Determination of the topology of the hydrophobic segment of mammalian diacylglycerol kinase epsilon in a cell membrane and its relationship to predictions from modeling.
The epsilon isoform of diacylglycerol kinase (DGKepsilon) is unique among mammalian DGKs in having a segment of hydrophobic amino acids comprising approximately residues 20 to 41. Several algorithms predict this segment to be a transmembrane (TM) helix. Using PepLook, we have performed an in silico analysis of the conformational preference of the segment in a hydrophobic environment comprising residues 18 to 42 of DGKepsilon. We find that there are two distinct groups of stable conformations, one corresponding to a straight helix that would traverse the membrane and the second corresponding to a bent helix that would enter and leave the same side of the membrane. Furthermore, the calculations predict that substituting the Pro32 residue in the hydrophobic segment with an Ala will cause the hydrophobic segment to favor a TM orientation. We have expressed the P32A mutant of DGKepsilon, with a FLAG tag (an N-terminal 3xFLAG epitope tag) at the amino terminus, in COS-7 cells. We find that this mutation causes a large reduction in both k(cat) and K(m) while maintaining k(cat)/K(m) constant. Specificity of the P32A mutant for substrates with polyunsaturated acyl chains is retained. The P32A mutant also has higher affinity for membranes since it is more difficult to extract from the membrane with high salt concentration or high pH compared with the wild-type DGKepsilon. We also evaluated the topology of the proteins with confocal immunofluorescence microscopy using NIH 3T3 cells. We find that the FLAG tag at the amino terminus of the wild-type enzyme is not reactive with antibodies unless the cell membrane is permeabilized with detergent. We also demonstrate that at least a fraction of the wild-type DGKepsilon is present in the plasma membrane and that comparable amounts of the wild-type and P32A mutant proteins are in the plasma membrane fraction. This indicates that in these cells the hydrophobic segment of the wild-type DGKepsilon is not TM but takes up a bent conformation. In contrast, the FLAG tag at the amino terminus of the P32A mutant is exposed to antibody both before and after membrane permeabilization. This modeling approach thus provides an explanation, not provided by simple predictive algorithms, for the observed topology of this protein in cell membranes. The work also demonstrates that the wild-type DGKepsilon is a monotopic protein.


[Contraceptive measures and HIV transmission protection among women with HIV/AIDS].
Sexual intercourse is currently the route of transmission among women that has most contributed to the feminization of the HIV/AIDS epidemic. As an ongoing effort to establish more appropriate standards for health counseling, the study's purpose was to investigate the use of contraceptive methods that would also prevent HIV/AIDS women against disease transmission. An exploratory study was developed in an outpatient clinic of a public university hospital, a reference center of HIV/AIDS patients in the mid-south region of the state of São Paulo, Brazil, during a 5-month-period (2000 and 2001). The study was carried out in 73 HIV/AIDS women. Data were collected using a semi-structured questionnaire exploring subjects' sociodemographics, contraception method used and HIV status of their sex partners. A descriptive data analysis was performed and the contents of open answers were grouped into themes. Fischer's exact test was applied for analyzing some variables at a 5% significance level. Content analysis was carried out according to Bardin's proposal.(2) Most women at reproductive age were married and had been infected almost exclusively through heterosexual contact. Of them, 35.4% reported having an HIV discordant partner and 13.7% used inadequate contraceptive methods that failed to protect them against HIV transmission. The study results call for the need of continuous education on safer sex among HIV/AIDS women to empower them to discuss with their partners alternative options of exercising their sexuality and to raise awareness on their contraceptive choices in a way to protect their own health, their partner's and even their unborn offspring's health.


A Natural Population Derived from Species Hybridizationin the Drosophila ananassae Species Complexon Penang Island, Malaysia.
We surveyed natural population of the Drosophila ananassae species complex on Penang Island, Malaysia. Analyses of phenotypic traits, chromosome arrangements, molecular markers, and reproductive isolation suggest the existence of two species: D. ananassae and D. cf. parapallidosa. Molecular marker analysis indicates that D. cf. parapallidosa carries chromosome Y and 4 introgressions from D. ananassae. Thus, D. cf. parapallidosa seems to be a hybrid descendant that recently originated from a natural D. parapallidosa♀× D. ananassae♂ cross. Furthermore, D. cf. parapallidosa behaves differently from authentic D. parapallidosa with respect to its reproductive isolation from D. ananassae. Premating isolation is usually seen in only the D. ananassae♀× D. parapallidosa♂ cross, but we observed it in crosses of both directions between D. ananassae and D. cf. parapallidosa. In addition, hybrid males from the D. ananassae♀× D. parapallidosa♂ cross are usually sterile, but they were fertile when D. ananassae♀ were mated with D. cf. parapallidosa ♂. We attempted an artificial reconstruction of the hybrid species to simulate the evolutionary process(es) that produced D. cf. parapallidosa. This is a rare case of natural hybrid population in Drosophila and may be a useful system for elucidating speciation with gene flow.


Origin and Evolution of Flavin-Based Electron Bifurcating Enzymes.
Twelve evolutionarily unrelated oxidoreductases form enzyme complexes that catalyze the simultaneous coupling of exergonic and endergonic oxidation-reduction reactions to circumvent thermodynamic barriers and minimize free energy loss in a process known as flavin-based electron bifurcation. Common to these 12 bifurcating (Bf) enzymes are protein-bound flavin, the proposed site of bifurcation, and the electron carrier ferredoxin. Despite the documented role of Bf enzymes in balancing the redox state of intracellular electron carriers and in improving the efficiency of cellular metabolism, a comprehensive description of the diversity and evolutionary history of Bf enzymes is lacking. Here, we report the taxonomic distribution, functional diversity, and evolutionary history of Bf enzyme homologs in 4,588 archaeal, bacterial, and eukaryal genomes and 3,136 community metagenomes. Bf homologs were primarily detected in the genomes of anaerobes, including those of sulfate-reducers, acetogens, fermenters, and methanogens. Phylogenetic analyses of Bf enzyme catalytic subunits (oxidoreductases) suggest they were not a property of the Last Universal Common Ancestor of Archaea and Bacteria, which is consistent with the limited and unique taxonomic distributions of enzyme homologs among genomes. Further, phylogenetic analyses of oxidoreductase subunits reveal that non-Bf homologs predate Bf homologs. These observations indicate that multiple independent recruitments of flavoproteins to existing oxidoreductases enabled coupling of numerous new electron Bf reactions. Consistent with the role of these enzymes in the energy metabolism of anaerobes, homologs of Bf enzymes were enriched in metagenomes from subsurface environments relative to those from surface environments. Phylogenetic analyses of homologs from metagenomes reveal that the earliest evolving homologs of most Bf enzymes are from subsurface environments, including fluids from subsurface rock fractures and hydrothermal systems. Collectively, these data suggest strong selective pressures drove the emergence of Bf enzyme complexes via recruitment of flavoproteins that allowed for an increase in the efficiency of cellular metabolism and improvement in energy capture in anaerobes inhabiting a variety of subsurface anoxic habitats where the energy yield of oxidation-reduction reactions is generally low.


Regulatory role of the N terminus of the vacuolar calcium-ATPase in cauliflower.
The vacuolar calmodulin (CaM)-stimulated Ca(2+)-ATPase, BCA1p, in cauliflower (Brassica oleracea) has an extended N terminus, which was suggested to contain a CaM-binding domain (S. Malmström, P. Askerlund, M.G. Palmgren [1997] FEBS Lett 400: 324-328). The goal of the present study was to determine the role of the N terminus in regulating BCA1p. Western analysis using three different antisera showed that the N terminus of BCA1p is cleaved off by trypsin and that the N terminus contains the CaM-binding domain. Furthermore, the expressed N terminus binds CaM in a Ca(2+)-dependent manner. A synthetic peptide corresponding to the CaM-binding domain of BCA1p (Ala-19 to Leu-43) strongly inhibited ATP-dependent Ca(2+) pumping by BCA1p in cauliflower low-density membranes, indicating that the CaM-binding region of BCA1p also has an autoinhibitory function. The expressed N terminus of BCA1p and a synthetic peptide (Ala-19 to Met-39) were good substrates for phosphorylation by protein kinase C. Sequencing of the phosphorylated fusion protein and peptide suggested serine-16 and/or serine-28 as likely targets for phosphorylation. Phosphorylation of serine-28 had no effect on CaM binding to the alanine-19 to methionine-39 peptide. Our results demonstrate the regulatory importance of the N terminus of BCA1p as a target for CaM binding, trypsin cleavage, and phosphorylation, as well as its importance as an autoinhibitory domain.


An expanded palette of dopamine sensors for multiplex imaging in vivo.
Genetically encoded dopamine sensors based on green fluorescent protein (GFP) enable high-resolution imaging of dopamine dynamics in behaving animals. However, these GFP-based variants cannot be readily combined with commonly used optical sensors and actuators, due to spectral overlap. We therefore engineered red-shifted variants of dopamine sensors called RdLight1, based on mApple. RdLight1 can be combined with GFP-based sensors with minimal interference and shows high photostability, permitting prolonged continuous imaging. We demonstrate the utility of RdLight1 for receptor-specific pharmacological analysis in cell culture, simultaneous assessment of dopamine release and cell-type-specific neuronal activity and simultaneous subsecond monitoring of multiple neurotransmitters in freely behaving rats. Dual-color photometry revealed that dopamine release in the nucleus accumbens evoked by reward-predictive cues is accompanied by a rapid suppression of glutamate release. By enabling multiplexed imaging of dopamine with other circuit components in vivo, RdLight1 opens avenues for understanding many aspects of dopamine biology.


