning adaptive changes in renal proximal tubular function after acute or chronic hyperglycemia. We examined hyperglycemia-induced changes in Na/MI transport, Na(+)-K(+)-ATPase activity, and PKC activity using three proximal tubule-like cell lines (JTC12, LLC-PK1, and OK/E cells) and primary cultures of human proximal tubular epithelium (HK cells) cultured for varying periods in low- or high-glucose media, myo-Inositol (MI) transport was mediated by a high-affinity (Km approximately 50 mumol/l) Na(+)-dependent saturable process in the four cell lines. Hyperglycemia produced a time-dependent and persistent increase in Na/MI transport in all cell lines. Chronic hyperglycemia increased the Km for MI transport in LLC-PK1 cells and increased the Vmax in both LLC-PK1 and JTC12 cells. Glucose competitively inhibited Na/MI transport in all low-glucose cells and in high-glucose HK, JTC12, and OK/E cells but had no effect on transport in high-glucose LLC-PK1 cells. Acute hyperglycemia also produced time-dependent increases in Na(+)-K(+)-ATPase activity in all cell lines, a change that persisted only in HK cells. A 24-h exposure to high glucose had no effect on PKC activity in any of the cell lines but increased Ca/phospholipid-dependent PKC activity in membrane fractions from chronically high-glucose LLC-PK1 and OK/E cells. These data suggest that hyperglycemia causes acute changes in proximal tubule function and long-lived adaptive responses in Na/MI transport and the PKC signaling pathway.


Bimusicalism: The Implicit Dual Enculturation of Cognitive and Affective Systems.
One prominent example of globalization and mass cultural exchange is bilingualism, whereby world citizens learn to understand and speak multiple languages. Music, similar to language, is a human universal, and subject to the effects of globalization. In two experiments, we asked whether bimusicalism exists as a phenomenon, and whether it can occur even without explicit formal training and extensive music-making. Everyday music listeners who had significant exposure to music of both Indian (South Asian) and Westerners traditions (IW listeners) and listeners who had experience with only Indian or Western culture (I or W listeners) participated in recognition memory and tension judgment experiments where they listened to Western and Indian music. We found that while I and W listeners showed an in-culture bias, IW listeners showed equal responses to music from both cultures, suggesting that dual mental and affective sensitivities can be extended to a nonlinguistic domain.


Similar slow down in running speed progression in species under human pressure.
Running speed in animals depends on both genetic and environmental conditions. Maximal speeds were here analysed in horses, dogs and humans using data sets on the 10 best performers covering more than a century of races. This includes a variety of distances in humans (200-1500 m). Speed has been progressing fast in the three species, and this has been followed by a plateau. Based on a Gompertz model, the current best performances reach 97.4% of maximal velocity in greyhounds to 100.3 in humans. Further analysis based on a subset of individuals and using an 'animal model' shows that running speed is heritable in horses (h(2) = 0.438, P = 0.01) and almost so in dogs (h(2) = 0.183, P = 0.08), suggesting the involvement of genetic factors. Speed progression in humans is more likely due to an enlarged population of runners, associated with improved training practices. The analysis of a data subset (40 last years in 800 and 1500 m) further showed that East Africans have strikingly improved their speed, now reaching the upper part of the human distribution, whereas that of Nordic runners stagnated in the 800 m and even declined in the 1500 m. Although speed progression in dogs and horses on one side and humans on the other has not been affected by the same genetic/environmental balance of forces, it is likely that further progress will be extremely limited.


Assessing the volume-outcome hypothesis and region-level quality improvement interventions: pancreas cancer surgery in two Canadian Provinces.
The volume-outcome hypothesis suggests that if increased provider procedure volume is associated with improved patient outcomes, then greater regionalization to high-volume providers should improve region-level outcomes. Quality improvement interventions for pancreas cancer surgery implemented in year 1999 in Ontario, Canada were designed to regionalize surgery to high-volume hospitals and decrease operative mortality. Similar interventions were not used in Quebec, Canada. We assessed the volume-outcome hypothesis and the impact of the Ontario quality improvement interventions. Administrative databases helped identify pancreatic resections from years 1994 to 2004 and relevant patient and hospital characteristics. Hospitals were high-volume if they provided ≥10 procedures in a given calendar year. Outcomes were regionalization of surgery to high-volume providers and rates of operative mortality. From 1994 to 2004 the percentage of cases in high-volume hospitals increased from 33 to 71% in Ontario and from 36 to 76% in Quebec. Annual rates of operative mortality dropped in Ontario (10.4-2.2% or less) and changed little in Quebec (7.2-9.8%). Changes in measures over time in both provinces were similar before and after year 1999. Regionalization was associated with improved operative mortality in Ontario but not in Quebec, undermining the volume-outcome hypothesis. The Ontario quality improvement interventions likely were of little influence since patterns in regionalization and operative mortality were similar before and after year 1999.


Intraluminal pressure modulates the magnitude and the frequency of induced vasomotion in rat arteries.
Arterial vasomotion and its relation to intraluminal pressure were investigated in vitro in isolated rat arteries. Femoral arteries (mean diameter = 768.2 +/- 25 microns, n = 5) and mesenteric arteries (mean diameter = 393.4 +/- 32 microns, n = 5) were used in this study. Arterial segments were excised, mounted on microcannulas and perfused with Tyrode's solution at a constant flow (100 microliters/min). After equilibration, intraluminal pressure was stepwise changed from 0 to 120 mm Hg. The changes in the outer diameter of the vessels were measured continuously over a period of 4 h after the equilibration. Vasomotion was induced by constrictor agonists (norepinephrine 10(-6) M for mesenteric arteries and norepinephrine 10(-6) M + Bay K8644 10(-7) M for femoral arteries) and was maintained only in the presence of the above-mentioned drugs. Both vasomotion magnitude and frequency are modulated by pressure. Vasomotion frequency increases with pressure increase. When intraluminal pressure varied between 0 and 120 mm Hg, vasomotion frequency varied between 0.19 and 0.49 Hz for mesenteric arteries and between 0.04 and 0.23 Hz for femoral arteries. Thus, vasomotion frequency differed clearly between the two vessel types. Vasomotion amplitude shows a biphasic relationship with a maximum occurring at about 40 mm Hg for mesenteric arteries and 50 mm Hg for femoral arteries. Based on these findings, it is hypothesized that vasomotion amplitude relates to the active mechanical properties of the artery and, in particular, to its contractile capacity.


