Plastic debris in Mediterranean seabirds.
Plastic debris is often ingested by marine predators and can cause health disorders and even death. We present the first assessment of plastic ingestion in Mediterranean seabirds. We quantified and measured plastics accumulated in the stomach of 171 birds from 9 species accidentally caught by longliners in the western Mediterranean from 2003 to 2010. Cory's shearwaters (Calonectris diomedea) showed the highest occurrence (94%) and large numbers of small plastic particles per affected bird (on average N = 15.3 ± 24.4 plastics and mass = 23.4 ± 49.6 mg), followed by Yelkouan shearwaters (Puffinus yelkouan, 70%, N = 7.0 ± 7.9, 42.1 ± 100.0 mg), Balearic shearwaters (Puffinus mauretanicus, 70%, N = 3.6 ± 2.9, 5.5 ± 9.7 mg) and the rest of species (below 33%, N = 2.7, 113.6 ± 128.4 mg). Plastic characteristics did not differ between sexes and were not related to the physical condition of the birds. Our results point out the three endemic and threatened shearwater species as being particularly exposed to plastic accumulation.


Sleep, Sleep Disorders, and Sexual Dysfunction.
Good sleep is necessary for good health. Sleep health is increasingly recognized as important for physical and mental health by both the medical profession and the general public, and there is great interest in how to avoid and treat sleep disorders and problems. Recent research indicates that insufficient sleep, disrupted sleep, and sleep disorders affect many aspects of human health including sexual function. In fact, patients with urological disorders or erectile dysfunction (ED) may have a sleep disorder that contributes to their urological or sexual dysfunction. Obstructive sleep apnea, insomnia, shift work disorder, and restless legs syndrome are all common sleep disorders and are associated with ED and/or other urological disorders. Therefore, careful attention should be paid to the diagnosis and treatment of concomitant sleep disorders in patients with sexual dysfunction. In this review, we provide an overview of what sleep is and how it is assessed in the clinic or laboratory; our current understanding of the functions of sleep and sleep health; a description of common sleep disorders, as well as how they are diagnosed and treated; and how sleep and its disorders are associated with male sexual dysfunction. Sleep is considered to be a 'third pillar of health', along with diet and exercise. With an understanding of common sleep disorders and how they can impact male sexual function, the urologist can ensure that sleep disorders are considered as a contributor to sexual dysfunction in their patients in order to provide them with the optimal treatment for overall health.


Identification of cytochrome P-450IIB1 as a cocaine-bioactivating isoform in rat hepatic microsomes and in cultured rat hepatocytes.
Induction of cytochrome P-450-dependent monooxygenases with phenobarbital (PB) or other hepatic drug-metabolizing enzyme inducers in the rat is associated with enhanced cocaine hepatotoxicity both in vivo and in cultured rat hepatocytes. To demonstrate whether the major PB-inducible P-450 subfamily (P-450IIB) could be involved in the metabolic activation of cocaine, rates of cocaine N-demethylation (the first step of cocaine bioactivation) and the rate of irreversible (covalent) binding of tritiated cocaine to hepatic microsomal proteins (a measure for the overall bioactivation) were determined in microsomes from saline or PB-pretreated rats. PB pretreatment augmented Vmax (6-fold), but not KM, of cocaine N-demethylation. Similarly, the rate of irreversible protein binding was 3-fold increased in microsomes from PB-pretreated rats as compared with those from saline controls. Addition of benzphetamine, a substrate of P-450IIB, markedly inhibited cocaine irreversible binding. In addition, various concentrations of cocaine inhibited microsomal pentoxyresorufin O-depentylase activity in a competitive-type pattern. A polyclonal antibody raised against purified rat P-450IIB1 markedly inhibited cocaine N-demethylation as compared with control incubations with preimmune IgG. Finally, pretreatment of rats with PB potentiated cocaine-induced cytotoxicity in primary, short-term cultured hepatocytes, assessed as lactate dehydrogenase release into the culture medium. This enhancing effect of PB became even more evident in glutathione-depleted cells. These results suggest that cocaine is metabolized and bioactivated by P-450IIB1 in the rat liver, and that induction of this isoform with various agents may be associated with enhanced lethal hepatocyte injury in the rat.


A semiquantitative metric for evaluating clinical actionability of incidental or secondary findings from genome-scale sequencing.
As genome-scale sequencing is increasingly applied in clinical scenarios, a wide variety of genomic findings will be discovered as secondary or incidental findings, and there is debate about how they should be handled. The clinical actionability of such findings varies, necessitating standardized frameworks for a priori decision making about their analysis. We established a semiquantitative metric to assess five elements of actionability: severity and likelihood of the disease outcome, efficacy and burden of intervention, and knowledge base, with a total score from 0 to 15. The semiquantitative metric was applied to a list of putative actionable conditions, the list of genes recommended by the American College of Medical Genetics and Genomics (ACMG) for return when deleterious variants are discovered as secondary/incidental findings, and a random sample of 1,000 genes. Scores from the list of putative actionable conditions (median = 12) and the ACMG list (median = 11) were both statistically different than the randomly selected genes (median = 7) (P < 0.0001, two-tailed Mann-Whitney test). Gene-disease pairs having a score of 11 or higher represent the top quintile of actionability. The semiquantitative metric effectively assesses clinical actionability, promotes transparency, and may facilitate assessments of clinical actionability by various groups and in diverse contexts.Genet Med 18 5, 467-475.


Phytotoxic effects of (+/-)-catechin in vitro, in soil, and in the field.
Exploring the residence time of allelochemicals released by plants into different soils, episodic exposure of plants to allelochemicals, and the effects of allelochemicals in the field has the potential to improve our understanding of interactions among plants. We conducted experiments in India and the USA to understand the dynamics of soil concentrations and phytotoxicity of (+/-)-catechin, an allelopathic compound exuded from the roots of Centaurea maculosa, to other plants in vitro and in soil. Experiments with single and pulsed applications into soil were conducted in the field. Experimental application of (+/-)-catechin to soils always resulted in concentrations that were far lower than the amounts added but within the range of reported natural soil concentrations. Pulses replenished (+/-)-catechin levels in soils, but consistently at concentrations much lower than were applied, and even pulsed concentrations declined rapidly. Different natural soils varied substantially in the retention of (+/-)-catechin after application but consistent rapid decreases in concentrations over time suggested that applied experimental concentrations may overestimate concentrations necessary for phytotoxicity by over an order of magnitude. (+/-)-Catechin was not phytotoxic to Bambusa arundinacea in natural Indian soil in a single pulse, but soil concentrations at the time of planting seeds were either undetectable or very low. However, a single dose of (+/-)-catechin suppressed the growth of bamboo in sand, in soil mixed with organic matter, and Koeleria macrantha in soils from Montana and Romania, and in field applications at 40 microg l(-1). Multiple pulses of (+/-)-catechin were inhibitory at very low concentrations in Indian soil. Our results demonstrate that (+/-)-catechin is highly dynamic in natural soils, but is phytotoxic well below natural concentrations measured in some soils and applied at low concentrations in the field. However, there is substantial conditionality in the effects of the allelochemical.


Conjugated linoleic acid increases skeletal muscle ceramide content and decreases insulin sensitivity in overweight, non-diabetic humans.
The effect of conjugated linoleic acid (CLA), a popular weight-loss supplement, on insulin sensitivity in humans is controversial and has not been extensively studied. To date no studies have examined the effects of CLA supplementation on human skeletal muscle metabolism or lipid content. It is also unknown whether CLA accumulates in human skeletal muscle with supplementation. In the present study, 9 overweight, non-diabetic individuals received 4 g/d of mixed CLA isomers in the form of 1 g supplements, for 12 weeks. CLA isomers significantly increased in both plasma and skeletal muscle following supplementation. Skeletal muscle ceramide content was also significantly increased, although there was no consistent change in muscle diacylglycerol or triacylglycerol content. Insulin sensitivity was significantly decreased (p<or=0.05) following CLA supplementation, with a significant increase in glucose and insulin area under the curve during an oral glucose tolerance test (p<or=0.005). The protein content of PPARalpha, for which CLA is a potent ligand, was unchanged following supplementation. Accordingly, there was no change in markers of muscle oxidative capacity. In conclusion, mixed-isomer CLA supplementation in overweight non-diabetic humans results in the accumulation of CLA and ceramide in skeletal muscle, and decreases insulin sensitivity.


In vivo genotoxicity assessment of aluminium oxide nanomaterials in rat peripheral blood cells using the comet assay and micronucleus test.
Advances in nanotechnology and its usage in various fields have led to the exposure of humans to engineered nanomaterials (NMs) and there is a need to tackle the potential human health effects before these materials are fully exploited. The main purpose of the current study was to assess whether aluminium oxide NMs (Al(2)O(3)-30 nm and Al(2)O(3)-40 nm) could cause potential genotoxic effects in vivo. Characterization of Al(2)O(3)-30 nm and Al(2)O(3)-40 nm was done with transmission electron microscopy, dynamic light scattering and laser Doppler velocimetry prior to their use in this study. The genotoxicity end points considered in this study were the frequency of micronuclei (MN) and the percentage of tail DNA (% Tail DNA) migration in rat peripheral blood cells using the micronucleus test (MNT) and the comet assay, respectively. Genotoxic effects were evaluated in groups of female Wistar rats (five per group) after single doses of 500, 1000 and 2000 mg/kg body weight (bw) of Al(2)O(3)-30 nm, Al(2)O(3)-40 nm and Al(2)O(3)-bulk. Al(2)O(3)-30 nm and Al(2)O(3)-40 nm showed a statistically significant dose-related increase in % Tail DNA for Al(2)O(3)-30 nm and Al(2)O(3)-40 nm (P < 0.05). However, Al(2)O(3)-bulk did not induce statistically significant changes over control values. The MNT also revealed a statistically significant (P < 0.05) dose-dependent increase in the frequency of MN, whereas Al(2)O(3)-bulk did not show any significant increase in frequency of MN compared to control. Cyclophosphamide (40 mg/kg bw) used as a positive control showed statistically significant (P < 0.001) increase in % Tail DNA and frequency of MN. The biodistribution of Al(2)O(3)-30 nm and Al(2)O(3)-40 nm and Al(2)O(3)-bulk in different rat tissues, urine and feces was also studied 14 days after treatment using inductively coupled plasma mass spectrometry. The data indicated that tissue distribution of Al(2)O(3) was size dependent. Our findings suggest that Al(2)O(3) NMs were able to cause size- and dose-dependent genotoxicity in vivo compared to Al(2)O(3)-bulk and control groups.


