Immunochromatographic strip test for detection of genus Cronobacter.
Members of the genus Cronobacter are opportunistic pathogens formerly known as Enterobacter sakazakii, which induce severe meningitis and sepsis in neonates and infants, with a high fatality rate. In this work, a simple and rapid immunochromatographic strip test for the detection of this pathogen was developed. Following the shortened bacteria cultivation and isolation of DNA, a specific gene sequence targeting 16S rRNA from Cronobacter spp. was amplified by PCR using 5'-end labelled specific primers. The PCR product, amplicon labelled with digoxigenin on one side and biotin on the other side, was directly added to the immunochromatographic strip test, composed of nitrocellulose membrane with bound antibody against digoxigenin in the test line. The visualization was mediated by colloidal carbon conjugated to neutravidin, and the appearance of grey/black line was indicative of the presence of specific amplicon. Colour intensity of the test line in pathogen-positive assay was visually distinguishable from that of negative sample within 10 min. The visual detection limit of PCR product was 8 ng. The specificity of the developed method was confirmed by standard microbiological techniques. Whole detection procedure with the incorporated immunostrip was applied to analysis of infant formulae samples, contaminated with less than 10 cells of Cronobacter spp. per 10 g. The results from immunochromatographic test indicated the absolute agreement with those from standard microbiological methods. Moreover, the developed procedure considerably reduced the total analysis time to 16 h whereas the reference microbiological method needs 6-7 days.


Strength of vital force in classical homeopathy: bio-psycho-social-spiritual correlates within a complex systems context.
To explore associations between a global rating for the classical homeopathic construct of vital force and clinician and patient ratings on previously validated bio-psycho-social-spiritual questionnaires. Sixty-two (62) community-recruited patients with fibromyalgia (FM) were assessed at baseline prior to a clinical trial of individualized homeopathy. Two homeopaths jointly performed case-taking interviews. A conventional medical provider independently evaluated patients with a standardized history and physical examination. Homeopaths rated each patient's vital force (five-point Likert scale, with 1 = very weak to 5 = very strong). Homeopaths and the conventional medical provider rated their Clinical Global Impression (CGI) of the severity of illness (1 = normal; 7 = among the most extremely ill). Patients completed self-rating scales on pain, global health, mood, quality of life, coping style, health locus of control, multidimensional well-being, spirituality, sense of coherence, positive states of mind, and social desirability. Greater vital force ratings (mean 2.9 standard deviation [SD] 0.6) correlated moderately (p < or = 0.005) with less severe CGI illness ratings by the homeopaths (r =-0.59), decreased patient-rated mental confusion (r =-0.43), higher vigor (r = 0.38), and greater positive states of mind (r = 0.36). Vital force also showed correlations (p < 0.05) with lower CGI ratings by the conventional medical provider (r =-0.32), better selfrated quality of life (r = 0.33), lesser fatigue (r =-0.31), better global health (r = 0.29), greater sense of coherence (r = 0.28), powerful-others health locus of control (r = 0.27), increased emotional well-being (r = 0.27), and higher social desirability (r = 0.27), but not with age, pain, or illness duration. Homeopathic vital force ratings reflect better perceived mental function, energy, and positive dimensions of the individual, beyond absence of disease.


Hormone-sensitive lipase in differentiated 3T3-L1 cells and its activation by cyclic AMP-dependent protein kinase.
Differentiation of 3T3-L1 fibroblasts to adipocyte-like cells was accompanied by a 19-fold increase in neutral triglyceride lipase activity, a 12-fold increase in diglyceride lipase activity, a 10-fold increase in monoglyceride lipase activity, and a 280-fold increase in cholesterol esterase activity. In contrast, acid acylhydrolase activities did not increase during differentiation. The rate of glycerol release from unstimulated intact cells increased by more than 1 order of magnitude upon differentiation. Isoproterenol (1 microM) and 1-methyl-3-isobutylxanthine (0.1 mM) further stimulated this rate of glycerol release 3-fold. The neutral triglyceride lipase activity in cell-free preparations of differentiated cells was activated 105% by cyclic AMP-dependent protein kinase. Neutral cholesterol esterase, diglyceride lipase, and monoglyceride lipase were also activated (117%, 10%, and 37+, respectively) by cyclic AMP-dependent protein kinase. In contrast, protein kinase had no effect on any of the four lysosomal acid acylhydrolase activities. Thus, hormone-sensitive lipase, the most characteristic and functionally important enzyme of adipose tissue, has been characterized in differentiated 3T3-L1 cells. The 3T3-L1 cell should be a valuable model system in which to study regulation of hormone-sensitive lipase, particularly its long-term regulation.


Cation-π interactions in protein-ligand binding: theory and data-mining reveal different roles for lysine and arginine.
We have studied the cation-π interactions of neutral aromatic ligands with the cationic amino acid residues arginine, histidine and lysine using ab initio calculations, symmetry adapted perturbation theory (SAPT), and a systematic meta-analysis of all available Protein Data Bank (PDB) X-ray structures. Quantum chemical potential energy surfaces (PES) for these interactions were obtained at the DLPNO-CCSD(T) level of theory and compared against the empirical distribution of 2012 unique protein-ligand cation-π interactions found in X-ray crystal structures. We created a workflow to extract these structures from the PDB, filtering by interaction type and residue pKa. The gas phase cation-π interaction of lysine is the strongest by more than 10 kcal mol-1, but the empirical distribution of 582 X-ray structures lies away from the minimum on the interaction PES. In contrast, 1381 structures involving arginine match the underlying calculated PES with good agreement. SAPT analysis revealed that underlying differences in the balance of electrostatic and dispersion contributions are responsible for this behavior in the context of the protein environment. The lysine-arene interaction, dominated by electrostatics, is greatly weakened by a surrounding dielectric medium and causes it to become essentially negligible in strength and without a well-defined equilibrium separation. The arginine-arene interaction involves a near equal mix of dispersion and electrostatic attraction, which is weakened to a much smaller degree by the surrounding medium. Our results account for the paucity of cation-π interactions involving lysine, even though this is a more common residue than arginine. Aromatic ligands are most likely to interact with cationic arginine residues as this interaction is stronger than for lysine in higher polarity surroundings.


Glutamine depletion induces murine neonatal melena with increased apoptosis of the intestinal epithelium.
To investigate the possible biological outcome and effect of glutamine depletion in neonatal mice and rodent intestinal epithelial cells. We developed three kinds of artificial milk with different amounts of glutamine; Complete amino acid milk (CAM), which is based on maternal mouse milk, glutamine-depleted milk (GDM), and glutamine-rich milk (GRM). GRM contains three-fold more glutamine than CAM. Eighty-seven newborn mice were divided into three groups and were fed with either of CAM, GDM, or GRM via a recently improved nipple-bottle system for seven days. After the feeding period, the mice were subjected to macroscopic and microscopic observations by immunohistochemistry for 5-bromo-2'-deoxyuridine (BrdU) and Ki-67 as markers of cell proliferation, and for cleaved-caspase-3 as a marker of apoptosis. Moreover, IEC6 rat intestinal epithelial cells were cultured in different concentrations of glutamine and were subject to a 4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate cell proliferation assay, flow cytometry, and western blotting to examine the biological effect of glutamine on cell growth and apoptosis. During the feeding period, we found colonic hemorrhage in six of 28 GDM-fed mice (21.4%), but not in the GRM-fed mice, with no differences in body weight gain between each group. Microscopic examination showed destruction of microvilli and the disappearance of glycocalyx of the intestinal wall in the colon epithelial tissues taken from GDM-fed mice. Intake of GDM reduced BrdU incorporation (the average percentage of BrdU-positive staining; GRM: 13.8%, CAM: 10.7%, GDM: 1.14%, GRM vs GDM: P < 0.001, CAM vs GDM: P < 0.001) and Ki-67 labeling index (the average percentage of Ki-67-positive staining; GRM: 24.5%, CAM: 22.4% GDM: 19.4%, GRM vs GDM: P = 0.001, CAM vs GDM: P = 0.049), suggesting that glutamine depletion inhibited cell proliferation of intestinal epithelial cells. Glutamine deprivation further caused the deformation of the nuclear membrane and the plasma membrane, accompanied by chromatin degeneration and an absence of fat droplets from the colonic epithelia, indicating that the cells underwent apoptosis. Moreover, immunohistochemical analysis revealed the appearance of cleaved caspase-3 in colonic epithelial cells of GDM-fed mice. Finally, when IEC6 rat intestinal epithelial cells were cultured without glutamine, cell proliferation was significantly suppressed after 24 h (relative cell growth; 4 mmol/L: 100.0% ± 36.1%, 0 mmol/L: 25.3% ± 25.0%, P < 0.05), with severe cellular damage. The cells underwent apoptosis, accompanied by increased cell population in sub-G0 phase (4 mmol/L: 1.68%, 0.4 mmol/L: 1.35%, 0 mmol/L: 5.21%), where dying cells are supposed to accumulate. Glutamine is an important alimentary component for the maintenance of intestinal mucosa. Glutamine deprivation can cause instability of the intestinal epithelial alignment by increased apoptosis.


Spectral Radiative Flux Divergence and Its Variability in the Troposphere in the 0.4-2.4-micro Region.
Within the program of the Complex Atmospheric Energetics Experiment (CAENEX-70) and with the help of an IL-18 aircraft flying laboratory, we have measured spectral (in the 0.4-2.4-microm interval) radiative fluxes, the energy balance, and flux divergence of energy in a free atmosphere (at 0.3-8.4-km heights) with Z = 55 degrees over a sandy surface. Simultaneously, the results of aerosol measurements gave the physical explanation to many peculiar features of optical observations. In particular, aerosol absorption of shortwave radiation has been isolated. The probable variability of aerosol absorption has been estimated on the basis of observational data obtained on other days.