Transfusion practice and safety: current status and possibilities for improvement.
Audits of practice and incident reporting, most notably to national haemovigilance schemes, indicate that poor hospital transfusion practice is frequent and occasionally results in catastrophic consequences for patients. Improvements in practice are needed and depend on a combined approach including a better understanding of the causes of errors; a reduction in the complexity of routine procedures taking advantage of new technology systems, which enforce agreed guidelines and policies; the setting and regular monitoring of performance standards for key aspects of the hospital transfusion process, improved organisation of transfusion in hospitals and staff training; and further research on the safe and effective use of blood and alternatives to donor blood. There needs to be a greater recognition that 'transfusion safety' applies to the hospital transfusion process as well as the contents of blood bags and that resources need to be provided for the improvement of transfusion safety and management in hospitals commensurate to their importance.


Serum and cerebrospinal fluid neuron-specific enolase for diagnosis of tuberculous meningitis.
Late diagnosis and treatment lead to high mortality and poor prognosis in tuberculous meningitis (TbM). A rapid and accurate diagnosis is necessary for a good prognosis. Neuron-specific enolase (NSE) has been investigated as a biochemical marker of nervous tissue damage. In the present study, the usefulness of NSE was evaluated, and a cut-off value for the differential diagnosis of TbM was proposed. Patient charts were reviewed for levels of serum and cerebrospinal fluid (CSF) NSE, obtained from a diagnostic CSF study of samples in age- and gender-matched TbM (n=15), aseptic meningitis (n=28) and control (n=37) patients. CSF/serum NSE ratio was higher in the TbM group than those of the control and aseptic groups (p=0.001). In binary logistic regression, CSF white blood cell count and CSF/serum NSE ratio were significant factors for diagnosis of TbM. When the cut-off value of the CSF/serum NSE ratio was 1.21, the sensitivity was 86.7% and the specificity was 75.4%. The CSF/serum NSE ratio could be a useful parameter for the early diagnosis of TbM. In addition, the authors of the present study suggest a cut-off value of 1.21 for CSF/serum NSE ratio.


Ultrasound mediated delivery of quantum dots from a proof of concept capsule endoscope to the gastrointestinal wall.
Biologic drugs, defined as therapeutic agents produced from or containing components of a living organism, are of growing importance to the pharmaceutical industry. Though oral delivery of medicine is convenient, biologics require invasive injections because of their poor bioavailability via oral routes. Delivery of biologics to the small intestine using electronic delivery with devices that are similar to capsule endoscopes is a promising means of overcoming this limitation and does not require reformulation of the therapeutic agent. The efficacy of such capsule devices for drug delivery could be further improved by increasing the permeability of the intestinal tract lining with an integrated ultrasound transducer to increase uptake. This paper describes a novel proof of concept capsule device capable of electronic application of focused ultrasound and delivery of therapeutic agents. Fluorescent markers, which were chosen as a model drug, were used to demonstrate in vivo delivery in the porcine small intestine with this capsule. We show that the fluorescent markers can penetrate the mucus layer of the small intestine at low acoustic powers when combining microbubbles with focused ultrasound during in vivo experiments using porcine models. This study illustrates how such a device could be potentially used for gastrointestinal drug delivery and the challenges to be overcome before focused ultrasound and microbubbles could be used with this device for the oral delivery of biologic therapeutics.


Increased Cerebrospinal Fluid Production as a Possible Mechanism Underlying Caffeine's Protective Effect against Alzheimer's Disease.
Alzheimer's disease (AD), the most common type of dementia among older people, is characterized by the accumulation of β-amyloid (Aβ) senile plaques and neurofibrillary tangles composed of hyperphosphorylated tau in the brain. Despite major advances in understanding the molecular etiology of the disease, progress in the clinical treatment of AD patients has been extremely limited. Therefore, new and more effective therapeutic approaches are needed. Accumulating evidence from human and animal studies suggests that the long-term consumption of caffeine, the most commonly used psychoactive drug in the world, may be protective against AD. The mechanisms underlying the suggested beneficial effect of caffeine against AD remain to be elucidated. In recent studies, several potential neuroprotective effects of caffeine have been proposed. Interestingly, a recent study in rats showed that the long-term consumption of caffeine increased cerebrospinal fluid (CSF) production, associated with the increased expression of Na(+)-K(+) ATPase and increased cerebral blood flow. Compromised function of the choroid plexus and defective CSF production and turnover, with diminished clearance of Aβ, may be one mechanism implicated in the pathogenesis of late-onset AD. If reduced CSF turnover is a risk factor for AD, then therapeutic strategies to improve CSF flow are reasonable. In this paper, we hypothesize that long-term caffeine consumption could exert protective effects against AD at least in part by facilitating CSF production, turnover, and clearance. Further, we propose a preclinical experimental design allowing evaluation of this hypothesis.


Nonlinear terahertz frequency conversion via graphene microribbon array.
By exploiting the interesting trait of graphene to have electrically tunable first- and third-order conductivities besides its capability to support plasmonic resonances at terahertz frequencies, here, through the nonlinear finite-difference time-domain numerical technique we developed, we demonstrate a noticeable improvement in the conversion efficiency of third-harmonic generation (THG) from a graphene microribbon array by more than five orders of magnitude compared to an infinite graphene sheet, under normal illumination of terahertz waves. As the Fermi level and period length of the ribbon array increase, the transmission obviously manifests a blue shift but denotes a red shift with an increase in ribbon width. The quality factor of resonance (and so the THG efficiency) also shows improvement with an increase in graphene Fermi level, carrier mobility and period length and is degraded by an increase in ribbon width. Generating new frequencies, terahertz signal processing, spectroscopy and so on are among the plethora of valuable potential applications envisioned to be developed based on the findings reported here.