Gender differences in the pharmacokinetics of rivastigmine in rats.
The effect of gender on the pharmacokinetics of rivastigmine (CAS 123441-03-2) was studied in male and female Wistar rats following intravenous bolus administration. The area under the plasma concentration-time curve (AUC), apparent volume of distribution (Vd), systemic clearance (CL), and terminal plasma halflife (t1/2) of rivastigmine were compared between male and female rats. Compared to male rats, female rats exhibited higher plasma rivastigmine levels showing significantly (p < 0.05) larger AUC (226.77 vs. 149.68 ng h/ml), Vd (6.70 vs. 4.13 L), t1/2 (0.84 vs. 0.34 h) and a lower CL (5.51 vs. 8.35 L/h). The male rats had a 2.5 fold greater elimination rate constant than female rats (2.02 vs. 0.82 h(-1)). Gender had a significant effect on the pharmacokinetics of rivastigmine. Gender differences were reported due to gonadal hormones, and the observed difference in pharmacokinetics of rivastigmine might be attributed to testosterone in male rats.


Chemiluminescent examination of abiotic oxidative stress of watercress.
Watercress (Nasturtium officinale) is an aquatic plant that readily bioaccumulates heavy metals that may be found in contaminated aquatic systems. Toxic effects of contaminants on the physiological processes cause changes in oxidase enzymatic activity in watercress, which can be measured with a luminometer. The luminometer uses the reaction produced when peroxidases break down hydrogen peroxide into water and an oxygen radical. The resulting oxyradical binds to and oxidizes phenolic groups, producing a measureable luminescent reaction. Nasturtium officinale plants were exposed to 3 different concentrations of heavy metals, including lead, nickel, copper, and manganese for 24 h, 48 h, and 72 h. Aquatic exposure to the 4 heavy metals caused a significant increase in oxidative enzyme production. Fluorometric and morphometric measurements were also conducted to compare plant stress with the oxidative enzyme analyses. Fluorometric measurements performed on plants stressed by exposure to heavy metals revealed no significant decreases in photosystem II efficiency. Morphometric measurements of root length showed decreased root growth resulting from exposures to Ni, Cu, and Mn.


Studies in porphyria. IV. Expression of the gene defect of acute intermittent porphyria in cultured human skin fibroblasts and amniotic cells: prenatal diagnosis of the porphyric trait.
The gene lesion of the porphyrin-heme synthetic pathway in acute intermittent porphyria (AIP) is reflected in a deficient level of activity of the cytosol enzyme uroporphyrinogen I synthetase (URO-S). A marked URO-S deficiency has been demonstrated in the liver and in circulating erythrocytes of individuals with both active and latent AIP. This enzymic abnormality accounts for the excessive production and excretion into urine of the porphyrin precursors, lamda-aminolevulinic acid (ALA) and porphobilinogen (PBG) in AIP subjects. In this study, utilizing cell culture techniques, a marked URO-S deficiency has also been demonstrated in skin fibroblasts from AIP patients and in cells derived through aminocentesis from an approximately 17-wk old fetus. The prenatal diagnosis of the AIP trait in this fetus was confirmed postnatally by the demonstration in the child of a deficient level of erythrocyte URO-S activity which was comparable to those found in her AIP mother and affected sibling and which was approximately one-half the levels characterizing her normal father and aunt and a second unaffected sibling. The identification of the URO-S deficiency in cultured human fibroblasts from AIP patients was facilitated by a newly developed, sensitive assay for the enzyme activity. In this assay, the ability of such cells to convert ALA to protoporphyrin was quantitated; in the sequence of reactions involved in this transformation, URO-S is limiting so that the gene defect of AIP could be simply and precisely determined by appropriate spectrofluorometry of cell extracts. The technique described has distinct advantages over the direct enzymatic assay for URO-S activity in cultured human skin fibroblasts and permits clear differentiation of AIP carrier from normal individuals.


Borrelia burgdorferi-associated cutaneous B cell lymphoma: clinical and immunohistologic characterization of four cases.
Four patients with low-grade malignant B cell lymphoma of the skin in association with chronic Borrelia burgdorferi infection are presented. Plaque-shaped or nodular erythematous lesions with ill-defined borders were seen. Clinical progression was slow; the skin tumors occurred for up to 7 to 15 years. Extracutaneous involvement was found in only one case. Immunohistologic investigations showed an expression of B cell markers with restriction to only one light chain type and absence of T cell antigens. The growth fraction was 5% to 30%, as shown with the monoclonal antibody Ki-67. The immunoarchitecture of the tumors in three patients was unusual compared with established criteria for cutaneous B cell lymphoma and revealed similarities to mucosa-associated B cell lymphoma. Some immunohistologic heterogeneity may indicate the development of monoclonal proliferation that originated from different phases of B lymphocytic differentiation. In three cases no clinical signs of B. burgdorferi infection were found; in one patient acrodermatitis chronica atrophicans was present. The occurrence of acrodermatitis chronica atrophicans and malignant lymphomas was frequently reported in the European literature before B. burgdorferi was recognized. These findings suggest a relation between B. burgdorferi infection and cutaneous B cell lymphoma. In geographic regions where infected ticks are present, borreliosis should be considered in patients with cutaneous B cell lymphoma.


Atherosclerosis in coronary artery and aorta in a semi-urban population by applying modified American Heart Association classification of atherosclerosis: An autopsy study.
Ischemic heart disease (IHD) following atherosclerosis is a giant killer and the incidence of atherosclerosis in coronary arteries is rapidly increasing among Indians. The study was formulated to assess the histomorphological atherosclerotic changes in aorta and coronary arteries at autopsy by applying the modified American Heart Association classification of atherosclerosis based on morphological descriptions to find out the age and sex related prevalence of atherosclerosis in the semi-urban population of Kolar, a district in Southern India. Autopsy was conducted on 113 cases whose age ranged from 8-85 years. Autopsy was conducted by the conventional technique; heart and the aorta were removed and fixed in 10% formalin. The heart was dissected along the direction of flow of blood and aorta along the posterior surface. Microscopic assessment of the three main coronary arteries and aorta was done using the modified American Heart Association classification of atherosclerosis. Proportions were analyzed using Chi-square test. The number of males was 78 (69%) and number of females was 35 (31%). Mean age was 37.11 ± 15.69 years. Increased incidence of intermediate lesions was noted in young individuals (15-34 yrs). Atherosclerotic lesions were more in left anterior descending artery compared to other coronary arteries and in abdominal aorta compared to thoracic and ascending aorta. Vulnerable plaques were more in right coronary artery. With cardiovascular disease attaining pandemic proportions, the study of subclinical atherosclerosis is the need of the hour to estimate the disease burden in the asymptomatic population. The increased amount of atherosclerosis (advanced and intermediate lesions) found in the young population in this study gives an indication that anti-atherogenic preventive measures need to be implemented in young individuals, so as to prevent coronary artery disease from causing premature death.


Refining prognosis and identifying targetable pathways for high-risk endometrial cancer; a TransPORTEC initiative.
This study aimed to investigate whether molecular analysis can be used to refine risk assessment, direct adjuvant therapy, and identify actionable alterations in high-risk endometrial cancer. TransPORTEC, an international consortium related to the PORTEC3 trial, was established for translational research in high-risk endometrial cancer. In this explorative study, routine molecular analyses were used to detect prognostic subgroups: p53 immunohistochemistry, microsatellite instability and POLE proofreading mutation. Furthermore, DNA was analyzed for hotspot mutations in 13 additional genes (BRAF, CDKNA2, CTNNB1, FBXW7, FGFR2, FGFR3, FOXL2, HRAS, KRAS, NRAS, PIK3CA, PPP2R1A, and PTEN) and protein expression of ER, PR, PTEN, and ARID1a was analyzed. Rates of distant metastasis, recurrence-free, and overall survival were calculated using the Kaplan-Meier method and log-rank test. In total, samples of 116 high-risk endometrial cancer patients were included: 86 endometrioid; 12 serous; and 18 clear cell. For endometrioid, serous, and clear cell cancers, 5-year recurrence-free survival rates were 68%, 27%, and 50% (P=0.014) and distant metastasis rates 23%, 64%, and 50% (P=0.001), respectively. Four prognostic subgroups were identified: (1) a group of p53-mutant tumors; (2) microsatellite instable tumors; (3) POLE proofreading-mutant tumors; and (4) a group with no specific molecular profile (NSMP). In group 3 (POLE-mutant; n=14) and group 2 (microsatellite instable; n=19) patients, no distant metastasis occurred, compared with 50% distant metastasis rate in group 1 (p53-mutant; n=36) and 39% in group 4 (NSMP; P<0.001). Five-year recurrence-free survival was 93% and 95% for group 3 (POLE-mutant) and group 2 (microsatellite instable) vs 42% (group 1, p53-mutant) and 52% (group 4, NSMP; P<0.001). Targetable FBXW7 and FGFR2 mutations (6%), alterations in the PI3K-AKT pathway (60%) and hormone receptor positivity (45%) were frequently found. In conclusion, molecular analysis of high-risk endometrial cancer identifies four distinct prognostic subgroups, with potential therapeutic implications. High frequencies of targetable alterations were identified and may serve as targets for individualized treatment.


Spatial priming in visual search: memory for body-centred information.
Spatial priming allows memory for target locations to be evaluated, whereby when a target appears in the same location across trials, participants become more efficient at locating it and consequently their search times decrease. Previously, we reported priming effects when the location of a target was repeated with respect to the participant's body but not when it was repeated relative to their eye position; therefore, suggesting that body-centred information is available after a delay of at least a few seconds (Ball et al. in Exp Brain Res 204:585-594, 2010). However, we were unable to rule out the possibility that stable allocentric cues within the room may have contributed to the priming effects that we observed. In this current study, we introduced a condition where despite participants moving to a new location between trials, their position relative to the target was maintained. This movement disrupted any potential room-based cues about the target location. While we replicated our previous finding of priming when the location of the target was repeated relative to the viewer when no movement was required, we also found robust priming effects when participants moved to a new location in between trials. Thus, we provide clear evidence that in our spatial priming task, the location of the target was being coded in a body-centred reference frame and that this information is available after a delay.