Valproic acid substantially downregulated genes folr1, IGF2R, RGS2, COL6A3, EDNRB, KLF6, and pax-3, N-acetylcysteine alleviated most of the induced gene alterations in chicken embryo model.
Valproic acid induced teratogenicity at genetic and somatic levels, the action mechanism is still unclear. We hypothesized that folate receptor gene (folr1) and others may be interacting to elicit neural tube defect (NTD), while N-acetylcysteine (NAC) may be beneficial for protection. In chicken embryo model, the experiment was conducted in two parts. The first part was carried out to test the optimum dose of VPA. The second part was conducted to test the protective effect of NAC at doses 10 and 20 mM. VPA induced dysvascularization, incomplete somite enclosure, histone deacetylase (HDAC) inhibition, folate deficiency, homocysteine accumulation, SOD inhibition, glutathione depletion, elevated MDA and hydrogen peroxide. NAC alleviated most of these adverse effects. The microarray analysis revealed 17 genes downregulated and four upregulated. The relevancy covered translation (23%), signal transduction (23%), transcription (16%), cell adhesion (16%), neural cell migration (8%), transport (7%), and organismal development (7%). The genes insulin-like growth factor 2 receptor gene (IGF2R), regulator of G-protein signaling 4 gene (RGS4), alpha 3 (VI) collagen gene (COL6A3), endothelin receptor type b gene (EDNRB), and Krüppel-like factor 6 gene (KLF6) substantially downregulated in reality were directly intermodulating and associated with NTD. VPA downregulated folr1 gene in a dose responsive manner without affecting pax-3 gene, which was ascribed to the metahypoxic state. Conclusively, VPA affects 21 genes: 17 downregulated and four upregulated. VPA dose responsively downregulates gene folr1 without affecting pax-3 gene. These adverse effects can be partially alleviated by N-acetylcysteine.


Formation of Monolithic Ion-Selective Transport Media Based on "Click" Cross-Linked Hyperbranched Polyglycerol.
In the emerging field of organic bioelectronics, conducting polymers and ion-selective membranes are combined to form resistors, diodes, transistors, and circuits that transport and process both electronic and ionic signals. Such bioelectronics concepts have been explored in delivery devices that translate electronic addressing signals into the transport and dispensing of small charged biomolecules at high specificity and spatiotemporal resolution. Manufacturing such "iontronic" devices generally involves classical thin film processing of polyelectrolyte layers and insulators followed by application of electrolytes. This approach makes miniaturization and integration difficult, simply because the ion selective polyelectrolytes swell after completing the manufacturing. To advance such bioelectronics/iontronics and to enable applications where relatively larger molecules can be delivered, it is important to develop a versatile material system in which the charge/size selectivity can be easily tailormade at the same time enabling easy manufacturing of complex and miniaturized structures. Here, we report a one-pot synthesis approach with minimal amount of organic solvent to achieve cationic hyperbranched polyglycerol films for iontronics applications. The hyperbranched structure allows for tunable pre multi-functionalization, which combines available unsaturated groups used in crosslinking along with ionic groups for electrolytic properties, to achieve a one-step process when applied in devices for monolithic membrane gel formation with selective electrophoretic transport of molecules.


The Fountain of Youth Will Not Last Forever: End of Life in Patients Receiving Mechanical Circulatory Support.
Health care providers face a unique set of decision-making, assessment, and equipment challenges at the end of life of patients supported with a left ventricular assist device. The aim for this article is to assist the multidisciplinary team in caring for patients with a left ventricular assist device in all phases of end-of-life care. This review includes common causes of death for patients with a left ventricular assist device, assessment at end of life, physiological and palliative care considerations, withdrawal of left ventricular assist device support, and equipment considerations.


Zebrafish Rho kinase 2 acts downstream of Wnt11 to mediate cell polarity and effective convergence and extension movements.
During vertebrate gastrulation convergence and extension (CE), movements narrow and lengthen embryonic tissues. In Xenopus and zebrafish, a noncanonical Wnt signaling pathway constitutes the vertebrate counterpart to the Drosophila planar cell polarity pathway and regulates mediolateral cell polarization underlying CE. Despite the identification of several signaling molecules required for normal CE, the downstream transducers regulating individual cell behaviors driving CE are only beginning to be elucidated. Moreover, how defective mediolateral cell polarity impacts CE is not understood. Here, we show that overexpression of zebrafish dominant-negative Rho kinase 2 (dnRok2) disrupts CE without altering cell fates, phenocopying noncanonical Wnt signaling mutants. Moreover, Rho kinase 2 (Rok2) overexpression partially suppresses the slb/wnt11 gastrulation phenotype, and ectopic expression of noncanonical Wnts modulates Rok2 intracellular distribution. In addition, time-lapse analyses associate defective dorsal convergence movements with impaired cell elongation, mediolateral orientation, and consequently failure to migrate along straight paths. Transplantation experiments reveal that dnRok2 cells in wild-type hosts neither elongate nor orient their axes. In contrast, wild-type cells are able to elongate their cell bodies in dnRok2 hosts, even though they fail to orient their axes. During zebrafish gastrulation, Rok2 acts downstream of noncanonical Wnt11 signaling to mediate mediolateral cell elongation required for dorsal cell movement along straight paths. Furthermore, elongation and orientation of the cell body are independent properties that require both cell-autonomous and nonautonomous Rok2 function.


Early expression of injury-induced neuropeptide Y in primary sensory neurons and the cuneate nucleus in diabetic rats with median nerve transection.
In this study we examined the temporal changes in neuropeptide Y (NPY) expression in dorsal root ganglion (DRG) neurons and cuneate nucleus (CN) in streptozotocin (STZ)-induced diabetic rats with or without median nerve transection (MNT). Numerous NPY-like immunoreactive (NPY-LI) neurons and fibers were detected in the DRG and CN of the diabetic MNT (DMNT) rats respectively, but not in those with diabetes-alone. Following MNT, the time-course of NPY expression pattern in the diabetic DRG and CN was similar and both peaked at 2 weeks, which was earlier than those in the non-diabetic MNT rats. Consequently, the expression of neurotrophin-3 (NT-3) immunoreactivity in DRG neurons was coincidentally decreased and reached the nadir at 2 weeks in the diabetic MNT rats, which was also earlier than that in the non-diabetic MNT rats. Following electrical stimulation of the injured nerves, the number of NPY-LI fibers became attenuated and the induced c-Fos-LI cells concurrently appeared in the ipsilateral CN. In the diabetic CN, the number of c-Fos-LI cells also peaked at 2 weeks after MNT, which was consistent with the temporal pattern of changes in NPY expression. The results suggest that in diabetes, MNT induced NPY expression via the reduction of NT-3, and electrical stimulation of the injured median nerve evoked the release of NPY and accordingly more c-Fos-LI cells were identified in the CN. Furthermore, this study demonstrated early NPY and c-Fos expression in the diabetic rats after MNT, suggesting that the development of neuropathic signs may be advanced in hyperglycemic rats.


Use of aldehyde fixatives to determine the rate of synaptic transmitter release.
Aldehyde fixation continues to be useful to prepare synapses for freeze-fracture, but it may increase the rate of transmitter release. The effects of different aldehyde fixatives on spontaneous quantal release (m.e.p.p.s), and on the corresponding synaptic vesicle exocytosis at frog nerve-muscle synapses were investigated with the hope of finding a way to minimize side effects of fixation. Increases in m.e.p.p.s of up to 50 S-1 occurred during fixation, despite the species of aldehyde used in the fixative, and this fixative effect decreased only slightly as aldehyde concentration was increased. Increases in m.e.p.p frequency were not blocked by tetrodotoxin, by lowering external calcium and raising external magnesium concentration, or by lowering the total osmotic strength of the fixative. The smallest increase in m.e.p.p. frequency was in 3% glutaraldehyde and corresponded to the lowest level of synaptic vesicle exocytosis seen by freeze-fracture, 0.15 per micrometers of active zone. The effects of aldehyde fixation on m.e.p.p. frequency and synaptic vesicle exocytosis could not be avoided, but this study suggests how its effect on morphological changes in synapses might be minimized.


Reference values of handgrip dynamometry of healthy adults: a population-based study.
Although maximal voluntary handgrip strength (HGS) is considered a reliable tool in nutritional assessment there are few reference data available. This paper presents reference values for handgrip strength of healthy adults (age > or = 20 years) from a household survey. Data were obtained from a representative sample of adults (1122 males and 1928 females) living in Niterói, Rio de Janeiro, Brazil. HGS was measured three times with a Jamar mechanical dynamometer in both hands and the highest value used in the analysis. The percentile distribution of HGS was calculated according to sex and age categories. Mean values of right and left HGS were 42.8 and 40.9 kg for males, and 25.3 and 24.0 kg for females, respectively. HGS increased with age and significantly decreased after 40 and 50 year-olds for women and men, respectively. Body mass index (BMI) was associated with HGS in both sexes but only underweight male subjects had significantly lower HGS values. The highest HGS values are observed at the 4th decade of life with significant declines thereafter. HGS is significantly associated with BMI. The reference values of HGS may be useful in assessing the nutritional status of similar adult urban population.