Pathoanatomy of first-time, traumatic, anterior glenohumeral subluxation events.
Relative to dislocations, glenohumeral subluxation events have received little attention in the literature, despite a high incidence in young athletes. The pathoanatomy of first-time, traumatic, anterior subluxation events has not been defined, to our knowledge. As part of a prospective evaluation of all cases of shoulder instability sustained during one academic year in a closed cohort of military academy cadets, a total of thirty-eight first-time, traumatic, anterior glenohumeral subluxation events were documented. Clinical subluxation events were defined as incomplete instability events that did not require a manual reduction maneuver. Twenty-seven of those events were evaluated with plain radiographs and magnetic resonance imaging within two weeks after the injury and constitute the cohort studied. Magnetic resonance imaging studies were independently evaluated by a musculoskeletal radiologist blinded to the clinical history. Arthroscopic findings were available for the fourteen patients who underwent arthroscopic surgery. Of the twenty-seven patients who sustained a first-time, traumatic, anterior subluxation, twenty-two were male and five were female, and their mean age was twenty years. Plain radiographs revealed three osseous Bankart lesions and two Hill-Sachs lesions. Magnetic resonance imaging revealed a Bankart lesion in twenty-six of the twenty-seven patients and a Hill-Sachs lesion in twenty-five of the twenty-seven patients. Of the fourteen patients who underwent surgery, thirteen had a Bankart lesion noted during the procedure. Of the thirteen patients who chose nonoperative management, four experienced recurrent instability. Two of the thirteen patients left the academy for nonmedical reasons and were lost to follow-up. The remaining seven patients continued on active-duty service and had not sought care for a recurrent instability event at the time of writing. First-time, traumatic, anterior subluxation events result in a high rate of labral and Hill-Sachs lesions. These findings suggest that clinical subluxation events encompass a broad spectrum of incomplete events, including complete separations of the articular surfaces with spontaneous reduction. A high index of suspicion for this injury in young athletes is warranted, and magnetic resonance imaging may reveal a high rate of pathologic changes, suggesting that a complete, transient luxation of the glenohumeral joint has occurred.


Correlation of susceptibility of Cryptococcus neoformans to amphotericin B with clinical outcome.
Testing of Cryptococcus neoformans for susceptibility to antifungal drugs by standard microtiter methods has not been shown to correlate with clinical outcomes. This report describes a modified quantitative broth macrodilution susceptibility method showing a correlation with both the patient's quantitative biological response in the cerebrospinal fluid (CSF) and the survival of 85 patients treated with amphotericin B (AMB). The Spearman rank correlation between the quantitative in vitro measure of susceptibility and the quantitative measure of the number of organisms in the patient's CSF was 0.37 (P < 0.01; 95% confidence interval [95% CI], 0.20, 0.60) for the first susceptibility test replicate and 0.46 (P < 0.001; 95% CI, 0.21, 0.62) for the second susceptibility test replicate. The median in vitro estimated response (defined as the fungal burden after AMB treatment) at 1.5 mg/liter AMB for patients alive at day 14 was 5 CFU (95% CI, 3, 8), compared to 57 CFU (95% CI, 4, 832) for those who died before day 14. These exploratory results suggest that patients whose isolates show a quantitative in vitro susceptibility response below 10 CFU/ml were more likely to survive beyond day 14.


Effects of deferasirox-deferoxamine on myocardial and liver iron in patients with severe transfusional iron overload.
Deferasirox (DFX) monotherapy is effective for reducing myocardial and liver iron concentrations (LIC), although some patients may require intensive chelation for a limited duration. HYPERION, an open-label single-arm prospective phase 2 study, evaluated combination DFX-deferoxamine (DFO) in patients with severe transfusional myocardial siderosis (myocardial [m] T2* 5-<10 ms; left ventricular ejection fraction [LVEF] ≥56%) followed by optional switch to DFX monotherapy when achieving mT2* >10 ms. Mean dose was 30.5 mg/kg per day DFX and 36.3 mg/kg per day DFO on a 5-day regimen. Geometric mean mT2* ratios (Gmeanmonth12/24/Gmeanbaseline) were 1.09 and 1.30, respectively, increasing from 7.2 ms at baseline (n = 60) to 7.7 ms at 12 (n = 52) and 9.5 ms at 24 months (n = 36). Patients (17 of 60; 28.3%) achieved mT2* ≥10 ms and ≥10% increase from baseline at month 24; 15 switched to monotherapy during the study based on favorable mT2*. LIC decreased substantially from a baseline of 33.4 to 12.8 mg Fe/g dry weight at month 24 (-52%). LVEF remained stable with no new arrhythmias/cardiac failure. Five patients discontinued with mT2* <5 ms and 1 died (suspected central nervous system infection). Safety was consistent with established monotherapies. Results show clinically meaningful improvements in mT2* in about one-third of patients remaining on treatment at month 24, alongside rapid decreases in LIC in this heavily iron-overloaded, difficult-to-treat population. Combination therapy may be useful when rapid LIC reduction is required, regardless of myocardial iron overload. This trial was registered at www.clinicaltrials.gov as #NCT01254227.


Open laparoscopy for laparoscopic cholecystectomy.
This report deals with the first 350 laparoscopic cholecystectomies performed at Creighton University, beginning in October 1989, by one of the authors (R.J.F., Jr.). An open technique was used for initial access to the abdominal cavity in 343 patients (98%). Patients included 274 females and 76 males, with an average age of 45.5 years. Weight ranged from 72 to 316 pounds (32-142.2 kg). The laparoscopic procedure was successfully completed in 338 (96.6%). Operative cholangiography was performed in 113 (32.3%), and 52 (14.7%) had acute cholecystitis as determined by the final pathological report. Median operating time was 80 min. There was no operative mortality. Bile duct injury was not seen. Postoperative complications occurred in 21 patients (6%), only two of which were directly related to the open technique (cellulitis of the umbilical wound requiring antibiotics). A third patient (0.3% of 343 successful laparoscopies) required reoperation on postoperative day 4 for a small bowel perforation; the exact cause could not be determined, but conceivably it was related to the umbilical minilaparotomy. We conclude that the open technique is safe and effective method for initial peritoneal access prior to laparoscopic cholecystectomy.


