Effect of arterial impedance changes on the end-systolic pressure-volume relation.
To study the end-systolic pressure-volume relationship of left ventricle ejection against physiological afterload, we imposed seven simulated arterial impedances on excised canine left ventricles connected to a newly developed servo-pump system. We set each of the impedance parameters (resistance, capacitance, and characteristic impedance) to 50, 100, and 200% of normal value (resistance: 3 mm Hg sec/ml; capacitance: 0.4 ml/mm Hg; characteristic impedance: 0.2 mm Hg sec/ml), while leaving the other parameters normal. Under a given impedance, the end-systolic pressure-volume relationship was determined by preloading the ventricle at four different end-diastolic volumes. There was no significant change in the slope of the end-systolic pressure-volume relationship with changes in any of the afterloading impedance parameters. However, the volume intercept of the end-systolic pressure-volume relationship decreased significantly with resistance from 5.5 +/- 1.0 (SE) ml at resistance equal to 1.5 mm Hg sec/ml to 0.6 +/- 1.8 ml at resistance equal to 6 mm Hg sec/ml (P less than 0.01). The volume axis intercept also decreased with characteristic impedance, from 5.9 +/- 2.0 ml at a characteristic impedance of 0.1 mm Hg sec/ml to 5.4 +/- 2.1 ml at a characteristic impedance of 0.4 mm Hg sec/ml, (P less than 0.05). We conclude that the slope of the end-systolic pressure-volume relationship is insensitive to a wide range of changes in afterload impedance, but its volume intercept is dependent on resistance and characteristic impedance.


Electron-induced dissociation of singly charged organic cations as a tool for structural characterization of pharmaceutical type molecules.
Collision-induced dissociation (CID) and electron-induced dissociation (EID) have been investigated for a selection of small, singly charged organic molecules of pharmaceutical interest. Comparison of these techniques has shown that EID carried out on an FTICR MS and CID performed on a linear ion trap MS produce complementary data. In a study of 33 molecule-cations, EID generated over 300 product ions compared to 190 product ions by CID with an average of only 3 product ions per precursor ion common to both tandem MS techniques. Even multiple stages of CID failed to generate many of the product ions observed following EID. The charge carrying species is also shown to have a very significant effect on the degree of fragmentation and types of product ion resulting from EID. Protonated species behave much like the ammonium adduct with suggestion of a hydrogen atom from the charge carrying species strongly affecting the fragmentation mechanism. Sodium and potassium are retained by nearly every product ion formed from [M + Na](+) or [M + K](+) and provide information to complement the EID of [M + H](+) or [M + NH(4)](+). In summary, EID is proven to be a fitting partner to CID in the structural elucidation of small singly charged ions and by studying EID of a molecule-ion holding different charge carrying species, an even greater depth of detail can be obtained for functional groups commonly used in synthetic chemistry.


Validation and Implementation of BRCA1/2 Variant Screening in Ovarian Tumor Tissue.
BRCA1/2 variant analysis in tumor tissue could streamline the referral of patients with epithelial ovarian, fallopian tube, or primary peritoneal cancer to genetic counselors and select patients who benefit most from targeted treatment. We investigated the sensitivity of BRCA1/2 variant analysis in formalin-fixed, paraffin-embedded tumor tissue using a combination of next-generation sequencing and copy number variant multiplex ligation-dependent probe amplification. After optimization using a training cohort of known BRCA1/2 mutation carriers, validation was performed in a prospective cohort in which screening of BRCA1/2 tumor DNA and leukocyte germline DNA was performed in parallel. BRCA1 promoter hypermethylation and pedigree analysis were also performed. In the training cohort, 45 of 46 germline BRCA1/2 variants were detected (sensitivity, 98%). In the prospective cohort (n = 62), all six germline variants were identified (sensitivity, 100%), together with five somatic BRCA1/2 variants and eight cases with BRCA1 promoter hypermethylation. In four BRCA1/2 variant-negative patients, surveillance or prophylactic management options were offered on the basis of positive family histories. We conclude that BRCA1/2 formalin-fixed, paraffin-embedded tumor tissue analysis reliably detects BRCA1/2 variants. When taking family history of BRCA1/2 variant-negative patients into account, tumor BRCA1/2 variant screening allows more efficient selection of epithelial ovarian cancer patients for genetic counseling and simultaneously selects patients who benefit most from targeted treatment.


Pharmacology of the metabotropic glutamate receptor mediated current at the climbing fiber to Purkinje cell synapse.
Different forms of synaptic plasticity in the cerebellum are mediated by metabotropic glutamate receptors (mGluRs). At parallel fiber (PF) to Purkinje cell (PC) synapses activation of mGluR gives rise to a well known slow synaptic current inhibited by antagonists of mGluR1. The distribution of mGluR types in the climbing fiber (CF) to PC synapses is not well known. However, a mGluR1alpha-mediated all-or-none postsynaptic current was also demonstrated at the CF-PC synapse (Dzubay and Otis, Neuron 36, 1159, 2002). Using whole cell patch-clamp recording from PCs in rat cerebellar slices with AMPA receptors blocked and glutamate uptake impaired we demonstrate a more complex pharmacology of a current obtained by single or train CF stimulation. The mGluR1 specific antagonist CPCCOEt in a group of cells suppressed this response while in a similar number of other cells it induced a potentiating effect. The antagonists of mGluR groups II and III (LY341495 and MSOP, respectively) predominantly suppressed the current. The ambiguous effect of CPCCOEt was checked by measuring the paired-pulse depression of the CF EPSC, which was not changed with the antagonist in normal as well as in low (0.5 mM) external Ca(2+) (used to prevent saturation of AMPARs), thus excluding a presynaptic effect. However, CPCCOEt induced a rise in the amplitude (by approximately 50%) as well as a prolongation (p<0.05) of the decay time of CF EPSCs at normal 2 mM Ca(2+), i.e. under conditions of AMPAR saturation, thus indicating an effect of postsynaptic origin. In 0.5 mM Ca(2+) the decay of CF EPSCs was longer but it was also significantly prolonged (p?0.01) by CPCCOEt. However, the CF EPSC amplitude was not significantly affected indicating an underlying Ca(2+)-dependent mechanism. Thus, the pharmacology of the PC mGluR-mediated response points to a dual postsynaptic role of mGluR1 giving rise to a slow postsynaptic current but also regulating other presumably mGluR-dependent currents via second messenger molecules and Ca(2+). The additional electrophysiological role of mGluR II & III types was also indicated. Such a complex regulatory mechanism may have an important role in the mGluR-dependent forms of homosynaptic plasticity and motor learning at the CF-PC synapse.


Chemotherapy-Regulated microRNA-125-HER2 Pathway as a Novel Therapeutic Target for Trastuzumab-Mediated Cellular Cytotoxicity in Small Cell Lung Cancer.
Small cell lung cancer (SCLC) accounts for 15% of all lung cancer cases and is a highly lethal disease. For the last several decades, the standard treatment for SCLC has been deadlocked, and new therapeutic strategies are urgently needed. HER2 is a member of the HER family and has been reported to be overexpressed in 30% of SCLC cases with poor prognosis. However, the clinical relevance of HER2-targeted therapy for SCLC remains unclear. Here, we first identify that cytotoxic drugs induce significant HER2 overexpression through microRNA-125a (miR-125a) and miR-125b downregulation, which in turn act as a novel therapeutic target for trastuzumab-mediated cellular cytotoxicity in SCLC. In this study, we showed that treatment of the HER2-positive SCLC cells, SBC-3 and SBC-5, with cytotoxic drugs induced a significant upregulation of HER2. Cisplatin (CDDP) treatment of SCLC cells resulted in a significant downregulation of miR-125a and miR-125b. We confirmed that miR-125a and miR-125b bound to the 3'-untranslated regions of HER2 mRNA, and that downregulation of miR-125a and miR-125b resulted in upregulation of HER2 in SCLC cells, suggesting a relationship between cytotoxic drug exposure and miR-125/HER2 dysregulation. Furthermore, using a calcein assay, we demonstrated a significantly enhanced cytotoxic effect of CDDP and trastuzumab that was mediated via antibody-dependent cellular cytotoxicity. Finally, we clearly demonstrated the enhanced antitumor effect of these agents in an orthotopic lung cancer model in vivo. Our results offer a novel therapeutic strategy for HER2-positive SCLCs by using trastuzumab combined with cytotoxic drugs.


High-intensity aerobic training lowers blood pressure and modulates the renal renin-angiotensin system in spontaneously hypertensive rats.
Background: This study aimed to verify the effects of high-intensity aerobic training (HIAT) on BP control and renin-angiotensin system (RAS) components in renal tissue of SHR. Ten SHRs received HIAT or control for 8-weeks. At the end of the training, the SBP showed a reduction of ~ 30mmHg (p < .01) in HIAT and increased by ~ 15 mmHg in the control group. HIAT resulted in a higher release of nitrite, IL-6, ACE2 and ATR2. These results indicated an association between BP, NO and renal RAS.Abbreviations: JAA: writing, carried out all experimental procedures, performed statistical analysis, original draft and revised manuscript DMS: data interpretation, formal analysis, writing, editing and revised manuscript BAP: carried all experimental procedures, revised manuscritpt CPCG: carried all experimental procedures, revised manuscritpt MEN: experimental procedures, revised manuscript and data interpretation RWP: drafted and revised manuscript RCA: writing, experimental procedures, revised manuscript JP: writing, data interpretation and revised manuscript OLF: writing, original draft and revised manuscript.


Clickable methionine as a universal probe for labelling intracellular bacteria.
Despite their clinical and biological importance, the cell biology of obligate intracellular bacteria is less well understood than that of many free-living model organisms. One reason for this is that they are mostly genetically intractable. As a consequence, it is not possible to engineer strains expressing fluorescent proteins and therefore fluorescence light microscopy - a key tool in host-pathogen cell biology studies - is difficult. Strain diversity also limits the universality of antibody-based immunofluorescence approaches. Here, we have developed a universal labelling protocol for intracellular bacteria based on a clickable methionine analog. Whilst we have applied this to obligate intracellular bacteria, we expect it to be useful for labelling free living bacteria as well as other intracellular pathogens.


