The role of red and processed meat in colorectal cancer development: a perspective.
This paper is based on a workshop held in Oslo, Norway in November 2013, in which experts discussed how to reach consensus on the healthiness of red and processed meat. Recent nutritional recommendations include reducing intake of red and processed meat to reduce cancer risk, in particular colorectal cancer (CRC). Epidemiological and mechanistic data on associations between red and processed meat intake and CRC are inconsistent and underlying mechanisms are unclear. There is a need for further studies on differences between white and red meat, between processed and whole red meat and between different types of processed meats, as potential health risks may not be the same for all products. Better biomarkers of meat intake and of cancer occurrence and updated food composition databases are required for future studies. Modifying meat composition via animal feeding and breeding, improving meat processing by alternative methods such as adding phytochemicals and improving our diets in general are strategies that need to be followed up.


Considerations for treatment duration in responders to immune checkpoint inhibitors.
Immune checkpoint inhibitors (ICIs) have improved overall survival for cancer patients, however, optimal duration of ICI therapy has yet to be defined. Given ICIs were first used to treat patients with metastatic melanoma, a condition that at the time was incurable, little attention was initially paid to how much therapy would be needed for a durable response. As the early immunotherapy trials have matured past 10 years, a significant per cent of patients have demonstrated durable responses; it is now time to determine whether patients have been overtreated, and if durable remissions can still be achieved with less therapy, limiting the physical and financial toxicity associated with years of treatment. Well-designed trials are needed to identify optimal duration of therapy, and to define biomarkers to predict who would benefit from shorter courses of immunotherapy. Here, we outline key questions related to health, financial and societal toxicities of over treating with ICI and present four unique clinical trials aimed at exposing criteria for early cessation of ICI. Taken together, there is a serious liability to overtreating patients with ICI and future work is warranted to determine when it is safe to stop ICI.


The Role of the Microenvironment in Prostate Cancer-Associated Bone Disease.
The bone is a common site for metastasis in patients with advanced prostate carcinoma, and provides a 'fertile' milieu which stimulates tumour growth and associated bone disease. For years, the concept of treatment strategies has remained targeting the tumour itself; however, the occurrence of chemoresistance remains a challenge now more than ever. The attraction of targeting the bone microenvironment in order to disrupt tumour localisation and proliferation stems from the idea that stromal cells are superiorly stable at a genetic level, thus decreasing the risk of resistance manifestation. In this review, we will discuss recent findings with regards to the pathogenesis of prostate cancer-induced bone disease and recent therapeutic strategies in an aim to evaluate the ever increasing role of the microenvironment in disease progression.


Liver-related mortality in human-immunodeficiency-virus-infected patients between 1995 and 2003 in the French GERMIVIC Joint Study Group Network (MORTAVIC 2003 Study).
The objective of the present study was to determine mortality because of end-stage liver disease (ESLD) in a nationwide population of HIV-infected patients, 7 years following the introduction of highly active antiretroviral therapy (HAART). All departments of internal medicine and infectious diseases from the GERMIVIC Study Group prospectively recorded all deaths in HIV-infected patients during 2003. Fifty-nine departments, following a total of 20 940 HIV-infected patients, participated in the study. Results were compared with those of previous surveys conducted using similar methodology in 1995, 1997 and 2001. Among 215 deaths observed during 2003, 101 (46.9%) were related to AIDS, 27 (12.6%) to ESLD and 87 (40.5%) to other causes. Mortality because of ESLD represented 23.7% of non-AIDS-related deaths. Patients dying from ESLD had chronic hepatitis because of hepatitis C virus (HCV) in 92.6% of cases and moderate (30-60 g) or high (>60 g) alcohol consumption (43.5% and 26.0%, respectively). In this population, deaths because of ESLD were 1.5% in 1995, 6.6% in 1997, 14.3% in 2001 and 12.6% in 2003. The prevalence of hepatocellular carcinoma as a cause of death remained high in 2003 but stable when compared with 2001 (25%vs 14.8%). Treatment of hepatitis C in patients who died from ESLD was more frequent in 2003 (44.4%) than in 2001 (26.3%). Seven years after the introduction of HAART, ESLD associated with HCV infections is a leading cause of mortality in HIV-infected patients, which did not increase between the years 2001 and 2003.


Monte Carlo simulation and patient dosimetry for a kilovoltage cone-beam CT unit.
The purpose of this work is to characterize the x-ray volume imager (XVI), the cone-beam computed tomography (CBCT) unit mounted on the Elekta Synergy linac, with F1 bowtie filter and to calculate the three-dimensional dose delivered to patients using volumetric acquisition. The XVI is modeled in detail using a new Monte Carlo (MC) code, BEAMPP, under development at the National Research Council Canada. In this investigation, a new component module is developed to accurately model the unit's bowtie filter used in conjunction with the available beam collimators at the clinical energy of 120 kV. The modeling is compared against percentage depth dose (PDD) and profile measurements. Kilovoltage radiation beams' phase space files are also analyzed. The authors also describe a method for the absolute dose calibration of the MC model of the CBCT unit when used in a clinical volumetric acquisition mode. Finally, they calculate three-dimensional patient dose from CBCT image acquisition in three clinical cases of interest: Pelvis, lung, and head and neck. The agreement between measurement and MC is shown to be very good: Within +/- 2% for the PDD and within +/- 3.5% inside the radiation field for all the collimators with the F1 bowtie filter. A full account of the absolute calibration method is given and dose calculation is validated against ion chamber measurements in different locations of a plastic phantom. Calculations and experiments agree within +/- 2% or better in both at the center and the periphery of the phantom, with worst agreement of 4.5% at the surface of the phantom and for one specific combination of collimator and filter. Patient dose from CBCT scan reveals that dose to tissue is between 2 and 2.5 cGy for a pelvis or a lung full acquisition. For H&N dose to tissue is 5 cGy, with the unit presets used in this work. Dose to bony structures can be two to three times higher than dose to tissue. The XVI CBCT unit has been fully modeled including the F1 bowtie filter. Absolute dose distribution from the unit has been successfully validated. Full MC patient dose calculation has shown that the three-dimensional dose distribution from CBCT is complex. Patient dose from CBCT exposure cannot be completely accounted for by using a numerical factor as an estimate of the dose at the center of the body. Furthermore, additional dose to bone should be taken into account when adopting any IGRT strategy and weighed vs the unquestionable benefits of the technique in order to optimize treatment. Full three-dimensional dose calculation is recommended if patient dose from CBCT is to be integrated in any adaptive planning strategy.


Dipolar coupling of nanoparticle-molecule assemblies: An efficient approach for studying strong coupling.
Strong light-matter interactions facilitate not only emerging applications in quantum and non-linear optics but also modifications of properties of materials. In particular, the latter possibility has spurred the development of advanced theoretical techniques that can accurately capture both quantum optical and quantum chemical degrees of freedom. These methods are, however, computationally very demanding, which limits their application range. Here, we demonstrate that the optical spectra of nanoparticle-molecule assemblies, including strong coupling effects, can be predicted with good accuracy using a subsystem approach, in which the response functions of different units are coupled only at the dipolar level. We demonstrate this approach by comparison with previous time-dependent density functional theory calculations for fully coupled systems of Al nanoparticles and benzene molecules. While the present study only considers few-particle systems, the approach can be readily extended to much larger systems and to include explicit optical-cavity modes.


Herpes Genitalis: Diagnosis, Treatment and Prevention.
Herpes genitalis is caused by the herpes simplex virus type 1 or type 2 and can manifest as primary or recurrent infection. It is one of the most common sexually transmitted infections and due to associated physical and psychological morbidity it constitutes a considerable, often underestimated medical problem. In addition to providing the reader with basic knowledge of the pathogen and clinical presentation of herpes genitalis, this review article discusses important aspects of the laboratory diagnostics, antiviral therapy and prophylaxis. The article is aimed at all health-care workers managing patients with herpes genitalis and attempts to improve the often suboptimal counselling, targeted use of laboratory diagnostics, treatment and preventive measures provided to patients.


Orotracheal fibreoptic intubation for rapid sequence induction of anaesthesia.
We assessed whether flexible fibreoptic-guided orotracheal intubation could be rapidly and successfully achieved during a simulated rapid sequence induction in 30 anaesthetised and paralysed patients. Rapid sequence induction was simulated by applying practised cricoid pressure. Using a flexible fibreoptic laryngoscope with camera and closed circuit television, an anaesthetist experienced with the technique performed orotracheal endoscopy and intubation with a cuffed 7.0-mm Portex tracheal tube through a VBM Bronchoscope Airway. Fibreoptic intubation was successful at the first attempt in 28 patients (93%); two patients required two attempts. Mean (SD) time from removal of the facemask from the patient's face to the appearance of carbon dioxide in the expired breath after intubation was 111 (46) s (median 100 s; range 54-195 s). There were one or more difficulties in 13 patients (43%). These difficulties were largely avoidable and included problems with fibreoptic equipment, the Bronchoscope Airway, copious secretions, cricoid pressure or railroading of the tracheal tube. Flexible fibreoptic-guided orotracheal intubation may have a place in the management of failed intubation during a rapid sequence induction.


Controlling superconducting spin flow with spin-flip immunity using a single homogeneous ferromagnet.
Spin transport via electrons is typically plagued by Joule heating and short decay lengths due to spin-flip scattering. It is known that dissipationless spin currents can arise when using conventional superconducting contacts, yet this has only been experimentally demonstrated when using intricate magnetically inhomogeneous multilayers, or in extreme cases such as half-metals with interfacial magnetic disorder. Moreover, it is unknown how such spin supercurrents decay in the presence of spin-flip scattering. Here, we present a method for generating a spin supercurrent by using only a single homogeneous magnetic element. Remarkably, the spin supercurrent generated in this way does not decay spatially, in stark contrast to normal spin currents that remain polarized only up to the spin relaxation length. We also expose the existence of a superconductivity-mediated torque even without magnetic inhomogeneities, showing that the different components of the spin supercurrent polarization respond fundamentally differently to a change in the superconducting phase difference. This establishes a mechanism for tuning dissipationless spin and charge flow separately, and confirms the advantage that superconductors can offer in spintronics.