Periodic Refresher Emails for Emergency Department Mass Casualty Incident Plans.
This mass casualty incident (MCI) curriculum is intended for use as refresher content in the months between more formal education, such as hands-on MCI training and drills. The target audience for each topic varies, but the majority of them apply to all disciplines such as direct patient care roles (emergency room technicians, nurses, paramedics, advanced practice practitioners, resident physicians, attending physicians, etc.) and emergency department clerks/coordinators. Topics intended for only one or more discipline are labeled as such. See curriculum chart or email schedule (Appendix AI) for details. This curriculum is intended for use as weekly refresher emails spanning up to a 30-week period. There have been an increasing number of mass casualty events occurring throughout the country in recent years, many of which involve penetrating trauma. Education surrounding response to an MCI is broad and has many complex and ever-changing aspects that require staff to be updated on the most current information. This curriculum is intended to maintain a knowledge base of MCI processes to mitigate degradation of necessary knowledge between hands-on MCI training. The educational strategies used in this curriculum include short weekly refresher emails and optional external links for further reading. This content was evaluated for efficacy by administering electronic knowledge tests at baseline, mid-way (at 16 weeks), and at the end of the curriculum (32 weeks) via email. Additionally, brief content questions were asked in person while staff members were on shift throughout the entire study period, and a post-study survey was administered in order to obtain staff opinions on email length and training processes in general. Scores for the knowledge tests were slightly higher at the end of the 32 weeks compared to baseline. Subjective feedback was positive overall at the end of the testing period. Training and maintaining knowledge of roles and concepts of mass casualty incidents is vital since such events will never happen when expected. Short refresher emails allow educators to use spaced repetition and interleaving methods which have been shown to be a helpful adjunct to maintain knowledge, skills, and attitudes learned in more formal training.1,2. Mass casualty incident, emergency department, decontamination, blast injury, media relations, biological agents, reprocessing, crisis standard of care, SALT (Sort, Assess, Lifesaving Interventions, Treatment/Transport) triage, personal protective equipment, disaster carts, airways, passive security, family reunification.


Global versus local conservation focus of U.S. state agency endangered bird species lists.
The development of species priorities for conservation at local or regional scales (for example, within a state or province) poses an interesting paradox. One the one hand, locally or regionally-derived species priorities may lead to greater interest in and resources directed to biodiversity conservation by local or regional institutions. On the other hand, locally or regionally-derived species priorities could overlook national or global priorities. We assessed U.S. state government agency endangered-threatened bird lists to determine the comparative representation of species of global versus local conservation significance on them. State lists tended to be represented primarily by species of low global risk-low global responsibility (range: 15-100%; mean 51%) and high global risk-high global responsibility (range: 0-73%; mean 35%). In 25 states, more than half of the species on the state lists were in the low global risk-low global responsibility category. Most U.S. state agency lists represent a combined strategy of highlighting species of both local and global conservation significance. Even with this combined local-global strategy, most state lists were predominated by species that represent local but not global conservation significance. Such a strategy could have profound negative consequences for many species that are not formally recognized under national endangered species protections but that are also left off of state-level endangered species lists.


Self-assembled hyaluronic acid nanoparticles for active tumor targeting.
Hyaluronic acid nanoparticles (HA-NPs), which are formed by the self-assembly of hydrophobically modified HA derivatives, were prepared to investigate their physicochemical characteristics and fates in tumor-bearing mice after systemic administration. The particle sizes of HA-NPs were controlled in the range of 237-424 nm by varying the degree of substitution of the hydrophobic moiety. When SCC7 cancer cells over-expressing CD44 (the receptor for HA) were treated with fluorescently labeled Cy5.5-HA-NPs, strong fluorescence signals were observed in the cytosol of these cells, suggesting efficient intracellular uptake of HA-NPs by receptor-mediated endocytosis. In contrast, no significant fluorescence signals were observed when Cy5.5-labeled HA-NPs were incubated with normal fibroblast cells (CV-1) or with excess free-HA treated SCC7 cells. Following systemic administration of Cy5.5-labeled HA-NPs with different particle sizes into a tumor-bearing mouse, their biodistribution was monitored as a function of time using a non-invasive near-infrared fluorescence imaging system. Irrespective of the particle size, significant amounts of HA-NPs circulated for two days in the bloodstream and were selectively accumulated into the tumor site. The smaller HA-NPs were able to reach the tumor site more effectively than larger HA-NPs. Interestingly, the concentration of HA-NPs in the tumor site was dramatically reduced when mice were pretreated with an excess of free-HA. These results imply that HA-NPs can accumulate into the tumor site by a combination of passive and active targeting mechanisms.


The KGBR viewpoint-lighting ambiguity.
We address the visual ambiguities that arise in estimating object and scene structure from a set of images when the viewpoint and lighting are unknown. We obtain a novel viewpoint-lighting ambiguity called the KGBR that corresponds to a group of three-dimensional affine transformations on the object or scene geometry combined with transformations on the object or scene albedo. Our analysis assumes orthographic projection with an affine camera model. We include photometric cues, such as shadowing and shading, that we model using Lambertian reflectance functions with shadows (cast and attached) and multiple light sources (but no interreflections). We relate the KGBR to affine ambiguities in estimating shape and to the generalized bas-relief (GBR) ambiguity.


Seroprevalence of Trypanosoma cruzi in raccoons from Tennessee.
Trypanosoma cruzi is the etiologic agent of Chagas' disease. Autochthonous human and canine transmission of T. cruzi has been documented in Tennessee, but little is known about its ecology, including the prevalence of T. cruzi among wildlife in Tennessee. Serum samples from 706 raccoons (Procyon lotor) from 10 counties in the Ridge and Valley and Blue Ridge Mountains ecoregions of eastern Tennessee were tested for antibodies reactive with T. cruzi using the indirect fluorescent antibody assay. Two hundred six (29.2%) samples were seropositive, with 9 counties yielding positive samples (range 14.6-63.6%). Significantly more raccoons from rural habitats (35.1%) were found positive for T. cruzi exposure than were those from suburban habitats (23.1%, P < 0.001). Land cover class was not associated with seropositivity status (P = 0.441), even though deciduous forest was the most common site from where raccoons were trapped and the most common site of positive raccoons in rural areas (42%). Interestingly, age was positively associated with seropositivity. Raccoons older than 1 yr (adults) were 40.1% seropositive compared to 12.2% of those less than 1 yr (juveniles; P < 0.001). Female adults were significantly more likely to be exposed to T. cruzi than were male adult raccoons (P < 0.001). No significant seroprevalence difference was seen among male and female juveniles. This study contributes to understanding the dynamics of T. cruzi exposure within raccoon populations in Tennessee. The importance of habitat (rural vs. suburban) and microhabitat (dens) in risk of exposure to these populations is also discussed.


Human histamine N-methyltransferase pharmacogenetics: common genetic polymorphisms that alter activity.
Histamine N-methyltransferase (HNMT) catalyzes a major pathway in histamine metabolism. Levels of HNMT activity in humans are regulated by inheritance. We set out to study the molecular basis for this genetic regulation. Northern blot analysis showed that HNMT is highly expressed in the kidney, so we determined levels of enzyme activity and thermal stability in 127 human renal biopsy samples. DNA was isolated from 12 kidney samples with widely different HNMT phenotypes, and exons of the HNMT gene were amplified with the polymerase chain reaction. In these 12 samples, we observed a C314T transition that resulted in a Thr105Ile change in encoded amino acid, as well as an A939G transition within the 3'-untranslated region. All remaining renal biopsy samples then were genotyped for these two variant sequences. Frequencies of the alleles encoding Thr105 and Ile105 in the 114 samples studied were 0.90 and 0.10, respectively, whereas frequencies for the nucleotide A939 and G alleles were 0.79 and 0.21, respectively. Kidney samples with the allele encoding Ile105 had significantly lower levels of HNMT activity and thermal stability than did those with the allele that encoded Thr105. These observations were confirmed by transient expression in COS-1 cells of constructs that contained all four alleles for these two polymorphisms. COS-1 cells transfected with the Ile105 allele had significantly lower HNMT activity and immunoreactive HNMT protein than did those transfected with the Thr105 allele. These observations will make it possible to test the hypothesis that genetic polymorphisms for HNMT may play a role in the pathophysiology of human disease.


Genome erosion in a nitrogen-fixing vertically transmitted endosymbiotic multicellular cyanobacterium.
An ancient cyanobacterial incorporation into a eukaryotic organism led to the evolution of plastids (chloroplasts) and subsequently to the origin of the plant kingdom. The underlying mechanism and the identities of the partners in this monophyletic event remain elusive. To shed light on this evolutionary process, we sequenced the genome of a cyanobacterium residing extracellularly in an endosymbiosis with a plant, the water-fern Azolla filiculoides Lam. This symbiosis was selected as it has characters which make it unique among extant cyanobacterial plant symbioses: the cyanobacterium lacks autonomous growth and is vertically transmitted between plant generations. Our results reveal features of evolutionary significance. The genome is in an eroding state, evidenced by a large proportion of pseudogenes (31.2%) and a high frequency of transposable elements (approximately 600) scattered throughout the genome. Pseudogenization is found in genes such as the replication initiator dnaA and DNA repair genes, considered essential to free-living cyanobacteria. For some functional categories of genes pseudogenes are more prevalent than functional genes. Loss of function is apparent even within the 'core' gene categories of bacteria, such as genes involved in glycolysis and nutrient uptake. In contrast, serving as a critical source of nitrogen for the host, genes related to metabolic processes such as cell differentiation and nitrogen-fixation are well preserved. This is the first finding of genome degradation in a plant symbiont and phenotypically complex cyanobacterium and one of only a few extracellular endosymbionts described showing signs of reductive genome evolution. Our findings suggest an ongoing selective streamlining of this cyanobacterial genome which has resulted in an organism devoted to nitrogen fixation and devoid of autonomous growth. The cyanobacterial symbiont of Azolla can thus be considered at the initial phase of a transition from free-living organism to a nitrogen-fixing plant entity, a transition process which may mimic what drove the evolution of chloroplasts from a cyanobacterial ancestor.