Efficacy of automated continuous positive airway pressure in children with sleep-related breathing disorders in an attended setting.
The purpose of this study was to evaluate the safety and efficacy of automated continuous positive airway pressure (Auto-CPAP) in children. Sleep-related breathing disorders (SRBDs) include the clinical spectrum of symptomatic chronic snoring, upper airway resistance syndrome, and obstructive sleep apnea. This spectrum occurs in adults and children. Less data are available for children despite recognition of the condition's prevalence. CPAP has been an established treatment for adults and children. Treatment with Auto-CPAP has been available for adults but has not been reported previously in children. A group of 14 children (8 months to 12 years old) was evaluated prospectively with baseline polysomnographic study and CPAP titration performed with Auto-CPAP under sleep technologist supervision. The results demonstrated that Auto-CPAP is sensitive and effective for children with obstructive sleep apnea in an attended setting. There was 1 subject who did not seem to tolerate Auto-CPAP, but when she was switched to conventional CPAP, she did not tolerate that either. In this subject, the mask never fit well. She was excluded from the analysis. All other patients had a decrease in the number of abnormal breathing events during sleep. The respiratory disturbance index decreased from a mean of 12.6 (SD: 12.4) to 2.6 (SD: 2.7) events per hour. The lowest oxygen saturation improved from a mean of 86% (SD: 10.8) to 93.6% (SD: 3.9). We conclude that Auto-CPAP is safe and effective in an attended environment. Auto-CPAP did not eliminate all the abnormal respiratory events. In subjects 1 and 14, the final respiratory index improved but remained >5 events per hour (5.9 and 7.7, respectively). We suspect that this was because of problems with the masks leaking, which illustrates the importance of follow-up and possible need for retitration in some patients. Proper mask fit is essential for successful treatment. Additional work is needed to evaluate its utility in the home setting. This study was designed to evaluate Auto-CPAP titration in an attended environment. It did not indicate information about the effectiveness in an unattended or home setting. We demonstrate that Auto-CPAP is able to detect abnormal breathing events during sleep in children and may provide the necessary pressure to correct these events. Auto-CPAP can be used safely for pressure titration in an attended setting. Auto-CPAP devices from different manufactures are commercially available for adults. These different devices may have different algorithms and sensitivities to detect abnormal breathing episodes. This study was performed with only 1 specific model of Auto-CPAP. Our results should not be extrapolated to other Auto-CPAP devices without empirical confirmation of the devices' ability to detect and correct events in children. Auto-CPAP can be an alternative treatment for SRBDS in the pediatric population. These results allow for speculation of possible applications for Auto-CPAP in children. A potential advantage of Auto-CPAP includes permitting the initiation of treatment while awaiting a standard CPAP titration. The variable pressure response of Auto-CPAP allows for treatment under different situations such as upper airway infections, different sleeping positions, and changes in weight. As the child grows, the amount of positive pressure needed to maintain airway patency may change. Auto-CPAP may be able to adjust to these changing pressure requirements. Auto-CPAP does not eliminate the need for periodic office visits and evaluations of the clinical course.


[Italian intersocietary consensus document on aspirin therapy in primary cardiovascular prevention].
The indications for the use of aspirin in primary cardiovascular prevention continue to be a source of intense debate, with major international guidelines providing conflicting advices. This document, written by delegates of the main Italian scientific societies dealing with cardiovascular prevention and modeled on a similar document by the European Society of Cardiology Working Group on Thrombosis, reviews the evidence in favor and against the use of aspirin therapy in primary prevention based on data cumulated so far, including recent data linking aspirin with cancer protection. While awaiting the results of several ongoing studies, this document argues for a pragmatic approach to the use of low-dose aspirin in primary cardiovascular prevention, and suggests its use in patients at high cardiovascular risk, defined as ≥2 major cardiovascular events (death, myocardial infarction, or stroke) projected per 100 person-years, who are not at increased risk of bleeding.


Effect of Apolipoprotein E ɛ4 Carrier Status on Cognitive Response to Acetylcholinesterase Inhibitors in Patients with Alzheimer's Disease: A Systematic Review and Meta-Analysis.
The apolipoprotein E ɛ4 (APOE ɛ4) genotype is the major genetic risk factor for Alzheimer's disease (AD). However, its effect on an individual's response to treatment is less well understood. Many studies have reported that the presence or absence of APOE ɛ4 may have influence on the therapeutic response for acetylcholinesterase inhibitors (AChEIs), but the results were inconsistent. This study performed a systematic review and meta-analysis to evaluate the association between response of treatment with AChEIs and the APOE ɛ4 carrier status. Clinical studies with AD patients reporting APOE ɛ4 genotype were included in the analysis. Cognitive outcome was measured by the change in Mini-Mental State Examination (MMSE), cognition subscales of the Alzheimer's Disease Assessment Scale (ADAS-cog), or Cognitive Abilities Screening Instrument (CASI). A random effects model was employed to calculate the standardized mean difference (SMD) and odds ratio (OR). Of the 284 screened abstracts, 38 studies were identified, 30 of which were included for meta-analysis. Continuous data for assessing the association between APOE ε4 and cognitive outcomes of AChEIs were available from 18 studies. The cognitive outcomes showed no significant difference between APOE ε4 carriers and APOE ε4 non-carriers (SMD = 0.022, 95% CI: -0.089∼0.133, p = 0.702, I2 = 55.3%). Twelve studies with binary data were included, also revealing insignificant difference between the two groups (OR = 1.164, 95% CI: 0.928∼1.459, p = 0.189, I2 = 16.4%). Subgroup analysis indicated that AChEIs were significantly more effective than placebo in both groups. APOE ɛ4 carrier status had no significant influence on the treatment response to AChEIs in patients with AD. AChEIs had a positive therapeutic effect compared with placebo regardless of APOE ε4 carrier status.


The Saccharomyces cerevisiae Nrd1-Nab3 transcription termination pathway acts in opposition to Ras signaling and mediates response to nutrient depletion.
The Saccharomyces cerevisiae Nrd1-Nab3 pathway directs the termination and processing of short RNA polymerase II transcripts. Despite the potential for Nrd1-Nab3 to affect the transcription of both coding and noncoding RNAs, little is known about how the Nrd1-Nab3 pathway interacts with other pathways in the cell. Here we present the results of a high-throughput synthetic lethality screen for genes that interact with NRD1 and show roles for Nrd1 in the regulation of mitochondrial abundance and cell size. We also provide genetic evidence of interactions between the Nrd1-Nab3 and Ras/protein kinase A (PKA) pathways. Whereas the Ras pathway promotes the transcription of genes involved in growth and glycolysis, the Nrd1-Nab3 pathway appears to have a novel role in the rapid suppression of some genes when cells are shifted to poor growth conditions. We report the identification of new mRNA targets of the Nrd1-Nab3 pathway that are rapidly repressed in response to glucose depletion. Glucose depletion also leads to the dephosphorylation of Nrd1 and the formation of novel nuclear speckles that contain Nrd1 and Nab3. Taken together, these results indicate a role for Nrd1-Nab3 in regulating the cellular response to nutrient availability.


Butterfly species richness and diversity in the Trishna Wildlife Sanctuary in South Asia.
Several wildlife sanctuaries in the world are home to the surviving populations of many endemic species. Trishna wildlife sanctuary in northeast India is protected by law, and is home to the last surviving populations of Asian bison (Bos gorus Smith), spectacle monkey (Trachypithecus phayrie Blyth), capped langur (Trachypithecus pileatus Blyth), slow loris (Nycticebus coucang Boddaert), wild cat (Felis chaus Schreber), and wild boars (Sus scrofa L.), among many other animals and plants. The sanctuary was explored for species richness and diversity of butterflies. A six-month-long study revealed the occurrence of 59 butterfly species that included 21 unique species and 9 species listed in the threatened category. The mixed moist deciduous mature forest of the sanctuary harbored greater species richness and species diversity (39 species under 31 genera) than other parts of the sanctuary, which is comprised of regenerated secondary mixed deciduous forest (37 species under 32 genera), degraded forests (32 species under 28 genera), and open grassland with patches of plantations and artificial lakes (24 species under 17 genera). The majority of these species showed a distribution range throughout the Indo-Malayan region and Australasia tropics, and eight species were distributed in the eastern parts of South Asia, including one species, Labadea martha (F.), which is distributed in the eastern Himalayas alone. Estimator Chao 2 provided the best-predicted value of species richness. The steep slope of the species accumulation curve suggested the occurrence of a large number of rare species, and a prolonged gentle slope suggested a higher species richness at a higher sample abundance. The species composition of vegetation-rich habitats showed high similarity in comparison to vegetation-poor habitats.


Molecular-scale thermoelectricity: a worst-case scenario.
This article highlights a novel strategy for designing molecules with high thermoelectric performance, which are resilient to fluctuations. In laboratory measurements of thermoelectric properties of single-molecule junctions and self-assembled monolayers, fluctuations in frontier orbital energies relative to the Fermi energy EF of electrodes are an important factor, which determine average values of transport coefficients, such as the average Seebeck coefficient 〈S〉. In a worst-case scenario, where the relative value of EF fluctuates uniformly over the HOMO-LUMO gap, a "worst-case scenario theorem" tells us that the average Seebeck coefficient will vanish unless the transmission coefficient at the LUMO and HOMO resonances take different values. This implies that junction asymmetry is a necessary condition for obtaining non-zero values of 〈S〉 in the presence of large fluctuations. This conclusion that asymmetry can drive high thermoelectric performance is supported by detailed simulations on 17 molecules using density functional theory. Importantly, junction asymmetry does not imply that the molecules themselves should be asymmetric. We demonstrate that symmetric molecules possessing a localised frontier orbital can achieve even higher thermoelectric performance than asymmetric molecules, because under laboratory conditions of slight symmetry breaking, such orbitals are 'silent' and do not contribute to transport. Consequently, transport is biased towards the nearest "non-silent" frontier orbital and leads to a high ensemble averaged Seebeck coefficient. This effect is demonstrated for a spatially-symmetric 1,2,3-triazole-based molecule, a rotaxane-hexayne macrocycle and a phthalocyanine.