An approach to evaluating bias and uncertainty in estimates of external dose obtained from personal dosimeters.
This paper describes an approach to quantifying errors in recorded estimates of external radiation dose obtained from personal dosimeters and applies the approach to dose estimates of workers at the Hanford site. Because a major objective of this evaluation is to provide the information needed for adjusting epidemiologic dose-response analyses of worker data for errors in dose estimates, the paper addresses the extent that errors for different workers are correlated, focuses on recorded doses as estimates of organ doses, and focuses on recorded doses as estimates of organ doses, and gives consideration to both annual and cumulative doses. The evaluation emphasizes errors resulting from the fact that dosimeters are limited in their ability to respond accurately to all radiation energies to which workers are exposed or to radiation coming from all directions. For each of several sources of error, systematic bias factors are estimated for two energy ranges (100--300 keV and 300--1,000 keV), two geometries (anterior-posterior and rotational), and four calendar year periods. These are then combined using information provided by health physicists on energies and geometries in Hanford exposure environments. Except for the period before 1958, deep dose, the objective of modern dosimetry systems, was found to be fairly accurately estimated. Lung dose was found to be overestimated by about 10%, and bone marrow dose was found to be overestimated by about 50%. However, many aspects of this evaluation relied heavily on subjective judgments, and, thus, these factors are subject to considerable uncertainty. Estimates of uncertainty in the bias factors and uncertainty reflecting random error are provided.


A transmission electron microscopy investigation: the membrane complex in spermatogenesis of Fenneropenaeus chinensis.
The transforming characteristics of the membrane complex in spermatogenesis of Fenneropenaeus chinensis have been studied by using transmission electron microscopy. Two types of membrane complex have been investigated based on their sources: one originating from nucleus and the other from cytoplasm. The first one, consisted of annular structures, monolayer membrane blebs, and double or multi-lamellar membrane vesicles, emerges in the primary spermatocyte, then diffuses with the nuclear membrane and finally enters the cytoplasm. This type of membrane complex seems to play an important role in the materials transfusion from nucleus to cytoplasm, and it mainly exists inside the primary spermatocyte with some inside the secondary spermatocyte. The latter, originated from cytoplasm, is formed during the anaphase of spermiogenesis. It also exists in mature sperm, locating at both sides of the nucleus under the acrosomal cap. This type of membrane complex mainly comprises rings of convoluted membrane pouches, together with mitochondria, annular lamina bodies, fragments of endoplasmic reticulum, nuclear membrane and some nuclear particles. It releases vesicles and particles into the acrosomal area during the formation of the perforatorium, suggesting a combined function of the endoplasmic reticulum, mitochondria and Golgi's mechanism.


Quantitation of human papillomavirus DNA in plasma of oropharyngeal carcinoma patients.
To determine whether human papillomavirus (HPV) DNA can be detected in the plasma of patients with HPV-positive oropharyngeal carcinoma (OPC) and to monitor its temporal change during radiotherapy. We used polymerase chain reaction to detect HPV DNA in the culture media of HPV-positive SCC90 and VU147T cells and the plasma of SCC90 and HeLa tumor-bearing mice, non-tumor-bearing controls, and those with HPV-negative tumors. We used real-time quantitative polymerase chain reaction to quantify the plasma HPV DNA in 40 HPV-positive OPC, 24 HPV-negative head-and-neck cancer patients and 10 non-cancer volunteers. The tumor HPV status was confirmed by p16(INK4a) staining and HPV16/18 polymerase chain reaction or HPV in situ hybridization. A total of 14 patients had serial plasma samples for HPV DNA quantification during radiotherapy. HPV DNA was detectable in the plasma samples of SCC90- and HeLa-bearing mice but not in the controls. It was detected in 65% of the pretreatment plasma samples from HPV-positive OPC patients using E6/7 quantitative polymerase chain reaction. None of the HPV-negative head-and-neck cancer patients or non-cancer controls had detectable HPV DNA. The pretreatment plasma HPV DNA copy number correlated significantly with the nodal metabolic tumor volume (assessed using (18)F-deoxyglucose positron emission tomography). The serial measurements in 14 patients showed a rapid decline in HPV DNA that had become undetectable at radiotherapy completion. In 3 patients, the HPV DNA level had increased to a discernable level at metastasis. Xenograft studies indicated that plasma HPV DNA is released from HPV-positive tumors. Circulating HPV DNA was detectable in most HPV-positive OPC patients. Thus, plasma HPV DNA might be a valuable tool for identifying relapse.


Dibenzo-p-dioxins in the environment from ceramics and pottery produced from ball clay mined in the United States.
Processed ball clay samples used in the production of ceramics and samples of the ceramic products were collected and analyzed for the presence and concentration of the 2,3,7,8-Cl substituted polychlorinated dibenzo-p-dioxins and -furans (PCDDs/PCDFs). The processed ball clay had average PCDD concentrations of 3.2 ng/g toxic equivalents, a congener profile, and isomer distribution consistent with those found previously in raw ball clay. The PCDF concentrations were below the average limit of detection (LOD) of 0.5 pg/g. The final fired ceramic products were found to be free of PCDDs/PCDFs at the LODs. A consideration of the conditions involved in the firing process suggests that the PCDDs, if not destroyed, may be released to the atmosphere and could represent an as yet unidentified source of dioxins to the environment. In addition, the PCDDs in clay dust generated during manufacturing operations may represent a potential occupational exposure.


Hippocampal network abnormalities explain amnesia after VGKCC-Ab related autoimmune limbic encephalitis.
Limbic encephalitis associated with antibodies to components of the voltage-gated potassium channel complex (VGKCC-Ab-LE) often leads to hippocampal atrophy and persistent memory impairment. Its long-term impact on regions beyond the hippocampus, and the relationship between brain damage and cognitive outcome, are poorly understood. We investigated the nature of structural and functional brain abnormalities following VGKCC-Ab-LE and its role in residual memory impairment. A cross-sectional group study was conducted. Twenty-four VGKCC-Ab-LE patients (20 male, 4 female; mean (SD) age 63.86 (11.31) years) were recruited post-acutely along with age- and sex-matched healthy controls for neuropsychological assessment, structural MRI and resting-state functional MRI (rs-fMRI). Structural abnormalities were determined using volumetry and voxel-based morphometry; rs-fMRI data were analysed to investigate hippocampal functional connectivity (FC). Associations of memory performance with neuroimaging measures were examined. Patients showed selective memory impairment. Structural analyses revealed focal hippocampal atrophy within the medial temporal lobes, correlative atrophy in the mediodorsal thalamus, and additional volume reduction in the posteromedial cortex. There was no association between regional volumes and memory performance. Instead, patients demonstrated reduced posteromedial cortico-hippocampal and inter-hippocampal FC, which correlated with memory scores (r = 0.553; r = 0.582, respectively). The latter declined as a function of time since the acute illness (r = -0.531). VGKCC-Ab-LE results in persistent isolated memory impairment. Patients have hippocampal atrophy with further reduced mediodorsal thalamic and posteromedial cortical volumes. Crucially, reduced FC of remaining hippocampal tissue correlates more closely with memory function than does regional atrophy.


[Genetic risk factors of ischemic stroke identified by a genome-wide association study].
Recent research suggests that many genetic variants and environmental exposures can influence the occurrence and progression of common diseases, including stroke. However, the genetic risk factors for ischemic stroke, the most common type of stroke in developed countries, are almost unknown. We conducted a genome-wide association study using 1112 Japanese subjects who has suffered from ischemic stroke. Age- and sex-matched healthy individuals were selected as controls from the participants of the health examination survey conducted in the Hisayama study in 2002-03. We used a two-stage screening method and identified 2 genes related to the susceptibility of ischemic stroke: PRKCH and AGTRL1. A non-synonymous single nucleotide polymorphism (SNP) in the PRKCH gene (rs2230500, V374I) was significantly associated with lacunar infarction in 2 independent case control samples. The nonsynonymous SNP altered the kinase activity of protein kinase C-eta (PKCeta). We also found that PKCeta; was mainly expressed in vascular endothelial cells and foamy macrophages in human atherosclerotic lesions and that its expression was enhanced as the lesion progressed. Four SNPs around the AGTRL1 gene were closely associated with ischemic stroke. An SNP in the promoter region (rs9943582) enhanced the expression level of AGTRL1 mRNA. A population-based cohort study with 14 years of follow-up revealed that functional SNPs of these genes were found to significantly increase the incidence of ischemic stroke. Therefore we suggest that genome-wide association study is a powerful tool for identifying new susceptibility genes and can provide new insights into the pathogenesis, prevention, and treatment of ischemic stroke.


Variations in Helicobacter pylori cytotoxin-associated genes and their influence in progression to gastric cancer: implications for prevention.
Helicobacter pylori (HP) is a bacterium that colonizes the human stomach and can establish a long-term infection of the gastric mucosa. Persistent Hp infection often induces gastritis and is associated with the development of peptic ulcer disease, atrophic gastritis, and gastric adenocarcinoma. Virulent HP isolates harbor the cag (cytotoxin-associated genes) pathogenicity island (cagPAI), a 40 kb stretch of DNA that encodes components of a type IV secretion system (T4SS). This T4SS forms a pilus for the injection of virulence factors into host target cells, such as the CagA oncoprotein. We analyzed the genetic variability in cagA and other selected genes of the HP cagPAI (cagC, cagE, cagL, cagT, cagV and cag Gamma) using DNA extracted from frozen gastric biopsies or from clinical isolates. Study subjects were 95 cagA+ patients that were histologically diagnosed with chronic gastritis or gastric cancer in Venezuela and Mexico, areas with high prevalence of Hp infection. Sequencing reactions were carried out by both Sanger and next-generation pyrosequencing (454 Roche) methods. We found a total of 381 variants with unambiguous calls observed in at least 10% of the originally tested samples and reference strains. We compared the frequencies of these genetic variants between gastric cancer and chronic gastritis cases. Twenty-six SNPs (11 non-synonymous and 14 synonymous) showed statistically significant differences (P<0.05), and two SNPs, in position 1039 and 1041 of cagE, showed a highly significant association with cancer (p-value = 2.07×10⁻⁶), and the variant codon was located in the VirB3 homology domain of Agrobacterium. The results of this study may provide preliminary information to target antibiotic treatment to high-risk individuals, if effects of these variants are confirmed in further investigations.