Potential roles and clinical utility of prebiotics in newborns, infants, and children: proceedings from a global prebiotic summit meeting, New York City, June 27-28, 2008.
Initial bacterial colonization, including colonization with health-positive bacteria, such as bifidobacteria and lactobacilli, is necessary for the normal development of intestinal innate and adaptive immune defenses. The predominance of beneficial bacteria in the gut microflora of breast-fed infants is thought to be, at least in part, supported by the metabolism of the complex mixture of oligosaccharides present in human breast milk, and a more adult-type intestinal microbiota is found in formula-fed infants. Inadequate gut colonization, dysbiosis, may lead to an increased risk of infectious, allergic, and autoimmune disorders later in life. The addition of appropriate amounts of selected prebiotics to infant formulas can enhance the growth of bifidobacteria or lactobacilli in the colonic microbiota and, thereby, might produce beneficial effects. Among the substrates considered as prebiotics are the oligosaccharides inulin, fructo-oligosaccharides, galacto-oligosaccharides, and lactulose. There are some reports that such prebiotics have beneficial effects on various markers of health. For example, primary prevention trials in infants have provided promising data on prevention of infections and atopic dermatitis. Additional well-designed prospective clinical trials and mechanistic studies are needed to advance knowledge further in this promising field.


Evaluating lignocellulosic biomass, its derivatives, and downstream products with Raman spectroscopy.
The creation of fuels, chemicals, and materials from plants can aid in replacing products fabricated from non-renewable energy sources. Before using biomass in downstream applications, it must be characterized to assess chemical traits, such as cellulose, lignin, or lignin monomer content, or the sugars released following an acid or enzymatic hydrolysis. The measurement of these traits allows researchers to gage the recalcitrance of the plants and develop efficient deconstruction strategies to maximize yields. Standard methods for assessing biomass phenotypes often have experimental protocols that limit their use for screening sizeable numbers of plant species. Raman spectroscopy, a non-destructive, non-invasive vibrational spectroscopy technique, is capable of providing qualitative, structural information and quantitative measurements. Applications of Raman spectroscopy have aided in alleviating the constraints of standard methods by coupling spectral data with multivariate analysis to construct models capable of predicting analytes. Hydrolysis and fermentation products, such as glucose and ethanol, can be quantified off-, at-, or on-line. Raman imaging has enabled researchers to develop a visual understanding of reactions, such as different pretreatment strategies, in real-time, while also providing integral chemical information. This review provides an overview of what Raman spectroscopy is, and how it has been applied to the analysis of whole lignocellulosic biomass, its derivatives, and downstream process monitoring.


TALEN-mediated somatic mutagenesis in murine models of cancer.
Cancer genome sequencing has identified numerous somatic mutations whose biologic relevance is uncertain. In this study, we used genome-editing tools to create and analyze targeted somatic mutations in murine models of liver cancer. Transcription activator-like effector nucleases (TALEN) were designed against β-catenin (Ctnnb1) and adenomatous polyposis coli (Apc), two commonly mutated genes in hepatocellular carcinoma (HCC), to generate isogenic HCC cell lines. Both mutant cell lines exhibited evidence of Wnt pathway dysregulation. We asked whether these TALENs could create targeted somatic mutations after hydrodynamic transfection into mouse liver. TALENs targeting β-catenin promoted endogenous HCC carrying the intended gain-of-function mutations. However, TALENs targeting Apc were not as efficient in inducing in vivo homozygous loss-of-function mutations. We hypothesized that hepatocyte polyploidy might be protective against TALEN-induced loss of heterozygosity, and indeed Apc gene editing was less efficient in tetraploid than in diploid hepatocytes. To increase efficiency, we administered adenoviral Apc TALENs and found that we could achieve a higher mutagenesis rate in vivo. Our results demonstrate that genome-editing tools can enable the in vivo study of cancer genes and faithfully recapitulate the mosaic nature of mutagenesis in mouse cancer models. Cancer Res; 74(18); 5311-21. ©2014 AACR.


Mycoplasma infection and hypoxia initiate succinate accumulation and release in the VM-M3 cancer cells.
Succinate is known to act as an inflammatory signal in classically activated macrophages through stabilization of HIF-1α leading to IL-1β production. Relevant to this, hypoxia is known to drive succinate accumulation and release into the extracellular milieu. The metabolic alterations associated with succinate release during inflammation and under hypoxia are poorly understood. Data are presented showing that Mycoplasma arginini infection of VM-M3 cancer cells enhances the Warburg effect associated with succinate production in mitochondria and eventual release into the extracellular milieu. We investigated how succinate production and release was related to the changes of other soluble metabolites, including itaconate and 2-HG. Furthermore, we found that hypoxia alone could induce succinate release from the VM-M3 cells and that this could occur in the absence of glucose-driven lactate production. Our results elucidate metabolic pathways responsible for succinate accumulation and release in cancer cells, thus identifying potential targets involved in both inflammation and hypoxia. This article is part of a Special Issue entitled 20th European Bioenergetics Conference, edited by László Zimányi and László Tretter.


Soil warming, carbon-nitrogen interactions, and forest carbon budgets.
Soil warming has the potential to alter both soil and plant processes that affect carbon storage in forest ecosystems. We have quantified these effects in a large, long-term (7-y) soil-warming study in a deciduous forest in New England. Soil warming has resulted in carbon losses from the soil and stimulated carbon gains in the woody tissue of trees. The warming-enhanced decay of soil organic matter also released enough additional inorganic nitrogen into the soil solution to support the observed increases in plant carbon storage. Although soil warming has resulted in a cumulative net loss of carbon from a New England forest relative to a control area over the 7-y study, the annual net losses generally decreased over time as plant carbon storage increased. In the seventh year, warming-induced soil carbon losses were almost totally compensated for by plant carbon gains in response to warming. We attribute the plant gains primarily to warming-induced increases in nitrogen availability. This study underscores the importance of incorporating carbon-nitrogen interactions in atmosphere-ocean-land earth system models to accurately simulate land feedbacks to the climate system.