Apoptosis of mouse hippocampal cells induced by Taenia crassiceps metacestode factor.
Seizures, headache, depression and neurological deficits are the signs and symptoms most frequently reported in human neurocysticercosis. However, the cause of the associated learning and memory deficits is unknown. Here, we used Taenia crassiceps infection in mice as a model of human cysticercosis. The effects of T. crassiceps metacestode infection or T. crassiceps metacestode factor (MF) treatment on mouse hippocampal cells were studied; control mice were included. At 45 days after infection or treatment of the mice with MF, all mice were anaesthetized and perfused transcardially with saline followed by phosphate-buffered 10% formalin. Then the brains were carefully removed. Coronal sections stained using several techniques were analysed. Extensive and significant apoptosis was found in the experimental animals, mainly in the dentate gyrus, CA1, CA2, CA3 and neighbouring regions, in comparison with the apparently intact cells from control mice (P < 0.01). These results suggest that neurological deficits, especially the learning and memory deficits, may be generated by extensive apoptosis of hippocampal cells.


Hippocampal synaptic metaplasticity requires inhibitory autophosphorylation of Ca2+/calmodulin-dependent kinase II.
Virtually all CNS synapses display the potential for activity-dependent long-term potentiation (LTP) and/or long-term depression (LTD). Intriguingly, the potential to exhibit LTP or LTD at many central synapses itself is powerfully modulated by previous synaptic activity. This higher-order form of plasticity has been termed metaplasticity. Here, we show that inhibitory autophosphorylation of Ca2+/calmodulin-dependent kinase II (CaMKII) is required for hippocampal metaplasticity at the lateral perforant path-dentate granule cell synapse. Brief 10 Hz priming, which does not affect basal synaptic transmission, caused a dramatic, pathway-specific and long-lasting (up to 18 h) reduction in subsequently evoked LTP at lateral perforant path synapses. In contrast, LTD was unaffected by priming. The induction of lateral perforant path metaplasticity required the activation of NMDA receptors during priming. In addition, metaplasticity was absent in knock-in mice expressing alphaCaMKII that cannot undergo inhibitory phosphorylation, indicating that inhibitory autophosphorylation of alphaCaMKII at threonines 305/306 is required for metaplasticity. Metaplasticity was not observed in the medial perforant pathway, consistent with the observation that CaMKII activity was not required for the induction of LTP at this synapse. Thus, modulation of alphaCaMKII activity via autophosphorylation at Thr305/Thr306 is a key mechanism for metaplasticity that may be of importance in the integration of temporally separated episodes of activity.


The bone resorption inhibitors odanacatib and alendronate affect post-osteoclastic events differently in ovariectomized rabbits.
Odanacatib (ODN) is a bone resorption inhibitor which differs from standard antiresorptives by its ability to reduce bone resorption without decreasing bone formation. What is the reason for this difference? In contrast with other antiresorptives, such as alendronate (ALN), ODN targets only the very last step of the resorption process. We hypothesize that ODN may therefore modify the remodeling events immediately following osteoclastic resorption. These events belong to the reversal phase and include recruitment of osteoblasts, which is critical for connecting bone resorption to formation. We performed a histomorphometric study of trabecular remodeling in vertebrae of estrogen-deficient rabbits treated or not with ODN or ALN, a model where ODN, but not ALN, was previously shown to preserve bone formation. In line with our hypothesis, we found that ODN treatment compared to ALN results in a shorter reversal phase, faster initiation of osteoid deposition on the eroded surfaces, and higher osteoblast recruitment. The latter is reflected by higher densities of mature bone forming osteoblasts and an increased subpopulation of cuboidal osteoblasts. Furthermore, we found an increase in the interface between osteoclasts and surrounding osteoblast-lineage cells. This increase is expected to favor the osteoclast-osteoblast interactions required for bone formation. Regarding bone resorption itself, we show that ODN, but not ALN, treatment results in shallower resorption lacunae, a geometry favoring bone stiffness. We conclude that, compared to standard antiresorptives, ODN shows distinctive effects on resorption geometry and on reversal phase activities which positively affect osteoblast recruitment and may therefore favor bone formation.


Afferent projections to the ventromedial hypothalamic nucleus in a lizard, Gekko gecko.
Afferent projections to the ventromedial hypothalamic nucleus (VMH) of a lizard, Gekko gecko, were studied with wheat germ agglutinin conjugated to horseradish peroxidase. Results indicated that the shell and core areas of the VMH are largely innervated by different telencephalic nuclei but similar brain-stem nuclei. The common brainstem projections include the superior raphe, two isthmal populations (the ventral isthmal nucleus and parvocellular isthmal nucleus), and a dorsal thalamic projection from the posteroventral nucleus. Brainstem projections to the shell but not to the core of the VMH arise from the laterodorsal tegmental nucleus. Telencephalic projections to the VMH core originate from the ventrolateral septum, centromedial DVR, lateral amygdala, medial amygdala, interstitial amygdala, and ventral anterior amygdala. Telencephalic projections to the VMH shell come from the ventral pallidum, the anterior septal nucleus, the dorsal septal nucleus, the striatoamygdalar area, and the ventral posterior amygdala. These results, combined with connectional and topological information from other studies in amphibians, reptiles, and mammals, will be used to suggest homologies between a number of limbic areas, including several amygdalar nuclei.


Proteomic-based approach to gain insight into reprogramming of THP-1 cells exposed to Leishmania donovani over an early temporal window.
Leishmania donovani, a protozoan parasite, is the causative agent of visceral leishmaniasis. It lives and multiplies within the harsh environment of macrophages. In order to investigate how intracellular parasite manipulate the host cell environment, we undertook a quantitative proteomic study of human monocyte-derived macrophages (THP-1) following infection with L. donovani. We used the isobaric tags for relative and absolute quantification (iTRAQ) method and liquid chromatography-tandem mass spectrometry (LC-MS/MS) to compare expression profiles of noninfected and L. donovani-infected THP-1 cells. We detected modifications of protein expression in key metabolic pathways, including glycolysis and fatty acid oxidation, suggesting a global reprogramming of cell metabolism by the parasite. An increased abundance of proteins involved in gene transcription, RNA splicing (heterogeneous nuclear ribonucleoproteins [hnRNPs]), histones, and DNA repair and replication was observed at 24 h postinfection. Proteins involved in cell survival and signal transduction were more abundant at 24 h postinfection. Several of the differentially expressed proteins had not been previously implicated in response to the parasite, while the others support the previously identified proteins. Selected proteomics results were validated by real-time PCR and immunoblot analyses. Similar changes were observed in L. donovani-infected human monocyte-derived primary macrophages. The effect of RNA interference (RNAi)-mediated gene knockdown of proteins validated the relevance of the host quantitative proteomic screen. Our findings indicate that the host cell proteome is modulated after L. donovani infection, provide evidence for global reprogramming of cell metabolism, and demonstrate the complex relations between the host and parasite at the molecular level.


CAVITATION OF THE GHON FOCUS IN AN HIV-INFECTED INFANT WHO ACQUIRED TUBERCULOSIS AFTER THE INITIATION OF HAART.
Tuberculosis immune reconstitution inflammatory syndrome (IRIS) may present as new or worsening cavitation. We present an HIV-infected infant in whom TB infection and subsequent cavitation of the Ghon focus appeared to coincide with immune reconstitution due to highly active antiretroviral therapy (HAART). TB-IRIS in response to infection that occurs after starting HAART has not previously been described.


Refinement of protein-protein complexes in contact map space with metadynamics simulations.
Accurate protein-protein complex prediction, to atomic detail, is a challenging problem. For flexible docking cases, current state-of-the-art docking methods are limited in their ability to exhaustively search the high dimensionality of the problem space. In this study, to obtain more accurate models, an investigation into the local optimization of initial docked solutions is presented with respect to a reference crystal structure. We show how physics-based refinement of protein-protein complexes in contact map space (CMS), within a metadynamics protocol, can be performed. The method uses 5 times replicated 10 ns simulations for sampling and ranks the generated conformational snapshots with ZRANK to identify an ensemble of n snapshots for final model building. Furthermore, we investigated whether the reconstructed free energy surface (FES), or a combination of both FES and ZRANK, referred to as CSα , can help to reduce snapshot ranking error.


Montane meadow change during drought varies with background hydrologic regime and plant functional group.
Climate change models for many ecosystems predict more extreme climatic events in the future, including exacerbated drought conditions. Here we assess the effects of drought by quantifying temporal variation in community composition of a complex montane meadow landscape characterized by a hydrological gradient. The meadows occur in two regions of the Greater Yellowstone Ecosystem (Gallatin and Teton) and were classified into six categories (M1-M6, designating hydric to xeric) based upon Satellite pour l'Observation de la Terre (SPOT) satellite imagery. Both regions have similar plant communities, but patch sizes of meadows are much smaller in the Gallatin region. We measured changes in the percent cover of bare ground and plants by species and functional groups during five years between 1997 and 2007. We hypothesized that drought effects would not be manifested evenly across the hydrological gradient, but rather would be observed as hotspots of change in some areas and minimally evident in others. We also expected varying responses by plant functional groups (forbs vs. woody plants). Forbs, which typically use water from relatively shallow s,oils compared to woody plants, were expected to decrease in cover in mesic meadows, but increase in hydric meadows. Woody plants, such as Artemisia, were expected to increase, especially in mesic meadows. We identified several important trends in our meadow plant communities during this period of drought: (1) bare ground increased significantly in xeric meadows of both regions (Gallatin M6 and Teton M5) and in mesic (M3) meadows of the Teton, (2) forbs decreased significantly in the mesic and xeric meadows in both regions, (3) forbs increased in hydric (M1) meadows of the Gallatin region, and (4) woody species showed increases in M2 and M5 meadows of the Teton region and in M3 meadows of the Gallatin region. The woody response was dominated by changes in Artemisia spp. and Chrysothamnus viscidiflorus. Thus, our results supported our expectations that community change was not uniform across the landscape, but instead could be predicted based upon functional group responses to the spatial and temporal patterns of water availability, which are largely a function of plant water use and the hydrological gradient.