Long-term prognosis of patients with ulcerative colitis treated with cytapheresis therapy.
Although accumulating studies in Japan show that cytapheresis (CAP) therapy is safe and effective for the induction of remission of moderate or severe ulcerative colitis (UC), the long-term prognosis of UC patients treated with CAP is unknown. The aim of this study was to determine the long-term prognosis of UC patients treated with CAP. Ninety patients treated previously with CAP and followed for more than 3 years were evaluated. The rates of operation, readmission, and use or dose-up of corticosteroid were analyzed as long-term prognosis. Following the first course of CAP treatment, 64% of patients showed clinical improvement (> 4-point decrease in the clinical activity index (CAI)), and 49% of patients achieved clinical remission (CAI ≤ 4). Longer disease duration and lower age at the first CAP treatment correlated significantly with the therapeutic effects of CAP (p = 0.003 and 0.035, respectively). The rates of operation and readmission were significantly lower in patients who showed previous clinical effects of CAP than in those who did not respond to CAP. The rates of operation and readmission were also significantly lower in patients whose treatment was combined with immunomodulators after the initiation of CAP than in patients who did not use immunomodulators. Importantly, the second course of CAP was also effective in most of the patients who showed a clinical response to the first CAP. Patients who achieve remission after the first CAP therapy may have a good long-term prognosis and a good response to a second CAP therapy even after relapse.


Inhibition of Thioredoxin/Thioredoxin Reductase Induces Synthetic Lethality in Lung Cancers with Compromised Glutathione Homeostasis.
Glutathione (GSH)/GSH reductase (GSR) and thioredoxin/thioredoxin reductase (TXNRD) are two major compensating thiol-dependent antioxidant pathways that maintain protein dithiol/disulfide balance. We hypothesized that functional deficiency in one of these systems would render cells dependent on compensation by the other system for survival, providing a mechanism-based synthetic lethality approach for treatment of cancers. The human GSR gene is located on chromosome 8p12, a region frequently lost in human cancers. GSR deletion was detected in about 6% of lung adenocarcinomas in The Cancer Genome Atlas database. To test whether loss of GSR sensitizes cancer cells to TXNRD inhibition, we knocked out or knocked down the GSR gene in human lung cancer cells and evaluated their response to the TXNRD inhibitor auranofin. GSR deficiency sensitized lung cancer cells to this agent. Analysis of a panel of 129 non-small cell lung cancer (NSCLC) cell lines revealed that auranofin sensitivity correlated with the expression levels of the GSR, glutamate-cysteine ligase catalytic subunit (GCLC), and NAD(P)H quinone dehydrogenase 1 (NQO1) genes. In NSCLC patient-derived xenografts with reduced expression of GSR and/or GCLC, growth was significantly suppressed by treatment with auranofin. Together, these results provide a proof of concept that cancers with compromised expression of enzymes required for GSH homeostasis or with chromosome 8p deletions that include the GSR gene may be targeted by a synthetic lethality strategy with inhibitors of TXNRD. SIGNIFICANCE: These findings demonstrate that lung cancers with compromised expression of enzymes required for glutathione homeostasis, including reduced GSR gene expression, may be targeted by thioredoxin/thioredoxin reductase inhibitors.


Motivation and physiologic responses of playing a physically interactive video game relative to a sedentary alternative in children.
While there is emerging research outlining the physiologic cost of the physically interactive Nintendo Wii, there are no evaluations of the relative reinforcing value (RRV) of the Wii versus a sedentary alternative. The purpose of this study is to evaluate the physiologic cost, RRV, and liking of playing Wii Sports Boxing (Wii) versus a traditional sedentary video game (SVG) in 11 lean and 13 overweight/obese 8- to 12-year-old children. Heart rate (HR) and VO(2) were assessed during rest, treadmill walking, and playing an SVG and Wii using a counterbalance design. Liking was assessed during treadmill walking and video game play. RRV was assessed for Wii versus SVG. Average HR, VO(2), and liking were significantly greater for Nintendo Wii (p < or = 0.001 for all) than all other conditions. Lean children displayed a greater (p < 0.001) peak responding for access to Wii relative to the SVG while overweight/obese children did not (p > or = 0.16). Wii was a well-liked activity of greater physiologic intensity than both the SVG and treadmill walking. Lean children were more motivated while overweight/obese children were equally as motivated to play Wii relative to the SVG.


Acanthamoeba and bacteria produce antimicrobials to target their counterpart.
In the microbial ecosystem, microbes compete for space and nutrients. Consequently, some have developed the ability to kill or inhibit the growth of other competing microbes by producing antimicrobial substances. As the 'producer' species are generally immune to these substances, their compounds act on the competing microbial species and give the producer more space and access to nutrients for growth. Many currently used antibiotics were developed by exploiting this potential of certain microbes. Here, the free-living amoeba, Acanthamoeba castellanii, was investigated for its antibacterial activity against representative Gram positive and Gram negative bacteria, while bacterial isolates were tested for their anti-amoebic properties. Conditioned medium from A. castellanii showed remarkable bactericidal properties against methicillin-resistant Staphylococcus aureus (MRSA) exhibiting almost 100% kill rate, but had limited effect against Acinetobacter sp., Pseudomonas aeruginosa and vancomycin-resistant Enterococcus faecalis (VRE). Similarly, the conditioned medium of E. coli K1 and Enterobacter sp., exhibited potent anti-Acanthamoebic effects in a concentration-dependent manner. Conditioned media of Acanthamoeba, E. coli K1 and Enterobacter sp. showed no cytotoxicity in vitro when tested against human brain microvascular endothelial cells. Active molecule/s in aforementioned amoebic and two bacterial conditioned media were 5 - 10 kDa, and <5 kDa respectively. A. castellanii conditioned medium showed potent bactericidal properties against MRSA. The active molecule(s) are heat- and pronase-resistant, and in the 5 to 10 kDa molecular mass range. Contrary to this, E. coli K1 and Enterobacter sp., conditioned medium showed anti-amoebic effects that are <5 kDa in molecular mass, suggestive of active metabolites.


Mesenteric vascular responsiveness in a rat model of pregnancy-induced hypertension.
Reduced perfusion to the placenta in early pregnancy is believed to be the initiating factor in the development of preeclampsia, triggering local ischemia and systemic vascular hyperresponsiveness. This sequence of events creates a predisposition to the development of altered vascular function and hypertension. This study was designed to determine the influence of placental insufficiency on the responsiveness of mesenteric resistance arteries in an animal model of preeclampsia. Placental insufficiency was induced by reduction in uteroplacental perfusion pressure (RUPP) in experimental Sprague-Dawley rat dams. The uterine branches of the ovarian arteries and the abdominal aortae of pregnant rats were surgically constricted on gestational Day 14. Dams in the control group underwent a sham procedure. Rats were euthanized on gestational Day 20, followed by removal of the small intestine and adjacent mesentery. First-order mesenteric resistance arteries were mounted on a small vessel wire myograph and challenged with incremental concentrations of vasoconstrictors and vasorelaxants. Mesenteric arteries in dams with placental insufficiency demonstrated an increased maximal tension to phenylephrine (7.15 +/- 0.15 vs. 5.4 +/- 0.27 mN/mm, P < 0.001); potassium chloride at 60 mM (3.43 +/- 0.11 vs. 2.77 +/- 0.14 mN/mm, P < 0.01) and 120 mM (3.92 +/- 0.18 vs. 2.97 +/- 0.16 mN/mm, P < 0.01); and angiotensin II (2.59 +/- 0.42 vs. 1.51 +/- 0.22 mN/mm, P < 0.05). Maximal relaxation to endothelium-dependent relaxants acetylcholine and calcium ionophore (A23187) was not significantly reduced. Data suggest that placental insufficiency leads to hyperresponsiveness to vasoconstrictor stimuli in mesenteric arteries.


Li substituent tuning of LED phosphors with enhanced efficiency, tunable photoluminescence, and improved thermal stability.
Solid-state phosphor-converted white light-emitting diodes (pc-WLEDs) are currently revolutionizing the lighting industry. To advance the technology, phosphors with high efficiency, tunable photoluminescence, and high thermal stability are required. Here, we demonstrate that a simple lithium incorporation in NaAlSiO4:Eu system enables the simultaneous fulfillment of the three criteria. The Li substitution at Al sites beside Na sites in NaAlSiO4:Eu leads to an enhanced emission intensity/efficiency owing to an effective Eu3+ to Eu2+ reduction, an emission color tuning from yellow to green by tuning the occupation of different Eu sites, and an improvement of luminescence thermal stability as a result of the interplay with Li-related defects. A pc-WLED using the Li-codoped NaAlSiO4:Eu as a green component exhibits improved performance. The phosphors with multiple activator sites can facilitate the positive synergistic effect on luminescence properties.


Health care utilization, somatic and mental health distress, and well-being among widowed and non-widowed female survivors of war.
The aim of the study was to assess levels of somatic and mental health distress, well-being, AS WELL AS utilization of primary and specialist health care services among war-related widowed and non-widowed female civilian survivors of war. 100 war-related widowed lone mothers and 106 non-widowed mothers who had experienced the Kosovo war ten years previously participated in the study. Measures of somatic, depressive, post-traumatic stress, anxiety, and grief complaints, subjective well-being, and utilization of health care services during the previous three months were used. Compared to non-widowed mothers, widowed lone mothers reported significantly higher levels of somatic, depressive, post-traumatic stress, and anxiety complaints. Further, they reported significantly lower levels of subjective well-being as composed of positive and negative affect and satisfaction with life. More than half of both widowed and non-widowed mothers reported utilization of health care services during the last three months, without significant differences between the groups. However, only three percent of widowed lone mothers and four percent of non-bereaved mothers reported utilization of mental health services during the last three months, despite high levels of mental health distress especially among widowed lone mothers. Among widowed lone mothers, severity of prolonged grief symptoms significantly predicted number of contacts of specialist health care use over and above sociodemographic variables, number of war-related events, and other psychopathology. War-related widowed lone mothers suffer from elevated somatic and mental distress even a decade after the war. The tiny proportion of widowed lone mothers in use of mental health services can be seen as a reflection of lack of previous and current mental health services to meet mental health needs of this population.