MECHANISMS IN ENDOCRINOLOGY: Medical consequences of doping with anabolic androgenic steroids: effects on reproductive functions.
Anabolic androgenic steroids (AASs) are appearance and performance-enhancing drugs (APEDs) used in competitive athletics, in recreational sports, and by body-builders. The global lifetime prevalence of AASs abuse is 6.4% for males and 1.6% for women. Many AASs, often obtained from the internet and dubious sources, have not undergone proper testing and are consumed at extremely high doses and in irrational combinations, also along with other drugs. Controlled clinical trials investigating undesired side effects are lacking because ethical restrictions prevent exposing volunteers to potentially toxic regimens, obscuring a causal relationship between AASs abuse and possible sequelae. Because of the negative feedback in the regulation of the hypothalamic-pituitary-gonadal axis, in men AASs cause reversible suppression of spermatogenesis, testicular atrophy, infertility, and erectile dysfunction (anabolic steroid-induced hypogonadism). Should spermatogenesis not recover after AASs abuse, a pre-existing fertility disorder may have resurfaced. AASs frequently cause gynecomastia and acne. In women, AASs may disrupt ovarian function. Chronic strenuous physical activity leads to menstrual irregularities and, in severe cases, to the female athlete triad (low energy intake, menstrual disorders and low bone mass), making it difficult to disentangle the effects of sports and AASs. Acne, hirsutism and (irreversible) deepening of the voice are further consequences of AASs misuse. There is no evidence that AASs cause breast carcinoma. Detecting AASs misuse through the control network of the World Anti-Doping Agency (WADA) not only aims to guarantee fair conditions for athletes, but also to protect them from medical sequelae of AASs abuse.


Stress increases putative gonadotropin inhibitory hormone and decreases luteinizing hormone in male rats.
The subjective experience of stress leads to reproductive dysfunction in many species, including rodents and humans. Stress effects on reproduction result from multilevel interactions between the hormonal stress response system, i.e., the hypothalamic-pituitary-adrenal (HPA) axis, and the hormonal reproductive system, i.e., the hypothalamic-pituitary-gonadal (HPG) axis. A novel negative regulator of the HPG axis known as gonadotropin-inhibitory hormone (GnIH) was recently discovered in quail, and orthologous neuropeptides known as RFamide-related peptides (RFRPs) have also been identified in rodents and primates. It is currently unknown, however, whether GnIH/RFRPs influence HPG axis activity in response to stress. We show here that both acute and chronic immobilization stress lead to an up-regulation of RFRP expression in the dorsomedial hypothalamus (DMH) of adult male rats and that this increase in RFRP is associated with inhibition of downstream HPG activity. We also show that adrenalectomy blocks the stress-induced increase in RFRP expression. Immunohistochemistry revealed that 53% of RFRP cells express receptors for glucocorticoids (GCs), indicating that adrenal GCs can mediate the stress effect through direct action on RFRP cells. It is thought that stress effects on central control of reproduction are largely mediated by direct or indirect effects on GnRH-secreting neurons. Our data show that stress-induced increases in adrenal GCs cause an increase in RFRP that contributes to hypothalamic suppression of reproductive function. This novel insight into HPA-HPG interaction provides a paradigm shift for work on stress-related reproductive dysfunction and infertility, and indicates that future work on stress and reproductive system interactions must include investigation of the role of GnIH/RFRP.


Interpreting temporal fluctuations in resting-state functional connectivity MRI.
Resting-state functional connectivity is a powerful tool for studying human functional brain networks. Temporal fluctuations in functional connectivity, i.e., dynamic functional connectivity (dFC), are thought to reflect dynamic changes in brain organization and non-stationary switching of discrete brain states. However, recent studies have suggested that dFC might be attributed to sampling variability of static FC. Despite this controversy, a detailed exposition of stationarity and statistical testing of dFC is lacking in the literature. This article seeks an in-depth exploration of these statistical issues at a level appealing to both neuroscientists and statisticians. We first review the statistical notion of stationarity, emphasizing its reliance on ensemble statistics. In contrast, all FC measures depend on sample statistics. An important consequence is that the space of stationary signals is much broader than expected, e.g., encompassing hidden markov models (HMM) widely used to extract discrete brain states. In other words, stationarity does not imply the absence of brain states. We then expound the assumptions underlying the statistical testing of dFC. It turns out that the two popular frameworks - phase randomization (PR) and autoregressive randomization (ARR) - generate stationary, linear, Gaussian null data. Therefore, statistical rejection can be due to non-stationarity, nonlinearity and/or non-Gaussianity. For example, the null hypothesis can be rejected for the stationary HMM due to nonlinearity and non-Gaussianity. Finally, we show that a common form of ARR (bivariate ARR) is susceptible to false positives compared with PR and an adapted version of ARR (multivariate ARR). Application of PR and multivariate ARR to Human Connectome Project data suggests that the stationary, linear, Gaussian null hypothesis cannot be rejected for most participants. However, failure to reject the null hypothesis does not imply that static FC can fully explain dFC. We find that first order AR models explain temporal FC fluctuations significantly better than static FC models. Since first order AR models encode both static FC and one-lag FC, this suggests the presence of dynamical information beyond static FC. Furthermore, even in subjects where the null hypothesis was rejected, AR models explain temporal FC fluctuations significantly better than a popular HMM, suggesting the lack of discrete states (as measured by resting-state fMRI). Overall, our results suggest that AR models are not only useful as a means for generating null data, but may be a powerful tool for exploring the dynamical properties of resting-state fMRI. Finally, we discuss how apparent contradictions in the growing dFC literature might be reconciled.


Retinoid treatment of experimental allergic encephalomyelitis. IL-4 production correlates with improved disease course.
Experimental allergic encephalomyelitis (EAE) is an autoimmune disease characterized by central nervous system inflammation and demyelination. Retinoids regulate cell differentiation and growth by binding to and activating retinoic acid receptors, which seem to be nuclear transcription factors. The effect of retinoids on chronic relapsing EAE produced by the transfer of myelin basic protein (MBP)-specific lymph node cells (LNC) was studied. All-trans-retinoic acid (tRA) inhibited the proliferation of MBP-specific LNC in vitro. However, the capacity of these cells to transfer EAE was markedly reduced by concentrations of tRA that only mildly inhibited T cell proliferation. The presence of tRA during in vitro MBP-specific LNC activation resulted in a considerable increase in IL-4 mRNA, whereas mRNA for IL-2, TNF-alpha, and IFN-gamma was decreased. Increased IL-4 also was detected in culture supernatants. However, the presence of a neutralizing Ab to IL-4 (11B11) during MBP-specific LNC activation in vitro did not reverse the inhibition of encephalitogenicity caused by tRA. The administration of retinoids in vivo resulted in an improved clinical course, even when given after disease onset. These findings suggest that T cell activation in the presence of tRA results in the development of T cells of the Th2 phenotype, which, in turn, might be responsible for the decrease in the encephalitogenicity of MBP-specific T cells. The modulation by retinoids of an immune response dominated by Th1-like T cells to one in which the protective cytokines of Th2-like cells predominate may have potential relevance for human demyelinating diseases such as multiple sclerosis.


Magnetic nanoparticle hyperthermia for treating locally advanced unresectable and borderline resectable pancreatic cancers: the role of tumor size and eddy-current heating.
Tumor volume largely determines the success of local control of borderline resectable and locally advanced pancreatic cancer with current therapy. We hypothesized that a tumor-mass normalized dose of magnetic nanoparticle hyperthermia (MNPH) with alternating magnetic fields (AMFs) reduces the effect of tumor volume for treatment. 18 female athymic nude mice bearing subcutaneous MiaPaCa02 human xenograft tumors were treated with MNPH following intratumor injections of 5.5 mg Fe/g tumor of an aqueous suspension of magnetic iron-oxide nanoparticles. Mice were randomly divided into control (n = 5) and treated groups having small (0.15 ± 0.03 cm3, n = 4) or large (0.30 ± 0.06 cm3, n = 5) tumors. We assessed the clinical feasibility of this approach and of pulsed AMF to minimize eddy current heating using a finite-element method to solve a bioheat equation for a human-scale multilayer model. Compared to the control group, both small and large MiaPaCa02 subcutaneous tumors showed statistically significant growth inhibition. Conversely, there was no significant difference in tumor growth between large and small tumors. Both computational and xenograft models demonstrated higher maximum tumor temperatures for large tumors compared to small tumors. Computational modeling demonstrates that pulsed AMF can minimize nonspecific eddy current heating. MNPH provides an advantage to treat large tumors because the MION dose can be adjusted to increase power. Pulsed AMF, with adjusted treatment time, can enhance MNPH in challenging cases such as low MION dose in the target tissue and/or large patients by minimizing nonspecific eddy current heating without sacrificing thermal dose to the target. Nanoparticle heterogeneity in tumors remains a challenge for continued research.


Effects of diphenyl diselenide on methylmercury toxicity in rats.
This study investigates the efficacy of diphenyl diselenide [(PhSe)2] in attenuating methylmercury- (MeHg-)induced toxicity in rats. Adult rats were treated with MeHg [5 mg/kg/day, intragastrically (i.g.)] and/ or (PhSe)2 [1 mg/kg/day, intraperitoneally (i.p.)] for 21 days. Body weight gain and motor deficits were evaluated prior to treatment, on treatment days 11 and 21. In addition, hepatic and cerebral mitochondrial function (reactive oxygen species (ROS) formation, total and nonprotein thiol levels, membrane potential (ΔΨm), metabolic function, and swelling), hepatic, cerebral, and muscular mercury levels, and hepatic, cerebral, and renal thioredoxin reductase (TrxR) activity were evaluated. MeHg caused hepatic and cerebral mitochondrial dysfunction and inhibited TrxR activity in liver (38,9%), brain (64,3%), and kidney (73,8%). Cotreatment with (PhSe)2 protected hepatic and cerebral mitochondrial thiols from depletion by MeHg but failed to completely reverse MeHg's effect on hepatic and cerebral mitochondrial dysfunction or hepatic, cerebral, and renal inhibition of TrxR activity. Additionally, the cotreatment with (PhSe)2 increased Hg accumulation in the liver (50,5%) and brain (49,4%) and increased the MeHg-induced motor deficits and body-weight loss. In conclusion, these results indicate that (PhSe)2 can increase Hg body burden as well as the neurotoxic effects induced by MeHg exposure in rats.