Salinimicrobium gaetbulicola sp. nov., isolated from tidal flat sediment.
A Gram-stain-negative, aerobic, non-flagellated, non-gliding and short rod- or rod-shaped bacterial strain, designated BB-My20(T), was isolated from tidal flat sediment taken from the southern coast of Korea. Strain BB-My20(T) grew optimally at 37 °C, at pH 7.0-7.5 and in the presence of 2 % (w/v) NaCl. A phylogenetic tree based on 16S rRNA gene sequences showed that strain BB-My20(T) fell within the clade comprising Salinimicrobium species, joining Salinimicrobium catena HY1(T), with which it had a 16S rRNA gene sequence similarity value of 97.4 %. It exhibited 95.4-96.9 % sequence similarity to the type strains of other members of the genus Salinimicrobium. Strain BB-My20(T) contained MK-6 as the predominant menaquinone and iso-C(15 : 0), anteiso-C(15 : 0) and iso-C(17 : 0) 3-OH as the major fatty acids. The major polar lipids detected in strain BB-My20(T) and S. catena JCM 14015(T) were phosphatidylethanolamine and one unidentified lipid. The DNA G+C content of strain BB-My20(T) was 45.1 mol% and its mean DNA-DNA relatedness value with S. catena JCM 14015(T) was 4.5 %. Differential phenotypic properties, together with its phylogenetic and genetic distinctiveness, revealed that strain BB-My20(T) can be distinguished from the four recognized species of the genus Salinimicrobium. On the basis of the data presented, strain BB-My20(T) is considered to represent a novel species of the genus Salinimicrobium, for which the name Salinimicrobium gaetbulicola sp. nov. is proposed; the type strain is BB-My20(T) ( = KCTC 23579(T) = CCUG 60898(T)).


One-year survival and renal function recovery of acute kidney injury patients with chronic heart failure.
To describe and analyze the clinical characteristics of acute kidney injury (AKI) patients with preexisting chronic heart failure (CHF) and to identify the prognostic factors of the 1-year outcome. A total of 120 patients with preexisting CHF who developed AKI between January 2005 and December 2010 were enrolled. CHF was diagnosed according to the European Society of Cardiology guidelines, and AKI was diagnosed using the RIFLE criteria. Clinical characteristics were recorded, and nonrecovery from kidney dysfunction as well as mortality were analyzed. The median age of the patients was 70 years, and 58.33% were male. 60% of the patients had an advanced AKI stage ('failure') and 90% were classified as NYHA class III/IV. The 1-year mortality rate was 35%. 25.83% of the patients progressed to end-stage renal disease after 1 year. Hypertension, anemia, coronary atherosclerotic heart disease and chronic kidney disease were common comorbidities. Multiple organ dysfunction syndrome (MODS; OR, 35.950; 95% CI, 4.972-259.952), arrhythmia (OR, 13.461; 95% CI, 2.379-76.161), anemia (OR, 6.176; 95% CI, 1.172-32.544) and RIFLE category (OR, 5.353; 95% CI, 1.436-19.952) were identified as risk factors of 1-year mortality. For 1-year nonrecovery from kidney dysfunction, MODS (OR, 8.884; 95% CI, 2.535-31.135) and acute heart failure (OR, 3.281; 95% CI, 1.026-10.491) were independent risk factors. AKI patients with preexisting CHF were mainly elderly patients who had an advanced AKI stage and NYHA classification. Their 1-year mortality and nonrecovery from kidney dysfunction rates were high. Identifying risk factors may help to improve their outcome.


Current drugs and medical treatment algorithms in the management of acute decompensated heart failure.
Acute decompensated heart failure (ADHF) is associated with increased hospitalization rates and high in-hospital mortality, and has emerged as a major public health problem over the past decade. In recent years, several new drugs and therapeutic approaches have failed to reduce short- and long-term morbidity and mortality in ADHF patients. New agents and strategies are under investigation in order to effectively reduce the mortality and morbidity in these patients. To review the recent experimental and clinical evidence on existing therapeutic algorithms and investigational drugs used for the treatment of ADHF. A systematic search of peer-reviewed publications was performed on Medline and EMBASE from January 1995 to January 2009. The results of unpublished trials were obtained from presentations at national and international meetings. Renal dysfunction and low systolic blood pressure (SBP) remain the main predictors of adverse clinical outcomes in ADHF patients. Thus, therapy should be tailored according to the level of SBP at admission, renal function and fluid retention. ADHF due to hypertensive disease should be treated with intravenous vasodilators and diuretics at low doses, while patients with low output syndrome need mainly inotropic support. However, few agents currently employed in the treatment of ADHF have been shown in large prospective randomized clinical trials to improve clinical outcomes. The calcium sensitizer levosimendan is superior than traditional inotropes in improving central hemodynamics and neurohormonal response in ADHF patients, without increasing their long-term survival. Vasopressin antagonists also seem to be promising and safe drugs in the treatment of ADHF patients, facilitating diuresis on top of standard-care therapy. Encouraging novel therapies include adenosine receptor antagonists, ularitide, istaroxime, cardiac myosin activators and relaxin. Clinical scenarios based on SBP are essential determinants of therapeutic approaches used for the management of ADHF. Traditional drugs (diuretics, dobutamine and milrinone) have several limitations in real clinical practice, and increase mortality rates. Investigational drugs targeting to novel pathophysiologic concepts are promising treatment approaches and ongoing trials will define their clinical efficacy and safety.


Fibroblast α11β1 integrin regulates tensional homeostasis in fibroblast/A549 carcinoma heterospheroids.
We have previously shown that fibroblast expression of α11β1 integrin stimulates A549 carcinoma cell growth in a xenograft tumor model. To understand the molecular mechanisms whereby a collagen receptor on fibroblast can regulate tumor growth we have used a 3D heterospheroid system composed of A549 tumor cells and fibroblasts without (α11+/+) or with a deletion (α11-/-) in integrin α11 gene. Our data show that α11-/-/A549 spheroids are larger than α11+/+/A549 spheroids, and that A549 cell number, cell migration and cell invasion in a collagen I gel are decreased in α11-/-/A549 spheroids. Gene expression profiling of differentially expressed genes in fibroblast/A549 spheroids identified CXCL5 as one molecule down-regulated in A549 cells in the absence of α11 on the fibroblasts. Blocking CXCL5 function with the CXCR2 inhibitor SB225002 reduced cell proliferation and cell migration of A549 cells within spheroids, demonstrating that the fibroblast integrin α11β1 in a 3D heterospheroid context affects carcinoma cell growth and invasion by stimulating autocrine secretion of CXCL5. We furthermore suggest that fibroblast α11β1 in fibroblast/A549 spheroids regulates interstitial fluid pressure by compacting the collagen matrix, in turn implying a role for stromal collagen receptors in regulating tensional hemostasis in tumors. In summary, blocking stromal α11β1 integrin function might thus be a stroma-targeted therapeutic strategy to increase the efficacy of chemotherapy.


Depressive symptoms are associated with (sub)clinical psychotic symptoms in patients with non-affective psychotic disorder, siblings and healthy controls.
Depression is a clinically relevant dimension, associated with both positive and negative symptoms, in patients with schizophrenia. However, in siblings it is unknown whether depression is associated with subclinical positive and negative symptoms. Method Depressive symptoms and their association with positive and negative symptoms were examined in 813 healthy siblings of patients with a non-affective psychotic disorder, 822 patients and 527 healthy controls. Depressive episodes meeting DSM-IV-TR criteria (lifetime) and depressed mood (lifetime) were assessed with the Comprehensive Assessment of Symptoms and History (CASH) in all three groups. In the patient group, the severity of positive and negative psychosis symptoms was assessed with the CASH. In the siblings and healthy controls, the severity of subclinical psychosis symptoms was assessed with the Community Assessment of Psychic Experiences (CAPE). Patients reported more lifetime depressed mood and more depressive episodes than both siblings and controls. Siblings had a higher chance of meeting lifetime depressive episodes than the controls; no significant differences in depressed mood were found between siblings and controls. In all three groups the number and duration of depressive symptoms were associated with (sub)clinical negative symptoms. In the patients and siblings the number of depressive symptoms was furthermore associated with (sub)clinical positive symptoms. Finally, lifetime depressed mood showed familial clustering but this clustering was absent for lifetime depressive episodes. These findings suggest that a co-occurring genetic vulnerability for both depressive and psychotic symptomatology exists on a clinical and a subclinical level.


New antifungal agents.
This article, rather than presenting an overview of all available antifungal agents, has provided an update on new information about older agents, as well as evolving information about new agents, including those currently undergoing clinical trials. Among the azoles, ketoconazole will continue to be used as a major antifungal agent in dermatology, but one must keep up with its side effects and drug interactions. The place of the new triazole fluconazole in the treatment of cutaneous fungal infections needs to be clarified by additional controlled studies. Other agents on the horizon which are still undergoing investigation include itraconazole, which should be especially useful for dermatophyte (including tinea unguium) and candidal infections, sporotrichosis, and unusual infections such as aspergillosis and phaeohyphomycosis; and terbinafine, a member of the new class of antifungals called allylamines, which is an orally and topically active fungicidal agent that should be very useful for all types of dermatophyte infections. Research continues into the effectiveness of members of other classes of antifungals, including piritetrate, cilofungin, and amorolfine. In the 1990s, dermatologists should have safer, more effective antifungal agents for treating cutaneous fungal infections.


A school intervention for mental health literacy in adolescents: effects of a non-randomized cluster controlled trial.
"Mental health for everyone" is a school program for mental health literacy and prevention aimed at secondary schools (13-15 yrs). The main aim was to investigate whether mental health literacy, could be improved by a 3-days universal education programme by: a) improving naming of symptom profiles of mental disorder, b) reducing prejudiced beliefs, and c) improving knowledge about where to seek help for mental health problems. A secondary aim was to investigate whether adolescent sex and age influenced the above mentioned variables. A third aim was to investigate whether prejudiced beliefs influenced knowledge about available help. This non-randomized cluster controlled trial included 1070 adolescents (53.9% boys, M age 14 yrs) from three schools in a Norwegian town. One school (n = 520) received the intervention, and two schools (n = 550) formed the control group. Pre-test and follow-up were three months apart. Linear mixed models and generalized estimating equations models were employed for analysis. Mental health literacy improved contingent on the intervention, and there was a shift towards suggesting primary health care as a place to seek help. Those with more prejudiced beliefs did not suggest places to seek help for mental health problems. Generally, girls and older adolescents recognized symptom profiles better and had lower levels of prejudiced beliefs. A low cost general school program may improve mental health literacy in adolescents. Gender specific programs and attention to the age and maturity of the students should be considered when mental health literacy programmes are designed and tried out. Prejudice should be addressed before imparting information about mental health issues.