Methods of purification of CTL-derived exosomes.
Exosomes are membrane nanovesicles (approximately <120 nm in size) released by most, if not all, living cells and in particular by leukocytes. They originate within the endocytic compartment by invagination of the endosome membrane. Therefore, they have a different biogenesis and molecular composition than microvesicles (>0.2 μm) shed from the plasma membrane. Although the functions of exosomes in vivo are beginning to be elucidated, increasing evidence suggests that exosomes constitute a mechanism of cell-to-cell communication, transferring antigens, proteins, mRNAs, and noncoding RNAs among cells. Interestingly, effector T cells including cytotoxic T lymphocytes (CTLs) release death-inducing molecules of the TNF superfamily through exosomes contained in their cytotoxic granules. The present chapter provides basic protocols for purification of exosomes secreted by CTLs.


Transfer standard for traceable dynamic calibration of stroboscopic scanning white light interferometer.
The reconstructed image of a moving sample always shows a distorted representation of reality. Therefore, one needs to calibrate, for example, out-of-plane nano-videos for quality control of nano-microelectromechanical systems (N-MEMS). Here we discuss how to calibrate and obtain confidence limits for stroboscopic scanning white light interferometry (SSWLI) data when there are differences in speed and amplitude across the field of view. Many N-MEMS devices rely on oscillating structures; consequently, one must calibrate movie recordings of these structures to have global standards and to allow inter-device comparison. We propose to use a quartz tuning fork driven off-resonance as a transfer standard. This approach allows a broad range of traceable frequencies and out-of-plane amplitudes to be introduced into selected parts of the field of view of the SSWLI device featuring similar optical surface properties to many N-MEMS devices without demanding an additional reference surface.


Sleep in the intensive care unit.
Abnormalities of sleep are extremely common in critically ill patients, but the mechanisms are poorly understood. About half of total sleep time occurs during the daytime, and circadian rhythm is markedly diminished or lost. Judgments based on inspection consistently overestimate sleep time and do not detect sleep disruption. Accordingly, reliable polygraphic recordings are needed to measure sleep quantity and quality in critically ill patients. Critically ill patients exhibit more frequent arousals and awakenings than is normal, and decreases in rapid eye movement and slow wave sleep. The degree of sleep fragmentation is at least equivalent to that seen in patients with obstructive sleep apnea. About 20% of arousals and awakenings are related to noise, 10% are related to patient care activities, and the cause for the remainder is not known; severity of underlying disease is likely an important factor. Mechanical ventilation can cause sleep disruption, but the precise mechanism has not been defined. Sleep disruption can induce sympathetic activation and elevation of blood pressure, which may contribute to patient morbidity. In healthy subjects, sleep deprivation can decrease immune function and promote negative nitrogen balance. Measures to improve the quantity and quality of sleep in critically ill patients include careful attention to mode of mechanical ventilation, decreasing noise, and sedative agents (although the latter are double-edged swords).


Intermodal Čerenkov radiation in a higher-order-mode fiber.
We demonstrate an intermodal Čerenkov radiation effect in a higher-order-mode (HOM) fiber with a mode crossing (i.e., two guided modes having the same propagation constant at the same wavelength). A frequency-shifted soliton in the vicinity of the mode-crossing wavelength emits a phase-matched dispersive wave in a different propagation mode. We develop a theoretical explanation for this nonlinear optical effect and demonstrate that the mode crossing in HOM fibers can be utilized to achieve simultaneous wavelength and mode conversion; the strength of this intermodal nonlinear interaction can be tuned by controlled fiber bending.


Key differentiating features between scrub typhus and hemorrhagic fever with renal syndrome in northern China.
Both scrub typhus and hemorrhagic fever with renal syndrome (HFRS) are severely epidemic in northern China and often present with acute undifferentiated fever. To correctly distinguish the two diseases at an early stage, we collected and compared clinical and routine laboratory data of 46 patients with confirmed scrub typhus and 49 patients with confirmed HFRS presenting to the outpatient departments of three town hospitals in northern China. Most patients with HFRS but none of the patients with scrub typhus had hemorrhagic manifestations. Retro-orbital pain, lumbar back pain, flank tenderness, proteinuria, and occult blood in urine often occurred in patients with HFRS. However, skin eschar, regional lymphadenopathy, and maculopapular rash were more commonly found in patients with scrub typhus. In addition, platelet counts in patients with HFRS were significantly lower than in patients with scrub typhus. These findings will be useful for physicians to distinguish scrub typhus from HFRS.


A missense mutation in the aggrecan C-type lectin domain disrupts extracellular matrix interactions and causes dominant familial osteochondritis dissecans.
Osteochondritis dissecans is a disorder in which fragments of articular cartilage and subchondral bone dislodge from the joint surface. We analyzed a five-generation family in which affected members had autosomal-dominant familial osteochondritis dissecans. A genome-wide linkage analysis identified aggrecan (ACAN) as a prime candidate gene for the disorder. Sequence analysis of ACAN revealed heterozygosity for a missense mutation (c.6907G > A) in affected individuals, resulting in a p.V2303M amino acid substitution in the aggrecan G3 domain C-type lectin, which mediates interactions with other proteins in the cartilage extracellular matrix. Binding studies with recombinant mutated and wild-type G3 proteins showed loss of fibulin-1, fibulin-2, and tenascin-R interactions for the V2303M protein. Mass spectrometric analyses of aggrecan purified from patient cartilage verified that V2303M aggrecan is produced and present in the tissue. Our results provide a molecular mechanism for the etiology of familial osteochondritis dissecans and show the importance of the aggrecan C-type lectin interactions for cartilage function in vivo.


A finite element model for direction-dependent mechanical response to nanoindentation of cortical bone allowing for anisotropic post-yield behavior of the tissue.
A finite element model was developed for numerical simulations of nanoindentation tests on cortical bone. The model allows for anisotropic elastic and post-yield behavior of the tissue. The material model for the post-yield behavior was obtained through a suitable linear transformation of the stress tensor components to define the properties of the real anisotropic material in terms of a fictitious isotropic solid. A tension-compression yield stress mismatch and a direction-dependent yield stress are allowed for. The constitutive parameters are determined on the basis of literature experimental data. Indentation experiments along the axial (the longitudinal direction of long bones) and transverse directions have been simulated with the purpose to calculate the indentation moduli and the tissue hardness in both the indentation directions. The results have shown that the transverse to axial mismatch of indentation moduli was correctly simulated regardless of the constitutive parameters used to describe the post-yield behavior. The axial to transverse hardness mismatch observed in experimental studies (see, for example, Rho et al. [1999, "Elastic Properties of Microstructural Components of Human Bone Tissue as Measured by Nanoindentation," J. Biomed. Mater. Res., 45, pp. 48-54] for results on human tibial cortical bone) can be correctly simulated through an anisotropic yield constitutive model. Furthermore, previous experimental results have shown that cortical bone tissue subject to nanoindentation does not exhibit piling-up. The numerical model presented in this paper shows that the probe tip-tissue friction and the post-yield deformation modes play a relevant role in this respect; in particular, a small dilatation angle, ruling the volumetric inelastic strain, is required to approach the experimental findings.


Thromboxane synthetase inhibition and thromboxane receptor blockade preserve pulmonary and circulatory function in a porcine burn sepsis model.
Thromboxane A2 (TxA2) is a key mediator in the pathophysiology of severe burns and sepsis. This study was performed to assess the potential benefits of TxA2 synthetase inhibition and TxA2 receptor blockade in sepsis after severe thermal injury. Pigs with left atrial, aortic, and pulmonary artery catheters received a 40 percent third-degree burn and, 24 hours later, 100 micrograms per kg Escherichia coli endotoxin. The antagonist treatment (BM) group was treated with the TxA2 receptor antagonist BM 13.177, the synthetase treatment (OKY) group with the TxA2 synthetase inhibitor OKY-046, and the control group received saline solution placebo. Another group without burn or endotoxin was used to assess the side effects of BM 13.177. Both drugs significantly attenuated the changes in pulmonary vascular resistance index, cardiac index, arterial PO2, shunt, oxygen delivery, and oxygen consumption seen after endotoxin. However, cardiac index was significantly decreased in group BM before endotoxin. In healthy pigs, BM 13.177 decreased cardiac index and oxygen delivery and increased the pulmonary vascular resistance index. TxA2 synthetase inhibitors and TxA2 receptor blockers are potentially useful in sepsis after severe burns. Comparison between drugs was complicated by the adverse effects of the antagonist, and further investigation with other antagonists is needed.


Characterization of cholecystokinin receptors on bovine gallbladder membranes.
Cholecystokinin (CCK) binding to its receptors on a muscularis membrane fraction of bovine gallbladder was characterized using a biologically active CCK-33-[125I]Bolton-Hunter conjugate. Receptor binding was localized to the muscularis layer of the gallbladder; no binding was seen on either mucosal or serosal membranes. At 24 degrees C and pH 6.5, binding was maximal after 60-90 min of incubation, remained at a plateau for at least 240 min, and was reversed by the addition of unlabeled CCK-8. Optimal binding was seen at pH of 5.5 and required the presence of magnesium. Gallbladder binding data, best fit by a two-parameter model using a nonlinear least-squares computer program, was consistent with a single order of binding sites with a Kd of 618 +/- 168 pM and a binding capacity of 100.5 +/- 15.7 fmol/mg prot (mean +/- SE, n = 5). CCK-8 and CCK-33 inhibited 125I-CCK binding to gallbladder membranes with similar potencies, whereas desulfated CCK-8, gastrin I and II, and CCK-4 were at least 500 times less potent than CCK-33. The CCK antagonists dibutyryl cGMP and proglumide inhibited 125I-CCK binding with an IC50 of 31 and 600 microM, respectively. The present studies therefore demonstrate the existence of a specific CCK receptor on bovine gallbladder muscularis membranes with a high degree of selectivity for CCK analogues.