Formulary Management Activities and Practice Implications Among Public Sector Hospital Pharmaceutical and Therapeutics Committees in a South African Province.
The World Health Organization identified Pharmaceutical and Therapeutics Committees (PTCs) at district and hospital levels as one of the pivotal models to promote rational use of medicines (RUM). This is endorsed by the Government in South Africa. Formulary development and management is one of the main functions of PTCs. This study aimed to describe the formulary management activities among PTCs in public hospitals in Gauteng Province, South Africa, following initiatives to promote RUM in South Africa. Qualitative, nonparticipatory, observational study, observing 26 PTC meetings. Data were coded and categorized using NVivo9® qualitative data analysis software. Themes and sub-themes were developed. The themes and sub-themes on formulary management are the principal focus of this paper. More than half of the observed PTCs reviewed their formulary lists. There was variation in the review process among institutions providing different levels of care. Various aspects were considered for formulary management especially requests for medicines to be added. These included cost considerations (mainly focusing on acquisition costs), evidence-based evaluation of clinical trials, patient safety, clinical experience and changes in the National Essential Medicines List (NEML). The tertiary PTCs mostly dealt with applications for new non-EML medicines, while PTCs in the other hospitals mainly requested removal or addition of EML medicines to the list. This is the first study from Gauteng Province, South Africa, reporting on how decisions are actually taken to include or exclude medicines onto formularies within public sector hospitals providing different levels of care. Various approaches are adopted at different levels of care when adding to- or removing medicines from the formulary lists. Future programs should strengthen PTCs in specialized aspects of formulary management. A more structured approach to formulary review at the local PTC level should be encouraged in line with the national approach when reviewing possible additions to the NEML.


Oxidative Stress and Metabolic Perturbations in Wooden Breast Disorder in Chickens.
This study was conducted to characterize metabolic features of the breast muscle (pectoralis major) in chickens affected with the Wooden Breast myopathy. Live birds from two purebred chicken lines and one crossbred commercial broiler population were clinically examined by manual palpation of the breast muscle (pectoralis major) at 47-48 days of age. Metabolite abundance was determined by gas chromatography/mass spectrometry (GC/MS) and liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) using breast muscle tissue samples from 16 affected and 16 unaffected chickens. Muscle glycogen content was also quantified in breast muscle tissue samples from affected and unaffected chickens. In total, levels of 140 biochemicals were significantly different (FDR<0.1 and fold-change A/U>1.3 or <0.77) between affected and unaffected chickens. Glycogen content measurements were considerably lower (1.7-fold) in samples taken from Wooden Breast affected birds when compared with samples from unaffected birds. Affected tissues exhibited biomarkers related to increased oxidative stress, elevated protein levels, muscle degradation, and altered glucose utilization. Affected muscle also showed elevated levels of hypoxanthine, xanthine, and urate molecules, the generation of which can contribute to altered redox homeostasis. In conclusion, our findings show that Wooden Breast affected tissues possess a unique metabolic signature. This unique profile may identify candidate biomarkers for diagnostic utilization and provide mechanistic insight into altered biochemical processes contributing to tissue hardening associated with the Wooden Breast myopathy in commercial chickens.


Lévy statistics of emission from a novel random amplifying medium: an optical realization of the Arrhenius cascade.
We report the observation of Lévy-like statistical configuration-to-configuration fluctuations in the intensity of emission from a novel system, the fiber-random amplifying medium, where active fiber segments are embedded randomly in a bulk of pointlike passive scatterers. Some rare configurations of fibers provide long, guided amplifying paths for the photons, leading to high jumps in the intensity, and thus to Lévy statistics. This system provides an optical realization of the Arrhenius cascade.


NEDL1, a novel ubiquitin-protein isopeptide ligase for dishevelled-1, targets mutant superoxide dismutase-1.
Approximately 20% of familial amyotrophic lateral sclerosis (FALS) arises from germ-line mutations in the superoxide dismutase-1 (SOD1) gene. However, the molecular mechanisms underlying the process have been elusive. Here, we show that a neuronal homologous to E6AP carboxyl terminus (HECT)-type ubiquitin-protein isopeptide ligase (NEDL1) physically binds translocon-associated protein-delta and also binds and ubiquitinates mutant (but not wild-type) SOD1 proportionately to the disease severity caused by that particular mutant. Immunohistochemically, NEDL1 is present in the central region of the Lewy body-like hyaline inclusions in the spinal cord ventral horn motor neurons of both FALS patients and mutant SOD1 transgenic mice. Two-hybrid screening for the physiological targets of NEDL1 has identified Dishevelled-1, one of the key transducers in the Wnt signaling pathway. Mutant SOD1 also interacted with Dishevelled-1 in the presence of NEDL1 and caused its dysfunction. Thus, our results suggest that an adverse interaction among misfolded SOD1, NEDL1, translocon-associated protein-delta, and Dishevelled-1 forms a ubiquitinated protein complex that is included in potentially cytotoxic protein aggregates and that mutually affects their functions, leading to motor neuron death in FALS.


Treatment efficiency and economic feasibility of biological oxidation, membrane filtration and separation processes, and advanced oxidation for the purification and valorization of olive mill wastewater.
Olive mill wastewater (OMW) is a major waste stream resulting from numerous operations that occur during the production stages of olive oil. The resulting effluent contains various organic and inorganic contaminants and its environmental impact can be notable. The present work aims at investigating the efficiency of (i) jet-loop reactor with ultrafiltration (UF) membrane system (Jacto.MBR), (ii) solar photo-Fenton oxidation after coagulation/flocculation pre-treatment and (iii) integrated membrane filtration processes (i.e. UF/nanofiltration (NF)) used for the treatment of OMW. According to the results, the efficiency of the biological treatment was high, equal to 90% COD and 80% total phenolic compounds (TPh) removal. A COD removal higher than 94% was achieved by applying the solar photo-Fenton oxidation process as post-treatment of coagulation/flocculation of OMW, while the phenolic fraction was completely eliminated. The combined UF/NF process resulted in very high conductivity and COD removal, up to 90% and 95%, respectively, while TPh were concentrated in the NF concentrate stream (i.e. 93% concentration). Quite important is the fact that the NF concentrate, a valuable and polyphenol rich stream, can be further valorized in various industries (e.g. food, pharmaceutical, etc.). The above treatment processes were found also to be able to reduce the initial OMW phytotoxicity at greenhouse experiments; with the effluent stream of solar photo-Fenton process to be the least phytotoxic compared to the other treated effluents. A SWOT (Strength, Weakness, Opportunities, Threats) analysis was performed, in order to determine both the strengths of each technology, as well as the possible obstacles that need to overcome for achieving the desired levels of treatment. Finally, an economic evaluation of the tested technologies was performed in an effort to measure the applicability and viability of these systems at real scale; highlighting that the cost cannot be regarded as a 'cut off criterion', since the most cost-effective option in not always the optimum one.


Incidence of treatment for postabortion complications in India, 2015.
Abortion has been legal under broad criteria in India since 1971. However, access to legal abortion services remains poor. In the past decade, medication abortion (MA) has become widely available in India and use of this method outside of health facilities accounts for over 70% of all abortions. Morbidity from unsafe abortion remains an important health issue. The informal providers who are the primary source of MA may have poor knowledge of the method and may offer inadequate or inaccurate advice on use of the method. Misuse of the method can result in women seeking treatment for true complications as well as during the normal processes of MA. An estimated 5% of all abortions are done using highly unsafe methods and performed by unskilled providers, also contributing to abortion morbidity. This paper provides new representative abortion-related morbidity measures at the national and subnational levels from a large-scale 2015 study of six Indian states-Assam, Bihar, Gujarat, Madhya Pradesh, Tamil Nadu and Uttar Pradesh. The outcomes include the number and treatment rates of women with complications resulting from induced abortion and the type of complications. The total number of women treated for abortion complications at the national level is 5.2 million, and the rate is 15.7 per 1000 women of reproductive age per year. In all six study states, a high proportion of all women receiving postabortion care were admitted with incomplete abortion from use of MA-ranging from 33% in Tamil Nadu to 65% in Assam. The paper fills an important gap by providing new evidence that can inform policy-makers and health planners at all levels and lead to improvements in the provision of postabortion care and legal abortion services-improvements that would greatly reduce abortion-related morbidity and its costs to Indian women, their families and the healthcare system.


MicroRNAs: From Female Fertility, Germ Cells, and Stem Cells to Cancer in Humans.
MicroRNAs are a family of naturally occurring small noncoding RNA molecules that play an important regulatory role in gene expression. They are suggested to regulate a large proportion of protein encoding genes by mediating the translational suppression and posttranscriptional control of gene expression. Recent findings show that microRNAs are emerging as important regulators of cellular differentiation and dedifferentiation, and are deeply involved in developmental processes including human preimplantation development. They keep a balance between pluripotency and differentiation in the embryo and embryonic stem cells. Moreover, it became evident that dysregulation of microRNA expression may play a fundamental role in progression and dissemination of different cancers including ovarian cancer. The interest is still increased by the discovery of exosomes, that is, cell-derived vesicles, which can carry different proteins but also microRNAs between different cells and are involved in cell-to-cell communication. MicroRNAs, together with exosomes, have a great potential to be used for prognosis, therapy, and biomarkers of different diseases including infertility. The aim of this review paper is to summarize the existent knowledge on microRNAs related to female fertility and cancer: from primordial germ cells and ovarian function, germinal stem cells, oocytes, and embryos to embryonic stem cells.


High intrinsic carrier mobility and photon absorption in the perovskite CH3NH3PbI3.
The carrier transport and optical properties of the hybrid organic-inorganic perovskite CH3NH3PbI3 are investigated using first-principles approaches. We found that the electron and hole mobilities could reach surprisingly high values of 7-30 × 10(3) and 1.5-5.5 × 10(3) cm(2) V(-1) s(-1), respectively, and both are estimated to be much higher than the current experimental measurements. The high carrier mobility is ascribed to the intrinsically small effective masses of anti-bonding band-edge states. The above results imply that there is still space to improve the performance of related solar cells. This material also has a sharp photon absorption edge and an absorption coefficient as high as 10(5) cm(-1), both of which contribute to effective utilization of solar radiation. Although band-edge states are mainly derived from the inorganic ions of Pb and I, thermal movement of the organic base has indirect influences on the bandgap and carrier effective masses, resulting in the temperature-dependent solar cell efficiencies.