Pollen ultrastructure in anther cultures of Datura innoxia. II. The generative-cell wall.
In young pollen grains of Datura innoxia, a wall of the usual hemispherical type separates the 2 gametophytic cells initially and, in the electron microscope, appears as an electron-translucent matrix which is contiguous with the intine. Before detachment of the generative cell from the intine, the matrix decreases in thickness and in places is dispersed altogether leaving the plasmalemmae on either side of it in close apposition. A particularly prominent zone, triangular in profile, is left where the wall joins with the intine. After detachment of the cell, remnants of the matrix can be seen distributed irregularly around the cell and it is supposed that these are partly derived from material in the triangular zone as the cell is drawn away from the intine. The wall residues persist throughout the maturation phase of the pollen and are considered to be either callose resulting from incomplete digestion of the initial wall, or some other polysaccharide material which is unevenly laid down along the wall and concentrated at the junction with the intine. In pollen induced into embryogenesis by anther culture, wall material is also distributed irregularly around the detached cell in a series of discrete zones, but these are more extensive than in vivo, closer together and in many instances highly dilated. The wall profiles thus have a beaded appearance, the 'beads' being connected together by short links of the 2 apposed plasmalemmae. The contents of the swollen zones have a similar electron density to that of the matrix in vivo but also show traces of a fibrillar component. It is postulated that this unusual swelling is a prelude to dispersal of the wall by disruption of the plasmalemmal links and to the establishment of cytoplasmic continuity between the 2 cells. The significance of such binucleate pollen grains in the formation of non-haploid embryos is discussed.


Age and Smoking Related Changes in Metal Ion Levels in Human Lens: Implications for Cataract Formation.
Age-related cataract formation is the primary cause of blindness worldwide and although treatable by surgical removal of the lens the majority of sufferers have neither the finances nor access to the medical facilities required. Therefore, a better understanding of the pathogenesis of cataract may identify new therapeutic targets to prevent or slow its progression. Cataract incidence is strongly correlated with age and cigarette smoking, factors that are often associated with accumulation of metal ions in other tissues. Therefore this study evaluated the age-related changes in 14 metal ions in 32 post mortem human lenses without known cataract from donors of 11 to 82 years of age by inductively coupled plasma mass spectrometry; smoking-related changes in 10 smokers verses 14 non-smokers were also analysed. A significant age-related increase in selenium and decrease in copper ions was observed for the first time in the lens tissue, where cadmium ion levels were also increased as has been seen previously. Aluminium and vanadium ions were found to be increased in smokers compared to non-smokers (an analysis that has only been carried out before in lenses with cataract). These changes in metal ions, i.e. that occur as a consequence of normal ageing and of smoking, could contribute to cataract formation via induction of oxidative stress pathways, modulation of extracellular matrix structure/function and cellular toxicity. Thus, this study has identified novel changes in metal ions in human lens that could potentially drive the pathology of cataract formation.


Preparation, characterization and evaluation of antibacterial activity of catechins and catechins-Zn complex loaded β-chitosan nanoparticles of different particle sizes.
This study used β-chitosan nanoparticles (β-CS NPs) of different particle sizes to encapsulate catechins (CAT) or CAT-Zn complex by ionic gelation technology. The antibacterial activity of CAT or CAT-Zn complex loaded β-CS NPs against Escherichia coli and Listeria innocua were investigated based on bacterial growth curve, minimum inhibitory concentration (MIC), and minimum bacterial concentration (MBC). Fourier transform infrared spectrometer (FT-IR) was employed to study the incorporation of CAT or CAT-Zn complex into β-CS NPs. The CAT-Zn complex loaded β-CS NPs had particle size of 208-591 nm, polydispersity index (PDI) of 0.377-0.395, and positive Zeta-potential of 39.17-45.62 mV. The CAT-Zn complex loaded β-CS NPs of smaller particle sizes showed higher antibacterial activity than that of larger particle size ones. The MIC and MBC of CAT-Zn complex loaded β-CS NPs of the smallest particle size against L. innocua and E. coli were 0.031 and 0.063 mg/mL, and 0.063 and 0.125 mg/mL, respectively. This study suggested that encapsulation of CAT-Zn complex in β-CS NPs improved the antibacterial activity of CAT and CAT-Zn complex, and the encapsulators have great potential to be used as antibacterial substances for food and other applications through either direct addition or incorporation into packaging materials.


Model Comparison for Abiotic versus Biotic Pollen Dispersal.
An agent-based model with a correlated random walk is used to explore pollination within a forest. For abiotic dispersal, say via the wind, we use a purely random walk where there is no correlation between consecutive steps and for biotic dispersal, say via insect, we use a moderate or highly correlated random walk. In particular, we examine the differences in a number of biological measurement between a purely random walk and a correlated random walk in terms of gene dispersal in low and high plant densities.


Homicide-suicide and other violent deaths: an international comparison.
Homicides followed by the suicide of the perpetrator constitute a serious form of interpersonal violence. Until now no study has directly compared homicide-suicides to other violent deaths from multiple countries, allowing for a better understanding of the nature of these violent acts. Using country-specific data, this study describes and compares the incidence and patterns of homicide-suicide as well as the relationship between homicide-suicide, homicide, suicide and domestic homicide in the Netherlands, Switzerland and the United States. The results indicate that cross-nationally, homicide-suicides are more likely than other types of lethal violence to involve a female victim, multiple victims, take place in a residential setting and to be committed by a firearm. Although homicide-suicides display many similarities across the different countries, differences exist regarding age and the use of firearms in the offence. This study indicates that homicides followed by suicides differ from both homicides and suicides in similar ways internationally. Cross-national differences in the availability of firearms may explain the international variation of homicide-suicide rates and patterns.