Chemotherapy-induced neutropenia does not correlate with DNA repair gene polymorphisms and treatment efficacy in advanced non-small-cell lung cancer patients.
Platinum doublets are standard chemotherapy for advanced non-small-cell lung cancer (NSCLC). The aim of this study was to assess whether neutropenia is: (1) an indicator for treatment efficacy, or (2) associated with specific polymorphisms. Four hundred ninety-four patients, treated with cisplatin-docetaxel were retrospectively analyzed. Relative dose intensity (RDI) was assessed for both drugs. Neutrophil counts were assessed only on Day 21 of each cycle. Genotyping was performed for 4 different polymorphisms in ERCC1, XRCC3, XPD-23, and XPD-10. The median overall survival was 9 months. The mean RDI was 0.94 for cisplatin and 0.93 for docetaxel. Four hundred three patients received ≥ 3 cycles of chemotherapy, and 239 received ≥ 6 cycles. Thirty-one percent developed neutropenia, and 19% had Grade (G)3-4 neutropenia. RDI was lower in patients with neutropenia (G1-4; 0.87-0.93) when compared with those without (G0; 0.94-0.95; P < .02). Male patients (P = .02) had inferior survival when compared with female patients, and ECOG (Eastern Cooperative Oncology Group) 1-2 patients (P < .001) had worse survival when compared with ECOG 0. There was no significant survival difference with respect to Grade of neutropenia (G0, 8.7 vs. G1-2, 11.6 vs. G3-4, 9.6 months; P = .41). In ECOG 0 patients, survival was significantly better for neutropenic G1-4 (hazard ratio [HR], 0.55; 95% confidence interval [CI], 0.31-0.96; P = .034) when compared with non-neutropenic (G0) patients. No association was observed between examined polymorphisms and neutropenia. RDI was significantly higher in patients who did not develop neutropenia during treatment, but as the nadir period was not explored in our study, the low occurrence of neutropenia in our cohort is considered underestimated. There was no significant survival difference with respect to grade of neutropenia. Finally, none of the examined single nucleotide polymorphisms (SNPs) were associated with the presence of neutropenia, disease characteristics, response rates, or survival.


Generalised pollination systems for three invasive milkweeds in Australia.
Because most plants require pollinator visits for seed production, the ability of an introduced plant species to establish pollinator relationships in a new ecosystem may have a central role in determining its success or failure as an invader. We investigated the pollination ecology of three milkweed species - Asclepias curassavica, Gomphocarpus fruticosus and G. physocarpus - in their invaded range in southeast Queensland, Australia. The complex floral morphology of milkweeds has often been interpreted as a general trend towards specialised pollination requirements. Based on this interpretation, invasion by milkweeds contradicts the expectation than plant species with specialised pollination systems are less likely to become invasive that those with more generalised pollination requirements. However, observations of flower visitors in natural populations of the three study species revealed that their pollination systems are essentially specialised at the taxonomic level of the order, but generalised at the species level. Specifically, pollinators of the two Gomphocarpus species included various species of Hymenoptera (particularly vespid wasps), while pollinators of A. curassavica were primarily Lepidoptera (particularly nymphalid butterflies). Pollinators of all three species are rewarded with copious amounts of highly concentrated nectar. It is likely that successful invasion by these three milkweed species is attributable, at least in part, to their generalised pollinator requirements. The results of this study are discussed in terms of how data from the native range may be useful in predicting pollination success of species in a new environment.


The effect of conceptual and contextual familiarity on transfer performance.
Applying a previously learned concept to a novel problem is an important but difficult process called transfer. It is suggested that a commonsense analogy aids in transfer by linking novel concepts to familiar ones. How the context of practice affects transfer when learning using analogies is still unclear. This study investigated the effect of a commonsense analogy and context familiarity for transfer of physiological concepts. First year psychology students (n = 24) learned three concepts: Starling's law, Laplace's law, and laminar-turbulent flow. The control group saw standard explanations while the intervention group saw an additional commonsense analogy. The context of learning was the organ system used for two practice clinical cases which differed for all concepts. Testing consisted of 12 new clinical cases. Starling's law cases used the organ system from practice while the other concepts presented in both novel and familiar organ systems. Half of the sample repeated testing after 1 week delay. The outcome was ratings of explanations of cases on a 0-3 scale. The effect of analogy was significant (Mean = 1.24 with, 0.86 without, F(1,22) = 4.26, p < 0.05) but not after delay (means of 1.08 and 0.75 respectively, F = (1,10), p = 0.06) There was significant effect for familiar context (Same = 1.23 (Starling), different = 0.68 (Laplace) and 0.73 (laminar-turbulent flow) (F(2,44) = 5.14, p < 0.01). Laplace's law and laminar turbulent flow cases in the familiar organ system had means of 1.65 and 1.77 respectively compared to novel cases with means of 0.74 and 0.68 (F(1,22) = 35.64, p < 0.0001). Similar effects were observed after delay. There was significant decay in performance after delay for all participants (immediate = 1.17, delayed = 0.91, F = 11.9 (1,10) p < 0.01). Common analogies aid conceptual understanding necessary for transfer. Despite conceptual aids, solving transfer problems is difficult.


Meditation or exercise for preventing acute respiratory infection: a randomized controlled trial.
This study was designed to evaluate potential preventive effects of meditation or exercise on incidence, duration, and severity of acute respiratory infection (ARI) illness. Community-recruited adults aged 50 years and older were randomized to 1 of 3 study groups: 8-week training in mindfulness meditation, matched 8-week training in moderate-intensity sustained exercise, or observational control. The primary outcome was area-under-the-curve global illness severity during a single cold and influenza season, using the Wisconsin Upper Respiratory Symptom Survey (WURSS-24) to assess severity. Health care visits and days of missed work were counted. Nasal wash collected during ARI illness was assayed for neutrophils, interleukin-8, and viral nucleic acid. Of 154 adults randomized into the study, 149 completed the trial (82% female, 94% white, mean age 59.3 ± 6.6 years). There were 27 ARI episodes and 257 days of ARI illness in the meditation group (n = 51), 26 episodes and 241 illness days in the exercise group (n = 47), and 40 episodes and 453 days in the control group (n = 51). Mean global severity was 144 for meditation, 248 for exercise, and 358 for control. Compared with control, global severity was significantly lower for meditation (P = .004). Both global severity and total days of illness (duration) trended toward being lower for the exercise group (P=.16 and P=.032, respectively), as did illness duration for the meditation group (P=.034). Adjusting for covariates using zero-inflated multivariate regression models gave similar results. There were 67 ARI-related days of-work missed in the control group, 32 in the exercise group (P = .041), and 16 in the meditation group (P <.001). Health care visits did not differ significantly. Viruses were identified in 54% of samples from meditation, 42% from exercise, and 54% from control groups. Neutrophil count and interleukin-8 levels were similar among intervention groups. Training in meditation or exercise may be effective in reducing ARI illness burden.


Determinants of adherence to recommendations for depressed elderly patients in primary care: a multi-methods study.
It is logical that tailoring implementation strategies to address identified determinants of adherence to clinical practice guidelines should improve adherence. This study aimed to identify and prioritize determinants of adherence to six recommendations for elderly patients with depression. Group and individual interviews and a survey were conducted in Norway. Individual and group interviews with healthcare professionals and patients, and a mailed survey of healthcare professionals. A generic checklist of determinants of practice was used to categorize suggested determinants. Physicians and nurses from primary and specialist care, psychologists, researchers, and patients. Determinants of adherence to recommendations for depressed elderly patients in primary care. A total of 352 determinants were identified, of which 99 were prioritized. The most frequently identified factors had to do with dissemination of guidelines, general practitioners' time constraints, the low prioritization of elderly patients with depression, and the patients' or relatives' wish for medication. Approximately three-quarters of the determinants were from three of the seven domains in the generic checklist: individual healthcare professional factors, patient factors, and incentives and resources. The survey did not provide useful information due to a low response rate and a lack of responses to open-ended questions. The list of prioritized determinants can inform the design of interventions to implement recommendations for elderly patients with depression. The importance of the determinants that were identified may vary across communities, practices. and patients. Interventions that address important determinants are necessary to improve practice.


Genomic history of the Italian population recapitulates key evolutionary dynamics of both Continental and Southern Europeans.
The cline of human genetic diversity observable across Europe is recapitulated at a micro-geographic scale by variation within the Italian population. Besides resulting from extensive gene flow, this might be ascribable also to local adaptations to diverse ecological contexts evolved by people who anciently spread along the Italian Peninsula. Dissecting the evolutionary history of the ancestors of present-day Italians may thus improve the understanding of demographic and biological processes that contributed to shape the gene pool of European populations. However, previous SNP array-based studies failed to investigate the full spectrum of Italian variation, generally neglecting low-frequency genetic variants and examining a limited set of small effect size alleles, which may represent important determinants of population structure and complex adaptive traits. To overcome these issues, we analyzed 38 high-coverage whole-genome sequences representative of population clusters at the opposite ends of the cline of Italian variation, along with a large panel of modern and ancient Euro-Mediterranean genomes. We provided evidence for the early divergence of Italian groups dating back to the Late Glacial and for Neolithic and distinct Bronze Age migrations having further differentiated their gene pools. We inferred adaptive evolution at insulin-related loci in people from Italian regions with a temperate climate, while possible adaptations to pathogens and ultraviolet radiation were observed in Mediterranean Italians. Some of these adaptive events may also have secondarily modulated population disease or longevity predisposition. We disentangled the contribution of multiple migratory and adaptive events in shaping the heterogeneous Italian genomic background, which exemplify population dynamics and gene-environment interactions that played significant roles also in the formation of the Continental and Southern European genomic landscapes.


Mouse models of estrogen receptor-positive breast cancer.
Breast cancer is the most frequent malignancy and second leading cause of cancer-related deaths among women. Despite advances in genetic and biochemical analyses, the incidence of breast cancer and its associated mortality remain very high. About 60 - 70% of breast cancers are Estrogen Receptor alpha (ER-α) positive and are dependent on estrogen for growth. Selective estrogen receptor modulators (SERMs) have therefore provided an effective targeted therapy to treat ER-α positive breast cancer patients. Unfortunately, development of resistance to endocrine therapy is frequent and leads to cancer recurrence. Our understanding of molecular mechanisms involved in the development of ER-α positive tumors and their resistance to ER antagonists is currently limited due to lack of experimental models of ER-α positive breast cancer. In most mouse models of breast cancer, the tumors that form are typically ER-negative and independent of estrogen for their growth. However, in recent years more attention has been given to develop mouse models that develop different subtypes of breast cancers, including ER-positive tumors. In this review, we discuss the currently available mouse models that develop ER-α positive mammary tumors and their potential use to elucidate the molecular mechanisms of ER-α positive breast cancer development and endocrine resistance.