Clinical application of standardized cognitive assessment using fMRI. I. Matrix reasoning.
Functional MRI is increasingly recognized for its potential as a powerful new tool in clinical neuropsychology. This is likely due to the fact that, with some degree of innovation, it is possible to convert practically any familiar cognitive test into one that can be performed in the MRI scanning environment. However, like any assessment approach, meaningful interpretation of fMRI data for the purpose of patient evaluation crucially requires normative data derived from a sample of unimpaired persons, against which individual patients may be compared. Currently, no such normative data are available for any fMRI-based cognitive testing protocol. In this paper, we report the first of a series of fMRI-compatible cognitive assessment protocols, a matrix reasoning test (f-MRT), for which normative samples of functional activation have been collected from unimpaired control subjects and structured in a manner that makes individual patient evaluation possible in terms of familiar z-score distributions. Practical application of the f-MRT is demonstrated via a contrastive case-study of two individuals with cognitive impairment in which fMRI data identifies subtleties in patient deficits otherwise missed by conventional measures of performance.


Micellar complexes of all-trans retinoic acid with polyvinylalcohol-nicotinoyl esters as new parenteral formulations in neuroblastoma.
All-trans-retinoic acid (ATRA) is now included in many antitumor therapeutic schemes for the treatment of acute promyelocytic leukemia, Kaposi's sarcoma, head and neck squamous cell carcinoma, ovarian carcinoma, bladder cancer, and neuroblastoma. Unfortunately, its poor aqueous solubility hampers its parenteral formulation, whereas oral administration of ATRA is associated with progressively diminishing drug levels in plasma, which is related to induction of retinoic acid-binding proteins and increased drug catabolism by cytochrome P450-mediated reactions. An ATRA formulation, obtained by complexation of the drug into polymeric micelles, might be suitable for parenteral administration overcoming these unwanted effects. To this purpose, amphiphilic polymers were prepared by polyvinylalcohol (PVA) partial esterification with nicotinoyl moieties and their functional properties evaluated with regard to ATRA complexation. The physicochemical characteristics of the polymers and the complexes were analyzed by 1H-NMR, Dynamic Light Scattering (DLS), Capillary Electophoresis (CE), and were correlated with the complex ability to improve the drug solubilization and release the free drug in an aqueous environment. Subsequently, the best complex, providing the highest ATRA solubilization and release, was evaluated in vitro to test its cytotoxicity towards neuroblastoma cell lines. The PVA substitution degree calculated from 1H-NMR was found to be 5.0%, 8.2%, 15.3% (nicotinoyl moiety:PVA monomer molar ratio), while capillary electrophoresis analysis on the complexes revealed that the drug loadings were 0.95%, 1.20%, 4.76% (ATRA:polymer w:w) for PVA substitution degrees of 5.0%, 8.2%, and 15.3%, respectively. Complexation strongly increased ATRA aqueous solubility, which reached 1.20 +/- 0.25 mg/mL. The DLS measurements of the polymers and the complexes in aqueous solutions revealed mean sizes always below 400 nm, low polydispersity (min 0.202 +/- 0.013, max 0.450 +/- 0.032), and size almost unaffected by concentration. Drug fractional release did not exceed 8% after 48 h. The cytotoxicity studies against neuroblastoma cell lines outlined a significant growth inhibition effect of complexed ATRA with respect to free ATRA. These data suggest that the systems analyzed may be suitable carriers for parenteral administration of ATRA and other hydrophobic antitumor drugs, where the carriers are required to improve drug aqueous solubility and delay drug release almost after their accumulation in solid tumors.


Formulation, stability, and delivery of live attenuated vaccines for human use.
The successful use of live attenuated viral and bacterial vaccines depends not only on the proper choice and delivery of the microorganisms, but also on maintaining the sufficient potency required for an immune response. The inherent lability of live organisms presents a particular formulation challenge in terms of stabilizing and preserving vaccine viability during manufacturing, storage, and administration. This review examines pharmaceutical approaches to the stabilization, formulation, and lyophilization of biological macromolecules in general, as well as the specific applicability of these principles to live attenuated viral and bacterial vaccines. Several formulation development case studies with live vaccines are presented. In addition, comparative stability data are summarized for many other live viral and bacterial preparations. Various pharmaceutical issues with conventional and novel delivery systems for administration of parenteral and oral live vaccines are also discussed.


Prevalence of Bartonella henselae and Bartonella clarridgeiae in stray cats.
The aim of the present work was to determine by blood culture the prevalence of blood infection with Bartonella species in a well-defined, European, urban stray cat population. Therefore, 94 stray cats were trapped from 10 cat colonies. Blood samples of these cats were cultured on both blood agar and liquid medium in order to raise the likelihood of bacterial detection. Fifty blood samples (53%) gave a positive culture result for Bartonella species. Isolate identification was performed by sequencing the first 430 bases of the 16S ribosomal DNA. Three types of sequences were thus obtained. The first type (17 isolates; 34%) was identical to that of B. henselae Houston-1 and the corresponding strains were referred as B. henselae type I. The second sequence type (18 isolates; 36%) was identical to that initially described as "BA-TF," and the corresponding strains were referred to as B. henselae type II. The third sequence type (15 isolates; 30%) was identical to that of the Bartonella clarridgeiae type strain (ATCC 51734). Our study points out the major role of stray cats as a reservoir of Bartonella spp. which can be transmitted to pet cats and, consequently, to humans. The study also highlights the high prevalence of B. clarridgeiae (16%) in the blood of stray cats.


Combination of PCR targeting the VD2 of omp1 and reverse line blot analysis for typing of urogenital Chlamydia trachomatis serovars in cervical scrape specimens.
In this study we developed and evaluated a new PCR-based typing assay, directed to the VD2 region of the omp1 gene, for the detection and typing of urogenital Chlamydia trachomatis infections. A nested VD2 PCR-reverse line blot (RLB) assay was developed for the typing of nine different urogenital serovars of C. trachomatis. The assay developed was tested with reference strains of C. trachomatis serovars and cervical scrapes of 86 Colombian women previously found to be positive for C. trachomatis by using plasmid PCR. Two sets of primers directed to the VD2 region of the omp1 gene of C. trachomatis were designed, and fragments of 220 and 166 bp were generated in the primary and nested PCRs, respectively. In addition, an RLB assay was developed to identify nine different urogenital serovars of C. trachomatis (Ba, D, E, F, G, H, I, J, and K) and group controls, including group B (Ba, D, and E), group C (I, J, K, and H), and an intermediate group (F and G). Using this assay, we were able to type 81 of the 86 samples (94.2%). Of these samples, 91.3% were single C. trachomatis infections, and 8.7% were multiple infections. The most common serovars identified were serovars D (22.2%), F (18.5%), G (13.6%), and E (12.3%). Of the women with multiple C. trachomatis infections, >50% contained both serovars D and E. The nested VD2 PCR-RLB developed is a simple, fast, and specific method for the identification of individual urogenital C. trachomatis serovars previously detected by using plasmid PCR. Moreover, it is an appropriate method for studying multiple C. trachomatis infections and for use in large epidemiological studies.


Telephone Transmission and Earwitnesses: Performance on Voice Parades Controlled for Voice Similarity.
The effect of telephone transmission on a listener's ability to recognise a speaker in a voice parade is investigated. A hundred listeners (25 per condition) heard 1 of 5 'target' voices, then returned a week later for a voice parade. The 4 conditions were: target exposure and parade both at studio quality; exposure and parade both at telephone quality; studio exposure with telephone parade, and vice versa. Fewer correct identifications followed from telephone exposure and parade (64%) than from studio exposure and parade (76%). Fewer still resulted for studio exposure/telephone parade (60%) and, dramatically, only 32% for telephone exposure/studio parade. Certain speakers were identified more readily than others across all conditions. Confidence ratings reflected this effect of speaker, but not the effect of exposure/parade condition.


Serum amyloid protein A concentration in cryopyrin-associated periodic syndromes patients treated with interleukin-1 beta antagonist.
Cryopyrin-associated periodic syndromes (CAPS) are a group of chronic, relapsing autoinflammatory disorders which may be complicated by systemic AA amyloidosis. The aim of our study was to evaluate serum amyloid protein A (SAA) level in CAPS patients treated with Interleukin-1beta (IL-1β) antagonist and to correlate its level with treatment response. All patients of CAPS Italian Register treated with IL-1β inhibitor were enrolled. SAA levels before starting therapy, and at last visit were evaluated. Patients were then divided in complete responders and partial responders. Twenty-five patients were enrolled. SAA level before starting therapy was increased (median 118.5 mg/L, IQR 96.4-252.8; normal value <6.4 mg/L), while at last visit SAA was significantly reduced (median 4.3 mg/L, IQR 2.3-12.7) (p<0.001). However 12 patients still presented SAA levels beyond normal range, 10/25 patients (40%) showed a complete response to treatment. Conversely, 15 patients presented only a partial response, of which 12 for increased SAA value and 3 for increased CRP value. Patients with partial response had SAA values significantly higher than patients with complete response (median 12.6 mg/L; IQR 8.3-20.0 vs. 2.7 mg/L; IQR 1.6-4.1, p<0.001). Our results confirm the long term efficacy of anti IL-1β treatment in CAPS and the decrease of SAA levels; however 48% of patients still presented SAA elevation despite treatment. The real risk of these patients in developing amyloidosis is not clear but the persistent increase of SAA needs a close follow-up.


Frequency and ethnic distribution of the common DHCR7 mutation in Smith-Lemli-Opitz syndrome.
Smith-Lemli-Opitz syndrome (SLOS) is an inherited multiple malformation syndrome caused by enzymatic deficiency of 3beta-hydroxysterol-Delta(7)-reductase (DHCR7). SLOS is thought to be most common among European Caucasians, with an incidence of 1 in 20,000 to 1 in 30,000 births. To define the carrier rate and ethnic distribution of SLOS, we screened DNA samples from 2,978 unrelated individuals for the most common SLOS mutation (IVS8-1G-->C). Twenty-four heterozygotes of the IVS8-1G-->C mutation were detected in 2,978 individuals of European Caucasian and Black backgrounds. For European Caucasians, the carrier rate for SLOS may be as high as 1 in 30, suggesting an incidence of 1 in 1,700 to 1 in 13,400. This high number is supported by the recent observation of newborn and prenatal incidence of 1 in 22,000 in the Caucasian population. Ours is the first report of the IVS8-1G-->C mutation in persons of African ancestry. Published 2001 Wiley-Liss, Inc.