Efficacy and Safety of Alogliptin in Patients With Type 2 Diabetes: Analysis of the ATTAK-J Study.
Dipeptidyl peptidase-4 (DPP-4) inhibitors have been shown to reduce hemoglobin A1c (HbA1c) in patients with type 2 diabetes, but the reduction varies between patients and adequate glycemic control may not be achieved. We evaluated the efficacy and safety of the DPP-4 inhibitor alogliptin in the real clinical setting, and analyzed factors associated with the improvement of HbA1c by alogliptin treatment. A retrospective observational study was performed in patients with type 2 diabetes attending hospitals or clinics belonging to the Kanagawa Physicians Association who received treatment with alogliptin for 1 year or longer. Patients using insulin were excluded from the study. The efficacy endpoints were HbA1c (National Glycohemoglobin Standardization Program value), blood glucose (fasting/postprandial), body weight, blood pressure (systolic/diastolic), liver function (glutamate oxaloacetate transaminase, glutamate pyruvate transaminase, and γ-glutamyl transpeptidase), kidney function (serum creatinine and estimated glomerular filtration rate), serum lipids (total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, and triglycerides), and serum amylase. Adverse events were compiled to assess safety. Of 330 patients whose case records were collected, 27 patients were excluded for protocol violations, leaving 303 patients to form the full analysis set. Compared with baseline, HbA1c showed a decrease by 0.54±1.22% (mean ± standard deviation) after 12 months of alogliptin treatment. Factor analysis demonstrated that the change of HbA1c after 12 months was significantly influenced by the baseline HbA1c level, duration of diabetes, concomitant use of sulfonylureas, and compliance with diet therapy. In addition, there was a significant reduction of total cholesterol, low-density lipoprotein cholesterol, and the estimated glomerular filtration rate after 12 months of alogliptin treatment, as well as a significant increase in serum creatinine. No significant changes of body weight, blood pressure, or liver function were observed. Symptoms of hypoglycemia occurred in two patients (0.6%). Alogliptin displayed a significant hypoglycemic effect and excellent safety in routine clinical use. Factors influencing the change of HbA1c with alogliptin therapy may include the HbA1c at the start of treatment, the duration of diabetes, use of sulfonylureas, and compliance with diet therapy.


Botsourcing and outsourcing: Robot, British, Chinese, and German workers are for thinking--not feeling--jobs.
Technological innovations have produced robots capable of jobs that, until recently, only humans could perform. The present research explores the psychology of "botsourcing"-the replacement of human jobs by robots-while examining how understanding botsourcing can inform the psychology of outsourcing-the replacement of jobs in one country by humans from other countries. We test four related hypotheses across six experiments: (1) Given people's lay theories about the capacities for cognition and emotion for robots and humans, workers will express more discomfort with botsourcing when they consider losing jobs that require emotion versus cognition; (2) people will express more comfort with botsourcing when jobs are framed as requiring cognition versus emotion; (3) people will express more comfort with botsourcing for jobs that do require emotion if robots appear to convey more emotion; and (4) people prefer to outsource cognition- versus emotion-oriented jobs to other humans who are perceived as more versus less robotic. These results have theoretical implications for understanding social cognition about both humans and nonhumans and practical implications for the increasingly botsourced and outsourced economy.


Characterisation of a new operon encoding a Zur-like protein and an associated ABC zinc permease in Listeria monocytogenes.
Metal ions uptake is mainly studied for iron, as it has often been implicated in bacterial virulence. Although Listeria monocytogenes virulence is expected to be controlled by the iron availability, little is known about such an uptake and its regulation. We describe the analysis of the first operon involved in metal ions uptake in L. monocytogenes. Its three ORFs encode respectively (1) an ABC protein, likely implicated in zinc uptake, (2) a hydrophobic membrane protein, generally associated with ABC proteins and (3) a ferric uptake regulator-like protein, that we named zinc uptake regulator, as it shows strong homologies with the zinc uptake regulator, a regulator of the zinc homeostasis in Bacillus subtilis. The expression of this operon is regulated by the zinc concentration.


Anomalies in the Fermi surface and band dispersion of quasi-one-dimensional CuO chains in the high-temperature superconductor YBa2Cu4O8.
We have investigated the electronic states in quasi-one-dimensional CuO chains by microprobe angle resolved photoemission spectroscopy. We find that the quasiparticle Fermi surface consists of six disconnected segments, consistent with recent theoretical calculations that predict the formation of narrow, elongated Fermi surface pockets for coupled CuO chains. In addition, we find a strong renormalization effect with a significant kink structure in the band dispersion. The properties of this latter effect [energy scale (∼40 meV), temperature dependence, and behavior with Zn-doping] are identical to those of the bosonic mode observed in CuO2 planes of high-temperature superconductors, indicating they have a common origin.


Characterization of IL-17A-producing cells in celiac disease mucosa.
Celiac disease (CD) is a gluten-sensitive enteropathy associated with a marked infiltration of the mucosa with IFN-gamma-secreting Th1 cells. Recent studies have shown that a novel subset of T cells characterized by expression of high levels of IL-17A, termed Th17 cells, may be responsible for pathogenic effects previously attributed to Th1 cells. In this study, we characterized the expression of IL-17A-producing cells in CD. By real-time PCR and ELISA, it was shown that expression of IL-17A RNA and protein is more pronounced in active CD biopsy specimens in comparison with inactive CD and normal mucosal biopsy specimens. Flow cytometry confirmed that IL-17A is overproduced in CD mucosa and that CD4(+) and CD4(+)CD8(+) cells were major sources. The majority of IL-17A-producing CD4(+) and CD4(+)CD8(+) cells coexpressed IFN-gamma but not CD161. The addition of a peptic-tryptic digest of gliadin to ex vivo organ cultures of duodenal biopsy specimens taken from inactive CD patients enhanced IL-17A production by both CD4(+) and CD4(+)CD8(+) cells. Because we previously showed that IL-21, a T cell-derived cytokine involved in the control of Th17 cell responses, is overproduced in CD, we next assessed whether IL-17A expression is regulated by IL-21. Blockade of IL-21 activity by a neutralizing IL-21 Ab reduced IL-17A expression in cultures of active CD and peptic-tryptic digest of gliadin-treated CD biopsy specimens. In conclusion, our data show that IL-17A is increased in CD and is produced by cells that also make IFN-gamma.


Low Dose Ketamine Infusion for Comorbid Posttraumatic Stress Disorder and Chronic Pain: A Randomized Double-Blind Clinical Trial.
To date, treatment options (i.e. psychotherapy, antidepressant medications) for patients with posttraumatic stress disorder (PTSD), are relatively few, and considering their limited efficacy, novel therapies have gained interest among researchers and treatment providers alike. Among patients with chronic pain (CP) about one third experience comorbid PTSD, which further complicates their already challenging pharmacological regimens. Low dose ketamine infusion has shown promise in PTSD, and in treatment of CP, however they have not been studied in comorbid population and under rigorous control conditions. We compared the effects of a single dose of either ketamine (0.5 mg/kg) or ketorolac (15 mg) over a 40-minute of IV infusion in CP patients with and without PTSD, in double blind, randomized study. Measures were collected before, during, one day and seven days after the infusion. A planned sample size of 40 patients randomly assigned to treatment order was estimated to provide 80% power to detect a hypothesized treatment difference after the infusion.Main Outcome and Measures: The primary outcome measures were change in PTSD symptom severity assessed with the Impact of Event Scale-Revised (IES-R) and Visual Analogue Scale (VAS) for pain administered by a study clinician 24 hours post infusion. Secondary outcome measures included Impact of Event Scale-Revised (IES-R), VAS and Brief Pain Inventory (Short Form) for pain 1 week after the infusion. Both treatments offered comparable improvement of PTSD and CP symptoms that persisted for 7 days after the infusion. Patients with comorbid PTSD and CP experienced less dissociative side effects compared to the CP group. Surprisingly, ketorolac infusion resulted in dissociative symptoms in CP patients only. This first prospective study comparing effects of subanesthetic ketamine versus ketorolac infusions for comorbid PTSD and CP, suggests that both ketamine and ketorolac might offer meaningful and durable response for both PTSD and CP symptoms.


A novel Acute Retroviral Syndrome Severity Score predicts the key surrogate markers for HIV-1 disease progression.
Best long-term practice in primary HIV-1 infection (PHI) remains unknown for the individual. A risk-based scoring system associated with surrogate markers of HIV-1 disease progression could be helpful to stratify patients with PHI at highest risk for HIV-1 disease progression. We prospectively enrolled 290 individuals with well-documented PHI in the Zurich Primary HIV-1 Infection Study, an open-label, non-randomized, observational, single-center study. Patients could choose to undergo early antiretroviral treatment (eART) and stop it after one year of undetectable viremia, to go on with treatment indefinitely, or to defer treatment. For each patient we calculated an a priori defined "Acute Retroviral Syndrome Severity Score" (ARSSS), consisting of clinical and basic laboratory variables, ranging from zero to ten points. We used linear regression models to assess the association between ARSSS and log baseline viral load (VL), baseline CD4+ cell count, and log viral setpoint (sVL) (i.e. VL measured ≥90 days after infection or treatment interruption). Mean ARSSS was 2.89. CD4+ cell count at baseline was negatively correlated with ARSSS (p = 0.03, n = 289), whereas HIV-RNA levels at baseline showed a strong positive correlation with ARSSS (p<0.001, n = 290). In the regression models, a 1-point increase in the score corresponded to a 0.10 log increase in baseline VL and a CD4+ cell count decline of 12/µl, respectively. In patients with PHI and not undergoing eART, higher ARSSS were significantly associated with higher sVL (p = 0.029, n = 64). In contrast, in patients undergoing eART with subsequent structured treatment interruption, no correlation was found between sVL and ARSSS (p = 0.28, n = 40). The ARSSS is a simple clinical score that correlates with the best-validated surrogate markers of HIV-1 disease progression. In regions where ART is not universally available and eART is not standard this score may help identifying patients who will profit the most from early antiretroviral therapy.


Ambulatory pediatric oncology CLABSIs: epidemiology and risk factors.
To compare the burden of central line-associated bloodstream infections (CLABSIs) in ambulatory versus inpatient pediatric oncology patients, and identify the epidemiology of and risk factors associated with ambulatory CLABSIs. We prospectively identified infections and retrospectively identified central line days and characteristics associated with CLABSIs from January 2009 to October 2010. A nested case-control design was used to identify characteristics associated with ambulatory CLABSIs. We identified 319 patients with central lines. There were 55 ambulatory CLABSIs during 84,705 ambulatory central line days (0.65 CLABSIs per 1,000 central line days (95% CI 0.49, 0.85)), and 19 inpatient CLABSIs during 8,682 inpatient central line days (2.2 CLABSIs per 1,000 central lines days (95% CI 1.3, 3.4)). In patients with ambulatory CLABSIs, 13% were admitted to an intensive care unit and 44% had their central lines removed due to the CLABSI. A secondary analysis with a sub-cohort, suggested children with tunneled, externalized catheters had a greater risk of ambulatory CLABSI than those with totally implantable devices (IRR 20.6, P < 0.001). Other characteristics independently associated with ambulatory CLABSIs included bone marrow transplantation within 100 days (OR 16, 95% CI 1.1, 264), previous bacteremia in any central line (OR 10, 95% CI 2.5, 43) and less than 1 month from central line insertion (OR 4.2, 95% CI 1.0, 17). In pediatric oncology patients, three times more CLABSIs occur in the ambulatory than inpatient setting. Ambulatory CLABSIs carry appreciable morbidity and have identifiable, associated factors that should be addressed in future ambulatory CLABSI prevention efforts.