Optimized mtDNA Control Region Primer Extension Capture Analysis for Forensically Relevant Samples and Highly Compromised mtDNA of Different Age and Origin.
The analysis of mitochondrial DNA (mtDNA) has proven useful in forensic genetics and ancient DNA (aDNA) studies, where specimens are often highly compromised and DNA quality and quantity are low. In forensic genetics, the mtDNA control region (CR) is commonly sequenced using established Sanger-type Sequencing (STS) protocols involving fragment sizes down to approximately 150 base pairs (bp). Recent developments include Massively Parallel Sequencing (MPS) of (multiplex) PCR-generated libraries using the same amplicon sizes. Molecular genetic studies on archaeological remains that harbor more degraded aDNA have pioneered alternative approaches to target mtDNA, such as capture hybridization and primer extension capture (PEC) methods followed by MPS. These assays target smaller mtDNA fragment sizes (down to 50 bp or less), and have proven to be substantially more successful in obtaining useful mtDNA sequences from these samples compared to electrophoretic methods. Here, we present the modification and optimization of a PEC method, earlier developed for sequencing the Neanderthal mitochondrial genome, with forensic applications in mind. Our approach was designed for a more sensitive enrichment of the mtDNA CR in a single tube assay and short laboratory turnaround times, thus complying with forensic practices. We characterized the method using sheared, high quantity mtDNA (six samples), and tested challenging forensic samples (n = 2) as well as compromised solid tissue samples (n = 15) up to 8 kyrs of age. The PEC MPS method produced reliable and plausible mtDNA haplotypes that were useful in the forensic context. It yielded plausible data in samples that did not provide results with STS and other MPS techniques. We addressed the issue of contamination by including four generations of negative controls, and discuss the results in the forensic context. We finally offer perspectives for future research to enable the validation and accreditation of the PEC MPS method for final implementation in forensic genetic laboratories.


Methadone during pregnancy: assessment of behavioral effects in the rat offspring.
Pregnant rats were administered methadone hydrochloride by gastric intubation. Beginning on day 8 of gestation, a drug group received 5 mg/kg/day; the daily maintenance dose was increased to 10 mg/kg after 4 days with the final dose given on day 22. An intubation control group received sterile water alone on the same gestation days and a nontreated control group was left undisturbed, All experimental and control litters were fostered at birth to untreated mothers, Body weights were low at birth among the methadone offspring but were similar to controls by weaning. During adulthood, offspring were tested on several operant tasks that included acquisition of a lever-pressing response, performance on a variable interval reinforcement schedule, acquisition and performance of an auditory-visual discrimination and response inhibition in a "punishment" paradigm. None of these measures revealed any learning or inhibitory impairments among the methadone-exposed offspring. The major behavioral effect produced by the treatment was a high response output which emerged for each sex in different phases of testing.


Role of chiral quantum Hall edge states in nuclear spin polarization.
Resistively detected NMR (RDNMR) based on dynamic nuclear polarization (DNP) in a quantum Hall ferromagnet (QHF) is a highly sensitive method for the discovery of fascinating quantum Hall phases; however, the mechanism of this DNP and, in particular, the role of quantum Hall edge states in it are unclear. Here we demonstrate the important but previously unrecognized effect of chiral edge modes on the nuclear spin polarization. A side-by-side comparison of the RDNMR signals from Hall bar and Corbino disk configurations allows us to distinguish the contributions of bulk and edge states to DNP in QHF. The unidirectional current flow along chiral edge states makes the polarization robust to thermal fluctuations at high temperatures and makes it possible to observe a reciprocity principle of the RDNMR response. These findings help us better understand complex NMR responses in QHF, which has important implications for the development of RDNMR techniques.


Cue-exposure therapy to decrease alcohol craving in virtual environment.
During abstinence from alcohol, craving is elicited by the cues and contexts previously associated with alcohol, which contribute to relapse. To prevent the craving and relapse experienced by alcoholics, cue-exposure therapy (CET) has been used to extinguish the association between alcohol and alcohol-related cues and contexts. This study applied CET, using a virtual reality (VR) system, to eight members of an Alcoholics Anonymous group for eight sessions. Cues and contexts most likely to elicit an urge to drink were selected through a preliminary survey in order to compose VR-CET scenarios: a glass, a bottle, food, and a bar were judged to be the most tempting for people in alcohol dependence and abstinence. Using these cues and contexts, a Japanese-style pub and a western bar were created. Each session was administered for 30 minutes by a psychiatrist and included an introduction, immersion, VR navigation, interviews about feelings, and self-report questionnaires about cravings. The eight sessions consisted of initial and closing sessions and person-, object-, and situation-focused sessions. As a result, a reduction in cue-elicited craving after VR-CET was reported. A mean score of 15.75 (SD = 10.91) on the Alcohol Urge Questionnaire in the first session decreased to 11.50 (SD = 5.76) in the final session. This study suggests that using virtual reality can enhance the effectiveness of CET.


Age benefits in everyday prospective memory: the influence of personal task importance, use of reminders and everyday stress.
The present diary study examined everyday prospective memory tasks in younger and old adults and explored the role of personal task importance, use of reminders and everyday stress as possible correlates of age-related prospective memory performance in everyday life. Results revealed an age benefit in everyday prospective memory tasks. In addition, task importance was identified as a critical moderator of age-related prospective memory performance. More frequent use of reminders and lower levels of stress, however, were associated with better prospective memory performance in general but did not contribute to age-related prospective memory performance. Exploring further possible correlates of prospective memory revealed that the strategy to reprioritize initially planned intentions was associated with age benefits in everyday prospective memory. Results suggest that the age-related benefit observed in experimenter-given tasks transfers to everyday prospective memory and varies in dependence of motivational and cognitive factors. Implications for theoretical models of prospective memory and aging are discussed.


Factors associated with adolescent initiation of injection drug use.
The purpose of this study was to evaluate the extent to which demographic, sexual, and non-injection drug use practices predict adolescent initiation of injection drug use. Street recruited injection drug users 15-30 years of age in Baltimore, Maryland, who initiated injection within five years of study enrollment, completed a questionnaire that included a year-by-year history regarding the five years prior to initiation of injection. Factors associated with initiation during adolescence (< or = 21 years of age) versus young adulthood (>21 ) were determined using logistic regression. Of 226 participants, most were female (61%) and African American (64%). Median age of participants was 25; median age at initiation of injection was 23. Factors significantly associated with adolescent initiation in multivariate analysis included race other than African American, and practices prior to initiating injection including condom use, lack of cocaine use, exclusive crack smoking just prior to initiation, and smoking marijuana. Adolescent initiates also had shorter durations of illicit drug use prior to initiating injection. Short-term non-injection drug use, particularly exclusive crack smoking, was associated with adolescent initiation of injection drug use. Early prevention efforts targeting this high-risk group of younger drug users are warranted in order to delay or prevent onset of injection drug use.


Genomic polymorphism in symbiotic populations of Photobacterium leiognathi.
Photobacterium leiognathi forms a bioluminescent symbiosis with leiognathid fishes, colonizing the internal light organ of the fish and providing its host with light used in bioluminescence displays. Strains symbiotic with different species of the fish exhibit substantial phenotypic differences in symbiosis and in culture, including differences in 2-D PAGE protein patterns and profiles of indigenous plasmids. To determine if such differences might reflect a genetically based symbiont-strain/host-species specificity, we profiled the genomes of P. leiognathi strains from leiognathid fishes using PFGE. Individual strains from 10 species of leiognathid fishes exhibited substantial genomic polymorphism, with no obvious similarity among strains; these strains were nonetheless identified as P. leiognathi by 16S rDNA sequence analysis. Profiling of multiple strains from individual host specimens revealed an oligoclonal structure to the symbiont populations; typically one or two genomotypes dominated each population. However, analysis of multiple strains from multiple specimens of the same host species, to determine if the same strain types consistently colonize a host species, demonstrated substantial heterogeneity, with the same genomotype only rarely observed among the symbiont populations of different specimens of the same host species. Colonization of the leiognathid light organ to initiate the symbiosis therefore is likely to be oliogoclonal, and specificity of the P. leiognathi/leiognathid fish symbiosis apparently is maintained at the bacterial species level rather than at the level of individual, genomotypically defined strain types.


Droplet manipulation on a structured shape memory polymer surface.
While methods for dynamic tuning of surface wettability to manipulate water droplets have been widely explored for many applications including digital microfluidics, those based on dynamically changeable surface morphology have remained challenging to achieve. In this work, we present a structured shape memory polymer (SMP) surface which shows dynamically tunable surface wettability through changeable surface morphology in order to manipulate water droplets. The structured SMP surface involves a SMP pillar array consisting of nanotextured small and large pillars which can change its morphology between permanent and temporary shapes upon thermomechanical loading. Specifically, the structured SMP surface dynamically creates a surface morphological gradient and changes its surface wettability during thermally induced shape recovery of the SMP pillar array. Different wetting characteristics of the structured SMP surface between permanent and temporary shapes are theoretically predicted and experimentally verified. Based on these measured wetting characteristics, the structured SMP surface is designed to demonstrate that the morphological difference between two shapes under a water droplet overcomes contact angle hysteresis, resulting in driving a water droplet, when combined with the thermal Marangoni effect.


Analysis of the components of cancer risk perception and links with intention and behaviour: A UK-based study.
Risk perception refers to how individuals interpret their susceptibility to threats, and has been hypothesised as an important predictor of intentions and behaviour in many theories of health behaviour change. However, its components, optimal measurement, and effects are not yet fully understood. The TRIRISK model, developed in the US, conceptualises risk perception as deliberative, affective and experiential components. In this study, we aimed to assess the replicability of the TRIRISK model in a UK sample by confirmatory factor analysis (CFA), explore the inherent factor structure of risk perception in the UK sample by exploratory factor analysis (EFA), and assess the associations of EFA-based factors with intentions to change behaviour and subsequent behaviour change. Data were derived from an online randomised controlled trial assessing cancer risk perception using the TRIRISK instrument and intention and lifestyle measures before and after communication of cancer risk. In the CFA analysis, the TRIRISK model of risk perception did not provide a good fit for the UK data. A revised model developed using EFA consisted of two separate "numerical" and "self-reflective" factors of deliberative risk perception, and a third factor combining affective with a subset of experiential items. This model provided a better fit to the data when cross-validated. Using multivariable regression analysis, we found that the self-reflective and affective-experiential factors of the model identified in this study were reliable predictors of intentions to prevent cancer. There were no associations of any of the risk perception factors with behaviour change. This study confirms that risk perception is clearly a multidimensional construct, having identified self-reflective risk perception as a new distinct component with predictive validity for intention. Furthermore, we highlight the practical implications of our findings for the design of interventions incorporating risk perception aimed at behaviour change in the context of cancer prevention.