Tuskegee redux: evolution of legal mandates for human experimentation.
Human health experiments systematically expose people to conditions beyond the boundaries of medical evidence. Such experiments have included legal-medical collaboration, exemplified in the U.S. by the Public Health Service (PHS) Syphilis Study (Tuskegee). That medical experiment was legal, conforming to segregationist protocols and specific legislative authorization which excluded a selected group of African Americans from any medical protection from syphilis. Subsequent corrective action outlawed unethical medical experiments but did not address other forms of collaboration, including PHS submission to laws which may have placed African American women at increased risk from AIDS and breast cancer. Today, anti-lobbying law makes it a felony for PHS workers to openly challenge legally anointed suspension of medical evidence. African Americans and other vulnerable populations may thereby face excess risks-not only from cancer, but also from motor vehicle crashes, firearm assault, end stage renal disease, and other problems-with PHS workers as silent partners.


Effects of Instructor Attractiveness on Learning.
Although a considerable body of research has examined the impact of student attractiveness on instructors, little attention has been given to the influence of instructor attractiveness on students. This study tested the hypothesis that persons would perform significantly better on a learning task when they perceived their instructor to be high in physical attractiveness. To test the hypothesis, participants listened to an audio lecture while viewing a photograph of instructor. The photograph depicted either a physically attractive instructor or a less attractive instructor. Following the lecture, participants completed a forced choice recognition task covering material from the lecture. Consistent with the predictions; attractive instructors were associated with more learning. Finally, we replicated previous findings demonstrating the role attractiveness plays in person perception.


Secondhand smoke and respiratory ill health in current smokers.
Numerous studies have concluded that secondhand smoke (SHS) is harmful to non-smokers but controversy persists regarding its effects on smokers. The impact of SHS exposure on the acute respiratory health of current active smokers was examined using a cross sectional design. 9923 uniformed staff in the Hong Kong Police Force completed a standardised questionnaire on current and past smoking, SHS exposure at home and at work, acute respiratory symptoms, and recent physician consultation. 3999 male current smokers were included in the analysis. About 5% of the smokers were exposed to SHS at home only, 53% were exposed at work only, and 30% were exposed both at home and at work. The prevalence ratios for respiratory symptoms (throat and nasal problems, cough, phlegm, and wheeze), physician consultation, and self medication were higher for those who were exposed to SHS at home or at work. The odds ratios of reporting one or more respiratory symptoms, for SHS exposures at home or at work, were 1.33 (95% confidence interval (CI) 1.12 to 1.59) and 1.66 (95% CI 1.36 to 2.02) respectively, after adjusting for age, marital status, education, rank and duties, exposure to self perceived dusty or polluted environment in previous job, and total dose of active smoking. The adjusted odds ratios showed significant positive dose-response gradients with SHS exposure at home, at work, and at both places combined. SHS exposure is strongly associated with increased acute respiratory symptoms and recent outpatient service utilisation in current smokers. If the association is causal, public health action to limit SHS exposure could also benefit smokers.


CysLT1 receptor inhibition in patients with Raynaud's phenomenon: capillaroscopic evidence of the role of leukotriene.
To assess the effect of the leukotriene receptor inhibitor (montelukast) on vascular alterations in fingers of patients with Raynaud's phenomenon. Patients with Raynaud's phenomenon of the hands secondary to inflammatory connective tissue disease were selected, and those with the following characteristics were excluded: smokers, arterial hypertension, and diabetes mellitus. All patients maintained their previous medications and started the use of montelukast, 10 mg/day, for 60 days. Naifold capillaroscopy of fingers was performed before the use of medication and after 30 and 60 days. Statistical analysis was performed with percentage, media, standard deviation, Fisher exact test, with 95% of confidence interval. The study assessed five Caucasian, female patients with Raynaud's phenomenon secondary to inflammatory connective tissue disease (three with scleroderma and two with mixed connective tissue disease), aged 42.4 ± 11.5 years, and with 9.6 ± 4.8 years of disease duration. Patients were on nifedipine and pentoxifylline, and those with mixed connective tissue disease were also on prednisone. The medications were maintained. After using montelukast for two months, nailfold capillaroscopy showed a reduction in edema and pallor, and normalization of capillary number, size, and distribution. The use of montelukast modified the capillary abnormalities observed on nailfold capillaroscopy of patients with Raynaud's phenomenon.


Economic evaluation of return-to-work interventions for mental disorder-related sickness absence: two years follow-up of a randomized clinical trial.
The objective was to (i) assess the long-term cost-effectiveness of acceptance and commitment therapy (ACT), a workplace dialog intervention (WDI), and ACT+WDI compared to treatment as usual (TAU) for common mental disorders and (ii) investigate any differences in cost-effectiveness between diagnostic groups. An economic evaluation from the healthcare and limited welfare perspectives was conducted alongside a randomized clinical trial with a two-year follow-up period. Persons with common mental disorders receiving sickness benefits were invited to the trial. We used registry data for cost analysis alongside participant data collected during the trial and the reduction in sickness absence days as treatment effect. A total of 264 participants with a diagnosis of depression, anxiety, or stress-induced exhaustion disorder participated in a two-year follow-up of a four-arm trial: ACT (N=74), WDI (N=60), ACT+WDI (N=70), and TAU (N=60). For all patients in general, there were no statistically significant differences between interventions in terms of costs or effect. The subgroup analyses suggested that from a healthcare perspective, ACT was a cost-effective option for depression or anxiety disorders and ACT+WDI for stress-induced exhaustion disorder. With a two-year time horizon, the probability of WDI to be cost-saving in terms of sickness benefits costs was 80% compared with TAU. ACT had a high probability of cost-effectiveness from a healthcare perspective for employees on sick leave due to depression or anxiety disorders. For participants with stress-induced exhaustion disorder, adding WDI to ACT seems to reduce healthcare costs, while WDI as a stand-alone intervention seems to reduce welfare costs.


Association between SNPs in Long Non-coding RNAs and the Risk of Female Breast Cancer in a Chinese Population.
Long non-coding RNAs (LncRNAs) have been reported to be involved in tumorigenesis and tumor progression. Single nucleotide polymorphisms (SNPs) in the lncRNAs also play a vital role in carcinogenesis. The aim of this study was to assess the relationships between the four selected tagSNPs (rs944289, rs3787016, rs1456315, rs7463708) in the lncRNAs and the risk of female breast cancer in a Chinese population. A case-control study was carried out involving in a total of 439 breast cancer patients and 439 age-matched healthy controls. The genotyping was performed with Sequenom MassARRAY and the expression of estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor-2 (HER-2) in tumor tissues was measured by the immunohistochemistry (IHC) assay. We found that rs3787016 TT genotype (adjusted odds ratio (OR) = 1.62, 95% confidence interval (CI) = 1.09-2.41, P = 0.018) was associated with an increased risk of female breast cancer, especially among the patients with premenopausal status (adjusted OR = 2.55, 95% CI = 1.30-4.97, P = 0.006). Moreover, a statistically significant increased risk of the rs3787016 TT genotype was observed among the patients with advanced tumor stage (Ⅲ and Ⅳ), poor histological grade (G3-G4), positive lymph node involvement, positive expression of ER and PR and negative expression of HER-2; rs7463708 GT and GT/GG genotype were associated with decreased risk of breast cancer in the subgroup of patients with postmenopausal status (GT versus (vs.) TT: adjusted OR = 0.67, 95% CI = 0.46-0.99, P = 0.043; GT/GG vs. TT: adjusted OR = 0.68, 95% CI = 0.47-0.98, P = 0.041) and tumor late-stage (GT vs. TT: adjusted OR = 0.65, 95% CI = 0.43-0.97, P = 0.037; GT/GG vs. TT: adjusted OR = 0.65, 95% CI = 0.44-0.96, P = 0.029). In short, rs3787016 TT genotype was associated with increased breast cancer risk and clinicopathologic features of the tumor, especially among premenopausal women.


DIO3 protects against thyrotoxicosis-derived cranio-encephalic and cardiac congenital abnormalities.
Maternal hyperthyroidism is associated with an increased incidence of congenital abnormalities at birth, but it is not clear which of these defects arise from a transient developmental excess of thyroid hormone and which depend on pregnancy stage, antithyroid drug choice, or unwanted subsequent fetal hypothyroidism. To address this issue, we studied a mouse model of comprehensive developmental thyrotoxicosis secondary to a lack of type 3 deiodinase (DIO3). Dio3-/- mice exhibited reduced neonatal viability on most genetic backgrounds and perinatal lethality on a C57BL/6 background. Dio3-/- mice exhibited severe growth retardation during the neonatal period and cartilage loss. Mice surviving after birth manifested brain and cranial dysmorphisms, severe hydrocephalus, choanal atresia, and cleft palate. These abnormalities were noticeable in C57BL/6J Dio3-/- mice at fetal stages, in addition to a thyrotoxic heart with septal defects and thin ventricular walls. Our findings stress the protecting role of DIO3 during development and support the hypothesis that human congenital abnormalities associated with hyperthyroidism during pregnancy are caused by transient thyrotoxicosis before clinical intervention. Our results also suggest thyroid hormone involvement in the etiology of idiopathic pathologies including cleft palate, choanal atresia, Chiari malformations, Kaschin-Beck disease, and Temple and other cranio-encephalic and heart syndromes.