Unwrapping noisy phase maps by use of a minimum-cost-matching algorithm.
An algorithm for unwrapping noisy phase maps by means of branch cuts has been proposed recently. These cuts join discontinuity sources that mark the beginning or end of a 2π phase discontinuity. After the placement of branch cuts, the unwrapped phase map is unique and independent of the unwrapping route. We show how a minimum-cost-matching graph-theory method can be used to find the set of cuts that has the global minimum of total cut length, in time approximately proportional to the square of the number of sources. The method enables one to unwrap unfiltered speckle-interferometry phase maps at higher source densities (0.1 sources pixel(-1)) than any previous branch-cut placement algorithm.


Clinical and biochemical characteristics of familial type 1 diabetes mellitus (FT1DM) compared to non-familial type 1 DM (NFT1DM).
Familial type 1 diabetes mellitus (FT1DM) comprises parent-offspring and sib-pair subgroups. The clinical and genetic characteristics of FT1DM cases with and without affected family members have been previously studied with varying results. Some investigators found similarity of presenting features whereas others reported significant differences between the two groups. To describe the clinical and biochemical characteristics of children with FT1DM in comparison with those with non-familial type 1 diabetes mellitus (NFT1DM). We performed a cross-sectional retrospective study in a cohort of children and adolescents with T1DM (n=424) aged between 6 months - 16 years attending to Hamad General Hospital Pediatric Diabetes Center, Doha (Qatar) from 2012-2016. They were divided into 2 groups. Group 1 consisted of 62 children and adolescent with FT1DM (parent-offspring or sib-pair). The other group (Group 2) consisted of 431 children and adolescents with NFT1DM. The clinical presentation and prevalence of β-cell autoimmunity (anti-glutamic acid decarboxylase (GAD) antibodies , anti-islet cell and anti-insulin antibodies), thyroid function (Free thyroxine: FT4 and thyroid-stimulating hormone: TSH), anti-thyroid peroxidase antibody (TPO) and anti-tissue transglutaminase (ATT) at their first presentation were recorded, described and analyzed. FT1 DM was more prevalent in boys versus girls (1.4:1, respectively) whereas the prevalence of NFT1DM did not differ between genders (1:1.1, respectively). F1DM occurred relatively early in childhood (40.7% before the age of 4 years and 72% before 9 years of age) versus NFT1DM which occurred relatively later in life (80% after the age of 4 years and 40% after the age of 9 years). 35.2% of FT1DM presented with diabetic ketoacidosis (DKA) versus 32.5% of T1DM patients. Anti-islet antibodies (Ab) were detected more frequently in FT1DM versus NFT1DM. The prevalence of positive anti-insulin and anti- GAD antibodies did not differ between the two groups. Anti TPO were detected in 27.2% of NFT1DM and 35.5% of FT1DM. A primary hypothyroidism, with positive ATPO, was more prevalent in FT1DM versus NFT1DM. ATT IgA was high in 5% of NFT1DM and 19.8% of FT1DM whereas ATT IgG was high in 4.4 % of NFT1DM and 15.4% of FT1DM. FT1DM is more prevalent in boys versus girls and occurs earlier in childhood compared to NFT1DM. Primary hypothyroidism was more prevalent in NFT1DM versus FT1DM. Anti-islet Ab and ATT antibodies were more prevalent in the FT1DM versus NFT1DM. The genetic background may explain some differences between FT1DM and NFT1DM including the age of onset, gender affection, as well as associated autoimmune disorders.


The level of salivary lactate dehydrogenase as an indicator of the association between gingivitis and related factors in Japanese university students.
The aim of this study was to investigate the association between the presence of gingivitis estimated using the salivary level of lactate dehydrogenase (LD) and related factors in young Japanese adults. Data from 1,915 participants (21.4 ± 2.5 years) were analyzed. Unstimulated saliva was collected from each participant and the salivary LD level was evaluated using a commercially available test kit with an integer scale ranging from 1 to 10. Gingivitis was defined as the LD level of ≥8. The number of permanent teeth, the simplified oral hygiene index (OHI-S), the presence of partially erupted molars and body mass index were recorded. Additionally, participants answered a questionnaire. The percentage of male participants, the number of permanent teeth, the OHI-S and the presence of partially erupted molars were higher, whereas the proportion receiving dental check-ups was lower in the gingivitis group (n = 88, 4.6%) than in the healthy group. Logistic regression analysis showed that gingivitis was significantly associated with OHI-S (OR: 2.68, 95% CI: 1.94-3.69) and receiving dental checkups (OR: 0.31, 95% CI: 0.10-0.99). The present findings indicated that the OHI-S and receiving dental checkups were significantly associated with gingivitis, as assessed by the salivary LD level, in this cohort.


A lake data set for the Tibetan Plateau from the 1960s, 2005, and 2014.
Long-term datasets of number and size of lakes over the Tibetan Plateau (TP) are among the most critical components for better understanding the interactions among the cryosphere, hydrosphere, and atmosphere at regional and global scales. Due to the harsh environment and the scarcity of data over the TP, data accumulation and sharing become more valuable for scientists worldwide to make new discoveries in this region. This paper, for the first time, presents a comprehensive and freely available data set of lakes' status (name, location, shape, area, perimeter, etc.) over the TP region dating back to the 1960s, including three time series, i.e., the 1960s, 2005, and 2014, derived from ground survey (the 1960s) or high-spatial-resolution satellite images from the China-Brazil Earth Resources Satellite (CBERS) (2005) and China's newly launched GaoFen-1 (GF-1, which means high-resolution images in Chinese) satellite (2014). The data set could provide scientists with useful information for revealing environmental changes and mechanisms over the TP region.


One of the giants of neurological surgery left us more than a decade ago, and neurosurgical literature did not show much interest.
One of the giants of neurological surgery left us over a decade ago. Charles George Drake died September 15, 1998 in London, Ontario after an extended bout with lung cancer. Although he will always be identified with taking posterior fossa aneurysm surgery from the realm of the daring to the domain of the routine, his contributions were much broader. Clinical neurosciences have been blessed in the past century by the life and works of Drake. In the neurosurgical world, the achievements of Drake are very well known and have been well recorded. Unfortunately, in the past decade since his passing, only one paper has been published about him and his contributions to neurosurgery. This is a historical paper regarding Charles George Drake that attempts to (1) remember Drake as a pioneer; (2) to evaluate lessons that we have learned from him; and (3) to address the question 'What made him great?'. As per Drake's teachings, this paper is meant to articulate the unique perspectives Charlie provided with respect to how we learn our craft, maintain the integrity of reporting, and implement suggestions as to how we may progress into the future. In conclusion, it is our hope that this paper will bring to life the unique character of Drake and his unprecedented blend of genius, creativity, technical skill, introspection, and ever-present humility for all international neurosurgeons to appreciate.


Protein kinase CK2 triggers cytosolic zinc signaling pathways by phosphorylation of zinc channel ZIP7.
The transition element zinc, which has recently been identified as an intracellular second messenger, has been implicated in various signaling pathways, including those leading to cell proliferation. Zinc channels of the ZIP (ZRT1- and IRT1-like protein) family [also known as solute carrier family 39A (SLC39A)] transiently increase the cytosolic free zinc (Zn(2+)) concentration in response to extracellular signals. We show that phosphorylation of evolutionarily conserved residues in endoplasmic reticulum zinc channel ZIP7 is associated with the gated release of Zn(2+) from intracellular stores, leading to activation of tyrosine kinases and the phosphorylation of AKT and extracellular signal-regulated kinases 1 and 2. Through pharmacological manipulation, proximity ligation assay, and mutagenesis, we identified protein kinase CK2 as the kinase responsible for ZIP7 activation. Together, the present results show that transition element channels in eukaryotes can be activated posttranslationally by phosphorylation, as part of a cell signaling cascade. Our study links the regulated release of zinc from intracellular stores to phosphorylation of kinases involved in proliferative responses and cell migration, suggesting a functional role for ZIP7 and zinc signals in these events. The connection with proliferation and migration, as well as the activation of ZIP7 by CK2, a kinase that is antiapoptotic and promotes cell division, suggests that ZIP7 may provide a target for anticancer drug development.


Hypoglycemic activity of the fungi Cordyceps militaris, Cordyceps sinensis, Tricholoma mongolicum, and Omphalia lapidescens in streptozotocin-induced diabetic rats.
Crude extracts were prepared from fruiting bodies and mycelia of the medicinal fungus Cordyceps militaris, and a polysaccharide-enriched fraction was obtained after extraction with hot water and ethanol precipitation. Polysaccharide-enriched fractions were similarly prepared from Cordyceps sinensis, Omphalia lapidescens, and Tricholoma mongolicum. The various aforementioned preparations were orally administered into different groups of adult rats 24 h before an intraperitoneal injection of streptozotocin (40 mg/kg body weight), and subsequently daily for another 4 days. The dosage used was 10 mg/kg body weight for polysaccharide-enriched preparations and 100 mg/kg body weight for crude extracts. Control rats received distilled water instead of crude extract or polysaccharide-enriched preparation. It was found in the control rats that plasma glucose level rose from about 90 mg/dl before streptozotocin injection to levels that were maintained at about 300 mg/dl postinjection. All preparations produced hypoglycemic effects. C. militaris polysaccharide-enriched fraction displayed a more prominent effect than that of C. sinensis polysaccharide-enriched fraction which in turn was more potent than that of O. lapidescens and T. mongolicum polysaccharide-enriched fractions. The hypoglycemic effect of C. militaris polysaccharide-enriched fraction was dose-dependent.


Metabolite profiling of the carnivorous pitcher plants Darlingtonia and Sarracenia.
Sarraceniaceae is a New World carnivorous plant family comprising three genera: Darlingtonia, Heliamphora, and Sarracenia. The plants occur in nutrient-poor environments and have developed insectivorous capability in order to supplement their nutrient uptake. Sarracenia flava contains the alkaloid coniine, otherwise only found in Conium maculatum, in which its biosynthesis has been studied, and several Aloe species. Its ecological role and biosynthetic origin in S. flava is speculative. The aim of the current research was to investigate the occurrence of coniine in Sarracenia and Darlingtonia and to identify common constituents of both genera, unique compounds for individual variants and floral scent chemicals. In this comprehensive metabolic profiling study, we looked for compound patterns that are associated with the taxonomy of Sarracenia species. In total, 57 different Sarracenia and D. californica accessions were used for metabolite content screening by gas chromatography-mass spectrometry. The resulting high-dimensional data were studied using a data mining approach. The two genera are characterized by a large number of metabolites and huge chemical diversity between different species. By applying feature selection for clustering and by integrating new biochemical data with existing phylogenetic data, we were able to demonstrate that the chemical composition of the species can be explained by their known classification. Although transcriptome analysis did not reveal a candidate gene for coniine biosynthesis, the use of a sensitive selected ion monitoring method enabled the detection of coniine in eight Sarracenia species, showing that it is more widespread in this genus than previously believed.