Increased temperature of malignant urinary bladder tumors in vivo: the application of a new method based on a catheter technique.
The aim of this study was to investigate the existence of any thermal difference between malignant tumors and inflammatory benign lesions of the human urinary bladder and to determine whether it correlates with tumor angiogenesis quantification. A new method, developed in our institute, is introduced to detect temperature in human urinary bladder, in vivo. This method is based on a thermography catheter. We calculated the differences of the temperature of the solid tumor and of a normal area (Delta T) on 20 subjects (mean age, 72.5 years; 95% confidence interval [CI], 68.5 to 76.4). According to the biopsy histology, Eight (40%) patients had benign tumors, and 12 (60%) had malignant tumors. We found significant differences of Delta T between patients with benign and malignant tumor (P <.001). Also, differences were found for the mean values of angiogenesis level between malignant and benign tumors (P =.0261), and a moderated positive correlation was estimated between the degree of angiogenesis and Delta T (P =.02). Based on logistic regression analysis, we found that a 1-degree increase of Delta T triples the odds of a patient having a malignant tumor (odds ratio = 2.91; 95% CI, 1.97 to 7.78; P <.001), adjusted for the degree of angiogenesis (P =.0236) and the grade of tumor (P <.001). A threshold point of Delta T = 0.7 degrees C was determined, with sensitivity 83% and specificity 75%. These findings suggest that the calculated difference of temperature between normal tissue and neoplastic area could be a useful criterion in the diagnosis of malignancy in tumors of the human urinary bladder.


Post-traumatic amnesia predicts intelligence impairment following traumatic brain injury: a meta-analysis.
Worldwide, millions of patients with traumatic brain injury (TBI) suffer from persistent and disabling intelligence impairment. Post-traumatic amnesia (PTA) duration is a promising predictor of intelligence following TBI. To determine (1) the impact of TBI on intelligence throughout the lifespan and (2) the predictive value of PTA duration for intelligence impairment, using meta-analytic methods. Electronic databases were searched for peer reviewed articles, published until February 2012. Studies reporting intelligence following TBI and injury severity by PTA duration were included. Meta-analytic methods generated effect sizes for full scale IQ (FSIQ), performance IQ (PIQ) and verbal IQ (VIQ), following mild TBI (PTA duration 1-24 h) and severe TBI (PTA duration >7 days), during the subacute phase of recovery (≤6 months post-injury) and the chronic phase (>6 months post-injury). Meta-regression elucidated the predictive value of PTA duration for intelligence impairment. Patients with severe TBI exhibited large depressions in FSIQ in the subacute phase of recovery (d = -1.07, 95% CI to 1.52 to -0.62; p<0.001), persisting into the chronic phase (d = -0.78, 95% CI -1.06 to -0.51; p<0.001). PIQ was more severely affected than VIQ in the subacute phase (Q1 =3.85; p<0.05) but not in the chronic phase (Q1 =0.03, p=0.87). Most importantly, longer PTA duration strongly predicted greater depressions of FSIQ and PIQ in the subacute phase (-0.76 ≤ βs ≤ -0.73, Ps<0.01) and FSIQ, PIQ and VIQ in the chronic phase (-0.80 ≤ βs ≤ -0.61, Ps<0.05). PTA duration is a valuable predictor of intelligence impairment following TBI. Results support the routine assessment of PTA duration in clinical settings.


Inheritance of chloroplast DNA in Chlamydomonas reinhardtii.
Two symmetrically located deletions of approximately 100 base pairs each have been identified in chloroplast DNA of Chlamydomonas reinhardtii. Although present in a mutant strain that requires acetate for growth, both deletions have been shown to be distinct from the nonphotosynthetic phenotype of this strain. These physical markers in the chloroplast genome and maternally inherited genetic markers showed strict cotransmission in reciprocal crosses. Thus, our results are consistent with the location of the well-characterized maternally inherited genetic markers in chloroplast DNA of C. reinhardtii.


Improving access to health care amongst vulnerable populations: a qualitative study of village malaria workers in Kampot, Cambodia.
There is growing interest in the expansion of community health workers programmes in low- and middle-income countries as a cost-effective approach to address shortages of health professionals. However, our understanding of the reception of large-scale programmes and how to improve them remains limited, with knowledge gaps about factors that may promote or discourage equitable access to services. This paper examines the case of the Village Malaria Workers (VMW) programme in Cambodia, an extensive community-based intervention for the management of malaria cases in remote rural areas. Fieldwork was conducted in Kampot province, in six case villages characterised by different programme configuration, population size, and distance to the nearest public health facility. In these locations, in-depth interviews (n = 71) with VMWs, village authorities, and residents were conducted to identify facilitators and challenges to service utilisation. Data analysis was informed by a conceptual framework based on five domains of access to services: awareness, accessibility, accommodation, availability, and acceptability. Factors that influenced the utilisation of VMW services in our research sites include: the nature of dissemination activities and their ability to reach different population groups; the village topography and the changing road infrastructure; the involvement of VMWs in other community roles and activities; perceptions about the type of disease after the onset of symptoms; the need for comprehensive diagnosis and care; perceptions about the status of VMWs as medical providers; length of VMW appointment. This study highlights the complexity and diversity of contextual factors that may influence the uptake of a community health programme. As in other countries, continued use of lay health workers in Cambodia to deliver diagnostic and curative services has the potential for great health and economic impact. However, further consideration should be given to the problem of access in different categories of residents and different contexts of implementation. In addition, a comprehensive mapping of changes in disease epidemiology, road infrastructure and the geography of access to services is crucial to inform policy development in this area.


Volatile metabolites produced by six fungal species compared with other indicators of fungal growth on cereal grains.
Six fungal species, Penicillium brevicompactum, P. glabrum, P. roqueforti, Aspergillus flavus, A. versicolor, and A. candidus, were inoculated on moistened and autoclaved wheat and oat grains. They were cultivated in glass vessels provided with an inlet and outlet for air. Air was passed through the vessels to collect volatile fungal metabolites on porous polymer adsorbents attached to the outlet. Samples were collected at two fungal growth stages. Adsorbed compounds were thermally desorbed, separated by gas chromatography, and identified by mass spectrometry. Differences in the production of volatile metabolites depended more on the fungal species than on the grain type. The fungal growth stage was not an important factor determining the composition of volatiles produced. 3-Methylfuran was produced in similar amounts regardless of the fungal species and substrate (oat versus wheat). The production of volatile metabolites was compared with the production of ergosterol and CO2 and the number of CFU. The production of volatile metabolites was more strongly correlated with accumulated CO2 production than with actual CO2 production and more strongly correlated with ergosterol contents of the grain than with numbers of CFU.


Oncostatin M in bronchoalveolar lavage correlates with the severity of sarcoidosis.
Recent studies have shown that oncostatin M (OSM) might have a role in T cell-mediated inflammatory processes in which mast cells are also involved. Patients with severe sarcoidosis might develop fibrotic changes in the lung. We assessed whether there was a correlation between mast cells expressing OSM in bronchoalveolar lavage (BAL) and the severity of sarcoidosis. Twelve consecutive patients with new diagnosis of sarcoidosis were eligible for the study. All underwent complete lung function tests, angiotensin converting enzyme (ACE), and bronchoscopy that included BAL and biopsies. Cytospins of BAL were prepared. All samples were incubated with the primary antibody rabbit anti-human c-kit, CD117 and stained for total mast cell count. The mouse anti-human Oncostatin M was applied and activated mast cells were counted. Clinical sarcoidosis parameters including ACE and lung functions were correlated with mast cells in BAL, as well as with OSM positive mast cells. FEV1 % was correlated with the percentage of activated mast cells, as well as with the percentage of OSM positive mast cells (r=0.61, p=0.033, 95% CI: 0.06-0.87; r=0.58, p=0.04, 95% CI: 0.015-0.86, respectively). FVC and FEV1/FVC correlated with activated mast cells (r=0.58, p=0.05; r=0.63, p=0.028, respectively). Direct correlation was found between clinical parameters including lung function tests (FEV1 and FVC) and OSM secretion from mast cells in patients with sarcoidosis. These findings suggest that mast cells and OSM have a role in sarcoidosis. Further studies to confirm these preliminary results are suggested.


Smith-Waterman peak alignment for comprehensive two-dimensional gas chromatography-mass spectrometry.
Comprehensive two-dimensional gas chromatography coupled with mass spectrometry (GC × GC-MS) is a powerful technique which has gained increasing attention over the last two decades. The GC × GC-MS provides much increased separation capacity, chemical selectivity and sensitivity for complex sample analysis and brings more accurate information about compound retention times and mass spectra. Despite these advantages, the retention times of the resolved peaks on the two-dimensional gas chromatographic columns are always shifted due to experimental variations, introducing difficulty in the data processing for metabolomics analysis. Therefore, the retention time variation must be adjusted in order to compare multiple metabolic profiles obtained from different conditions. We developed novel peak alignment algorithms for both homogeneous (acquired under the identical experimental conditions) and heterogeneous (acquired under the different experimental conditions) GC × GC-MS data using modified Smith-Waterman local alignment algorithms along with mass spectral similarity. Compared with literature reported algorithms, the proposed algorithms eliminated the detection of landmark peaks and the usage of retention time transformation. Furthermore, an automated peak alignment software package was established by implementing a likelihood function for optimal peak alignment. The proposed Smith-Waterman local alignment-based algorithms are capable of aligning both the homogeneous and heterogeneous data of multiple GC × GC-MS experiments without the transformation of retention times and the selection of landmark peaks. An optimal version of the SW-based algorithms was also established based on the associated likelihood function for the automatic peak alignment. The proposed alignment algorithms outperform the literature reported alignment method by analyzing the experiment data of a mixture of compound standards and a metabolite extract of mouse plasma with spiked-in compound standards.