Practice and payment preferences of newly practising family physicians in British Columbia.
To examine the remuneration model preferences of newly practising family physicians. Mixed-methods study comprising a cross-sectional, Web-based survey, as well as qualitative content analysis of answers to open-ended questions. British Columbia. University of British Columbia family practice residents who graduated between 2000 and 2009. Preferred remuneration models of newly practising physicians. The survey response rate was 31% (133 of 430). Of respondents, 71% (93 of 132) preferred non-fee-for-service practice models and 86% (110 of 132) identified the payment model as very or somewhat important in their choice of future practice. Three principal themes were identified from content analysis of respondents' open-ended comments: frustrations with fee-for-service billing, which encompassed issues related to aggravations with "the business side of things" and was seen as impeding "the freedom to focus on medicine"; quality of patient care, which embraced the importance of a payment model that supported "comprehensive patient care" and "quality rather than quantity"; and freedom to choose, which supported the plurality of practice preferences among providers who strived to provide quality care for patients, "whatever model you happen to be working in." Newly practising physicians in British Columbia preferred alternatives to fee-for-service payment models, which were perceived as contributing to fewer frustrations with billing systems, improved quality of work life, and better quality of patient care.


Radial arm maze performance of rats following repeated low level microwave radiation exposure.
We examined the possibility of changes in "working" memory of rats following whole body exposure to microwave (MW) radiation. During each of 10 days, we exposed rats within circularly polarized waveguides for 45 min to 2450 MHz fields at whole body SARs of 0.6 W/kg (2 micros pulses, 500 pps), followed by testing in a 12 arm, radial arm maze (RAM). Rats received a preexposure injection of one of three psychoactive compounds or saline, to determine whether a compound would interact with MW exposure to affect performance in the maze. Error rate, i.e., reentry into arms already visited, and time to criterion data for 10 consecutive days of testing were analyzed by a three way analysis of variance (ANOVA) using main effects of "exposure" and "drug" and a repeated factor of "test day." Our alpha limit for significance was P <.05. Analyzes of error rates revealed no significant exposure effect, no significant drug effect and no significant interaction between the two main factors. There was a significant difference in test days, as expected, with repeated test-trial days, which indicates that learning was accomplished. There was no significant interaction of test day and the other two factors. The results of our analyzes of time to criterion data included no significant exposure effect, a significant drug effect, a significant test day effect, and a significant interaction between drug and test day factors. Post hoc analyzes of the drug factor revealed that rats treated with either physostigmine or nalrexone hydrochloride, took significantly longer to complete the maze task than rats pretreated with saline or with naloxone methodide. We conclude that there is no evidence from the current study that exposure to of MW radiation under parameters examined caused decrements in the ability of rats to learn the spatial memory task.


Preparation of layered graphene and tungsten oxide hybrids for enhanced performance supercapacitors.
Tungsten oxide (WO3), which was originally poor in capacitive performance, is made into an excellent electrode material for supercapacitors by dispersing it on graphene (Gr). The obtained Gr-WO3 hybrids are characterized by X-ray diffraction, Raman spectroscopy, high-resolution transmission electron microscopy and scanning electron microscopy techniques, and evaluated as electrode materials for high-performance supercapacitors by cyclic voltammetry, galvanostatic charge-discharge curves and electrochemical impedance spectroscopy. A great improvement in specific capacitance is achieved with the present hybrids, from 255 F g-1 for WO3 nanoparticles to 580 F g-1 for Gr-WO3 hybrids (scanned at 1 A g-1 in 2 M KOH over a potential window of 0 to 0.45 V). The Gr-WO3 hybrid exhibits an excellent high rate capability and good cycling stability with more than 92% capacitance retention over 1000 cycles at a current density of 5 A g-1. The enhancement in supercapacitor performance of Gr-WO3 is not only attributed to its unique nanostructure with large specific surface area, but also its excellent electro-conductivity, which facilitates efficient charge transport and promotes electrolyte diffusion. As a whole, this work indicates that Gr-WO3 hybrids are a promising electrode material for high-performance supercapacitors.


Hybridization and population structure of the Culex pipiens complex in the islands of Macaronesia.
The Culex pipiens complex includes two widespread mosquito vector species, Cx. pipiens and Cx. quinquefasciatus. The distribution of these species varies in latitude, with the former being present in temperate regions and the latter in tropical and subtropical regions. However, their distribution range overlaps in certain areas and interspecific hybridization has been documented. Genetic introgression between these species may have epidemiological repercussions for West Nile virus (WNV) transmission. Bayesian clustering analysis based on multilocus genotypes of 12 microsatellites was used to determine levels of hybridization between these two species in Macaronesian islands, the only contact zone described in West Africa. The distribution of the two species reflects both the islands' biogeography and historical aspects of human colonization. Madeira Island displayed a homogenous population of Cx. pipiens, whereas Cape Verde showed a more intriguing scenario with extensive hybridization. In the islands of Brava and Santiago, only Cx. quinquefasciatus was found, while in Fogo and Maio high hybrid rates (∼40%) between the two species were detected. Within the admixed populations, second-generation hybrids (∼50%) were identified suggesting a lack of isolation mechanisms. The observed levels of hybridization may locally potentiate the transmission to humans of zoonotic arboviruses such as WNV.


Comparison of two ancient DNA extraction protocols for skeletal remains from tropical environments.
The tropics harbor a large part of the world's biodiversity and have a long history of human habitation. However, paleogenomics research in these climates has been constrained so far by poor ancient DNA yields. Here we compare the performance of two DNA extraction methods on ancient samples of teeth and petrous portions excavated from tropical and semi-tropical sites in Tanzania, Mexico, and Puerto Rico (N = 12). All samples were extracted twice, built into double-stranded sequencing libraries, and shotgun sequenced on the Illumina HiSeq 2500. The first extraction protocol, Method D, was previously designed for recovery of ultrashort DNA fragments from skeletal remains. The second, Method H, modifies the first by adding an initial EDTA wash and an extended digestion and decalcification step. No significant difference was found in overall ancient DNA yields or post-mortem damage patterns recovered from samples extracted with either method, irrespective of tissue type. However, Method H samples had higher endogenous content and more mapped reads after quality-filtering, but also higher clonality. In contrast, samples extracted with Method D had shorter average DNA fragments. Both methods successfully recovered endogenous ancient DNA. But, since surviving DNA in ancient or historic remains from tropical contexts is extremely fragmented, our results suggest that Method D is the optimal choice for working with samples from warm and humid environments. Additional optimization of extraction conditions and further testing of Method H with different types of samples may allow for improvement of this protocol in the future.


Breastfeeding, Antidepressants, and Depression in the Mercy Pregnancy and Emotional Well-Being Study.
Depression is consistently shown to predict lower rates of breastfeeding. In a handful of studies, breastfeeding has predicted lower depression symptoms. However, studies demonstrating the latter are limited in their measurement of both depression and breastfeeding and have not followed participants from pregnancy across the postpartum period. The primary aim of this study was to describe breastfeeding intentions and behaviors for the first 12 months postpartum among nonmedicated depressed, antidepressant-exposed, and control participants. The secondary aim was to examine group differences in the association between depressive symptoms and breastfeeding duration up to 12 months postpartum. First-trimester women ( N = 212) were recruited into a prospective longitudinal study. Depressive disorders at baseline were diagnosed using the Structured Clinical Interview for DSM-IV Axis I Disorders, and depressive symptoms were measured at the first and second trimesters and 6 and 12 months postpartum using the Edinburgh Postnatal Depression Scale. Breastfeeding duration, support from family and employers, and perceptions of participants' experience were measured. Depressed women and antidepressant-exposed women reported a trend toward lower rates of intention, initiation, and duration, but this did not reach statistical significance. There was a statistically significant difference on depressive symptoms for women taking antidepressants during pregnancy, compared with controls, when they continued to breastfeed for 12 months postpartum. This study did not find a strong association between depression or antidepressant use and intention to breastfeed, partner breastfeeding support, or initiation or duration of breastfeeding. However, for women who took antidepressants, there was evidence that breastfeeding for 12 months was associated with lower depressive symptoms.


Antithrombotic treatment of cerebrovascular disease.
The most common type of cerebrovascular disease is ischaemia or infarction from atherothrombosis or cardiac embolism. Antithrombotic treatment with an antiplatelet agent or anticoagulant assumes a prior clinical classification into categories of transient ischaemic attack (TIA) or minor stroke, acute partial stable stroke, stroke-in-progression, and completed stroke. Aspirin reduces the risk of stroke, myocardial infarction, and death after TIA or minor stroke secondary to atherothrombosis. Aspirin is effective in both sexes at a dose of 300 or 1200 mg/day. Ticlopidine (500 mg/day), a new antiplatelet agent, is more effective than aspirin in preventing stroke and death in patients with TIA or minor stroke. Ticlopidine (500 mg/day) is effective in preventing recurrent stroke, myocardial infarction, or vascular death in patients with completed stroke. Aspirin has not been directly shown to be effective after completed stroke. No clear evidence exists for the use of anticoagulants in atherothrombotic cerebral vascular disease in patients presenting with TIA or minor stroke, acute partial stable stroke, stroke-in-progression, or completed stroke. Anticoagulation for rheumatic valvular heart disease is effective in preventing recurrent embolism. Long-term anticoagulation of patients with mechanical prosthetic valves protects against initial embolism and prevents recurrent embolism. The addition of aspirin (500-1000 mg/day) to warfarin reduces the rate of cerebral embolism from mechanical prosthetic heart valves but is associated with increased bleeding. The addition of dipyridamole (400 mg/day) to warfarin may be more effective than aspirin in reducing the rate of cerebral embolism from mechanical prosthetic heart valves and has fewer bleeding side-effects. Anticoagulation during the hospital phase of myocardial infarction reduces the incidence of systemic embolism/stroke. Long-term anticoagulation of patients after the hospital phase of myocardial infarction reduces the incidence of systemic embolism/stroke, recurrent myocardial infarction and death. Prophylactic anticoagulant treatment of patients with non-valvular atrial fibrillation reduces the incidence of embolism, but the optimal duration of treatment is not known. Immediate anticoagulation of patients with completed cardioembolic stroke is safe and effective in preventing recurrent embolism.