The complexity of HLA class II (DRB1, DQB1, DM) associations with disseminated Mycobacterium avium complex infection among HIV-1-seropositive whites.
Earlier associations of polymorphism in classic HLA class II (DRB1 and DQB1) genes have been extended to include the accessory genes DMA and DMB as determinants of disseminated Mycobacterium avium complex (DMAC) infection among HIV-1-seropositive whites. From the Multicenter AIDS Cohort study, 176 DMAC cases were matched with 176 controls in a nested case-control study. PCR-based HLA genotyping techniques were used to resolve variants of DRB1 and DQB1 to their four-digit or five-digit alleles, and single-strand conformation polymorphism was used to resolve sequences in exon 3 at each DM locus. The DMA*0102 allele occurred less frequently among DMAC cases than among controls (OR = 0.46, p =.02). Combinations of DRB1 alleles with or without specific DMA and DMB variants showed significant differences in distributions between the cases and controls, but both of the previously associated class II alleles (DRB1*1501 and DRB1*0701) showed stronger positive associations with DMAC in the absence than in the presence of DMA*0102. Apparent joint effects of DRB1 and DM allelic combinations on occurrence and timing of DMAC suggest that class II disease relationships may be better predicted by biologically plausible interactive combinations than by polymorphisms in individual genes.


Topical MMP beacon enabled fluorescence-guided resection of oral carcinoma.
Each year almost 300,000 individuals worldwide are diagnosed with oral cancer, more than 90% of these being oral carcinoma [N. Engl. J. Med.328, 1841993]. Surgical resection is the standard of care, but accurate delineation of the tumor boundaries is challenging, resulting in either under-resection with risk of local recurrence or over-resection with increased functional loss and negative impact on quality of life. This study evaluates, in two pre-clinical in vivo tumor models, the potential of fluorescence-guided resection using molecular beacons activated by metalloproteinases, which are frequently upregulated in human oral cancer. In both models there was rapid (<15 min) beacon activation upon local application, allowing clear fluoresecence imaging in vivo and confirmed by ex vivo fluorescence microscopy and HPLC, with minimal activation in normal oral tissues. Although the tissue penetration was limited using topical application, these findings support further development of this approach towards translation to first-in-human trials.


Characterization of a helical protein designed from first principles.
The question of how the primary amino acid sequence of a protein determines its three-dimensional structure is still unanswered. One approach to this problem involves the de novo design of model peptides and proteins that should adopt desired three-dimensional structures. A systematic approach was aimed at the design of a four-helix bundle protein. The gene encoding the designed protein was synthesized and the protein was expressed in Escherichia coli and purified to homogeneity. The protein was shown to be monomeric, highly helical, and very stable to denaturation by guanidine hydrochloride (GuHCl). Thus a globular protein has been designed that is capable of adopting a stable, folded structure in aqueous solution.


INDUCED RESISTANCE OF THE CENTRAL NERVOUS SYSTEM TO EXPERIMENTAL INFECTION WITH EQUINE ENCEPHALOMYELITIS VIRUS : III. ABORTIVE INFECTION WITH WESTERN VIRUS AND SUBSEQUENT INTERFERENCE WITH THE ACTION OF HETEROLOGOUS VIRUSES.
Although vaccination of guinea pigs with formalin-inactivated Western equine encephalomyelitis virus rendered them specifically immune to an intracerebral challenge dose of 1,000 M.L.D. of Western virus, it failed to protect their central nervous system against the initial effects of the virus: the intracerebral challenge dose was followed by an abortive infection of 20 to 30 hours' duration characterized by fever and histopathological changes which simulated the response at that early stage of non-vaccinated control animals. During the abortive infection of immune animals, virus could occasionally be demonstrated in their brains; indeed, it was detected with about the same frequency it was isolated from brains of similarly inoculated, non-immune guinea pigs during corresponding early phases of the infection. About one week after the abortive infection there was found a marked transitory accumulation of specific neutralizing antibody in the brain tissue. See PDF for Equation equalled at this time 1:1 to 1:10 instead of the value of about 1:300 found under physiological conditions. Guinea pigs which had recovered from an abortive infection with Western virus were resistant for a limited period of time to the effects of intracerebral inoculations of the immunologically distinct viruses of Eastern equine encephalomyelitis or vesicular stomatitis.


Sex chromosome trisomies in Europe: prevalence, prenatal detection and outcome of pregnancy.
This study aims to assess prevalence and pregnancy outcome for sex chromosome trisomies (SCTs) diagnosed prenatally or in the first year of life. Data held by the European Surveillance of Congenital Anomalies (EUROCAT) database on SCT cases delivered 2000-2005 from 19 population-based registries in 11 European countries covering 2.5 million births were analysed. Cases included were livebirths diagnosed to 1 year of age, fetal deaths from 20 weeks gestation and terminations of pregnancy for fetal anomaly (TOPFA). In all, 465 cases of SCT were diagnosed between 2000 and 2005, a prevalence of 1.88 per 10,000 births (95% CI 1.71-2.06). Prevalence of XXX, XXY and XYY were 0.54 (95% CI 0.46-0.64), 1.04 (95% CI 0.92-1.17) and 0.30 (95% CI 0.24-0.38), respectively. In all, 415 (89%) were prenatally diagnosed and 151 (36%) of these resulted in TOPFA. There was wide country variation in prevalence (0.19-5.36 per 1000), proportion prenatally diagnosed (50-100%) and proportion of prenatally diagnosed resulting in TOPFA (13-67%). Prevalence of prenatally diagnosed cases was higher in countries with high prenatal detection rates of Down syndrome. The EUROCAT prevalence rate for SCTs diagnosed prenatally or up to 1 year of age represents 12% of the prevalence expected from cytogenetic studies of newborn babies, as the majority of cases are never diagnosed or are diagnosed later in life. There is a wide variation between European countries in prevalence, prenatal detection and TOPFA proportions, related to differences in screening policies as well as organizational and cultural factors.


Use of mobile learning by resident physicians in Botswana.
With the growth of mobile health in recent years, learning through the use of mobile devices (mobile learning [mLearning]) has gained recognition as a potential method for increasing healthcare providers' access to medical information and resources in resource-limited settings. In partnership with the University of Botswana School of Medicine (SOM), we have been exploring the role of smartphone-based mLearning with resident (physicians in specialty training) education. The SOM, which admitted its first class of medical students and residents in 2009, is committed to providing high-level on-site educational resources for resident physicians, even when practicing in remote locations. Seven residents were trained to use an Android-based myTouch 3G smartphone equipped with data-enabled subscriber identity module (SIM) cards and built-in camera. Phones contained locally loaded point-of-care and drug information applications, a telemedicine application that allows for the submission of cases to local mentors, and e-mail/Web access. Surveys were administered at 4 weeks and 8 weeks following distribution of phones. We found that smartphones loaded with point-of-care tools are effectively utilized by resident physicians in resource-limited settings, both for accessing point-of-care medical information at the bedside and engaging in self-directed learning at home.


The genetics of clinical outcome in multiple sclerosis.
Multiple sclerosis (MS) is a common inflammatory disease of the central nervous system (CNS), the clinical course of which varies considerably between patients. Genetic complexity and interactions with as yet unknown environmental factors have hindered researchers from fully elucidating the aetiology of the disease. In addition to influencing disease susceptibility, epidemiological evidence suggests that genetic factors may affect phenotypic expression of the disease. Genes that affect clinical outcome may be more effective therapeutic targets than those which determine susceptibility. We present in this review a comprehensive survey of the genes (both MHC- and non-MHC-related) that have been investigated for their role in disease outcome in MS. Recent studies implicating the role of the genotype and epistatic interactions in the MHC in determining outcome are highlighted.


Harnessing the potential of practice-based clinical optometry research in the United Kingdom.
Research is the core of evidence-based practice across all healthcare, in order to ensure optimum patient care. The College of Optometrists is a national standard setting institution for optometric practice in the United Kingdom. However, the standards are only as good as the available evidence, and currently there is little evidence relating directly to optometric practice. The National Institute of Health and Care Research, the General Medical Council and The College of Optometrists, amongst others, have published research strategies describing ambitious plans to expand the scope of healthcare research. The aim of this article is to raise awareness of these government initiatives and consider how they may relate to optometric practice. To improve optometrist research engagement, we need to address the barriers to research and implement strategies to overcome them. There are many opportunities to support research, with different degrees of involvement, from signposting patients to research studies, supporting recruitment or collecting data for a multicentre clinical trial, as well as undertaking an individual research project. Healthcare research is changing and there is scope for more practice-based research activities in optometry. Research should not be a solo endeavour but a multi-disciplinary effort. Greater collaborations across all stakeholders, including primary care, secondary care, academia, regulators and industry is needed to make this possible.


Structure of Ralstonia eutropha flavohemoglobin in complex with three antibiotic azole compounds.
Flavohemoglobins (flavoHbs) are enzymes that operate primarily as nitric oxide dioxygenases and shuttle thereby electrons among NAD(P)H, FAD, heme, and a ligated redox-active substrate such as O(2). They function in the bacterial defense against nitrosative stress and are therefore considered as targets for new antibiotic drugs. Recently, azole derivatives were proven to be attractive nitric oxide dioxygenase inhibitors, and to explore their binding characteristics, we determined the X-ray structure of the flavoHb from Ralstonia eutropha in a complex with miconazole (FHP(M)), econazole (FHP(E)), and ketoconazole (FHP(K)). In agreement with UV-vis spectroscopic data, one azole compound binds inside the distal heme pocket and ligates to the heme iron by its imidazole substituent. The two additional substituents, mostly chlorinated phenyl groups, form a series of van der Waals contacts with the protein matrix. Both interactions explain their high affinity for flavoHbs, the binding constants being 2.6, 1.2, and 11.6 μM for miconazole, econazole, and ketoconazole, respectively. The FHP(M) and FHP(Lip) (flavoHbs originally loaded with a phospholipid) structures share an "open" state and the FHP(E) and FHP(K) structures a "closed" state. Although the azole compounds were able to push the lipid out of its binding site, a fatty acid fragment is still bound inside the heme pocket of FHP(E) and FHP(K) and dictates the state of the protein. The ligand-induced open-to-closed transition involves a reorientation of the NADH domain accompanied by conformational changes in the C-terminal arm, helix E, and the CE loop resulting in an encapsulation of the heme-binding pocket. Implications of the observed open-to-closed process on the catalytic cycle are discussed.