Mandibular Chondrosarcoma Reconstructed Using Custom Implant and Radial Forearm Free Flap.
We describe the case of a 40-year-old male presenting with an enlarging left facial and temporal chondrosarcoma centered on the left mandibular coronoid process resulting in pathologic fractures of the zygomatic arch and focal erosion of the temporal bone. The patient underwent surgical resection via a combined hemicoronal and parotidectomy approach. The zygomatic arch was reconstructed with a patient-specific polyetheretherketone implant, and temporal contour was restored with a buried de-epithelialized radial forearm free flap. Final pathology demonstrated conventional central chondrosarcoma. This case demonstrates an unusual presentation of a rare head and neck chondrosarcoma that originated from the mandibular coronoid process with the unique use of a custom polyetheretherketone implant and vascularized fat and fascial reconstruction.


Biphasic Malignant Pleural Mesothelioma Masquerading as a Primary Skeletal Tumor.
Biphasic malignant pleural mesothelioma is a rare malignant tumor, usually presenting as a pleural-based mass in a patient with history of chronic asbestos exposure. We herein report a case of a 41-year-old man who presented with chest pain and had a chest computed tomography (CT) scan suggestive of a primary skeletal tumor originating from the ribs (chondrosarcoma or osteosarcoma), with no history of asbestos exposure. CT-guided core needle biopsies were diagnosed as malignant sarcomatoid mesothelioma. Surgical resection and chest wall reconstruction were performed, confirming the diagnosis and revealing a secondary histologic component (epithelioid), supporting the diagnosis of biphasic malignant mesothelioma.


Infrasonic hearing in birds: a review of audiometry and hypothesized structure-function relationships.
The perception of airborne infrasound (sounds below 20 Hz, inaudible to humans except at very high levels) has been documented in a handful of mammals and birds. While animals that produce vocalizations with infrasonic components (e.g. elephants) present conspicuous examples of potential use of infrasound in the context of communication, the extent to which airborne infrasound perception exists among terrestrial animals is unclear. Given that most infrasound in the environment arises from geophysical sources, many of which could be ecologically relevant, communication might not be the only use of infrasound by animals. Therefore, infrasound perception could be more common than currently realized. At least three bird species, each of which do not communicate using infrasound, are capable of detecting infrasound, but the associated auditory mechanisms are not well understood. Here we combine an evaluation of hearing measurements with anatomical observations to propose and evaluate hypotheses supporting avian infrasound detection. Environmental infrasound is mixed with non-acoustic pressure fluctuations that also occur at infrasonic frequencies. The ear can detect such non-acoustic pressure perturbations and therefore, distinguishing responses to infrasound from responses to non-acoustic perturbations presents a great challenge. Our review shows that infrasound could stimulate the ear through the middle ear (tympanic) route and by extratympanic routes bypassing the middle ear. While vibration velocities of the middle ear decline towards infrasonic frequencies, whole-body vibrations - which are normally much lower amplitude than that those of the middle ear in the 'audible' range (i.e. >20 Hz) - do not exhibit a similar decline and therefore may reach vibration magnitudes comparable to the middle ear at infrasonic frequencies. Low stiffness in the middle and inner ear is expected to aid infrasound transmission. In the middle ear, this could be achieved by large air cavities in the skull connected to the middle ear and low stiffness of middle ear structures; in the inner ear, the stiffness of round windows and cochlear partitions are key factors. Within the inner ear, the sizes of the helicotrema and cochlear aqueduct are expected to play important roles in shunting low-frequency vibrations away from low-frequency hair-cell sensors in the cochlea. The basilar papilla, the auditory organ in birds, responds to infrasound in some species, and in pigeons, infrasonic-sensitive neurons were traced back to the apical, abneural end of the basilar papilla. Vestibular organs and the paratympanic organ, a hair cell organ outside of the inner ear, are additional untested candidates for infrasound detection in birds. In summary, this review brings together evidence to create a hypothetical framework for infrasonic hearing mechanisms in birds and other animals.


Pancreatic acinar metaplasia in the distal oesophagus and the gastric cardia: prevalence, predictors and relation to GORD.
The nature of pancreatic acinar metaplasia (PAM) in the gastro-oesophageal junction (GOJ) remains obscure. We aimed to estimate its prevalence and investigate into its risk factors in a population-based series of first-time endoscopy patients. We investigated consecutive patients, endoscoped for the first time, representing defined catchment area populations. Biopsies were taken immediately below the GOJ and from the distal oesophagus. Endoscopy room-based cross-sectional clinical data were supplemented with exposure data from 160 population controls. Associations, expressed as odds ratios (OR), were modelled with multivariable logistic regression. A subsample of 26 patients underwent oesophageal pH monitoring. Among 644 patients (mean age 53 years, 43% men), PAM was found in 121 patients (19%), exclusively above the GOJ in 40 (6%), below GOJ in 67 (10%), and both above and below GOJ in 14 (2%). PAM exclusively above the GOJ and PAM exclusively below the GOJ were both borderline associated with age (2% increase in prevalence per year). PAM exclusively above the GOJ was significantly associated with female gender (OR 2.8, 95% CI 1.3-6.3) and presence of Helicobacter pylori immediately below the GOJ (OR 2.6, 95% CI 1.3-5.4). Out of 21 patients with Barrett's oesophagus (BO), 8 (38%) had PAM above the GOJ. The mean value for percentage time with oesophageal pH < 4.0 was 7.3% (95% CI 4.3-10.2%) among patients who had PAM above the GOJ (reference value 3.4%). Pancreatic acinar metaplasia might be an age-dependent lesion, associated with H. pylori, female gender and gastro-oesophageal reflux if located above the GOJ.


Optimizing spatial healthcare assets with Internet of Things.
Six percent of the total cost of healthcare delivery in Australia is from buying, building and maintaining physical assets. Current practice does not measure the efficient use of existing clinical spaces prior to making funding decisions for service expansion, remodeling or relocation. Healthcare service delivery can be increased through existing assets by optimizing the use of clinical space. The wait times for healthcare service consumers and capital expenditure pressures could be reduced, which would result in increased funds available for frontline services. Sensor technology has been used to study aspects of time in ambulatory outpatient clinics using Infra-red Tags or Radio Frequency Identification tags. This paper proposes the use of Internet of Things (IoT) technology to assist in the optimization of high-value clinical spaces and presents phase one of the project where a trial was held in a non-clinical location to evaluate sensor performance. In Phase two, sensors will be installed to count people across an ambulatory outpatient clinic in a live public healthcare environment to understand clinical space utilization and inform decision-makers. The data produced by the sensors on room use is processed for visualization in "dashboard" format so frontline and executive staff have evidence-based decision-making support for space optimization strategies. This paper presents the phase one trial and preliminary results that show the disparity space utilization patterns between the IoT sensed occupancy data with the current room reservation system in a non-clinical space.


The physical boundaries of public goods cooperation between surface-attached bacterial cells.
Bacteria secrete a variety of compounds important for nutrient scavenging, competition mediation and infection establishment. While there is a general consensus that secreted compounds can be shared and therefore have social consequences for the bacterial collective, we know little about the physical limits of such bacterial social interactions. Here, we address this issue by studying the sharing of iron-scavenging siderophores between surface-attached microcolonies of the bacterium Pseudomonas aeruginosa Using single-cell fluorescence microscopy, we show that siderophores, secreted by producers, quickly reach non-producers within a range of 100 µm, and significantly boost their fitness. Producers in turn respond to variation in sharing efficiency by adjusting their pyoverdine investment levels. These social effects wane with larger cell-to-cell distances and on hard surfaces. Thus, our findings reveal the boundaries of compound sharing, and show that sharing is particularly relevant between nearby yet physically separated bacteria on soft surfaces, matching realistic natural conditions such as those encountered in soft tissue infections.


Analysis of correlation between estradiol and fracture of femur neck.
Osteoporosis is a major public health challenge all over the world. Estrogen hormone was cited amongst other hormones to be an efficient hormone for the production and maintenance of bone density. This study was designed with the purpose of evaluating and analyzing the estradiol effect on fractures of femur neck in the Iranian society. This study evaluated men over 50 years of age suffering with mild trauma (falling off the same level height or lower) and with a fracture on their femur neck. Also, their serum level of estradiol was measured with an ELISA method. Using this procedure, the patients were assigned into groups with either normal estradiol serum level (10pg/ml and higher) or with lower than normal level (lower than 10 pg/ml). A control group including 50-year-old and older men without hip fracture, or its history, was chosen to access their estradiol serum level. Data collected from these two groups were statistically compared. A total of 120 patients were evaluated (60 in the control and 60 in the test group). The mean age of patients in the control and test groups were 67.9±10.22 and 69.5±8.84 years, respectively (p=0.376). Smoker patients' percentages in the control and test groups were 35% and 31.7%, respectively (p=0.699). On the basis of the serum estradiol level, patients' percentages with low estradiol level in control and test groups were 10% and 16.7%, respectively (p=0.283). The only significant factor in predicting serum estradiol level was smoking. In conclusion, in this study it was observed that fractures of the femoral neck following a mild trauma were not correlated to low level of serum estradiol.


Effect of alkali ions on optical properties of flavins: vibronic spectra of cryogenic M+lumiflavin complexes (M = Li-Cs).
Flavin compounds are frequently used by nature in photochemical processes because of their unique optical properties which can be strongly modulated by the surrounding environment such as solvation or coordination with metal ions. Herein, we employ vibronic photodissociation spectroscopy of cryogenic M+LF complexes composed of lumiflavin (LF, C13H12N4O2), the parent molecule of the flavin family, and alkali ions (M = Li-Cs) to characterize the strong impact of metalation on the electronic properties of the LF chromophore. With the aid of time-dependent density functional theory calculations (PBE0/cc-pVDZ) coupled to multidimensional Franck-Condon simulations, the visible photodissociation (VISPD) spectra of M+LF ions recorded in the 500-570 nm range are assigned to the S1 ← S0 (ππ*) transitions into the first optically bright S1 state of the lowest-energy M+LF(O4+) isomers. In this O4+ structure, M+ binds in a bent chelate to the lone pairs of both the O4 and the N5 atom of LF. Charge reorganization induced by S1 excitation strongly enhances the interaction between M+ and LF at this binding site, leading to substantial red shifts in the S1 absorption of the order of 10-20% (e.g., from 465 nm in LF to 567 nm in Li+LF). This strong change in M+LF interaction strength in M+LF(O4+) upon ππ* excitation can be rationalized by the orbitals involved in the S1 ← S0 transition and causes strong vibrational activity. In particular, progressions in the intermolecular bending and stretching modes provide an accurate measure of the strength of the M+LF bond. In contrast to the experimentally identified O4+ ions, the predicted S1 origins of other low-energy M+LF isomers, O2+ and O2, are slightly blue-shifted from the S1 of LF, demonstrating that the electronic properties of metalated LF not only drastically change with the size of the metal ion but also with its binding site.