Therapeutic exploitation of the physiological and molecular genetic alterations in head and neck cancer.
Despite improvements in the diagnosis and management of head and neck squamous cell carcinomas, there has been minimal increase in the long-term survival in these patients over the last 30 years. Treatment intensification with concurrent chemoradiotherapy has been shown to increase survival and improve organ preservation over radiotherapy alone in patients with locally advanced tumor; however, at a cost of increased long-term toxicity. Recent advances in molecular technology have ushered in a new age of targeted therapy, which holds promise for a better outcome for these patients with potentially less normal tissue toxicity. Some of the new approaches aim to specifically inhibit tumor growth and metastasis by targeting the tumor microenvironment or vasculature, whereas others focus on specific protein or signal transduction pathways. This review will summarize these new molecular and physiological based strategies that can be used for both treatment and chemoprevention of head and neck squamous cell carcinoma.


Chemical and biophysical insights into the propagation of prion strains.
Transmissible spongiform encephalopathies (TSEs) are lethal infectious neurodegenerative diseases. TSEs are caused by prions, infectious agents lacking informational nucleic acids, and possibly identical with higher-order aggregates of the cellular glycolipoprotein PrP(C). Prion strains are derived from TSE isolates that, even after inoculation into genetically identical hosts, cause disease with distinct patterns of protein aggregate deposition, incubation times, morphology of the characteristic brain damage, and cellular tropism. Most of these traits are relatively stable across serial passages. Here we review current techniques for studying prion strain differences in vivo and in cells, and discuss the strain phenomena in the general context of the knowledge gained from modeling prion fibril growth in vitro and in simple organisms.


Effects of ethanol on the proteasome interacting proteins.
Proteasome dysfunction has been repeatedly reported in alcoholic liver disease. Ethanol metabolism end-products affect the structure of the proteasome, and, therefore, change the proteasome interaction with its regulatory complexes 19S and PA28, as well as its interacting proteins. Chronic ethanol feeding alters the ubiquitin-proteasome activity by altering the interaction between the 19S and the 20S proteasome interaction. The degradation of oxidized and damaged proteins is thus decreased and leads to accumulation of insoluble protein aggregates, such as Mallory-Denk bodies. Ethanol also affects the immunoproteasome formation. PA28a/b interactions with the 20S proteasome are decreased in the proteasome fraction isolated from the liver of rats fed ethanol chronically, thus affecting the cellular antigen presentation and defense against pathogenic agents. Recently, it has been shown that ethanol also affects the proteasome interacting proteins (PIPs). Interaction of the proteasome with Ecm29 and with deubiquitinating enzymes Rpn11, UCH37, and Usp14 has been found to decrease. However, the two UBL-ubiquitin-associated domain (UBA) PIPs p62 and valosin-containing protein are upregulated when the proteasome is inhibited. The increase of these UBL-UBA proteins, as well as the increase in Hsp70 and Hsp25 levels, compensated for the proteasome failure and helped in the unfolding/docking of misfolded proteins. Chronic alcohol feeding to rats causes a significant inhibition of the proteasome pathway and this inhibition results from a decreases of the interaction between the 20S proteasome and the regulatory complexes, PIPs, and the ubiquitin system components.


Mitotic separation of two human X-linked genes in man--mouse somatic cell hybrids.
SIX INTERSPECIFIC SOMATIC HYBRID CELL LINES WERE DERIVED FROM A MOUSE LINE DEFICIENT IN HYPOXANTHINE: guanine phosphoribosyltransferase (HGPRT) and human diploid cells with normal enzyme activity. Human HGPRT was present in all six hybrids and the clones derived from them. However, in two of the six, and in some clones from another two, human glucose-6-phosphate dehydrogenase (G6PD) was absent. Since the structural loci for both these enzymes are X-linked in man, these findings suggest that these two loci have separated quite frequently through chromosome breakage and that they must be rather far apart on the X chromosome.


Trauma experience among homeless female veterans: correlates and impact on housing, clinical, and psychosocial outcomes.
This study examined lifetime exposure to traumatic events as reported by 581 homeless female veterans enrolled in a Homeless Women Veterans Program across 11 sites to characterize the types of trauma they experienced; their correlation with baseline characteristics; and their association with housing, clinical outcomes, and psychosocial functioning over a 1-year treatment period. Almost all participants endorsed multiple types and episodes of traumatic events. Among the most common were having someone close experience a serious or life-threatening illness (82%) and rape (67%). Exploratory factor analysis revealed 6 potential trauma categories: being robbed, experiencing accident or disasters, illness or death of others, combat, sexual assault, and physical assault. At baseline, trauma from sexual assault was associated with more days homeless (β = .18, p < .001), trauma from accidents or disasters was associated with poorer physical health (β = -.23, p < .001), and trauma from being robbed was related to greater use of drugs (β = .22, p < .001). Trauma reported at baseline, however, was not predictive of 1-year outcomes, suggesting type and frequency of trauma does not negatively affect the housing gains homeless women veterans can achieve through homeless services.


Nonlinear dynamics of Airy-vortex 3D wave packets: emission of vortex light waves.
The dynamics of 3D Airy-vortex wave packets is studied under the action of strong self-focusing Kerr nonlinearity. Emissions of nonlinear 3D waves out of the main wave packets with the topological charges were demonstrated. Because of the conservation of the total angular momentum, charges of the emitted waves are equal to those carried by the parental light structure. The rapid collapse imposes a severe limitation on the propagation of multidimensional waves in Kerr media. However, the structure of the Airy beam carrier allows the coupling of light from the leading, most intense peak into neighboring peaks and consequently strongly postpones the collapse. The dependence of the critical input amplitude for the appearance of a fast collapse on the beam width is studied for wave packets with zero and nonzero topological charges. Wave packets carrying angular momentum are found to be much more resistant to the rapid collapse.


The AP-2 family of transcription factors.
The AP-2 family of transcription factors consists of five different proteins in humans and mice: AP-2alpha, AP-2beta, AP-2gamma, AP-2delta and AP-2epsilon. Frogs and fish have known orthologs of some but not all of these proteins, and homologs of the family are also found in protochordates, insects and nematodes. The proteins have a characteristic helix-span-helix motif at the carboxyl terminus, which, together with a central basic region, mediates dimerization and DNA binding. The amino terminus contains the transactivation domain. AP-2 proteins are first expressed in primitive ectoderm of invertebrates and vertebrates; in vertebrates, they are also expressed in the emerging neural-crest cells, and AP-2alpha-/- animals have impairments in neural-crest-derived facial structures. AP-2beta is indispensable for kidney development and AP-2gamma is necessary for the formation of trophectoderm cells shortly after implantation; AP-2alpha and AP-2gamma levels are elevated in human mammary carcinoma and seminoma. The general functions of the family appear to be the cell-type-specific stimulation of proliferation and the suppression of terminal differentiation during embryonic development.


Hepatocellular carcinoma: consensus recommendations of the National Cancer Institute Clinical Trials Planning Meeting.
Hepatocelluar carcinoma (HCC) is the most common primary malignancy of the liver in adults and the third most common cause of cancer death worldwide. The incidence of HCC in the United States is rising steadily because of the prevalence of hepatitis C viral infection and other causes of hepatic cirrhosis. The majority of patients have underlying hepatic dysfunction, which complicates patient management and the search for safe and effective therapies. The Clinical Trials Planning Meeting (CTPM) in HCC was convened by the National Cancer Institute's Gastrointestinal Cancer Steering Committee to identify the key knowledge gaps in HCC and define clinical research priorities. The CTPM structured its review according to current evidence-based treatment modalities in HCC and prioritized the recommendations on the basis of the patient populations representing the greatest unmet medical need.


Major adverse cardiovascular event (MACE) after percutaneous coronary intervention in one-year follow-up study.
Percutaneous coronary intervention (PCI) is the most common revascularization procedure, with over 1 million performed each year, worldwide. Over the past 20 years, the increasing experience of operators coupled with the advent of newer technologies, including coronary stents and a variety of adjuvant drug therapies, have permitted more successful procedures and decreased the morbidity associated with PCIs. To identify the incidence, predictors, and clinical implications of Major Adverse Cardiovascular Events (MACE) after PCIs. This descriptive cross-sectional study was done in Bandar Abbas in Iran in 2015. All patients which treated with PCI in Shahid Mohammadi Hospital during a one-year period were employed. A total of 192 patients were included. At one-year follow-up in this study, incidence and predictors of MACE were evaluated in a prospective study. The data were analyzed by SPSS 19.0 and descriptive tests included frequency and percentage and mean and standard deviation. Also, Chi-square test was used for data analysis. A p value <0.05 was determined as significant. Of the 192 patients, 126 (65.6%) were men and 66 patients were female. Stent had been implemented in 93.8% of patients. Sixty two percent of patients were treated with only one stent, two stents were deployed in 29.7% of patients and 3.6% of patients were treated with three or more stents. Of these patients, 46.9% were treated with Drug Eluted Stent (DES) and 40.1% were treated with Bare Metal Stent (BMS). Both types of stents were used in 8.3% of patients. Also, 4.7% of the patients were treated with balloon angioplasty (POBA). Angioplasty success rate was 95.3% and procedural success rate defined as achieving TIMI III flow with residual coronary stenosis under 30%. No in-hospital mortality or emergency CABG was reported. Re-admission in first year after PCI was required in 34 (17.7%) patients of which, 20 of them (10.4%) needed target vessel revascularization (TVR). Readmission was significantly higher (p=0.034) in the group with BMS compared to those who had DES. MI occurred in 8 patients. Our study showed the superiority of DES in comparison with BMS in decreasing readmission and less TVR, but no effect on long term mortality. We recommend more studies in this setting because patients in special groups may benefit more from DES or BMS.