Pharmaceutical Treatment for Alzheimer's Disease and Related Dementias: Utilization and Disparities.
Four prescription drugs (donepezil, galantamine, memantine, and rivastigmine) are approved by the US FDA to treat symptoms of Alzheimer's disease (AD). Even modest effectiveness could potentially reduce the population-level burden of AD and related dementias (ADRD), especially for women and racial/ethnic minorities who have higher incidence of ADRD. Describe the prevalence of antidementia drug use and timing of initiation relative to ADRD diagnosis among a nationally representative group of older Americans, and if there are disparities in prevalence and timing by sex and race/ethnicity. Descriptive analyses and logistic regressions of Medicare claims (2008-2016) for beneficiaries who had an ADRD or dementia-related symptom diagnosis, or use of an FDA approved drug for AD. We investigate prevalence of use and timing of treatment initiation relative to ADRD diagnosis across time and beneficiary characteristics (age, sex, race/ethnicity, socioeconomic status, comorbidities). Among persons diagnosed with ADRD or related symptoms, 33.3% used an approved drug over the study period. Odds of use was higher among Whites than non-Whites. Among ADRD drug users, 40% initiated use within 6 months of the initial ADRD or related symptoms diagnosis, and 16% initiated prior to a diagnosis. We observed disparities by race/ethnicity: 28% of Asians, 24% of Hispanics, 16% of Blacks, and 15% of Whites initiated prior to diagnosis. The use of antidementia drugs is relatively low and varies widely by race/ethnicity. Heterogeneity in timing of initiation and use may affect health and cost outcomes, but these effects merit further study.


Exploring the immunomodulatory and anticancer properties of zerumbone.
Plant-derived immunomodulators and anti-cancer agents have attracted a lot of interest from natural product scientists for their efficacy and safety and their significant contribution towards understanding targeted drug action and drug delivery mechanisms. Zerumbone, the main constituent of Zingiber zerumbet rhizomes, has been investigated for its wide-spectrum role in treating multitargeted diseases. The rhizomes have been used as food flavoring agents in various cuisines and in herbal medicine. Many in vivo and in vitro studies have provided evidence of zerumbone as a potent immunomodulator as well as a potential anti-cancer agent. This review is an interesting compilation of all those significant outcomes from investigations carried out to date to explore the immunomodulatory and anticancer properties of zerumbone. The ultimate objective of this comprehensive review is to provide updated information and a critical assessment on zerumbone including its chemistry and immunomodulating and anticancer properties, which may be of paramount importance to provide a new path for ensuing research to discover new agents to treat cancers and immune-related diseases. In addition, updated information on the toxicology of zerumbone has also been summarized to provide its safety profile.


Pesticides and pollinators: A socioecological synthesis.
The relationship between pesticides and pollinators, while attracting no shortage of attention from scientists, regulators, and the public, has proven resistant to scientific synthesis and fractious in matters of policy and public opinion. This is in part because the issue has been approached in a compartmentalized and intradisciplinary way, such that evaluations of organismal pesticide effects remain largely disjoint from their upstream drivers and downstream consequences. Here, we present a socioecological framework designed to synthesize the pesticide-pollinator system and inform future scholarship and action. Our framework consists of three interlocking domains-pesticide use, pesticide exposure, and pesticide effects-each consisting of causally linked patterns, processes, and states. We elaborate each of these domains and their linkages, reviewing relevant literature and providing empirical case studies. We then propose guidelines for future pesticide-pollinator scholarship and action agenda aimed at strengthening knowledge in neglected domains and integrating knowledge across domains to provide decision support for stakeholders and policymakers. Specifically, we emphasize (1) stakeholder engagement, (2) mechanistic study of pesticide exposure, (3) understanding the propagation of pesticide effects across levels of organization, and (4) full-cost accounting of the externalities of pesticide use and regulation. Addressing these items will require transdisciplinary collaborations within and beyond the scientific community, including the expertise of farmers, agrochemical developers, and policymakers in an extended peer community.


Spiritual stress and coping model of divorce: a longitudinal study.
This study represents the first longitudinal effort to use a spiritual stress and coping model to predict adults' psychosocial adjustment following divorce. A community sample of 89 participants completed measures at the time of their divorce and 1 year later. Though the sample endorsed slightly lower levels of religiosity than the general U.S. population, most reported spiritual appraisals and positive and negative religious coping tied to divorce. Hierarchical regression analyses controlling general religiousness and nonreligious forms of coping indicated that (a) appraising divorce as a sacred loss or desecration at the time it occurred predicted more depressive symptoms and dysfunctional conflict tactics with the ex-spouse 1 year later; (b) positive religious coping reported about the year following divorce predicted greater posttraumatic growth 1 year after divorce; and (c) negative religious coping reported about the year following divorce predicted more depressive symptoms 1 year after the divorce. Bootstrapping mediation analyses indicated that negative religious coping fully mediated links between appraising the divorce as a sacred loss or desecration at the time it occurred and depressive symptoms 1 year later. In addition, moderation analyses revealed that negative religious coping is more strongly associated with depressive symptoms among those who form high versus low appraisals of their divorce as a sacred loss or desecration. These findings are relevant to divorce education and intervention provided by professionals in legal, family, mental health, and clerical roles. Implications are discussed for clinical and counseling psychology and religious communities.


Climate feedbacks at the tundra-taiga interface.
Feedbacks, or internal interactions, play a crucial role in the climate system. Negative feedback will reduce the impact of an external perturbation, a positive feedback will amplify the effect and could lead to an unstable system. Many of the feedbacks found in the climate system are positive; thus, for example, increasing CO2 levels will increase temperature, reduce the snow cover, increase the absorption of radiation and hence increase temperature further. The most obvious feedbacks, such as the snow example quoted above, are already included within our models of the climate and earth system. Others, such as the impact of increasing forest cover due to global warming, are only just being included. Others, such as, the impact of global warming on the northern peatlands and the impact of freshwater flows on the Arctic Ocean are not yet considered. The contrast in surface characteristics between low tundra vegetation to high taiga forest is considerable. The contrast is greatest in the winter, when the tundra is snow covered but the trees of the taiga protrude through the snow pack, and is probably the greatest contrast found on the land surface anywhere. This variation causes massive changes in the energy fluxes at the surface and hence the temperature conditions on the ground and within the atmosphere. There will be large resultant changes in the vegetation development, the carbon fluxes, the permafrost and the hydrology. The Arctic is already experiencing change and it is essential for us to understand the basic processes, and how these interact, to be confident of our predictions of environmental change in the future.


Sex differences in the persistence of natal environmental effects on life histories.
"Silver spoon" effects of early environments on adult characteristics and life-history trajectories are thought to be widespread among vertebrates and are important because they imply that environments can have cascading transgenerational effects. Here we test for such effects by using a sample of 5425 great tits (Parus major) for which both natal and breeding environments were known in detail. Female life histories were strongly coupled with breeding conditions, but we found little evidence for any persistent effects of the natal environment. In contrast, life span and breeding success in males were influenced by the quality of the environment in which they were raised. Potential explanations for persistent environmental effects in males, but not females, include differential sex allocation benefiting males from high-quality environments and phenotypic plasticity allowing females to optimize reproductive decisions to current conditions, hence masking any residual natal effects. Thus, in contrast to findings of much recent work, persistent effects of the early environment are not all-pervasive and may differ between the sexes within a single species, potentially leading to sex-differential selection. Further work should attempt to understand the conditions under which these transgenerational environmental effects are likely to occur.


Timely Diagnosis of Acute Kidney Injury Using Kinetic eGFR and the Creatinine Excretion to Production Ratio, E/eG - Creatinine Can Be Useful!
Post transplant repeated measurements of urine volume and serum creatinine (sCr) are used to assess kidney function. Under non-steady state conditions, repeated measurement of sCr allows calculation of the kinetic estimated GFR (KeGFR). Additional measurement of urinary creatinine allows the calculation of the creatinine excretion to (estimated) production ratio (E/eG). We hypothesized that post-transplant KeGFR and E/eG would predict delayed graft function (DGF), as early as 4 h and outperform a validated clinical model at 12 h. This was a retrospective analysis of prospectively acquired data in a study of 56 recipients of deceased-donor kidney transplant. We assessed predictive performance with the area under the receiver operator characteristic curve (AUC) and the added value to a clinical model with integrated discrimination improvement analysis. At 4 h, the AUC for E/eG was 0.87 (95% CI 0.77-0.96) and for KeGFR 0.69 (95% CI 0.56-0.83). Both E/eG and KeGFR improved the risk prediction of a clinical model for DGF by 32 and 18%, and for non-DGF by 17 and 10%, respectively. While E/eG had better predictive performance of DGF than KeGFR, KeGFR might also facilitate perioperative management including drug dosing after kidney transplantation. Together these measurements may facilitate the possibility of conducting trials of early intervention to ameliorate the adverse effects of ischaemia-reperfusion injury on long-term DGF.