Fatty Acid uptake and incorporation into phospholipids in the rat lens.
Phospholipids are a major component of lens fiber cells and influence the activity of membrane proteins. Previous investigations of fatty acid uptake by the lens are limited. The purpose of the present study was thus to determine whether exogenous fatty acids could be taken up by the rat lens and incorporated into molecular phospholipids. Lenses were incubated with fluorescently labeled palmitic acid and then analyzed by confocal microscopy. Concurrently, lenses incubated with either fluorescently labeled palmitic acid or the more physiologically relevant (13)C(18)-oleic acid were sectioned into nuclear and cortical regions and analyzed by highly sensitive and structurally selective electrospray ionization tandem mass spectrometry techniques. The detection of fluorescently labeled palmitic acid, even after 16 hours of incubation, was limited to approximately the outer 25% to 30% of the rat lens. Mass spectrometry also revealed the presence of free (13)C(18)-oleic acid in the cortex but not the nucleus. No evidence could be found for incorporation of fluorescently labeled palmitic acid into phospholipids; however, a low level of (13)C(18)-oleic acid incorporation into phosphatidylethanolamine (PE), specifically PE (PE 16:0/(13)C(18) 18:1) was detected in the lens cortex after 16 hours. These data demonstrate that uptake of exogenous (e.g., dietary fatty acids) by the lens and their incorporation into phospholipids is minimal, most likely occurring only during de novo synthesis in the outermost region of the lens. This finding adds support to the hypothesis that once synthesized there is no active remodeling or turnover of fiber cell phospholipids.


Evolution of HL-A and other major histocompatibility systems.
The HL-A region is a complex highly polymorphic system containing many genes with interrelated functions on the cell surface. Population data suggest the action of natural selection, perhaps via linked immune-response genes. Major histocompatibility systems, such as HL-A, are presumed to have their counterparts in at least all mammalian species. The complexity of the region suggests evolution by gene duplication. The integrity of the region may even extend to the human equivalent of the complex T locus of the mouse.


Mortality (1968-2008) in a French cohort of uranium enrichment workers potentially exposed to rapidly soluble uranium compounds.
Until recently, enrichment of uranium for civil and military purposes in France was carried out by gaseous diffusion using rapidly soluble uranium compounds. We analysed the relationship between exposure to soluble uranium compounds and exposure to external γ-radiation and mortality in a cohort of 4688 French uranium enrichment workers who were employed between 1964 and 2006. Data on individual annual exposure to radiological and non-radiological hazards were collected for workers of the AREVA NC, CEA and Eurodif uranium enrichment plants from job-exposure matrixes and external dosimetry records, differentiating between natural, enriched and depleted uranium. Cause-specific mortality was compared with the French general population via standardised mortality ratios (SMR), and was analysed via Poisson regression using log-linear and linear excess relative risk models. Over the period of follow-up, 131 161 person-years at risk were accrued and 21% of the subjects had died. A strong healthy worker effect was observed: all causes SMR=0.69, 95% CI 0.65 to 0.74. SMR for pleural cancer was significantly increased (2.3, 95% CI 1.06 to 4.4), but was only based on nine cases. Internal uranium and external γ-radiation exposures were not significantly associated with any cause of mortality. This is the first study of French uranium enrichment workers. Although limited in statistical power, further follow-up of this cohort, estimation of internal uranium doses and pooling with similar cohorts should elucidate potential risks associated with exposure to soluble uranium compounds.


Changes in the relative occupancy of metal-binding sites in the profile structure of the sarcoplasmic reticulum membrane induced by phosphorylation of the Ca2+ATPase enzyme in the presence of terbium: a time-resolved, resonance x-ray diffraction study.
Time-resolved, terbium resonance x-ray diffraction experiments have provided the locations of three different high-affinity metal-binding/transport sites on the Ca2+ATPase enzyme in the profile structure of the sarcoplasmic reticulum (SR) membrane. By considering these results in conjunction with the known, moderate-resolution profile structure of the SR membrane (derived from nonresonance x-ray and neutron diffraction studies), it was determined that the three metal-binding sites are located at the "headpiece/stalk" junction in the Ca2+ATPase profile structure, in the "transbilayer" portion of the enzyme profile near the center of the membrane phospholipid bilayer, and at the intravesicular surface of the membrane profile. All three metal-binding sites so identified are simultaneously occupied in the unphosphorylated enzyme conformation. Phosphorylation of the ATPase causes a redistribution of metal density among the sites, resulting in a net movement of metal density toward the intravesicular side of the membrane, i.e., in the direction of calcium active transport. We propose that this redistribution of metal density is caused by changes in the relative binding affinities of the three sites, mediated by local structural changes at the sites resulting from the large-scale (i.e., long-range) changes in the profile structure of the Ca2+ATPase induced by phosphorylation, as reported in an accompanying paper. The implications of these results for the mechanism of calcium active transport by the SR Ca2+ATPase are discussed briefly.


Neuroprotective and Cognitive Enhancement Potentials of Baicalin: A Review.
Neurodegenerative diseases are a heterogeneous group of disorders that are characterized by the gradual loss of neurons. The development of effective neuroprotective agents to prevent and control neurodegenerative diseases is specifically important. Recently, there has been an increasing interest in selecting flavonoid compounds as potential neuroprotective agents, owing to their high effectiveness with low side effects. Baicalin is one of the important flavonoid compounds, which is mainly isolated from the root of Scutellaria baicalensis Georgi (an important Chinese medicinal herb). In recent years, a number of studies have shown that baicalin has a potent neuroprotective effect in various in vitro and in vivo models of neuronal injury. In particular, baicalin effectively prevents neurodegenerative diseases through various pharmacological mechanisms, including antioxidative stress, anti-excitotoxicity, anti-apoptotic, anti-inflammatory, stimulating neurogenesis, promoting the expression of neuronal protective factors, etc. This review mainly focuses on the neuroprotective and cognitive enhancement effects of baicalin. The aim of the present review is to compile all information in relation to the neuroprotective and cognitive enhancement effects of baicalin and its molecular mechanisms of action in various in vitro and in vivo experimental models.