STIM is a Ca2+ sensor essential for Ca2+-store-depletion-triggered Ca2+ influx.
Ca(2+) signaling in nonexcitable cells is typically initiated by receptor-triggered production of inositol-1,4,5-trisphosphate and the release of Ca(2+) from intracellular stores. An elusive signaling process senses the Ca(2+) store depletion and triggers the opening of plasma membrane Ca(2+) channels. The resulting sustained Ca(2+) signals are required for many physiological responses, such as T cell activation and differentiation. Here, we monitored receptor-triggered Ca(2+) signals in cells transfected with siRNAs against 2,304 human signaling proteins, and we identified two proteins required for Ca(2+)-store-depletion-mediated Ca(2+) influx, STIM1 and STIM2. These proteins have a single transmembrane region with a putative Ca(2+) binding domain in the lumen of the endoplasmic reticulum. Ca(2+) store depletion led to a rapid translocation of STIM1 into puncta that accumulated near the plasma membrane. Introducing a point mutation in the STIM1 Ca(2+) binding domain resulted in prelocalization of the protein in puncta, and this mutant failed to respond to store depletion. Our study suggests that STIM proteins function as Ca(2+) store sensors in the signaling pathway connecting Ca(2+) store depletion to Ca(2+) influx.


The Vi capsular polysaccharide enables Salmonella enterica serovar typhi to evade microbe-guided neutrophil chemotaxis.
Salmonella enterica serovar Typhi (S. Typhi) causes typhoid fever, a disseminated infection, while the closely related pathogen S. enterica serovar Typhimurium (S. Typhimurium) is associated with a localized gastroenteritis in humans. Here we investigated whether both pathogens differ in the chemotactic response they induce in neutrophils using a single-cell experimental approach. Surprisingly, neutrophils extended chemotactic pseudopodia toward Escherichia coli and S. Typhimurium, but not toward S. Typhi. Bacterial-guided chemotaxis was dependent on the presence of complement component 5a (C5a) and C5a receptor (C5aR). Deletion of S. Typhi capsule biosynthesis genes markedly enhanced the chemotactic response of neutrophils in vitro. Furthermore, deletion of capsule biosynthesis genes heightened the association of S. Typhi with neutrophils in vivo through a C5aR-dependent mechanism. Collectively, these data suggest that expression of the virulence-associated (Vi) capsular polysaccharide of S. Typhi obstructs bacterial-guided neutrophil chemotaxis.


Enhanced desorption of persistent organic pollutants from microplastics under simulated physiological conditions.
Microplastics have the potential to uptake and release persistent organic pollutants (POPs); however, subsequent transfer to marine organisms is poorly understood. Some models estimating transfer of sorbed contaminants to organisms neglect the role of gut surfactants under differing physiological conditions in the gut (varying pH and temperature), examined here. We investigated the potential for polyvinylchloride (PVC) and polyethylene (PE) to sorb and desorb (14)C-DDT, (14)C-phenanthrene (Phe), (14)C-perfluorooctanoic acid (PFOA) and (14)C-di-2-ethylhexyl phthalate (DEHP). Desorption rates of POPs were quantified in seawater and under simulated gut conditions. Influence of pH and temperature was examined in order to represent cold and warm blooded organisms. Desorption rates were faster with gut surfactant, with a further substantial increase under conditions simulating warm blooded organisms. Desorption under gut conditions could be up to 30 times greater than in seawater alone. Of the POP/plastic combinations examined Phe with PE gave the highest potential for transport to organisms.


Coffee consumption and prostate cancer risk: a meta-analysis of cohort studies.
This meta-analysis was conducted to assess the association between coffee consumption and prostate cancer risk. Thirteen cohort studies with 34,105 cases and 539,577 participants were included in the meta-analysis. The summary relative risks (RRs) with 95% confidence intervals (CIs) for different coffee intake levels were calculated. Dose-response relationship was assessed using generalized least square trend estimation. The pooled RR for the highest vs. lowest coffee intake was 0.90 (95% CI: 0.85-0.95), with no significant heterogeneity across studies (P = 0.267; I(2) = 17.5%). The dose-response analysis showed a lower cancer risk decreased by 2.5% (RR = 0.975; 95% CI: 0.957-0.995) for every 2 cups/day increment in coffee consumption. Stratifying by geographic region, there was a statistically significant protective influence of coffee on prostate cancer risk among European populations. In subgroup analysis of prostate cancer grade, the summary RRs were 0.89 (95% CI: 0.83-0.96) for nonadvanced, 0.82 (95% CI: 0.61-1.10) for advanced and 0.76 (95% CI: 0.55-1.06) for fatal diseases. Our findings suggest that coffee consumption may be associated with a reduced risk of prostate cancer and it also has an inverse association with nonadvanced prostate cancer. Because of the limited number of studies, more prospective studies with large sample size are needed to confirm this association.


Depression and BMI influences the serum vascular endothelial growth factor level.
Recent studies suggest that the angiogenic cytokine vascular endothelial growth factor (VEGF) is involved in the pathogenesis of depression. However, only a few studies have investigated serum VEGF levels in individuals with depression, or the possible association between genetic variants within the VEGF gene and depression. The purpose of the present study was to investigate differences between serum VEGF levels in individuals with depression vs. control individuals, and associations between genetic markers located within VEGF and depression. In addition, determinants of the serum VEGF levels were identified. One-hundred and fifty-five depressed subjects and 280 controls were included in the study. All individuals returned a questionnaire and participated in a semi-structured diagnostic interview. Eleven single nucleotide polymorphisms were successfully analysed. VEGF levels were measured in serum by immunoassay and independent determinants of the serum VEGF level were assessed by generalized linear models.The main findings were that depression, severity of depression, previous depressive episodes, age and body mass index (BMI) were associated with higher serum VEGF levels. The genetic marker rs10434 was significantly associated with depression after correction for multiple testing, but not with the serum VEGF level. Our final model included depression and BMI as predictors of serum VEGF levels. Our study suggests a role for circulating serum VEGF in depression. Furthermore, our data also demonstrate that other factors than a diagnosis of depression influence the serum VEGF level. The importance of these factors should be emphasized when studies are compared.


Correlates of dependence and beliefs about the use of hypnotics among zolpidem and zopiclone users.
Zolpidem and zopiclone are the two most commonly prescribed Z-drugs approved to treat insomnia. To examine the demographic and clinical correlates of dependence and beliefs about hypnotic use among long-term zolpidem and zopiclone users in psychiatric treatment for insomnia. A total of 392 psychiatric outpatients who received zolpidem or zopiclone treatment for at least 3 months for insomnia were studied. Participants' severity of hypnotic dependence and beliefs about the use of hypnotics to treat sleep problems were assessed. The correlation of dependence and beliefs about zolpidem and zopiclone treatment with demographic characteristics, hypnotic-using behaviors, co-use of addictive substances, and depressive symptoms were analyzed using multiple regression analysis models. Zolpidem users reported more severe dependence and a lower level of necessity regarding the use of hypnotics than zopiclone users did. High equivalent doses of hypnotics and long duration of use were significantly associated with severe dependence and a low level of necessity. Severe depressive symptoms were signiciantly associated with severe dependence, a low level of necessity, and a low level of concern. Educational level was also associated with the levels of concern and necessity. Conclusions/Importance: There were differences in the level of dependence and belief about hypnotic use between zolpidem and zopiclone users. The correlates of dependence and belief identified in this study can serve as the basis for prevention and intervention programs.


Patterns of cognitive development in very low birth weight children during the first six years of life.
Cognitive development in very low birth weight (VLBW, < or = 1500 g) infants typically has been reported based on mean endpoints in cross-sectional studies. These overall group means mask individual patterns of cognitive development. Given the heterogeneity of VLBW infants, it is important to identify individual patterns of development and the factors associated with the different patterns. We sought to determine individual patterns of cognitive development over the first 6 years in VLBW children and to examine the relative influence of selected biomedical and sociodemographic factors on these patterns. VLBW infants (N = 203) were followed from birth to six years. Cognitive scores were obtained at four yearly intervals, and biomedical and social data were obtained beginning with the neonatal period. Cluster analysis was used to identify individual patterns of cognitive development. Five developmental patterns were identified: average-stable (13% of the sample); average-declined to low average (24% of the sample); average-declined to below average (43% of the sample); very low-increased to low average (8% of the sample); and very low-stable (12% of the sample). The patterns could be differentiated by several biomedical factors, including birth weight, gestational age, neonatal health, and 1-year assessments of neurological status and head circumference, as well as by level of maternal education. In particular, abnormal neurological status at 1 year was associated with a pattern of very low stable scores, and a suspicious status was associated with a pattern of improving cognitive development. Maternal education was influential among children born at the upper end of the VLBW range, who had a more favorable set of biomedical factors. Biomedical factors are of major importance for the cognitive development of VLBW infants, and their influence increases as birth weight declines. Differences in neurological integrity at 1 year were an important indicator of different patterns of cognitive development, especially for infants at the lower end of the VLBW range.


An intervention to reduce care-resistant behavior in persons with dementia during oral hygiene: a pilot study.
The primary purpose of this pilot study was to test the feasibility of an intervention designed to reduce care-resistant behaviors (CRBs) in persons with moderate-to-severe dementia during oral hygiene activities. The intervention, Managing Oral Hygiene Using Threat Reduction (MOUTh), combined best oral hygiene practices with CRB reduction techniques. Oral health was operationalized as the total score obtained from the Oral Health Assessment Tool (OHAT). CRB was measured using a refinement of the Resistiveness to Care Scale. Seven nursing home residents with dementia received twice daily mouth care for 14 days. The baseline OHAT mean score of 7.29 (SD = 1.25) improved to 1.00 (SD = 1.26, p < .001); CRB improved from 2.43 CRBs/minute (SD = 4.26) to 1.09 CRBs/minute (SD = 1.56, t = 1.97, df 41, p= .06). The findings from this pilot study suggest that the MOUTh intervention is feasible and reduced CRBs, thus allowing more effective oral care.