FOXM1 induces a global methylation signature that mimics the cancer epigenome in head and neck squamous cell carcinoma.
The oncogene FOXM1 has been implicated in all major types of human cancer. We recently showed that aberrant FOXM1 expression causes stem cell compartment expansion resulting in the initiation of hyperplasia. We have previously shown that FOXM1 regulates HELLS, a SNF2/helicase involved in DNA methylation, implicating FOXM1 in epigenetic regulation. Here, we have demonstrated using primary normal human oral keratinocytes (NOK) that upregulation of FOXM1 suppressed the tumour suppressor gene p16(INK4A) (CDKN2A) through promoter hypermethylation. Knockdown of HELLS using siRNA re-activated the mRNA expression of p16(INK4A) and concomitant downregulation of two DNA methyltransferases DNMT1 and DNMT3B. The dose-dependent upregulation of endogenous FOXM1 (isoform B) expression during tumour progression across a panel of normal primary NOK strains (n = 8), dysplasias (n = 5) and head and neck squamous cell carcinoma (HNSCC) cell lines (n = 11) correlated positively with endogenous expressions of HELLS, BMI1, DNMT1 and DNMT3B and negatively with p16(INK4A) and involucrin. Bisulfite modification and methylation-specific promoter analysis using absolute quantitative PCR (MS-qPCR) showed that upregulation of FOXM1 significantly induced p16(INK4A) promoter hypermethylation (10-fold, P<0.05) in primary NOK cells. Using a non-bias genome-wide promoter methylation microarray profiling method, we revealed that aberrant FOXM1 expression in primary NOK induced a global hypomethylation pattern similar to that found in an HNSCC (SCC15) cell line. Following validation experiments using absolute qPCR, we have identified a set of differentially methylated genes, found to be inversely correlated with in vivo mRNA expression levels of clinical HNSCC tumour biopsy samples. This study provided the first evidence, using primary normal human cells and tumour tissues, that aberrant upregulation of FOXM1 orchestrated a DNA methylation signature that mimics the cancer methylome landscape, from which we have identified a unique FOXM1-induced epigenetic signature which may have clinical translational potentials as biomarkers for early cancer screening, diagnostic and/or therapeutic interventions.


Living side by side with a virus: characterization of two novel plasmids from Thermococcus prieurii, a host for the spindle-shaped virus TPV1.
Microbial cells often serve as an evolutionary battlefield for different types of mobile genetic elements, such as viruses and plasmids. Here, we describe the isolation and characterization of two new archaeal plasmids which share the host with the spindle-shaped Thermococcus prieurii virus 1 (TPV1). The two plasmids, pTP1 and pTP2, were isolated from the hyperthermophilic archaeon Thermococcus prieurii (phylum Euryarchaeota), a resident of a deep-sea hydrothermal vent located at the East Pacific Rise at 2,700-m depth (7°25'24 S, 107°47'66 W). pTP1 (3.1 kb) and pTP2 (2.0 kb) are among the smallest known plasmids of hyperthermophilic archaea, and both are predicted to replicate via the rolling-circle mechanism. The two plasmids and the virus TPV1 do not have a single gene in common and stably propagate in infected cells without any apparent antagonistic effect on each other. The compatibility of the three genetic elements and the high copy number of pTP1 and pTP2 plasmids (50 copies/cell) might be useful for developing new genetic tools for studying hyperthermophilic euryarchaea and their viruses.


En bloc resection for lung cancer with chest wall invasion.
The aim of this study was to retrospectively assess the results of en bloc chest wall plus lung resection for patients with non-small cell lung cancer (NSCLC) invading the chest wall. From January 1986 to December 2000, of 1,820 patients having surgery for NSCLC, 42 (2.3%) patients with neoplasms involving the chest wall underwent en bloc chest wall and lung resection. Patient demographics, preoperative symptoms, operative procedures, tumor cell type and size, removed nodal status, and pathologic stage were summarized. The 5-year survival rates of the groups were compared. Postoperative staging revealed 28 were T3N0M0, 4 were T3N1M0, and 10 were T3N2M0. The in-hospital mortality rate was 11.9% (5/42). The mean age was 79.0 +/- 2.8 years in the patients who died of complications, which was significantly older than the mean age of 67.9 +/- 8.1 years in the patients who survived the surgery (p = 0.005). The overall 5-year survival was 28.4%. The 5-year survival was significantly longer in the patients with negative (N0) nodal metastasis than in those with N1 and/or N2 nodal metastasis (39.6% versus 7.1%, p = 0.01). Eleven patients had tumor involvement of the parietal pleura. Thirty-one patients had tumor involvement of the soft tissue and/or bone. There was no significant difference of 5-year survival rate between the patients with involvement of the parietal pleura only and the patients with involvement of the parietal pleura and the soft tissue and/or bone (10.9% versus 33.5%, p = 0.94). En bloc resection for bronchogenic carcinoma invading the chest wall provides a favorable prognosis in cases without nodal metastasis. Significant postoperative mortality is associated with old age (> 80 years). The 5-year survival rate is not significantly different between the patients with involvement of the parietal pleura only and the patients with involvement of the parietal pleura and the soft tissue and/or bone.


Biosynthetic characterization and chemoenzymatic assembly of the cryptophycins. Potent anticancer agents from cyanobionts.
The lichen cyanobacterial symbiont Nostoc sp. ATCC 53789 and its close relative Nostoc sp. GSV 224 are prolific producers of natural products, generating >25 derivatives of the cryptophycin class of secondary metabolites. Cryptophycin 1, the prototypic member of the class, is a potent tubulin-depolymerizing agent, and several semisynthetic derivatives are being developed as anticancer therapeutics. Here we provide a detailed characterization of the cryptophycin metabolic pathway by stable-isotope labeling experiments and through cloning, sequencing, and annotating the cryptophycin biosynthetic gene cluster. A comparative secondary metabolomic analysis based on polyketide (PK)/non-ribosomal peptide gene clusters from the phylogenetically related, non-cryptophycin producing cycad symbiont, Nostoc punctiforme ATCC 29133, was used to identify the cryptophycin biosynthetic genes that encompass approximately 40 kb within the lichen symbiont Nostoc sp. ATCC 53789 genome. The pathway encodes a collinear set of enzymes, including three modular PK synthases, two non-ribosomal peptide synthetase modules, and an integrated adenylation/ketoreductase didomain for elaboration of the leucic acid subunit. In addition, genes encoding key tailoring steps, including a FAD-dependent halogenase and CYP450 epoxidase, were identified. The inherent flexibility of the cryptophycin biosynthetic enzymes was harnessed to generate a suite of new analogues by altering the pool of PK starter units and selected amino acid extender groups. Characterization of the cryptophycin CYP450 enabled development of the first stereospecific synthesis of cryptophycin 2, through a tandem chemoenzymatic synthesis from the natural seco-cryptophycin 4 chain elongation intermediate.


Cutaneous microdialysis: cytokine evidence for altered innate reactivity in the skin of psoriasis patients?
Cutaneous microdialysis demonstrates cytokine production in living human skin. In the present study, microdialysis samples taken from uninvolved and lesional skin in three test subjects with psoriasis over 24 h have been investigated for cytokine content with a bead-based multiplex immunoassay from Luminex. Concentration curves for a set of Th1/Th2 and pro-inflammatory cytokines measured differed from a reference group of ten subjects without psoriasis. The time to return to near baseline values after innate insertion reactivity is between 9 and 16 h. Post-equilibration levels (17-24 h) for the three main cytokines elevated in the reference group were differentially elevated outside the range of the reference group for interleukin-1β (IL1β) and IL8 but not so for IL6. Two further cytokines, granulocyte-macrophage colony-stimulating factor and tumor necrosis factor-α not generally elevated in the reference group, showed elevated values in the test subjects. Multivariate time series analysis (chemometry) showed that cytokine patterns for the individual test subjects often fell outside the 99% confidence intervals of a model generated from the reference group. In a clinical research situation, cutaneous microdialysis is feasible, gives generally higher cytokine levels than in the blood and generates interpretable data on an individual's reactivity compared with a reference group. This may well prove useful in delineation of pathogenetic issues, selection of appropriate therapy and monitoring of subsequent response in inflammatory dermatoses such as psoriasis.


Non-invasive assessment of vasospasm following aneurysmal SAH using C-arm FDCT parenchymal blood volume measurement in the neuro-interventional suite: Technical feasibility.
Cerebral vasospasm is the leading cause of morbidity and mortality in patients with aneurysmal subarachnoid haemorrhage (SAH) surviving the initial ictus. Commonly used techniques for vasospasm assessment are digital subtraction angiography and transcranial Doppler sonography. These techniques can reliably identify only the major vessel spasm and fail to estimate its haemodynamic significance. To overcome these issues and to enable comprehensive non-invasive assessment of vasospasm inside the interventional suite, a novel protocol involving measurement of parenchymal blood volume (PBV) using C-arm flat detector computed tomography (FDCT) was implemented. Patients from the neuro-intensive treatment unit (ITU) with suspected vasospasm following aneurysmal SAH were scanned with a biplane C-arm angiography system using an intravenous contrast injection protocol. The PBV maps were generated using prototype software. Contemporaneous clinically indicated MR scan including the diffusion- and perfusion-weighted sequences was performed. C-arm PBV maps were compared against the MR perfusion maps. Distribution of haemodynamic impairment on C-arm PBV maps closely matched the pattern of abnormality on MR perfusion maps. On visual comparison between the two techniques, the extent of abnormality indicated PBV to be both cerebral blood flow and cerebral blood volume weighted. C-arm FDCT PBV measurements allow an objective assessment of the severity and localisation of cerebral hypoperfusion resulting from vasospasm. The technique has proved feasible and useful in very sick patients after aneurysmal SAH. The promise shown in this early study indicates that it deserves further evaluation both for post-SAH vasospasm and in other relevant clinical settings.