Challenges in using medicaid claims to ascertain child maltreatment.
Medicaid data contain International Classification of Diseases, Clinical Modification (ICD-9-CM) codes indicating maltreatment, yet there is a little information on how valid these codes are for the purposes of identifying maltreatment from health, as opposed to child welfare, data. This study assessed the validity of Medicaid codes in identifying maltreatment. Participants (n = 2,136) in the first National Survey of Child and Adolescent Well-Being were linked to their Medicaid claims obtained from 36 states. Caseworker determinations of maltreatment were compared with eight sets of ICD-9-CM codes. Of the 1,921 children identified by caseworkers as being maltreated, 15.2% had any relevant ICD-9-CM code in any of their Medicaid files across 4 years of observation. Maltreated boys and those of African American race had lower odds of displaying a maltreatment code. Using only Medicaid claims to identify maltreated children creates validity problems. Medicaid data linkage with other types of administrative data is required to better identify maltreated children.


Improving and sustaining core measure performance through effective accountability of clinical microsystems in an academic medical center.
Evidence-based performance measures, known as core measures, have been established by The Joint Commission to improve the quality of care for patient populations, such as those with acute myocardial infarction (AMI), heart failure, and community-acquired pneumonia (CAP), as well as to improve the quality of surgical care--the Surgical Care Improvement Project (SCIP) measures. Hospital administrators have traditionally held academic and community physicians and hospital clinicians accountable for integrating the core measures into daily practice. Such efforts have often led to suboptimal results because of the belief that the "organization" (macrosystem) is the appropriate level at which to work to improve quality. Stanford Hospital and Clinics (Stanford, California) has instead held leaders of clinical microsystems--the clinical units where care is provided--accountable to improve performance on the core measures. The strategic approaches taken for this initiative include engagement of the hospital's board of directors; clear assignment of accountability among interdisciplinary care teams to drive the change; implementation of a unit-based medical director program; transparency of core measure performance at the microsystem, mesosystem, and macrosystem levels; and concurrent monitoring with rapid feedback of results. In 2007, the first year of this initiative, the 24-metric composite compliance score for all four core measures increased from 64% to 82%. The composite score was sustained at a minimum of 90% during 2009 and Quarter 1 of 2010. Holding clinical microsystems accountable for improving unit performance proved beneficial to macrosystem performance of the Joint Commission core measures.


In search of Methuselah: estimating the upper limits to human longevity.
Estimates of the upper limits to human longevity have important policy implications that directly affect forecasts of life expectancy, active life expectancy, population aging, and social and medical programs tied to the size and health status of the elderly population. In the past, investigators have based speculations about the upper limits of human longevity on observations of past trends in mortality. Here the estimate of the upper bound is based on hypothesized reductions in current mortality rates necessary to achieve a life expectancy at birth from 80 to 120 years and an expectation of life at age 50 from 30 to 70 years. With the use of conditional probabilities of death from complete life tables for the United States, reductions in mortality required to achieve extreme longevity (that is, 80 to 120 years) were compared with those resulting from hypothetical cures for all cardiovascular diseases, ischemic heart disease, diabetes, and cancer. Results indicate that in order for life expectancy at birth to increase from present levels to what has been referred to as the average biological limit to life (age 85), mortality rates from all causes of death would need to decline at all ages by 55%, and at ages 50 and over by 60%. Given that hypothetical cures for major degenerative diseases would reduce overall mortality by 75%, it seems highly unlikely that life expectancy at birth will exceed the age of 85.


Prevalence and predictors of vitamin D deficiency in non-supplemented women with systemic lupus erythematosus in the Mediterranean region: a cohort study.
Over the past few years researchers have suggested that vitamin D plays a diverse role in autoimmune diseases such as systemic lupus erythematosus (SLE). We sought to determine the prevalence and predictors of vitamin D deficiency in a cohort of non-supplemented female SLE patients from the Mediterranean region. We carried out a prospective cohort study on all SLE patients who had visited the Department of Rheumatology at the Parc de Salut MAR (Barcelona, Spain) between June 2007 and December 2008, excluding those who had been taking vitamin D supplements (total: 73 patients, all female). For each patient, demographic information was collected; scores were measured for disease severity [SLE Disease Activity Index (SLEDAI)] and structural damage [Systemic Lupus International Collaborating Clinic/American College of Rheumatology, (SLICC/ACR) Damage Index]; pharmacological treatment was recorded; analytical variables were analysed; and plasma levels of 25-hydroxy vitamin D [25(OH)D] were quantified. Among the patients in our cohort, 68.5% [95% confidence interval (CI) 60.3-79.2] exhibited vitamin D deficiency [plasma level of 25(OH)D < 30 ng/mL]. The predictors for vitamin D deficiency were daily sunscreen use [odds ratio (OR) 1.67, p = 0.02] and high body mass index (BMI) (OR 1.32 when adjusted for seasons and patient age, p = 0.04). We did not find any correlation between vitamin D deficiency and SLEDAI score (p = 0.31), SLICC/ACR score (p = 0.82), or any other of the variables. Vitamin D insufficiency is highly prevalent among SLE patients, even in southern regions. Sunscreen use and obesity increase the risk. Clinicians should be aware of these factors and supplement SLE patients at risk of vitamin D deficiency accordingly.


Effect of high-fat diet on rat myometrium during pregnancy-isolated myometrial mitochondria are not affected.
Laboring women with elevated body mass index (BMI) have an increased risk of inefficient uterine labor contractions, and despite the significance of mitochondria in the production of energy to drive uterine contractions, mitochondrial function in the myometrium with reference to the BMI has not been explored. The objective of this study was to determine whether obesity prior to and during gestation affects oxidative capacity and/or morphology of mitochondria in the myometrium at term in an animal model. Rat dams were fed for 47 days prior to impregnation and during gestation with either (1) a regular chow diet, (2) a low-fat high-carbohydrate diet, or (3) a high-fat low-carbohydrate diet (n = 10 in each group). On day 20 of gestation, corresponding to term pregnancy, total hysterectomy was performed with subsequent examination of the function and morphology of myometrial mitochondria. Body composition was regularly assessed by quantitative magnetic resonance imaging, and blood sampling was done prior to diet assignment, impregnation, and hysterectomy. Dams on the high-fat low-carbohydrate diet achieved higher fat percentage compared to rats on the regular chow diet (p < 0.05). Maximal oxygen consumption, phosphate/oxygen ratio, or the amount of mitochondria per gram of myometrium did not differ between the three feeding groups. Electron microscopic examinations did not reveal any morphological differences in mitochondria between groups; however, a previously undescribed subsarcolemmal localization of the mitochondria in the myocyte was identified. We did not find evidence of altered myometrial mitochondrial function or morphology in this animal model of obesity prior to and during pregnancy.


Absolute and relative quantification of RNA modifications via biosynthetic isotopomers.
In the resurging field of RNA modifications, quantification is a bottleneck blocking many exciting avenues. With currently over 150 known nucleoside alterations, detection and quantification methods must encompass multiple modifications for a comprehensive profile. LC-MS/MS approaches offer a perspective for comprehensive parallel quantification of all the various modifications found in total RNA of a given organism. By feeding (13)C-glucose as sole carbon source, we have generated a stable isotope-labeled internal standard (SIL-IS) for bacterial RNA, which facilitates relative comparison of all modifications. While conventional SIL-IS approaches require the chemical synthesis of single modifications in weighable quantities, this SIL-IS consists of a nucleoside mixture covering all detectable RNA modifications of Escherichia coli, yet in small and initially unknown quantities. For absolute in addition to relative quantification, those quantities were determined by a combination of external calibration and sample spiking of the biosynthetic SIL-IS. For each nucleoside, we thus obtained a very robust relative response factor, which permits direct conversion of the MS signal to absolute amounts of substance. The application of the validated SIL-IS allowed highly precise quantification with standard deviations<2% during a 12-week period, and a linear dynamic range that was extended by two orders of magnitude.


Silicone wristbands detect individuals' pesticide exposures in West Africa.
We detected between 2 and 10 pesticides per person with novel sampling devices worn by 35 participants who were actively engaged in farming in Diender, Senegal. Participants were recruited to wear silicone wristbands for each of two separate periods of up to 5 days. Pesticide exposure profiles were highly individualized with only limited associations with demographic data. Using a 63-pesticide dual-column gas chromatography-electron capture detector (GC-ECD) method, we detected pyrethoid insecticides most frequently, followed by organophosphate pesticides which have been linked to adverse health outcomes. This work provides the first report of individualized exposure profiles among smallholder farmers in West Africa, where logistical and practical constraints have prevented the use of more traditional approaches to exposure assessment in the past. The wristbands and associated analytical method enabled detection of a broad range of agricultural, domestic, legacy and current-use pesticides, including esfenvalerate, cypermethrin, lindane, DDT and chlorpyrifos. Participants reported the use of 13 pesticide active ingredients while wearing wristbands. All six of the pesticides that were both reportedly used and included in the analytical method were detected in at least one wristband. An additional 19 pesticide compounds were detected beyond those that were reported to be in use, highlighting the importance of measuring exposure in addition to collecting surveys and self-reported use records. The wristband method is a candidate for more widespread use in pesticide exposure and health monitoring, and in the development of evidence-based policies for human health protection in an area where food security concerns are likely to intensify agricultural production and pesticide use in the near future.