Dynamic Causal Modelling of Active Vision.
In this paper, we draw from recent theoretical work on active perception, which suggests that the brain makes use of an internal (i.e., generative) model to make inferences about the causes of sensations. This view treats visual sensations as consequent on action (i.e., saccades) and implies that visual percepts must be actively constructed via a sequence of eye movements. Oculomotor control calls on a distributed set of brain sources that includes the dorsal and ventral frontoparietal (attention) networks. We argue that connections from the frontal eye fields to ventral parietal sources represent the mapping from "where", fixation location to information derived from "what" representations in the ventral visual stream. During scene construction, this mapping must be learned, putatively through changes in the effective connectivity of these synapses. Here, we test the hypothesis that the coupling between the dorsal frontal cortex and the right temporoparietal cortex is modulated during saccadic interrogation of a simple visual scene. Using dynamic causal modeling for magnetoencephalography with (male and female) human participants, we assess the evidence for changes in effective connectivity by comparing models that allow for this modulation with models that do not. We find strong evidence for modulation of connections between the two attention networks; namely, a disinhibition of the ventral network by its dorsal counterpart.SIGNIFICANCE STATEMENT This work draws from recent theoretical accounts of active vision and provides empirical evidence for changes in synaptic efficacy consistent with these computational models. In brief, we used magnetoencephalography in combination with eye-tracking to assess the neural correlates of a form of short-term memory during a dot cancellation task. Using dynamic causal modeling to quantify changes in effective connectivity, we found evidence that the coupling between the dorsal and ventral attention networks changed during the saccadic interrogation of a simple visual scene. Intuitively, this is consistent with the idea that these neuronal connections may encode beliefs about "what I would see if I looked there", and that this mapping is optimized as new data are obtained with each fixation.


White Paper AGA: Drug Development for Eosinophilic Esophagitis.
Since first characterized in 2 small case series in the early 1990s, eosinophilic esophagitis (EoE) has emerged as a commonly identified cause of esophageal symptoms in children and adults.1,2 Although several highly effectively dietary, pharmacologic, and endoscopic therapies have been reported, none is currently approved by either the US Food and Drug Administration (FDA) or European regulatory authorities. Evolving diagnostic criteria have challenged drug development, in particular the recognition of complex interactions with the most prevalent esophageal disorder, gastroesophageal reflux disease (GERD). Heterogeneity in the clinical presentations of affected children and adults has created difficulties with uniform inclusion criteria and the development of disease-specific, patient-reported outcome (PRO) instruments. Furthermore, controversies regarding the appropriate therapeutic endpoints of EoE have impeded the design of clinical trials. Despite these obstacles, collaborative efforts by investigators, industry, the FDA, and patient advocacy groups have resulted in substantial progress in drug development in EoE over the past 2 decades.3 The purpose of this article is to summarize discussions on EoE based on the 2016 Drug Development Conference sponsored by the Center for Diagnostics and Therapeutics of the American Gastroenterological Association.


Digital teaching tools and global learning communities.
In 2009, we started a project to support the teaching and learning of university-level plant sciences, called Teaching Tools in Plant Biology. Articles in this series are published by the plant science journal, The Plant Cell (published by the American Society of Plant Biologists). Five years on, we investigated how the published materials are being used through an analysis of the Google Analytics pageviews distribution and through a user survey. Our results suggest that this project has had a broad, global impact in supporting higher education, and also that the materials are used differently by individuals in terms of their role (instructor, independent learner, student) and geographical location. We also report on our ongoing efforts to develop a global learning community that encourages discussion and resource sharing.


Newspaper coverage of food insecurity in UK, 2016-2019: a multi-method analysis.
Food insecurity is a growing concern in the UK. Newspaper coverage can reflect and shape public and political views. We examined how frequently food insecurity was reported on in UK newspapers, how the problem and its drivers were described, and which solutions were proposed. Using Factiva, we searched for news articles that were substantively about food insecurity and published in national UK newspapers between 01 January 2016 and 11 June 2019. We examined whether the number of articles differed over the study period, and conducted a thematic analysis to theoretical saturation using a random sample of articles. Overall, 436 articles met our inclusion criteria and 132 (30%) were analysed thematically. Reporting was more prevalent in the summer, with mentions of 'holiday hunger' among children, and leading up to Christmas, when charity was encouraged. Articles often contained views from advocacy groups and charities, who appeared to play an important role in maintaining news interest in food insecurity. From the thematic analysis, we developed themes related to the problems ('definitions of food insecurity' and 'consequences of food insecurity for individuals'), drivers ('insufficient income as an immediate driver' and 'government versus individual responsibility'), and solutions ('charitable food aid' and 'calls for government action'). The problem of food insecurity was often defined by food bank use or hunger, but other definitions and a range of consequences for individuals were acknowledged. Articles identified government as a driver of food insecurity, especially in relation to the roll-out of Universal Credit. Few articles proposed individual failings as a driver of food insecurity. The reported existing solutions predominantly focused on food banking and redistributing 'food waste'. The public, charities, and individuals experiencing food insecurity were generally portrayed as supportive of government action to tackle food insecurity. However, contention within government regarding the extent of food insecurity, governmental responsibility and potential solutions was reported. Food insecurity was a topic of significant interest within UK newspapers. Newspapers were used to call for government action and advocate for structural, income-based solutions.


Natural history and expansive clinical profile of stress (tako-tsubo) cardiomyopathy.
This study was designed to define more completely the clinical spectrum and consequences of stress cardiomyopathy (SC) beyond the acute event. Stress cardiomyopathy is a recently recognized condition characterized by transient cardiac dysfunction with ventricular ballooning. Clinical profile and outcome were prospectively assessed in 136 consecutive SC patients. Patients were predominantly women (n = 130; 96%), but 6 were men (4%). Ages were 32 to 94 years (mean age 68 +/- 13 years); 13 (10%) were <or=50 years of age. In 121 patients (89%), SC was precipitated by intensely stressful emotional (n = 64) or physical (n = 57) events, including 22 associated with sympathomimetic drugs or medical/surgical procedures; 15 other patients (11%) had no evident stress trigger. Twenty-five patients (18%) were taking beta-blockers at the time of SC events. Three diverse ventricular contraction patterns were defined by cardiovascular magnetic resonance (CMR) imaging, usually with rapid return to normal systolic function, although delayed >2 months in 5%. Right and/or left ventricular thrombi were identified in 5 patients (predominantly by CMR imaging), including 2 with embolic events. Three patients (2%) died in-hospital and 116 (85%) have survived, including 5% with nonfatal recurrent SC events. All-cause mortality during follow-up exceeded a matched general population (p = 0.016) with most deaths occurring in the first year. In this large SC cohort, the clinical spectrum was heterogeneous with about one-third either male, <or=50 years of age, without a stress trigger, or with in-hospital death, nonfatal recurrence, embolic stroke, or delayed normalization of ejection fraction. Beta-blocking drugs were not absolutely protective and SC was a marker for increased noncardiac mortality. These data support expanded management and surveillance strategies including CMR imaging and consideration for anticoagulation.


Erythropoietic defect associated with reduced cell proliferation in mice lacking the 26S proteasome shuttling factor Rad23b.
Rad23a and Rad23b proteins are linked to nucleotide excision DNA repair (NER) via association with the DNA damage recognition protein xeroderma pigmentosum group C (XPC) are and known to be implicated in protein turnover by the 26S proteasome. Rad23b-null mice are NER proficient, likely due to the redundant function of the Rad23b paralogue, Rad23a. However, Rad23b-null midgestation embryos are anemic, and most embryos die before birth. Using an unbiased proteomics approach, we found that the majority of Rad23b-interacting partners are associated with the ubiquitin-proteasome system (UPS). We tested the requirement for Rad23b-dependent UPS activity in cellular proliferation and more specifically in the process of erythropoiesis. In cultured fibroblasts derived from embryos lacking Rad23b, proliferation rates were reduced. In fetal livers of Rad23b-null embryos, we observed reduced proliferation, accumulation of early erythroid progenitors, and a block during erythroid maturation. In primary wild-type (WT) erythroid cells, knockdown of Rad23b or chemical inhibition of the proteasome reduced survival and differentiation capability. Finally, the defects linked to Rad23b loss specifically affected fetal definitive erythropoiesis and stress erythropoiesis in adult mice. Together, these data indicate a previously unappreciated requirement for Rad23b and the UPS in regulation of proliferation in different cell types.


A comparison of the therapeutic efficacy and tolerability of etoricoxib and diclofenac in patients with osteoarthritis.
To evaluate and compare the efficacy and tolerability of etoricoxib and diclofenac in patients with osteoarthritis of the knee or hip. In this 6-week double-blind, active comparator controlled, parallel-group study eligible osteoarthritis patients were randomised to receive either etoricoxib 60 mg once daily (n = 256) or diclofenac 50 mg three times daily (n = 260). The primary study endpoint was the Western Ontario McMaster osteoarthritis index (WOMAC) pain subscale. Other endpoints included were the WOMAC stiffness and physical function subscales, and the Patient's Global Assessment of Response to Therapy (PGART) questionnaire. Early efficacy was evaluated using WOMAC first question (pain walking on a flat surface) and PGART 4 h after the morning dose of each drug on days 1 and 2. Rescue medication (paracetamol) used was also recorded. The study was designed to show comparable efficacy between etoricoxib 60 mg once daily and diclofenac 50 mg three times daily with respect to the primary endpoint and was conducted outside the United States at 67 centres in 29 countries. Etoricoxib (60 mg once daily) was comparable in efficacy to diclofenac (150 mg daily) on all the above parameters. The one exception was in the assessment of early efficacy where etoricoxib demonstrated significantly greater benefit within 4 h of taking the first dose on the first day of therapy (p = 0.007) as evaluated by the percentage of patients with good or excellent (PGART) responses. The treatment effects of both drugs were similar by the time day 2 was reached and were sustained throughout the 6 weeks of therapy. Both treatments were generally well tolerated. Etoricoxib is clinically effective in the therapy of osteoarthritis providing a magnitude of effect comparable to that of the maximum recommended daily dose of diclofenac. The onset of clinical benefit with etoricoxib on day one is more rapid than that of diclofenac. Both drugs were generally well tolerated.


Developing stochastic models for spatial inference: bacterial chemotaxis.
Biological systems are inherently inhomogeneous and spatial effects play a significant role in processes such as pattern formation. At the cellular level proteins are often localised either through static attachment or via a dynamic equilibrium. As well as spatial heterogeneity many cellular processes exhibit stochastic fluctuations and so to make inferences about the location of molecules there is a need for spatial stochastic models. A test case for spatial models has been bacterial chemotaxis which has been studied extensively as a model of signal transduction. By creating specific models of a cellular system that incorporate the spatial distributions of molecules we have shown how the fit between simulated and experimental data can be used to make inferences about localisation, in the case of bacterial chemotaxis. This method allows the robust comparison of different spatial models through alternative model parameterisations. By using detailed statistical analysis we can reliably infer the parameters for the spatial models, and also to evaluate alternative models. The statistical methods employed in this case are particularly powerful as they reduce the need for a large number of simulation replicates. The technique is also particularly useful when only limited molecular level data is available or where molecular data is not quantitative.