Mobile Telephone Text Messaging for Medication Adherence in Chronic Disease: A Meta-analysis.
Adherence to long-term therapies in chronic disease is poor. Traditional interventions to improve adherence are complex and not widely effective. Mobile telephone text messaging may be a scalable means to support medication adherence. To conduct a meta-analysis of randomized clinical trials to assess the effect of mobile telephone text messaging on medication adherence in chronic disease. MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials, PsycINFO, and CINAHL (from database inception to January 15, 2015), as well as reference lists of the articles identified. The data were analyzed in March 2015. Randomized clinical trials evaluating a mobile telephone text message intervention to promote medication adherence in adults with chronic disease. Two authors independently extracted information on study characteristics, text message characteristics, and outcome measures as per the predefined protocol. Odds ratios and pooled data were calculated using random-effects models. Risk of bias and study quality were assessed as per Cochrane guidelines. Disagreement was resolved by consensus. Sixteen randomized clinical trials were included, with 5 of 16 using personalization, 8 of 16 using 2-way communication, and 8 of 16 using a daily text message frequency. The median intervention duration was 12 weeks, and self-report was the most commonly used method to assess medication adherence. In the pooled analysis of 2742 patients (median age, 39 years and 50.3% [1380 of 2742] female), text messaging significantly improved medication adherence (odds ratio, 2.11; 95% CI, 1.52-2.93; P < .001). The effect was not sensitive to study characteristics (intervention duration or type of disease) or text message characteristics (personalization, 2-way communication, or daily text message frequency). In a sensitivity analysis, our findings remained robust to change in inclusion criteria based on study quality (odds ratio, 1.67; 95% CI, 1.21-2.29; P = .002). There was moderate heterogeneity (I2 = 62%) across clinical trials. After adjustment for publication bias, the point estimate was reduced but remained positive for an intervention effect (odds ratio, 1.68; 95% CI, 1.18-2.39). Mobile phone text messaging approximately doubles the odds of medication adherence. This increase translates into adherence rates improving from 50% (assuming this baseline rate in patients with chronic disease) to 67.8%, or an absolute increase of 17.8%. While promising, these results should be interpreted with caution given the short duration of trials and reliance on self-reported medication adherence measures. Future studies need to determine the features of text message interventions that improve success, as well as appropriate patient populations, sustained effects, and influences on clinical outcomes.


The diversity of subunit composition in nAChRs: evolutionary origins, physiologic and pharmacologic consequences.
Nicotinic acetylcholine receptors are made up of homologous subunits, which are encoded by a large multigene family. The wide number of receptor oligomers generated display variable pharmacological properties. One of the main questions underlying research in molecular pharmacology resides in the actual role of this diversity. It is generally assumed that the observed differences between the pharmacology of homologous receptors, for instance, the EC(50) for the endogenous agonist, or the kinetics of desensitization, bear some kind of physiologic relevance in vivo. Here we develop the quite challenging point of view that, at least within a given subfamily of nicotinic receptor subunits, the pharmacologic variability observed in vitro would not be directly relevant to the function of receptor proteins in vivo. In vivo responses are not expected to be sensitive to mild differences in affinities, and several examples of functional replacement of one subunit by another have been unravelled by knockout animals. The diversity of subunits might have been conserved through evolution primarily to account for the topologic diversity of subunit distribution patterns, at the cellular and subcellular levels. A quantitative variation of pharmacological properties would be tolerated within a physiologic envelope, as a consequence of a near-neutral genetic drift. Such a "gratuitous" pharmacologic diversity is nevertheless of practical interest for the design of drugs, which would specifically tackle particular receptor oligomers with a defined subunit composition among the multiple nicotinic receptors present in the organism.


[The effect of hypoxia on the branchial vascular system of the perfused trout head].
Measurements of inflow (VE) and arterial (Vsa) and venous (VSv) outflow in trout perfused head preparations show significant decreases of VE and VSa during low PO2 saline perfusion (20 torr). Comparable results are gained during adenosine perfusion (1.10(-8) to (1.10(-6) M). Theophylline (10(-4) M) inhibits both hypoxia and adenosine effects.


Factorial comparison of working memory models.
Three questions have been prominent in the study of visual working memory limitations: (a) What is the nature of mnemonic precision (e.g., quantized or continuous)? (b) How many items are remembered? (c) To what extent do spatial binding errors account for working memory failures? Modeling studies have typically focused on comparing possible answers to a single one of these questions, even though the result of such a comparison might depend on the assumed answers to both others. Here, we consider every possible combination of previously proposed answers to the individual questions. Each model is then a point in a 3-factor model space containing a total of 32 models, of which only 6 have been tested previously. We compare all models on data from 10 delayed-estimation experiments from 6 laboratories (for a total of 164 subjects and 131,452 trials). Consistently across experiments, we find that (a) mnemonic precision is not quantized but continuous and not equal but variable across items and trials; (b) the number of remembered items is likely to be variable across trials, with a mean of 6.4 in the best model (median across subjects); (c) spatial binding errors occur but explain only a small fraction of responses (16.5% at set size 8 in the best model). We find strong evidence against all 6 documented models. Our results demonstrate the value of factorial model comparison in working memory.


The relationship between leaf rolling and ascorbate-glutathione cycle enzymes in apoplastic and symplastic areas of Ctenanthe setosa subjected to drought stress.
The ascorbate-glutathione (ASC-GSH) cycle has an important role in defensive processes against oxidative damage generated by drought stress. In this study, the changes that take place in apoplastic and symplastic ASC-GSH cycle enzymes of the leaf and petiole were investigated under drought stress causing leaf rolling in Ctenanthe setosa (Rose.) Eichler (Marantaceae). Apoplastic and symplastic extractions of leaf and petiole were performed at different visual leaf rolling scores from 1 to 4 (1 is unrolled, 4 is tightly rolled and the others are intermediate forms). Glutathione reductase (GR), a key enzyme in the GSH regeneration cycle, and ascorbate (ASC) were present in apoplastic spaces of the leaf and petiole, whereas dehydroascorbate reductase (DHAR), which uses glutathione as reductant, monodehydroascorbate reductase (MDHAR), which uses NAD(P)H as reductant, and glutathione were absent. GR, DHAR and MDHAR activities increased in the symplastic and apoplastic areas of the leaf. Apoplastic and symplastic ASC and dehydroascorbate (DHA), the oxidized form of ascorbate, rose at all scores except score 4 of symplastic ASC in the leaf. On the other hand, while reduced glutathione (GSH) content was enhanced, oxidized glutathione (GSSG) content decreased in the leaf during rolling. As for the petiole, GR activity increased in the apoplastic area but decreased in the symplastic area. DHAR and MDHAR activities increased throughout all scores, but decreased to the score 1 level at score 4. The ASC content of the apoplast increased during leaf rolling. Conversely, symplastic ASC content increased at score 2, however decreased at the later scores. While the apoplastic DHA content declined, symplastic DHA rose at score 2, but later was down to the level of score 1. While GSH content enhanced during leaf rolling, GSSG content did not change except at score 2. As well, there were good correlations between leaf rolling and ASC-GSH cycle enzyme activities in the leaf (GR and DHAR) and leaf rolling and GSSG. These results showed that in apoplastic and symplastic areas, ASC-GSH cycle enzymes leading ROS detoxification may have a role in controlling leaf rolling.


Using geographical information systems to identify populations in need of improved accessibility to antivenom treatment for snakebite envenoming in Costa Rica.
Snakebite accidents are an important health problem in rural areas of tropical countries worldwide, including Costa Rica, where most bites are caused by the pit-viper Bothrops asper. The treatment of these potentially fatal accidents is based on the timely administration of specific antivenom. In many regions of the world, insufficient health care systems and lack of antivenom in remote and poor areas where snakebites are common, means that efficient treatment is unavailable for many snakebite victims, leading to unnecessary mortality and morbidity. In this study, geographical information systems (GIS) were used to identify populations in Costa Rica with a need of improved access to antivenom treatment: those living in areas with a high risk of snakebites and long time to reach antivenom treatment. Populations living in areas with high risk of snakebites were identified using two approaches: one based on the district-level reported incidence, and another based on mapping environmental factors favoring B. asper presence. Time to reach treatment using ambulance was estimated using cost surface analysis, thereby enabling adjustment of transportation speed by road availability and quality, topography and land use. By mapping populations in high risk of snakebites and the estimated time to treatment, populations with need of improved treatment access were identified. This study demonstrates the usefulness of GIS for improving treatment of snakebites. By mapping reported incidence, risk factors, location of existing treatment resources, and the time estimated to reach these for at-risk populations, rational allocation of treatment resources is facilitated.


Prevalence of herpes simplex virus type 1 glycoprotein G (gG) and gI genotypes in patients with herpetic keratitis.
Recent phylogenetic analyses on the herpes simplex virus type 1 (HSV-1) genes US4, encoding glycoprotein G (gG) and US7, encoding gI, of clinical HSV-1 isolates have led to the classification of HSV-1 into three genotypes, arbitrarily designated as A, B and C. The prevalence of the HSV-1 gG and gI genotypes and their potential disease association was determined in a large cohort of patients with herpetic keratitis (HK). Primary corneal HSV-1 isolates of 178 HK patients were genotyped by a PCR-based restriction fragment length polymorphism method targeting the viral genes US4 and US7. Genotype B was more frequently expressed by the corneal HSV-1 isolates compared with genotypes A and C. Fifty-five of 178 corneal isolates (31%) had different genotypes in both loci. No clinically relevant associations were observed between the HSV-1 genotypes and disease outcome in the HK patients studied. The data presented demonstrate a high frequency of recombinant corneal HSV-1 isolates and suggest that clinical outcome of HSV-1-induced keratitis is independent of a gG or gI genotype.


TNF blockers inhibit spinal radiographic progression in ankylosing spondylitis by reducing disease activity: results from the Swiss Clinical Quality Management cohort.
To analyse the impact of tumour necrosis factor inhibitors (TNFis) on spinal radiographic progression in ankylosing spondylitis (AS). Patients with AS in the Swiss Clinical Quality Management cohort with up to 10 years of follow-up and radiographic assessments every 2 years were included. Radiographs were scored by two readers according to the modified Stoke Ankylosing Spondylitis Spine Score (mSASSS) with known chronology. The relationship between TNFi use before a 2-year radiographic interval and progression within the interval was investigated using binomial generalised estimating equation models with adjustment for potential confounding and multiple imputation of missing values. Ankylosing Spondylitis Disease Activity Score (ASDAS) was regarded as mediating the effect of TNFi on progression and added to the model in a sensitivity analysis. A total of 432 patients with AS contributed to data for 616 radiographic intervals. Radiographic progression was defined as an increase in ≥2 mSASSS units in 2 years. Mean (SD) mSASSS increase was 0.9 (2.6) units in 2 years. Prior use of TNFi reduced the odds of progression by 50% (OR 0.50, 95% CI 0.28 to 0.88) in the multivariable analysis. While no direct effect of TNFi on progression was present in an analysis including time-varying ASDAS (OR 0.61, 95% CI 0.34 to 1.08), the indirect effect, via a reduction in ASDAS, was statistically significant (OR 0.75, 95% CI 0.59 to 0.97). TNFis are associated with a reduction of spinal radiographic progression in patients with AS. This effect seems mediated through the inhibiting effect of TNFi on disease activity.