Structure of a putative ClpS N-end rule adaptor protein from the malaria pathogen Plasmodium falciparum.
The N-end rule pathway uses an evolutionarily conserved mechanism in bacteria and eukaryotes that marks proteins for degradation by ATP-dependent chaperones and proteases such as the Clp chaperones and proteases. Specific N-terminal amino acids (N-degrons) are sufficient to target substrates for degradation. In bacteria, the ClpS adaptor binds and delivers N-end rule substrates for their degradation upon association with the ClpA/P chaperone/protease. Here, we report the first crystal structure, solved at 2.7 Å resolution, of a eukaryotic homolog of bacterial ClpS from the malaria apicomplexan parasite Plasmodium falciparum (Pfal). Despite limited sequence identity, Plasmodium ClpS is very similar to bacterial ClpS. Akin to its bacterial orthologs, plasmodial ClpS harbors a preformed hydrophobic pocket whose geometry and chemical properties are compatible with the binding of N-degrons. However, while the N-degron binding pocket in bacterial ClpS structures is open and accessible, the corresponding pocket in Plasmodium ClpS is occluded by a conserved surface loop that acts as a latch. Despite the closed conformation observed in the crystal, we show that, in solution, Pfal-ClpS binds and discriminates peptides mimicking bona fide N-end rule substrates. The presence of an apicoplast targeting peptide suggests that Pfal-ClpS localizes to this plastid-like organelle characteristic of all Apicomplexa and hosting most of its Clp machinery. By analogy with the related ClpS1 from plant chloroplasts and cyanobacteria, Plasmodium ClpS likely functions in association with ClpC in the apicoplast. Our findings open new venues for the design of novel anti-malarial drugs aimed at disrupting parasite-specific protein quality control pathways.


Fabrication of microstructured binary polymer brush "corrals" with integral pH sensing for studies of proton transport in model membrane systems.
Binary brush structures consisting of poly(cysteine methacrylate) (PCysMA) "corrals" enclosed within poly(oligoethylene glycol methyl ether methacrylate) (POEGMA) "walls" are fabricated simply and efficiently using a two-step photochemical process. First, the C-Cl bonds of 4-(chloromethyl)phenylsilane monolayers are selectively converted into carboxylic acid groups by patterned exposure to UV light through a mask and POEGMA is grown from unmodified chlorinated regions by surface-initiated atom-transfer radical polymerisation (ATRP). Incorporation of a ratiometric fluorescent pH indicator, Nile Blue 2-(methacryloyloxy)ethyl carbamate (NBC), into the polymer brushes facilitates assessment of local changes in pH using a confocal laser scanning microscope with spectral resolution capability. Moreover, the dye label acts as a radical spin trap, enabling removal of halogen end-groups from the brushes via in situ dye addition during the polymerisation process. Second, an initiator is attached to the carboxylic acid-functionalised regions formed by UV photolysis in the patterning step, enabling growth of PCysMA brushes by ATRP. Transfer of the system to THF, a poor solvent for PCysMA, causes collapse of the PCysMA brushes. At the interface between the collapsed brush and solvent, selective derivatisation of amine groups is achieved by reaction with excess glutaraldehyde, facilitating attachment of aminobutyl(nitrile triacetic acid) (NTA). The PCysMA brush collapse is reversed on transfer to water, leaving it fully expanded but only functionalized at the brush-water interface. Following complexation of NTA with Ni2+, attachment of histidine-tagged proteorhodopsin and lipid deposition, light-activated transport of protons into the brush structure is demonstrated by measuring the ratiometric response of NBC in the POEGMA walls.


White matter integrity in adolescents with histories of marijuana use and binge drinking.
Structural brain abnormalities have been observed in adolescents with alcohol use disorders but less is known about neuropathological brain characteristics of teens with sub-diagnostic binge drinking or the common pattern of binge drinking combined with marijuana use. The goal of this study was to examine white matter integrity in adolescents with histories of binge drinking and marijuana use. Diffusion tensor imaging (DTI) was conducted with 42 adolescents (ages 16-19) classified as controls, binge drinkers, or binge drinkers who are also heavy marijuana users. Tract based spatial analysis identified shared fiber structure across individuals and facilitated voxelwise comparisons of fractional anisotropy (FA) and mean diffusivity (MD) between groups. Significant between group differences were found in FA in eight white matter regions (ps < or = .016) between the binge drink-only group and controls, including superior corona radiata, inferior longitudinal fasciculus, inferior fronto-occipital fasciculus, and superior longitudinal fasciculus. Interestingly, in 4 of these same regions, binge drinkers who are also heavy marijuana users had higher FA than binge drinkers who did not use marijuana (ps<.05). MD did not differ between groups. Findings are largely consistent with research suggesting less neuropathology in adolescents without histories of substance use. However, binge drinkers who also use marijuana did not show as consistent a divergence from non-users as did the binge drink-only group. Detection of white matter alterations may have implications in identifying early cognitive dysfunction in substance using adolescents.


Potato leafroll virus: a classic pathogen shows some new tricks.
SUMMARY Taxonomy: PLRV is the type species of the genus Polerovirus, in the family Luteoviridae. Isolates are known from most continents, presumably all spread in potato material derived from the Andean region of South America. Physical properties: PLRV particles are isometric and c. 25 nm in diameter. They contain one major (c. 23 kDa) and one minor (c. 80 kDa) protein. The genome is a single 5.8 kb positive sense RNA that has neither a 5'-cap nor 3' poly(A) but carries a VPg. PLRV has a limited host range; about 20 largely solanaceous species have been infected experimentally. PLRV is a common pathogen of potato, and closely related isolates are occasionally found in tomato, but no other crops are affected. Infection, especially from infected seed potato stocks, causes leafrolling and stunting, the extent depending on the potato cultivar. Biological properties: The biology of PLRV is that of a classic luteovirus. Its isometric particles are persistently transmitted by aphids in a non-propagative manner, it multiplies largely in phloem tissue and disease symptoms reflect this localization. A decade or so of molecular study has revealed the many features of PLRV that are characteristic of its family. Key attractions: In recent years some interesting features of PLRV have emerged that are the focus of further investigation. These are, its phloem confinement, its movement in infected plants, its ability to suppress gene silencing and new ideas about the structure of its particles. This review describes the background to PLRV and points towards these new developments.