The automatic and the expected self: separating self- and familiarity biases effects by manipulating stimulus probability.
Attentional control over prepotent responses has previously been shown by manipulating the probability with which stimuli appear. Here, we examined whether prepotent responses to self-associated stimuli can be modulated by their frequency of occurrence. Participants were instructed to associate geometric shapes with the self, their mother, or a stranger before having to judge whether the sequential shape-label pairs matched or mismatched the instruction. The probability of the different shape-label pairs was varied. There was a robust advantage to self-related stimuli in all cases. Reducing the proportion of matched self pairs did not weaken performance with self-related stimuli, whereas reducing the frequency of either matched mother or stranger pairs hurt performance, relative to when the different match trials were equiprobable. In addition, while mother and stranger pairs jointly benefitted when they both occurred frequently, there were benefits only to self pairs when the frequency of self trials increased along with either mother or stranger trials. The results suggest that biases favoring self-related stimuli occur automatically, even when self-related stimuli have a low probability of occurrence, and that expectations to frequent, self-related stimuli operate in a relatively exclusive manner, minimizing biases to high-probability stimuli related to other people. In contrast, biases to high-familiarity stimuli (mother pairs) can be reduced when the items occur infrequently and they do not dominate expectations over other high-frequency stimuli.


Coxiella burnetii acid phosphatase inhibits the release of reactive oxygen intermediates in polymorphonuclear leukocytes.
Coxiella burnetii, the etiological agent of Q fever, is a small, Gram-negative, obligate intracellular bacterium. Replication of C. burnetii during infection has been shown to be increased by decreasing oxidative stress using p47(phox -/-) and iNOS(-/-) mice in vivo and by pharmacologic inhibitors in vitro. Building upon this model, we investigated the role polymorphonuclear leukocytes (PMN) play in the control of infection, since NADPH oxidase-mediated release of reactive oxygen intermediates (ROI) is a primary bactericidal mechanism for these cells that is critical for early innate clearance. Earlier studies suggested that C. burnetii actively inhibited release of ROI from PMN through expression of an unidentified acid phosphatase (ACP). Recent genomic annotations identified one open reading frame (CBU0335) which may encode a Sec- and type II-dependent secreted ACP. To test this model, viable C. burnetii propagated in tissue culture host cells or axenic media, C. burnetii extracts, or purified recombinant ACP (rACP) was combined with human PMN induced with 4-phorbol 12-myristate 13-acetate (PMA). The release of ROI was inhibited when PMN were challenged with viable C. burnetii, C. burnetii extracts, or rACP but not when PMN were challenged with electron beam-inactivated C. burnetii. C. burnetii extracts and rACP were also able to inhibit PMA-induced formation of NADPH oxidase complex on PMN membranes, suggesting a molecular mechanism responsible for this inhibition. These data support a model in which C. burnetii eludes the primary ROI killing mechanism of activated PMN by secreting at least one acid phosphatase.


Patient and Provider Use of Electronic Care Plans Generated From Patient-Reported Outcomes.
To determine if patients and providers perceived improved care processes through the delivery of personalized, electronic care plans (CPs) generated from the Carevive Care Planning System™. 121 women (51 with gynecologic cancer from Billings Clinic and 70 with breast cancer from Moffit Cancer Center) completed electronic patient-reported outcome assessments and were given electronically generated, personalized supportive CPs tailored to individual symptoms and local healthcare resources. Quantitative instruments evaluated feasibility, usability, acceptability, and satisfaction of the CPs from patient and provider perspectives. Qualitative interviews described patient perceptions of the CPs. Patients with cancer reported the CPs to be useful. Most perceived that CPs improved team communication, helped find needed resources, and helped manage symptoms. Provider satisfaction was highest with the platform's ability to customize patient recommendations. Interviews indicated that patients with cancer used their CP as a resource, preferred delivery at treatment initiation, and valued information to manage symptoms. Nurses play an integral role in patient education and in discussing individual care. Tailored CPs can be used as a teaching tool that patients with cancer can refer to for self-care.


[Relationship between depression and cancer].
One of the most serious and painful diseases all over the world is cancer. Despite the progress in preventing, diagnosing and treating cancer, the need for specialised services for people that are diagnosed or that have survived cancer appears to have increased. On the other hand, depressive disorders have been found to be of the most important causes of burden of disease. Depression is not only the most important cause of functional impairment in individuals that suffer from it, but also that especially in cancer survivors it can be a very important cause of non return to work. Approximately half of the cancer patients seem to develop some psychiatric symptomatology. Most commonly they present with reactive depression or a major depressive episode and some times they can even present with delirium. Most of the symptoms, especially following the diagnosis of cancer, seem to be reactive to the diagnosis of the chronic illness. Depression and cancer quite often coexist. The symptoms of the psychiatric disorder can precede, follow or coincide with the diagnosis of cancer. Surveys have shown that this could at times be explained by common biological factors that precipitate the appearance of both diseases. At the same time this could also be explained by psychological or environmental factors that could facilitate the appearance or exacerbate the symptoms of both illnesses. It has also been shown that the presentation of the symptoms of the cancer can complicate the manifestation of the depressive symptoms and vice versa. There are occasions where the cancer can cause depressive symptomatology like for example tumours of the central nervous system. On other occasions the treatment for cancer could cause or exacerbate existing depressive symptomatology. The prognosis of this comorbid condition appears to remain better if both disorders are taken into account and treated simultaneously. Therefore health services should take a holistic approach in the care of the cancer patient.


High-resolution fluorescence microscopy based on a cyclic sequential multiphoton process.
We demonstrate high-resolution fluorescence microscopy based on a cyclic sequential multiphoton (CSM) process, which gives rise to fluorescence emission following a sequence of cyclic transitions between the bright and dark states of a fluorophore induced by pump and reverse light. By temporally modulating the reverse intensity, we can extract the fluorescence signal generated through the CSM process. We show that the demodulated fluorescence signal is nonlinearly proportional to the excitation intensities and it gives a higher spatial resolution than that of a confocal microscope.


Efficacy of ProTaper Retreatment System in Root Canals Obturated with Gutta-Percha Using Two Different Sealers and GuttaFlow.
Objective. To compare the efficacy of ProTaper retreatment files in removing three different obturating materials. Study Design. Forty-five human, single-rooted premolars were divided into three experimental groups. Group 1 was obturated with gutta-percha and AH Plus sealer, Group 2 was obturated with gutta-percha and zinc oxide eugenol sealer, and Group 3 was obturated with GuttaFlow. Retreatment was done using the ProTaper universal rotary retreatment files. Root halves were visualized using magnifying loops at 3X magnification and optical stereomicroscope at 10X magnification. Images were analyzed using AutoCAD 2004 software to calculate area of the remaining debris in the canal. For statistical analysis were used variance test and ANOVA. Results. Total debris/canal area ratio between the three groups showed a statistically significant difference (P < 0.001). Conclusion. ProTaper retreatment system did not produce completely clean canals in any of the groups. However, it had the best efficacy towards removing silicon based obturating material GuttaFlow.


Arm-trunk coordination in the absence of proprioception.
During trunk-assisted reaching to targets placed within arm's length, the influence of trunk motion on the hand trajectory is compensated for by changes in the arm configuration. The role of proprioception in this compensation was investigated by analyzing the movements of 2 deafferented and 12 healthy subjects. Subjects reached to remembered targets (placed approximately 80 degrees ipsilateral or approximately 45 degrees contralateral to the sagittal midline) with an active forward movement of the trunk produced by hip flexion. In 40% of randomly selected trials, trunk motion was mechanically blocked. No visual feedback was provided during the experiment. The hand trajectory and velocity profiles of healthy subjects remained invariant whether or not the trunk was blocked. The invariance was achieved by changes in arm interjoint coordination that, for reaches toward the ipsilateral target, started as early as 50 ms after the perturbation. Both deafferented subjects exhibited considerable, though incomplete, compensation for the effects of the perturbation. Compensation was more successful for reaches to the ipsilateral target. Both deafferented subjects showed invariance between conditions (unobstructed or blocked trunk motion) in their hand paths to the ipsilateral target, and one did to the contralateral target. For the other deafferented subject, hand paths in the two types of trials began to deviate after about 50% into the movement, because of excessive elbow extension. In movements to the ipsilateral target, when deafferented subjects compensated successfully, the changes in arm joint angles were initiated as early as 50 ms after the trunk perturbation, similar to healthy subjects. Although the deafferented subjects showed less than ideal compensatory control, they compensated to a remarkably large extent given their complete loss of proprioception. The presence of partial compensation in the absence of vision and proprioception points to the likelihood that not only proprioception but also vestibulospinal pathways help mediate this compensation.


Lacrimal gland extranodal marginal zone B-cell lymphoma of MALT-type.
To evaluate the clinical features and outcome of extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue type (MALT lymphoma) in the lacrimal gland. Retrospective, noncomparative, observational case series in an academic referral setting. A consecutive series of 13 histologically verified MALT lymphoma in the lacrimal gland at presentation was studied. Clinical characteristics, treatment, and prognosis were analyzed. Eight males and five females with a median age of 64 years and a median follow-up time of 48 months were included. All patients had no prior lymphoma and initially presented as MALT lymphoma in the lacrimal gland. Extraorbital involvement at diagnosis was noted in six patients (46.2%). Two patients had autoimmune disease, and both had Stage IV disease at presentation. Treatment consisted of surgical resection in one patient, radiotherapy in four, chemotherapy in four, and combined radiotherapy and chemotherapy in four. Complete remissions were obtained in eight patients (61.5%). Patients with bilateral disease (61.5%) had a higher rate of advanced-stage disease and a poor outcome. Recurrence was noted in two patients. At the last follow-up, eight patients were free of disease, three were alive with disease, one died of sepsis as a complication of chemotherapy, and one died of lymphoma. MALT lymphoma in the lacrimal gland has a high rate of extraorbital involvement and synchronous bilateral lacrimal gland involvement at presentation. The prognosis is relatively poor, especially in patients with advanced disease and bilateral involvement. Extensive staging and long-term follow-up are warranted for these patients.