Understanding evolutionary potential in virtual CPU instruction set architectures.
We investigate fundamental decisions in the design of instruction set architectures for linear genetic programs that are used as both model systems in evolutionary biology and underlying solution representations in evolutionary computation. We subjected digital organisms with each tested architecture to seven different computational environments designed to present a range of evolutionary challenges. Our goal was to engineer a general purpose architecture that would be effective under a broad range of evolutionary conditions. We evaluated six different types of architectural features for the virtual CPUs: (1) genetic flexibility: we allowed digital organisms to more precisely modify the function of genetic instructions, (2) memory: we provided an increased number of registers in the virtual CPUs, (3) decoupled sensors and actuators: we separated input and output operations to enable greater control over data flow. We also tested a variety of methods to regulate expression: (4) explicit labels that allow programs to dynamically refer to specific genome positions, (5) position-relative search instructions, and (6) multiple new flow control instructions, including conditionals and jumps. Each of these features also adds complication to the instruction set and risks slowing evolution due to epistatic interactions. Two features (multiple argument specification and separated I/O) demonstrated substantial improvements in the majority of test environments, along with versions of each of the remaining architecture modifications that show significant improvements in multiple environments. However, some tested modifications were detrimental, though most exhibit no systematic effects on evolutionary potential, highlighting the robustness of digital evolution. Combined, these observations enhance our understanding of how instruction architecture impacts evolutionary potential, enabling the creation of architectures that support more rapid evolution of complex solutions to a broad range of challenges.


Accumulation of multiple copies of maize minichromosomes.
Multiple copies of B chromosomes in maize (Zea mays) can accumulate in the genome using the B chromosome's accumulation mechanism, specifically nondisjunction at the second pollen mitosis and preferential fertilization of the egg. Using this mechanism, we accumulated 4 different-sized minichromosomes derived from the B chromosome to test the chromosome limits of the cell. The accumulation of normal B chromosomes is associated with multiple phenotypes including white stripes and asymmetric leaf blades, but when minichromosomes are accumulated these symptoms are absent. We also found that multiple B chromosome-derived minichromosomes can coexist with A chromosome-derived minichromosomes. During the years that these experiments were conducted, we found many B chromosome rearrangements and fragments, 2 recoverable A chromosome fragments, and observed a minichromosome breakage-fusion-bridge cycle in roots.


Superior conjunction of pioneer 6.
Spectrograms of the radio signals from Pioneer 6 were taken as the spacecraft was occulted by the sun. The spectral bandwidths increased slowly at first, then very rapidly at 1 degree from the sun. In addition, six solar "events" produced marked increases of bandwidth lasting for several hours. The received signal power seemed unaffected by the solar corona.


The location of the high- and low-affinity bilirubin-binding sites on serum albumin: ligand-competition analysis investigated by circular dichroism.
The locations of three bilirubin (BR)-binding sites with different affinities were identified as subdomains IB, IIA and IIIA for five mammalian serum albumins (SAs): human (HSA), bovine (BSA), rat, (RSA), rabbit (RbSA) and sheep (SSA). The stereoselectivity of a high-affinity BR-binding site was identified in the BR/SA=1/1 system by circular dichroism (CD) spectroscopy, the sites with low affinity to BR were analyzed using difference CD. Site-specific ligand-competition experiments with ibuprofen (marker for subdomain IIIA) and hemin (marker for subdomain IB) did not reveal any changes for the BR/SA=1/1 system and showed a decrease of the bound BR at BR/SA=3/1. Both sites were identified as sites with low affinity to BR. The correlation between stereoselectivity and the arrangement of Arg-Lys residues indicated similarity between the BR-binding sites in subdomain IIIA for all of the SAs studied. Subdomain IB in HSA, BSA, SSA and RbSA has P-stereoselectivity while in RSA it has M-selectivity toward BR. A ligand-competition experiment with gossypol shows a decrease of the CD signal of bound BR for the BR/SA=1/1 system as well as for BR/SA=3/1. Subdomain IIA was assigned as a high-affinity BR-binding site. The P-stereoselectivity of this site in HSA (and RSA, RbSA) was caused by the right-hand localization of charged residues R257/R218-R222, whereas the left-hand orientation of R257/R218-R199 led to the M-stereoselectivity of the primary binding site in BSA (and SSA).


The acute respiratory exposure by intratracheal instillation of Sprague-Dawley rats with diesel particulate matter induces retinal thickening.
Adverse health effects of ambient particulate matter (PM) have been demonstrated in humans, mostly in terms of respiratory and cardiovascular events. However, whether ambient particle could affect the eyes had not been fully revealed. This study investigated the effect of acute respiratory exposure to PM on eyes. Diesel exhaust product (DEP) of 200 mg/l was given endotracheally in Sprague-Dawley rats for 1 h (n = 12) and compared to normal control (n = 4). We enucleated eyes and histologically evaluated. Immunohistochemical stains for CD34 (Dako, Glostrup, Denmark, 1:50) and Ki-67 (DakoCytomation, Glostrup, Denmark, 1:150) were performed to evaluate new vessel formation and proliferation activity. After endotracheal DEP exposure, the thickness of retina was significantly increased to 258 ± 96 μm in DEP group, while that of control was 113 ± 9 μm (p = 0.025). Among the retinal structure, inner plexiform, inner and outer nuclear and rod/cone cell layers were significantly thickened (p = 0.00, 0.017, 0.004, 0.001, respectively). The outer plexiform layer of DEP group showed a tendency of thickening, but statistically insignificant. The afferent fiber and ganglion cell layer showed no thickness difference between two groups, but prominent capillaries with congestion were noted in DEP group. Neither neovascularization nor increased proliferation was demonstrated on CD34 and Ki-67 immunohistochemical staining in DEP group. This study shows that the acute respiratory exposure of ambient PM increased retinal thickness, especially inner plexiform, inner and outer nuclear and rod/cone cell layers. We thought that increase of retinal thickness in DEP group resulted in hypoxia-induced edema.


First-Principles Quantum Dynamics of Singlet Fission: Coherent versus Thermally Activated Mechanisms Governed by Molecular π Stacking.
Singlet excitons in π-stacked molecular crystals can split into two triplet excitons in a process called singlet fission that opens a route to carrier multiplication in photovoltaics. To resolve controversies about the mechanism of singlet fission, we have developed a first principles nonadiabatic quantum dynamical model that reveals the critical role of molecular stacking symmetry and provides a unified picture of coherent versus thermally activated singlet fission mechanisms in different acenes. The slip-stacked equilibrium packing structure of pentacene derivatives is found to enhance ultrafast singlet fission mediated by a coherent superexchange mechanism via higher-lying charge transfer states. By contrast, the electronic couplings for singlet fission strictly vanish at the C(2h) symmetric equilibrium π stacking of rubrene. In this case, singlet fission is driven by excitations of symmetry-breaking intermolecular vibrations, rationalizing the experimentally observed temperature dependence. Design rules for optimal singlet fission materials therefore need to account for the interplay of molecular π-stacking symmetry and phonon-induced coherent or thermally activated mechanisms.


3D printing of milk-based product.
We developed a method to perform direct ink writing (DIW) three-dimensional (3D) printing of milk products at room temperature by changing the rheological properties of the printing ink. 3D printing of food products has been demonstrated by different methods such as selective laser sintering (SLS) and hot-melt extrusion. Methods requiring high temperatures are, however, not suitable to creating 3D models consisting of temperature-sensitive nutrients. Milk is an example of such foods rich in nutrients such as calcium and protein that would be temperature sensitive. Cold-extrusion is an alternative method of 3D printing, but it requires the addition of rheology modifiers and the optimization of the multiple components. To address this limitation, we demonstrated DIW 3D printing of milk by cold-extrusion with a simple formulation of the milk ink. Our method relies on only one milk product (powdered milk). We formulated 70 w/w% milk ink and successfully fabricated complex 3D structures. Extending our method, we demonstrated multi-material printing and created food with various edible materials. Given the versatility of the demonstrated method, we envision that cold extrusion of food inks will be applied in creating nutritious and visually appealing food, with potential applications in formulating foods with various needs for nutrition and materials properties, where food inks could be extruded at room temperature without compromising the nutrients that would be degraded at elevated temperatures.


Surgery in the elderly: is age a risk factor?
This prospective, observational one-year study analyzed 623 patients who were 60 years and older, out of a cohort of 2375 patients who were admitted consecutively to the general surgery wards of the University Hospital of the West Indies (UHWI). Even though only 9.7% of the Jamaican population are 60 years and older, this age group accounted for 26.2% of total admissions. Comparison of elderly and non-elderly patients showed no differences in gender, but less elderly patients were emergency admissions (52% vs 64%, p < 0.001), more underwent surgery (68% vs 60%, p < 0.001), their mean hospital stay was longer (11.5 vs 8.0 days, p < 0.001) and their mortality rate was higher (8.8% vs 1.9%, p < 0.001). Emergency admissions (52%) exceeded elective admissions in the elderly. Forty-four (80%) of the 55 deaths in the elderly group were admitted as emergencies compared to elective admissions (p < 0.001). There were 11 deaths among the 296 elective admissions (3.7%) but 44 deaths among the 327 emergency admissions (13.5%), a significant difference in mortality rates (p < 0.001). Overall, the death rate for males was higher. Cancer was the commonest admission diagnosis (21%) and that amongst mortalities. Steps to improve the opportunities for earlier admission and optimization of care of elderly surgical patients would not only benefit them but would be an important step towards a more efficient use of already scarce resources.


Health and re-employment in a two year follow up of long term unemployed.
The aim was to examine re-employment and changes in health during a two year follow up of a representative sample of long term unemployed. This was a cross sectional study and a two year follow up. Health was measured by psychometric testing, Hopkins symptom checklist, General health questionnaire, and medical examination. Health related selection to continuous unemployment and recovery by re-employment was estimated by logistic regression with covariances deduced from the labour market theories of human capital and segmented labour market. Four municipalities in Greenland, southern Norway. Participants were a random sample of 17 to 63 year old people registered as unemployed for more than 12 weeks. In the cross sectional study, the prevalence of depression, anxiety, and somatic illness was from four to 10 times higher than in a control group of employed people. In the follow up study, there was considerable health related selection to re-employment. A psychiatric diagnosis was associated with a 70% reduction in chances of obtaining a job. Normal performance on psychometric testing showed a two to three times increased chance of re-employment. Recovery of health following re-employment was less than expected from previous studies. Health related selection to long term unemployment seems to explain a substantial part of the excess mental morbidity among unemployed people. An increased proportion of the long term unemployed will be vocationally handicapped as years pass, putting a heavy burden on social services.