Animal Bite Injuries in Children: Review of Literature and Case Series.
Maxillofacial region in children is particularly vulnerable to animal bite injuries. These injuries may range from insignificant scratches to life-threatening neck and facial injuries. Children are the common victims, particularly of dog bites. Three cases of animal bite injuries in children with their clinical presentation and their management are being presented along with review of literature. Surgical management included cleansing and primary closure of the wound. Rabies and tetanus prophylaxis were given. The most common site of injury was the face. For the facial injuries, the most frequently affected area was the middle third (55%), also called as the "central target area." The small stature of children, the disproportionate size of the head relative to the body, their willingness to bring their faces close to the animal, and limited motor skills to provide defense are believed to account for this. The resulting soft-tissue injuries can vary in relation to their extent. Treatment involved initial surgical exploration, and secondary repair later depending on the severity of the injury. Prompt assessment and treatment can prevent most bite wound complications. Early management of such injuries usually guarantees satisfactory outcome. Prevention strategies include close supervision of child-dog interactions, better reporting of bites, etc. Agrawal A, Kumar P, Singhal R, Singh V, Bhagol A. Animal Bite Injuries in Children: Review of Literature and Case Series. Int J Clin Pediatr Dent 2017;10(1):67-72.


Amyloid related cerebral microbleed and plasma Aβ40 are associated with cognitive decline in Parkinson's disease.
Cerebral microbleeds (MBs) have been found in patients with cognitive decline. We aimed to examine whether MBs are associated with motor or cognitive decline in patients with Parkinson's disease (PD). We enrolled 135 PD patients and 34 healthy controls. All participants underwent brain MRI and plasma biomarker assays, including tau, Aβ42, Aβ40, and α-synuclein. PD with dementia (PDD) was operationally defined as Mini-Mental State Examination (MMSE) score < 26 and advanced motor stage was defined as Hoehn-Yahr stage ≥ 3 during "on" status. The association between MBs and disease severity was examined using multivariate logistic regression models. More lobar MBs were observed in PD patients than controls (20.7% vs. 3.3%, p = 0.031). PDD patients had more lobar MBs (33.3% vs. 15.6%, p = 0.034), more white matter hyperintensity (p = 0.021) and reduced hippocampal volume (p = 0.001) than PD with normal cognition. The presence of lobar MB (odds ratio = 2.83 [95% confidence interval 1.04-7.70], p = 0.042) and severe white matter hyperintensity (3.29 [1.21-8.96], p = 0.020) was independently associated with PDD after adjusting for vascular risk factors and other confounders. Furthermore, plasma Aβ40 levels were associated the MMSE score (p = 0.004) after adjusting for age and sex. Our findings demonstrated that lobar MBs, reduced hippocampal volume, and elevated plasma Aβ40 levels are associated with PDD.


Influence of interface-dipole interactions on the efficiency of fluorescence light collection near surfaces.
Recently Török and colleagues published in a series of papers a general wave-optical approach to calculating the light-collection efficiency function (CEF) for confocal imaging of dipole emitters [J. Mod. Opt. 45, 1681 (1998); J. Microec. 194, 127 (1999); Opt. Lett. 25, 1463 (2000)]. In their theory they did not address the possibility that the lifetimes of fluorescing molecules can change significantly near interfaces, which has a direct effect on the CEF. The research of Török and colleagues is extended here to include this effect, which may become important for imaging near surfaces.


A distinctive interaction between memory and chronic daytime somnolence in asymptomatic APOE e4 homozygotes.
To correlate memory measures with a trait measure of chronic daytime somnolence in cognitively normal individuals with different gene doses of the apolipoprotein E (APOE) e4 allele, a common Alzheimer's disease susceptibility gene. Cross-sectional, exploratory study of cognitive abilities in APOE e4 homozygotes (HMZ) (n=42), heterozygotes (HTZ), (n=42) and noncarriers (NC) (n=42) who are matched for age, gender, educational level, and family history of dementia. Tertiary care academic medical center. Cognitively normal residents of Maricopa County, Arizona who are 30-70 years of age, genotyped for APOE, and have no history of a sleep disorder N/A. Epworth Sleepiness Scale (ESS) and a battery of neuropsychological tests Age, education, gender, and insomnia complaints did not significantly differ among groups. Despite normal baseline memory scores, memory declined with increasing ESS on all eight memory measures in the HMZ, two of eight in the HTZ, and one of eight in the NC. Differences between HMZ and NC on the slope of memory decline with increasing ESS reached statistical significance on two verbal memory measures, AVLT Long-Term Memory (p=0.048) and Percent Delayed Recall (p=0.035). Chronic daytime somnolence is associated with a distinctive decline in verbal memory in cognitively normal APOE e4 HMZ, a group at particularly high risk of Alzheimer's disease. Additional studies are needed to confirm these exploratory findings and to determine the effects of acute somnolence on cognition in these genetic subgroups.


Managing the Unimaginable: Biobank Participant Views on Reconsent for Whole Genome Sequencing of Stored Biospecimens.
Background: DNA biobanks frequently obtain broad permissions from sample donors, who agree to allow their biospecimens to be used for a variety of future purposes. A limitation of this approach is that it may not be possible to discuss or anticipate all potential uses of biospecimens at the time patient consent is obtained. We surveyed biobank participants to clarify their views regarding the need to be informed about research involving whole genome sequencing (WGS). Methods: We invited 1200 participants in the Mayo Clinic Biobank to complete a survey inquiring about their support for WGS; their interest in being recontacted before WGS of their biospecimens; whether they would consent to WGS if asked; and the acceptability of proceeding with WGS if sample donors could not be reached. Results: Six hundred eighty-seven biobank participants returned completed surveys (57% response). More than 96% of biobank participants were supportive of WGS and would give permission for WGS of their sample, if asked. Nonetheless, 61% of biobank participants felt they should be recontacted before WGS was done. Participants were divided regarding the permissibility of conducting WGS if efforts to recontact sample donors were unsuccessful. Discussion: Our findings highlight a potential discrepancy between the broad permissions granted by biobank participants at the time they donated biospecimens and their views about the application of WGS to their samples. Biobank participants appear to value the ability to confirm their commitment to genetic research when the studies in question involve WGS, a technological capacity they may not have anticipated at the time they donated their biospecimens. Efforts to reevaluate biobank participants' views about the acceptability of new technologies may help to ensure alignment of participants' current beliefs and research applications that would have been difficult to anticipate at the time biospecimens were collected.


Tolerizing CTL by Sustained Hepatic PD-L1 Expression Provides a New Therapy Approach in Mouse Sepsis.
Cytotoxic T lymphocyte (CTL) activation contributes to liver damage during sepsis, but the mechanisms involved are largely unknown. Understanding the underlying principle will permit interference with CTL activation and thus, provide a new therapeutic option. Methods: To elucidate the mechanism leading to CTL activation we used the Hepa1-6 cell line in vitro and the mouse model of in vivo polymicrobial sepsis, following cecal-ligation and -puncture (CLP) in wildtype, myeloid specific NOX-2, global NOX2 and NOX4 knockout mice, and their survival as a final readout. In this in vivo setting, we also determined hepatic mRNA and protein expression as well as clinical parameters of liver damage - aspartate- and alanine amino-transaminases. Hepatocyte specific overexpression of PD-L1 was achieved in vivo by adenoviral infection and transposon-based gene transfer using hydrodynamic injection. Results: We observed downregulation of PD-L1 on hepatocytes in the murine sepsis model. Adenoviral and transposon-based gene transfer to restore PD-L1 expression, significantly improved survival and reduced the release of liver damage, as PD-L1 is a co-receptor that negatively regulates T cell function. Similar protection was observed during pharmacological intervention using recombinant PD-L1-Fc. N-acetylcysteine blocked the downregulation of PD-L1 suggesting the involvement of reactive oxygen species. This was confirmed in vivo, as we observed significant upregulation of PD-L1 expression in NOX4 knockout mice, following sham operation, whereas its expression in global as well as myeloid lineage NOX2 knockout mice was comparable to that in the wild type animals. PD-L1 expression remained high following CLP only in total NOX2 knockouts, resulting in significantly reduced release of liver damage markers. Conclusion: These results suggest that, contrary to common assumption, maintaining PD-L1 expression on hepatocytes improves liver damage and survival of mice during sepsis. We conclude that administering recombinant PD-L1 or inhibiting NOX2 activity might offer a new therapeutic option in sepsis.


Abnormal expression of the E2 component of the pyruvate dehydrogenase complex on the luminal surface of biliary epithelium occurs before major histocompatibility complex class II and BB1/B7 expression.
Primary biliary cirrhosis (PBC) is a chronic autoimmune liver disease characterized histologically by nonsuppurative destructive cholangitis. Sera from patients with PBC react with a series of intramitochondrial enzymes with the immunodominant response directed against the E2 component of the pyruvate dehydrogenase complex (PDC-E2). Recently, using tissue sections of late-stage PBC, we showed that there is increased expression in biliary epithelial cells of patients with PDC-E2 or a molecule cross-reactive with PDC-E2. Previous work has shown that biliary epithelial cells of patients with PBC express an increased amount of class II. To address the sequence of events in the evolution of PBC, we have focused our attention in this study on early biliary epithelial lesions. In particular, we have studied the liver of 22 female patients with PBC that was diagnosed as either stage I or stage II using both a mouse monoclonal antibody that has reactivity similar to human autoantibodies as well as a human Fab combinatorial prepared from the lymph node of a PBC patient. Tissues were simultaneously stained using antibodies to PDC-E2, class II, and BB1/B7. As a positive control, tissues from late-stage PBC were studied concurrently. By determining the order of expression among the three molecules, PDC-E2, class II, and BB1/B7, we report that the expression of PDC-E2 or a PDC-E2-like molecule on biliary duct epithelium of patients with PBC precedes the expression of BB1/B7 and major histocompatibility complex (MHC) class II molecules. The alteration of an autoantigen in biliary duct epithelium may be the earliest lesion in PBC.