An agent-based model of signal transduction in bacterial chemotaxis.
We report the application of agent-based modeling to examine the signal transduction network and receptor arrays for chemotaxis in Escherichia coli, which are responsible for regulating swimming behavior in response to environmental stimuli. Agent-based modeling is a stochastic and bottom-up approach, where individual components of the modeled system are explicitly represented, and bulk properties emerge from their movement and interactions. We present the Chemoscape model: a collection of agents representing both fixed membrane-embedded and mobile cytoplasmic proteins, each governed by a set of rules representing knowledge or hypotheses about their function. When the agents were placed in a simulated cellular space and then allowed to move and interact stochastically, the model exhibited many properties similar to the biological system including adaptation, high signal gain, and wide dynamic range. We found the agent based modeling approach to be both powerful and intuitive for testing hypotheses about biological properties such as self-assembly, the non-linear dynamics that occur through cooperative protein interactions, and non-uniform distributions of proteins in the cell. We applied the model to explore the role of receptor type, geometry and cooperativity in the signal gain and dynamic range of the chemotactic response to environmental stimuli. The model provided substantial qualitative evidence that the dynamic range of chemotactic response can be traced to both the heterogeneity of receptor types present, and the modulation of their cooperativity by their methylation state.


Cytotoxic T-cell abundance and virus load in human immunodeficiency virus type 1 and human T-cell leukaemia virus type 1.
The correlation between virus load and specific cytotoxic T-lymphocyte (CTL) frequency during the chronic phase in human immunodeficiency virus type 1 (HIV-1) infection has been found to be negative in cross-sectional studies. We report here that, in infection with the related retrovirus human T-cell leukaemia virus type 1 (HTLV-1), the correlation is positive in asymptomatic carriers and zero in patients with the associated inflammatory disease HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). We demonstrate that the direction of the correlation may depend on the efficacy of the CTL response using mathematical models. We conclude that the CTL response is effective in asymptomatic carriers of HTLV-1, but ineffective in patients with HAM/TSP. Virus-mediated impairment of specific CTL production in HIV-1 infection can account for the negative correlation observed.


Characterization of a monoclonal antibody that induces the acrosome reaction of sea urchin sperm.
A monoclonal antibody, J18/29, induces the acrosome reaction (AR) in spermatozoa of the sea urchin Strongylocentrotus purpuratus. J18/29 induces increases in both intracellular Ca2+ and intracellular pH similar to those occurring upon induction of the AR by the natural inducer, the fucose sulfate-rich glycoconjugate of egg jelly. Lowering the Ca2+ concentration or the pH of the seawater inhibits the J18/29-induced AR, as does treatment with Co2+, an inhibitor of Ca2+ channels. The J18/29-induced AR is also inhibited by verapamil, tetraethylammonium chloride, and elevated K+. All these treatments cause similar inhibition of the egg jelly-induced AR. J18/29 reacts with a group of membrane proteins ranging in molecular mass from 340 to 25 kD, as shown by immunoprecipitation of lysates of 125I-labeled sperm and Western blots. The most prominent reacting proteins are of molecular masses of 320, 240, 170, and 58 kD. The basis of the multiple reactivity appears to reside in the polypeptide chains of these proteins, as J18/29 binding is sensitive to protease digestion but resistant to periodate oxidation. There are approximately 570,000 sites per cell for J18/29 binding. J18/29 is the only reagent of known binding specificity that induces the AR; it identifies a subset of sperm membrane proteins whose individual characterization may lead to the isolation of the receptors involved in the triggering of the AR at fertilization.


Relationship between the increase of secretion of sTNF alpha induced by lipopolysaccharides and the enhanced expression of TACE mRNA in HL-60 cells and adhesive cells from human spleen.
In order to find out the potential modulators which influence the secretion of TNF alpha, the relationship between the amount of secreted TNF alpha (sTNF alpha) and the level of TNF alpha converting enzyme (TACE) gene expression was studied before and after the stimulation by lipopolysaccharides (LPS) to HL-60 cells and adhered cells isolated from human spleen using cytological and molecular biology techniques and methods (RT-PCR, Dot-blot hybridization, etc.). The experimental results showed that: (1) LPS could induce the increase of expression of TNF alpha mRNA and TACE mRNA, reaching the peak value at 6 h and 10 h respectively after addition of LPS into cell culture medium; (2) The anti-sense oligodeoxyribonucleotide (A-ODN) complementary to TACE mRNA sequence really inhibited the secretion of TNF alpha as a result of blocked translation of TACE gene; (3) Furthermore, it was also observed that RDQ, a kind of injection derived from Chinese traditional herb, had strongly inhibitory effects on the expression of TACE mRNA and secretion of sTNF alpha stimulated by LPS. The above results suggested that the TACE indeed involved in delivering and processing of pro-TNF alpha during the period of LPS stimulation and the study about the regulator/inhibitors of TACE gene expression would be very important to develop new types of therapy agents against toxic and side effects of sTNF alpha during the period of infection/inflammation to human body.


Premorbid anxiety and depression and baseline neurocognitive, ocular-motor and vestibular performance: A retrospective cohort study.
Concussion has become a growing concern among sport and healthcare practitioners. Experts continue to investigate ways to advance the quality of concussion evaluation, diagnosis and management. Psychological conditions have been reported to influence concussion assessment outcomes at baseline and post-concussion; however, little evidence has examined psychological conditions and their effect on multifaceted measures of concussion. A retrospective cohort design was employed to examine differences between those with and without a premorbid psychological condition for high school and collegiate athletes who completed a preseason baseline battery, consisting of symptom reporting, computerized neurocognitive assessment, Vestibular-Ocular Motor Screening (VOMS), and the King-Devick (KD) test. Forty athletes within the sample self-reported a diagnosed psychological risk factor, consisting of depression and/or anxiety, and each were matched with a discordant control. Controls were matched on sex, age, sport, concussion history and ocular history. Athletes with psychological conditions reported higher symptom severity and had worse visual motor speed than controls. There were no differences between groups on other neurocognitive domains, VOMS, or KD. These results suggest that vestibular-ocular tools may be more consistent or less likely to vary between those with and without a premorbid psychological diagnosis, adding value to tools such as the KD and VOMS.


A two-tiered model for simulating the ecological and evolutionary dynamics of rapidly evolving viruses, with an application to influenza.
Understanding the epidemiological and evolutionary dynamics of rapidly evolving pathogens is one of the most challenging problems facing disease ecologists today. To date, many mathematical and individual-based models have provided key insights into the factors that may regulate these dynamics. However, in many of these models, abstractions have been made to the simulated sequences that limit an effective interface with empirical data. This is especially the case for rapidly evolving viruses in which de novo mutations result in antigenically novel variants. With this focus, we present a simple two-tiered 'phylodynamic' model whose purpose is to simulate, along with case data, sequence data that will allow for a more quantitative interface with observed sequence data. The model differs from previous approaches in that it separates the simulation of the epidemiological dynamics (tier 1) from the molecular evolution of the virus's dominant antigenic protein (tier 2). This separation of phenotypic dynamics from genetic dynamics results in a modular model that is computationally simpler and allows sequences to be simulated with specifications such as sequence length, nucleotide composition and molecular constraints. To illustrate its use, we apply the model to influenza A (H3N2) dynamics in humans, influenza B dynamics in humans and influenza A (H3N8) dynamics in equine hosts. In all three of these illustrative examples, we show that the model can simulate sequences that are quantitatively similar in pattern to those empirically observed. Future work should focus on statistical estimation of model parameters for these examples as well as the possibility of applying this model, or variants thereof, to other host-virus systems.


Adherence to cervical and breast cancer programs is crucial to improving screening performance.
Cervical and breast cancer are the most common malignancies among women worldwide. Effective screening can facilitate early detection and dramatically reduce mortality rates. The interface between those screening patients and patients most needing screening is complex, and women in remote areas of rural counties face additional barriers that limit the effectiveness of cancer prevention programs. This study compared various methods to improve compliance with mass screening for breast and cervical cancer among women in a remote, rural region of Brazil. In 2003, a mobile unit was used to perform 10,156 mammograms and Papanicolaou smear tests for women living in the Barretos County region of São Paulo state, Brazil (consisting of 19 neighbouring cities). To reach the women, the following community outreach strategies were used: distribution of flyers and pamphlets; media broadcasts (via radio and car loudspeakers); and community healthcare agents (CHCAs) making home visits. The most useful intervention appeared to be the home visits by healthcare agents or CHCAs. These agents of the Family Health Programme of the Brazilian Ministry of Health reached an average of 45.6% of those screened, with radio advertisements reaching a further 11.9%. The great majority of the screened women were illiterate or had elementary level schooling (80.9%) and were of 'poor' or 'very poor' socioeconomic class (67.2%). Use of a mobile screening unit is a useful strategy in developing countries where local health systems have inadequate facilities for cancer screening in underserved populations. A multimodal approach to community outreach strategies, especially using CHCAs and radio advertisements, can improve the uptake of mass screening in low-income, low-educational background female populations.


Humic substances increase survival of freshwater shrimp Caridina sp. D to acid mine drainage.
Humic substances (HS) are known to decrease the toxicity of heavy metals to aquatic organisms, and it has been suggested that they can provide buffering protection in low pH conditions. Despite this, little is known about the ability for HS to increase survival to acid mine drainage (AMD). In this study, the ability of HS to increase survival of the freshwater shrimp (Caridina sp. D sensu Page et al. in Biol Lett 1:139-142, 2005) to acid mine drainage was investigated using test waters collected from the Mount Morgan open pit in Central Queensland with the addition of Aldrich humic acid (AHA). The AMD water from the Mount Morgan open pit is highly acidic (pH 2.67) as well as contaminated with heavy metals (1780 mg/L aluminum, 101 mg/L copper [Cu], 173 mg/L manganese, 51.8 mg/L zinc [Zn], and 51.8 mg/L iron). Freshwater shrimp were exposed to dilutions in the range of 0.5 % to 5 % AMD water with and without the addition of 10 or 20 mg/L AHA. In the absence of HS, all shrimp died in the 2.5 % AMD treatment. In contrast, addition of HS increased survival in the 2.5 % AMD treatment by ≤66 % as well as significantly decreased the concentration of dissolved Cu, cobalt, cadmium, and Zn. The decreased toxicity of AMD in the presence of HS is likely to be due to complexation and precipitation of heavy metals with the HS; it is also possible that HS caused changes to the physiological condition of the shrimp, thus increasing their survival. These results are valuable in contributing to an improved understanding of potential role of HS in ameliorating the toxicity of AMD environments.