Nucleic acid scavenging polymers inhibit extracellular DNA-mediated innate immune activation without inhibiting anti-viral responses.
Toll-like receptor (TLR) family members, 3, 7 and 9 are key components in initiation and progression of autoimmune disorders such as systemic lupus erythematosus (SLE). These TLRs are often referred to as nucleic acid-sensing TLRs based on their ability to recognize DNAs or RNAs produced by pathogens or damaged cells. During autoimmune disease progression these receptors recognize self nucleic acids as well as self nucleic acid-containing complexes and contribute to inflammatory cytokine production and subsequent enhancement of serum autoantibody levels. We have recently discovered that nucleic-acid scavenging polymers (NASPs) can neutralize the proinflammatory effects of nucleic acids. Here, we begin to explore what effects such NASPs have on normal immune function. We show that such NASPs can inhibit TLR activation without affecting nucleic acid-independent T cell activation. Moreover, we observe that stimulation of immune cells by encapsulated nucleic acids, such as those found in viral particles, is unaffected by NASPs. Thus NASPs only limit the activation of the immune system by accessible extra-cellular nucleic acid and do not engender non-specific immune suppression. These important findings suggest that NASPs represent a new approach toward anti-inflammatory drug development as these agents can potentially be utilized to block overt autoimmune disorders and inflammation while allowing normal immune responses to occur.


Improving Information on Maternal Medication Use by Linking Prescription Data to Congenital Anomaly Registers: A EUROmediCAT Study.
Research on associations between medication use during pregnancy and congenital anomalies is significative for assessing the safe use of a medicine in pregnancy. Congenital anomaly (CA) registries do not have optimal information on medicine exposure, in contrast to prescription databases. Linkage of prescription databases to the CA registries is a potentially effective method of obtaining accurate information on medicine use in pregnancies and the risk of congenital anomalies. We linked data from primary care and prescription databases to five European Surveillance of Congenital Anomalies (EUROCAT) CA registries. The linkage was evaluated by looking at linkage rate, characteristics of linked and non-linked cases, first trimester exposure rates for six groups of medicines according to the prescription data and information on medication use registered in the CA databases, and agreement of exposure. Of the 52,619 cases registered in the CA databases, 26,552 could be linked. The linkage rate varied between registries over time and by type of birth. The first trimester exposure rates and the agreements between the databases varied for the different medicine groups. Information on anti-epileptic drugs and insulins and analogue medicine use recorded by CA registries was of good quality. For selective serotonin reuptake inhibitors, anti-asthmatics, antibacterials for systemic use, and gonadotropins and other ovulation stimulants, the recorded information was less complete. Linkage of primary care or prescription databases to CA registries improved the quality of information on maternal use of medicines in pregnancy, especially for medicine groups that are less fully registered in CA registries.


The SUMO-targeted ubiquitin ligase subunit Slx5 resides in nuclear foci and at sites of DNA breaks.
The Slx5/Slx8 protein complex, a heterodimeric SUMO-targeted ubiquitin ligase, plays an important role in genomic integrity. Slx5/Slx8 is believed to interact with sumoylated proteins that reside in the nuclei of budding yeast cells. In this complex, Slx5, owing to at least two SUMO interacting motifs (SIMs), has been proposed to be the targeting subunit of the Slx8 ubiquitin ligase. However, little is known about the exact subnuclear localization and targets of Slx5/Slx8. In this study we show that Slx5, but not Slx8, forms prominent nuclear foci. The formation of these foci depends on SUMO and a SIM in Slx5. Therefore, we investigated the subnuclear localization and potential chromatin association of Slx5. Using co-localization studies in live cells and fixed chromatin, we were able to localize Slx5 to DNA damage induced foci of Rad52 and Rad9, two proteins involved in the cellular response to DNA damage. Subsequent chromatin immunoprecipitation (ChIP) studies revealed that Slx5 is associated with HO endonuclease induced chromosome breaks. Surprisingly, real-time PCR analysis of Slx5 ChIPs revealed that the level of Slx5 at HO breaks in an slx8 deletion background is reduced about 4-fold. These results indicate that the DNA-damage targeting of Slx5/Slx8 depends on formation of the heterodimer and that this occurs at a subset of nuclear foci also containing DNA damage repair and checkpoint factors.


NolR regulates diverse symbiotic signals of Sinorhizobium fredii HH103.
We have investigated in Sinorhizobium fredii HH103-1 (=HH103 Str(r)) the influence of the nolR gene on the production of three different bacterial symbiotic signals: Nod factors, signal responsive (SR) proteins, and exopolysaccharide (EPS). The presence of multiple copies of nolR (in plasmid pMUS675) repressed the transcription of all the flavonoid-inducible genes analyzed: nodA, nodD1, nolO, nolX, noeL, rhcJ, hesB, and y4pF. Inactivation of nolR (mutant SVQ517) or its overexpression (presence of pMUS675) altered the amount of Nod factors detected. Mutant SVQ517 produced Nod factors carrying N-methyl residues at the nonreducing N-acetyl-glucosamine, which never have been detected in S. fredii HH103. Plasmid pMUS675 increased the amounts of EPS produced by HH103-1 and SVQ517. The flavonoid genistein repressed EPS production of HH103-1 and SVQ517 but the presence of pMUS675 reduced this repression. The presence of plasmid pMUS675 clearly decreased the secretion of SR proteins. Inactivation, or overexpression, of nolR decreased the capacity of HH103 to nodulate Glycine max. However, HH103-1 and SVQ517 carrying plasmid pMUS675 showed enhanced nodulation capacity with Vigna unguiculata. The nolR gene was positively identified in all S. fredii strains investigated, S. xinjiangense CCBAU110, and S. saheli USDA4102. Apparently, S. teranga USDA4101 does not contain this gene.