LifeTime and improving European healthcare through cell-based interceptive medicine.
Here we describe the LifeTime Initiative, which aims to track, understand and target human cells during the onset and progression of complex diseases, and to analyse their response to therapy at single-cell resolution. This mission will be implemented through the development, integration and application of single-cell multi-omics and imaging, artificial intelligence and patient-derived experimental disease models during the progression from health to disease. The analysis of large molecular and clinical datasets will identify molecular mechanisms, create predictive computational models of disease progression, and reveal new drug targets and therapies. The timely detection and interception of disease embedded in an ethical and patient-centred vision will be achieved through interactions across academia, hospitals, patient associations, health data management systems and industry. The application of this strategy to key medical challenges in cancer, neurological and neuropsychiatric disorders, and infectious, chronic inflammatory and cardiovascular diseases at the single-cell level will usher in cell-based interceptive medicine in Europe over the next decade.


Consequences of traumatic brain injury for human vergence dynamics.
Traumatic brain injury involving loss of consciousness has focal effects in the human brainstem, suggesting that it may have particular consequences for eye movement control. This hypothesis was investigated by measurements of vergence eye movement parameters. Disparity vergence eye movements were measured for a population of 123 normally sighted individuals, 26 of whom had suffered diffuse traumatic brain injury (dTBI) in the past, while the remainder served as controls. Vergence tracking responses were measured to sinusoidal disparity modulation of a random-dot field. Disparity vergence step responses were characterized in terms of their dynamic parameters separately for the convergence and divergence directions. The control group showed notable differences between convergence and divergence dynamics. The dTBI group showed significantly abnormal vergence behavior on many of the dynamic parameters. The results support the hypothesis that occult injury to the oculomotor control system is a common residual outcome of dTBI.


An Evolving Choice in a Diverse Water Market: A Quality Comparison of Sachet Water with Community and Household Water Sources in Ghana.
Packaged water, particularly bagged sachet water, has become an important drinking water source in West Africa as local governments struggle to provide safe drinking water supplies. In Ghana, sachet water has become an important primary water source in urban centers, and a growing literature has explored various dimensions of this industry, including product quality. There is very little data on sachet water quality outside of large urban centers, where smaller markets often mean less producer competition and less government regulation. This study analyzes the microbiological quality of sachet water alongside samples of other common water sources at point-of-collection (POC) and point-of-use (POU) in 42 rural, peri-urban, and small-town Ghanaian communities using the IDEXX Colilert® 18 (Westbrook, ME). Levels of coliform bacteria and Escherichia coli detected in sachet water samples were statistically and significantly lower than levels detected in all other water sources at POU, including public taps and standpipes, and statistically similar or significantly lower at POC. In diverse waterscapes where households regularly patch together their water supply from different sources, sachet water appears to be an evolving alternative for safe drinking water despite many caveats, including higher unit costs and limited opportunities to recycle the plastic packaging.


[Smoking policy in mental health care].
People with mental illnesses tend to smoke more often and more heavily than other members of the public and their addiction to tobacco also has harmful effects on their physical health. So far, however, limited priority was given to smoking cessation in mental health care settings. To provide insight into the formal and informal smoking policies of Dutch mental health care organisations and into the nature and extent of the smoking cessation support they offer, and, additionally, to look at the opportunities for improvement in clinical settings. Document research on formal policies of 61 mental health care facilities, interviews with workers directly involved (n = 10), and a survey on policy implementation among staff members of treatment facilities (n = 600). One-third of the facilities did not have a formalised smoking policy document, and there was a marked difference between the smoking policies at the rest of the facilities. Treatment provision was limited, strongly dependent on the individual staff member, and was often not the most effective form of care (like medication). Many mental health patients really do want to give up smoking and often respond well to treatment. Psychiatrists play a key role in integrating and implementing an anti-smoking policy which will benefit their patients.


Epidemiological study on porcine circovirus type 2 (PCV2) infection in the European wild boar (Sus scrofa).
Porcine circovirus type 2 (PCV2) is considered as the causative agent of postweaning multisystemic wasting syndrome (PMWS) in domestic pigs, where the virus is ubiquitous as evidenced by serological surveys. We present the results of the first nationwide sero-survey on the presence of PCV2 antibodies in European wild boars, and report the first PMWS case in a wild boar from Spain. Sera from 656 hunter harvested wild boars from 45 different geographical sites and 22 additional imported animals were analysed by means of an immunoperoxidase monolayer assay (IPMA). We also examined the tissues from 55 healthy and one diseased wild boars for the presence of PCV2 nucleic acid and PMWS lesions by in situ hybridisation and histopathology, respectively. Additionally, abundance estimates of wild boars and field interviews were carried out on 30 sampling sites. The prevalence of medium to high PCV2 serological titres among the examined wild boars was 47.89 +/- 1.9%. Seropositive wild boars appeared in all but one of the geographical regions analysed. Seroprevalence and titre of PCV2 antibodies were closely related to the management of the wild boar populations. Wild boars from intensively managed, farm-like populations had higher prevalence than wild boars living in more natural situations. The effect of wild boar abundance and management on PCV2 antibody prevalence was further evidenced by the high correlation existing between the relative abundance estimates of animals and the percentage of wild boars with medium to high levels of PCV2 antibodies. PCV2 nucleic acid was detected in the tissues of three wild boars. One of these was diagnosed as PMWS. The results, in addition to information on piglet mortalities, suggest a potential role of PMWS in piglet mortality in intensively managed wild boar populations.


Integrative modeling defines the Nova splicing-regulatory network and its combinatorial controls.
The control of RNA alternative splicing is critical for generating biological diversity. Despite emerging genome-wide technologies to study RNA complexity, reliable and comprehensive RNA-regulatory networks have not been defined. Here, we used Bayesian networks to probabilistically model diverse data sets and predict the target networks of specific regulators. We applied this strategy to identify approximately 700 alternative splicing events directly regulated by the neuron-specific factor Nova in the mouse brain, integrating RNA-binding data, splicing microarray data, Nova-binding motifs, and evolutionary signatures. The resulting integrative network revealed combinatorial regulation by Nova and the neuronal splicing factor Fox, interplay between phosphorylation and splicing, and potential links to neurologic disease. Thus, we have developed a general approach to understanding mammalian RNA regulation at the systems level.


Minimum data set for mass-gathering health research and evaluation: a discussion paper.
This paper discusses the need for consistency in mass-gathering data collection and biomedical reporting. Mass gatherings occur frequently throughout the world, and having an understanding of the complexities of mass gatherings is important to inform health services about the possible required health resources. Factors within the environmental, psychosocial and biomedical domains influence the usage of health services at mass gatherings. The biomedical domain includes the categorization of presenting injury or illness, and rates such as patient presentation rate, transferred to hospital rate and referred to hospital rate. These rates provide insight into the usage of onsite health services, prehospital ambulance services. and hospital emergency department services. Within the literature, these rates are reported in a manner that is varied, haphazard and author dependent. This paper proposes moving away from an author-dependent practice of collection and reporting of data. An expert consensus approach is proposed as a means of further developing mass-gathering theory and moving beyond the current situation of reporting on individual case studies. To achieve this, a minimum data set with a data dictionary is proposed in an effort to generate conversation about a possible agreed minimum amount and type of information that should be collected consistently for research and evaluation at mass gatherings. Finally, this paper outlines future opportunities that will emerge from the consistent collection and reporting of mass-gathering data, including the possibility for meta-analysis, comparison of events across societies and modeling of various rates to inform health services.


Isolated left ventricular non-compaction as a cause of thrombo-embolic stroke: a case report and review.
Isolated left ventricular non-compaction is the result of incomplete myocardial morphogenesis, leading to persistence of the embryonic myocardium. The condition is recognised by an excessively prominent trabecular meshwork and deep intertrabecular recesses of the left ventricle. These intertrabecular recesses are prone to thrombus formation, with resultant embolic sequelae. We describe a case of cerebral thrombo-embolism in a young woman due to isolated left ventricular non-compaction.


Plasticity of thermal tolerance and metabolism but not water loss in an invasive reed frog.
Phenotypic plasticity may buffer the selection pressures on organisms that inhabit novel or rapidly-changing environments. We investigated plasticity of thermal tolerance, energetic and water loss traits and their interaction with behaviour in a small-bodied, arboreal anuran (Hyperolius marmoratus Rapp, Hyperoliidae) undergoing rapid range expansion into the winter rainfall region of South Africa. After short-term exposure to three temperatures (acclimation treatments) commonly encountered in their historical and novel ranges, frogs exhibited a broad thermal tolerance range (mean±s.d.: 42.1±2.9 °C) and higher plasticity in CTmax than in CTmin. Resting metabolic rate was lowest in cold-acclimated animals, while active metabolic rates were lowest in warm-acclimated frogs, likely reflecting compensation towards energy conservation. Evaporative water loss was not significantly altered by the acclimation treatments in either resting or active animals, indicating limited plasticity in this trait compared to metabolism. Our results suggest that plasticity of temperature limits and metabolism may benefit this species in variable environments such as those encountered in its expanded range. Lack of plasticity in water loss during resting and activity suggests that these frogs rely on their high cutaneous resistance and behavioural means to buffer climate variation. This study highlights the importance of synergistic interactions between physiology and behaviour in determining amphibian responses to temperature variation.


Vaccination against IL-33 Inhibits Airway Hyperresponsiveness and Inflammation in a House Dust Mite Model of Asthma.
In several clinical and experimental studies IL-33 and its receptor have been found to play important roles in the development of asthma and allergic airway inflammation. We evaluated the effects of vaccination against IL-33 in a mouse model of airway inflammation induced by house dust mite (HDM) allergen. Balb/c mice received the IL-33 vaccine subcutaneously, followed by intranasal administration of HDM for up to six weeks. Vaccination against IL-33 induced high titers of specific anti-IL-33 IgG antibodies that inhibited HDM-induced airway hyperresponsiveness (AHR) in the conducting airways and tissue damping. The vaccination also attenuated the HDM-induced elevation in the numbers of eosinophils in bronchoalveolar lavage fluid (BALF) and suppressed the accumulation of inflammatory cells in the airways. Furthermore, the levels of IL-17A, IL-25, IL-33 and TSLP in lung tissue homogenates were reduced by vaccination against IL-33. These observations demonstrate that vaccination against IL-33 inhibits HDM-induced development of AHR, airway inflammation and production of inflammatory cytokines. The results also indicate an important role of IL-33 in the regulation of AHR of the distal lung compartments. Thus, administration of such a vaccine is potentially an effective therapeutic tool for treating allergic asthma.