Aromatic residues at the edge of the antibody combining site facilitate viral glycoprotein recognition through membrane interactions.
The broadly neutralizing anti-HIV antibody 4E10 recognizes an epitope very close to the virus membrane on the glycoprotein gp41. It was previously shown that epitope recognition improves in a membrane context and that 4E10 binds directly, albeit weakly, to lipids. Furthermore, a crystal structure of Fab 4E10 complexed to an epitope peptide revealed that the centrally placed, protruding H3 loop of the antibody heavy chain does not form peptide contacts. To investigate the hypothesis that the H3 loop apex might interact with the viral membrane, two Trp residues in this region were substituted separately or in combination with either Ala or Asp by site-directed mutagenesis. The resultant IgG variants exhibited similar affinities for an epitope peptide as WT 4E10 but lower apparent affinities for both viral membrane mimetic liposomes and Env(-) virus. Variants also exhibited lower apparent affinities for Env(+) virions and failed to significantly neutralize a number of 4E10-sensitive viruses. For the extremely sensitive HXB2 virus, variants did neutralize, but at 37- to >250-fold lower titers than WT 4E10, with Asp substitutions exerting a greater effect on neutralization potency than Ala substitutions. Because reductions in lipid binding reflect trends in neutralization potency, we conclude that Trp residues in the antibody H3 loop enable membrane proximal epitope recognition through favorable lipid interactions. The requirement for lipophilic residues such as Trp adjacent to the antigen binding site may explain difficulties in eliciting 4E10-like neutralizing antibody responses by immunization and helps define a unique motif for antibody recognition of membrane proximal antigens.


Fire-driven vegetation type conversion in Southern California.
One consequence of global change causing widespread concern is the possibility of ecosystem conversions from one type to another. A classic example of this is vegetation type conversion (VTC) from native woody shrublands to invasive annual grasslands in the biodiversity hotspot of Southern California. Although the significance of this problem is well recognized, understanding where, how much, and why this change is occurring remains elusive owing to differences in results from studies conducted using different methods, spatial extents, and scales. Disagreement has arisen particularly over the relative importance of short-interval fires in driving these changes. Chronosequence approaches that use space for time to estimate changes have produced different results than studies of changes at a site over time. Here we calculated the percentage woody and herbaceous cover across Southern California using air photos from ~1950 to 2019. We assessed the extent of woody cover change and the relative importance of fire history, topography, soil moisture, and distance to human infrastructure in explaining change across a hierarchy of spatial extents and regions. We found substantial net decline in woody cover and expansion of herbaceous vegetation across all regions, but the most dramatic changes occurred in the northern interior and southern coastal areas. Variables related to frequent, short-interval fire were consistently top ranked as the explanation for shrub to grassland type conversion, but low soil moisture and topographic complexity were also strong correlates. Despite the consistent importance of fire, there was substantial geographical variation in the relative importance of drivers, and these differences resulted in different mapped predictions of VTC. This geographical variation is important to recognize for management decision-making and, in addition to differences in methodological design, may also partly explain differences in previous study results. The overwhelming importance of short-interval fire has management implications. It suggests that actions should be directed away from imposing fires to preventing fires. Prevention can be controlled through management actions that limit ignitions, fire spread, and the damage sustained in areas that do burn. This study also demonstrates significant potential for changing fire regimes to drive large-scale, abrupt ecological change.


A comparison of routine and case-managed pathways for recovery from musculoskeletal disorders in people in employment.
To compare outcomes in employed people from an enhanced routine management pathway for musculoskeletal disorders within National Health Service Scotland with an existing active case-management system, Working Health Services Scotland. The study comprised a service evaluation using anonymised routinely collected data from all currently employed callers presenting with musculoskeletal disorder to the two services. Baseline demographic and clinical data were collected. EuroQol EQ-5DTM scores at the start and end of treatment were compared for both groups, overall and by age, sex, socio-economic status, and anatomical site, and the impact of mental health status at baseline was evaluated. Active case-management resulted in greater improvement than enhanced routine care. Case-managed service users entered the programme earlier in the recovery pathway; there was evidence of spontaneous improvement during the longer waiting time of routine service clients but only if they had good baseline mental health. Those most disadvantaged through mental health co-morbidity showed the greatest benefit. People with musculoskeletal disorders who have poor baseline mental health status derive greatest benefit from active case-management. Case-management therefore contributes to reducing health inequalities and can help to minimise long-term sickness absence. Shorter waiting times contributed to better outcomes in the case-managed service. Implications for RehabilitationMusculoskeletal disorders are a major cause of inability to work.Case-management is effective in helping people with musculoskeletal disorders to return to work.People who have the poorest mental health are likely to gain the greatest benefit from case-management of their musculoskeletal disorders.


Increased susceptibility to primary infection with Listeria monocytogenes in germfree mice may be due to lack of accumulation of L-selectin+ CD44+ T cells in sites of inflammation.
The host defense of germfree (GF) mice against primary infection with Listeria monocytogenes was compared with that of specific-pathogen-free (SPF) mice. In SPF mice, the numbers of bacteria in the peritoneal cavity, liver, and spleen decreased gradually to undetectable levels by day 8 after intraperitoneal infection with a sublethal dose (2 X 10(3) CFU) of L. monocytogenes. On the other hand, the elimination of bacteria in these organs of GF mice was significantly impaired at this stage after inoculation. We have reported previously that T cells coexpressing L-selectin and CD44 play an important role in protection against L. monocytogenes through trafficking to sites of inflammation. Consistent with our previous findings, the number of unique L-selectin+ CD44+ T cells in the peritoneal cavity was remarkably increased on day 8 after infection in SPF mice, whereas such an increase was not evident in GF mice at this stage. Listeria-specific T-cell proliferation was normally detected in the lymph node cells of GF mice inoculated with L. monocytogenes, whereas the T-cell-proliferative response of the peritoneal exudate cells of GF mice was significantly impaired compared with that of SPF mice. These results suggest that the priming of T cells against listerial antigens normally occurs in the peripheral lymphoid organs of GF mice but the trafficking of the activated T cells to the inflamed sites may be severely impaired in GF mice, resulting in increased susceptibility to infection with L. monocytogenes.


The unfolded protein response is triggered by a plant viral movement protein.
Infection with Potato virus X (PVX) in Nicotiana benthamiana plants leads to increased transcript levels of several stress-related host genes, including basic-region leucine zipper 60 (bZIP60), SKP1, ER luminal binding protein (BiP), protein disulfide isomerase (PDI), calreticulin (CRT), and calmodulin (CAM). bZIP60 is a key transcription factor that responds to endoplasmic reticulum (ER) stress and induces the expression of ER-resident chaperones (BiP, PDI, CRT, and CAM). SKP1 is a component of SCF (for SKP1-Cullin-F box protein) ubiquitin ligase complexes that target proteins for proteasomal degradation. Expression of PVX TGBp3 from a heterologous vector induces the same set of genes in N. benthamiana and Arabidopsis (Arabidopsis thaliana) leaves. Virus-induced gene silencing was employed to knock down the expression of bZIP60 and SKP1, and the number of infection foci on inoculated leaves was reduced and systemic PVX accumulation was altered. Silencing bZIP60 led to the suppression of BiP and SKP1 transcript levels, suggesting that bZIP60 might be an upstream signal transducer. Overexpression of TGBp3 led to localized necrosis, but coexpression of TGBp3 with BiP abrogated necrosis, demonstrating that the unfolded protein response alleviates ER stress-related cell death. Steady-state levels of PVX replicase and TGBp2 (which reside in the ER) proteins were unaltered by the presence of TGBp3, suggesting that TGBp3 does not contribute to their turnover. Taken together, PVX TGBp3-induced ER stress leads to up-regulation of bZIP60 and unfolded protein response-related gene expression, which may be important to regulate cellular cytotoxicity that could otherwise lead to cell death if viral proteins reach high levels in the ER.


Assessing the accuracy of satellite derived global and national urban maps in Kenya.
Ninety percent of projected global urbanization will be concentrated in low income countries (United-Nations, 2004). This will have considerable environmental, economic and public health implications for those populations. Objective and efficient methods of delineating urban extent are a cross-sectoral need complicated by a diversity of urban definition rubrics world-wide. Large-area maps of urban extents are becoming increasingly available in the public domain, as are a wide-range of medium spatial resolution satellite imagery. Here we describe the extension of a methodology based on Landsat ETM and Radarsat imagery to the production of a human settlement map of Kenya. This map was then compared with five satellite imagery-derived, global maps of urban extent at Kenya national-level, against an expert opinion coverage for accuracy assessment. The results showed the map produced using medium spatial resolution satellite imagery was of comparable accuracy to the expert opinion coverage. The five global urban maps exhibited a range of inaccuracies, emphasising that care should be taken with use of these maps at national and sub-national scale.


Lay perceptions of type 2 diabetes in Scotland: bringing health services back in.
The growing prevalence of type 2 diabetes is placing Scottish health services under considerable strain. Consequently, diabetes services are undergoing a major process of reorganisation, including the devolvement of routine diabetes care/diabetic review from secondary to primary healthcare settings. This qualitative study was devised to explore newly diagnosed type 2 diabetes patients' perceptions of their disease and the health services they receive at a time when this restructuring of services is taking place. The sample comprised 40 patients resident in Lothian, Scotland, who had diverse experiences of services, some receiving GP-based care only, others having varying contact with hospital diabetes clinics. In-depth interviews were undertaken with patients, three times at six monthly intervals over 1 year, enabling their experiences to be tracked at critical junctures during the post-diagnostic period. Disease perceptions and health service delivery were found to be mutually informing and effecting. Not only did (different types of) health service delivery influence the ways in which patients thought about and self-managed their disease, over time patients' disease perceptions also informed their expectations of, and preferences for, diabetes services. We thus argue that there is a need for a reconceptualisation within the medical social sciences to take into account the context of healthcare and the economic/policy factors that inform health service delivery when looking at patients' disease perceptions. We also discuss the logistical and ethical challenges of drawing upon patients' perspectives, preferences and views in the design and delivery of future health services.