Clinical and pathological analysis of malignant carotid body tumour: a report of nine cases.
Malignant carotid body tumour (MCBT) is a clinically rare disease that often invades the carotid artery and cranial nerves. Diagnosis of malignant tumour should be based on extensive invasion of neighbouring organs and distant metastasis. Extensive resection should be undertaken early. Radiotherapy is effective, whereas chemotherapy is uncertain. To summarize the clinical pathological and prognostic characteristics of MCBT and explore methods for diagnosis and treatment. The study material comprised clinical, pathological, therapeutic and follow-up data concerning nine patients (four males, five females) with MCBT, treated at Tianjin Cancer Hospital between January 1956 and June 2006. The material was analysed retrospectively. Disease duration averaged 6.4 years. Shamblin classification was: one case, type II; 8 cases, type III. All nine patients underwent ultrasound examination, four underwent digital subtraction arteriography (DSA) and three had magnetic resonance angiography (MRA). Five patients underwent preoperative training of compression of the carotid (Matas test). Extensive resection was performed in all nine cases. The carotid artery was blocked in three patients. In one of these the artery was reconstructed with a vascular prosthesis, while two underwent carotid ligation. Eight patients suffered from a cranial nerve dysfunction (defect) and two suffered postoperatively from a hoarse voice, four had a glossal deviation, five had Horner's syndrome and one had a deviation of the lip angle. One patient had a congestive cough. The histopathological diagnosis in all nine cases was MCBT. One patient had metastases to a cervical lymph node and lung and another had liver metastasis. The median follow-up period was 3 years (range 6 months to 14 years). Six patients survived surgery, of whom two underwent radiotherapy. Two patients died and one could not be traced.


Morphometric characterization of microglial phenotypes in human cerebral cortex.
Microglia can adopt different morphologies, ranging from a highly ramified to an amoeboid-like phenotype. Although morphological properties of microglia have been described in rodents, little is known about their fine features in humans. The aim of this study was to characterize the morphometric properties of human microglia in gray and white matter of dorsal anterior cingulate cortex (dACC), a region implicated in behavioral adaptation to neuroinflammation. These properties were compared to those of murine microglia in order to gain a better appreciation of the differences displayed by these cells across species. Postmortem dACC samples were analyzed from 11 individuals having died suddenly without any history of neuroinflammatory, neurodegenerative, nor psychiatric illness. Tissues were sectioned and immunostained for the macrophage marker Ionized calcium binding adaptor molecule 1 (IBA1). Randomly selected IBA1-immunoreactive (IBA1-IR) cells displaying features corresponding to commonly accepted microglial phenotypes (ramified, primed, reactive, amoeboid) were reconstructed in 3D and all aspects of their morphologies quantified using the Neurolucida software. The relative abundance of each morphological phenotype was also assessed. Furthermore, adult mouse brains were similarly immunostained, and IBA1-IR cells in cingulate cortex were compared to those scrutinized in human dACC. In human cortical gray and white matter, all microglial phenotypes were observed in significant proportions. Compared to ramified, primed microglia presented an average 2.5 fold increase in cell body size, with almost no differences in branching patterns. When compared to the primed microglia, which projected an average of six primary processes, the reactive and amoeboid phenotypes displayed fewer processes and branching points, or no processes at all. In contrast, the majority of microglial cells in adult mouse cortex were highly ramified. This was also the case following a postmortem interval of 43 hours. Interestingly, the morphology of ramified microglia was strikingly similar between species. This study provides fundamental information on the morphological features of microglia in the normal adult human cerebral cortex. These morphometric data will be useful for future studies of microglial morphology in various illnesses. Furthermore, this first direct comparison of human and mouse microglia reveals that these brain cells are morphologically similar across species, suggesting highly conserved functions.


'The little things count': delivering dignified care.
In the wake of recent care failings there has been increasing demand for health professionals to provide "good" or dignified care. But what does this mean? This article summarises a study that aimed to ascertain what health professionals think they need to do to provide such care to their patients.


Urban impacts across realms: Making the case for inter-realm monitoring and management.
Burgeoning populations and the increasing concentration of humans in urban areas have resulted in extensive and increasing degradation and destruction of natural ecosystems. The multitude of impacts and their drivers in urban areas across realms are often studied at local scales, but there is regularly a mismatch between the spatial extent of the impacts and that of the pressures driving those impacts. For example, most human activities occur on land and therefore disturb terrestrial habitats (intrinsic impacts), but their impacts can also extend to the atmosphere and aquatic realms (extrinsic impacts). Management of urban impacts is often designed at local scales and aims to control local pressures, mostly overlooking pressures originating outside the 'managed' area. This is often due to jurisdictional barriers but can also result from the lack of knowledge and recognition among scientists and managers of larger scale pressures. With the aim to highlight the importance of ameliorating extrinsic impacts for holistic management of urban areas, this paper discusses the range and extent of extrinsic impacts produced by the most common pressures in urban environments. We discuss that the terrestrial realm is a 'net-donor' of impacts, as most human activities occur on land and the resulting impacts are transferred to aquatic and atmospheric realms. However, activities in aquatic realms can result in impacts on land. We conclude that, to achieve effective management strategies, greater collaboration is needed between scientists and managers focussing on different realms and regions and we present suggestions for approaches to achieve this.


Developmental toxicity of lead-contaminated sediment to mallard ducklings.
Sediment ingestion has been identified as an important exposure route for toxicants in waterfowl. The toxicity of lead-contaminated sediment from the Coeur d'Alene River Basin (CDARB) in Idaho was examined on posthatching development of mallard (Anas platyrhynchos) ducklings for 6 weeks. Day-old ducklings received either untreated control diet, clean sediment (24%) supplemented control diet, CDARB sediment (3,449 microg/g lead) supplemented diets at 12% or 24%, or a positive control diet containing lead acetate equivalent to that found in 24% CDARB. The 12% CDARB diet resulted in a geometric mean blood lead concentration of 1.41 ppm (WW) with over 90% depression of red blood cell ALAD activity and over threefold elevation of free erythrocyte protoporphyrin concentration. The 24% CDARB diet resulted in blood lead of 2.56 ppm with over sixfold elevation of protoporphyrin and lower brain weight. In this group the liver lead concentration was 7.92 ppm (WW), and there was a 40% increase in hepatic reduced glutathione concentration. The kidney lead concentration in this group was 7.97 ppm, and acid-fast inclusion bodies were present in the kidneys of four of nine ducklings. The lead acetate positive control group was more adversely affected in most respects than the 24% CDARB group. With a less optimal diet (mixture of two thirds corn and one third standard diet), CDARB sediment was more toxic; blood lead levels were higher, body growth and liver biochemistry (TBARS) were more affected, and prevalence of acid-fast inclusion bodies increased. Lead from CDARB sediment accumulated more readily in duckling blood and liver than reported in goslings, but at given concentrations was generally less toxic to ducklings. Many of these effects are similar to ones reported in wild mallards and geese within the CDARB.


Evaluation of novel imidazotetrazine analogues designed to overcome temozolomide resistance and glioblastoma regrowth.
The cellular responses to two new temozolomide (TMZ) analogues, DP68 and DP86, acting against glioblastoma multiforme (GBM) cell lines and primary culture models are reported. Dose-response analysis of cultured GBM cells revealed that DP68 is more potent than DP86 and TMZ and that DP68 was effective even in cell lines resistant to TMZ. On the basis of a serial neurosphere assay, DP68 inhibits repopulation of these cultures at low concentrations. The efficacy of these compounds was independent of MGMT and MMR functions. DP68-induced interstrand DNA cross-links were demonstrated with H2O2-treated cells. Furthermore, DP68 induced a distinct cell-cycle arrest with accumulation of cells in S phase that is not observed for TMZ. Consistent with this biologic response, DP68 induces a strong DNA damage response, including phosphorylation of ATM, Chk1 and Chk2 kinases, KAP1, and histone variant H2AX. Suppression of FANCD2 expression or ATR expression/kinase activity enhanced antiglioblastoma effects of DP68. Initial pharmacokinetic analysis revealed rapid elimination of these drugs from serum. Collectively, these data demonstrate that DP68 is a novel and potent antiglioblastoma compound that circumvents TMZ resistance, likely as a result of its independence from MGMT and mismatch repair and its capacity to cross-link strands of DNA.


The solution structure of the MANEC-type domain from hepatocyte growth factor activator inhibitor-1 reveals an unexpected PAN/apple domain-type fold.
A decade ago, motif at N-terminus with eight-cysteines (MANEC) was defined as a new protein domain family. This domain is found exclusively at the N-terminus of >400 multi-domain type-1 transmembrane proteins from animals. Despite the large number of MANEC-containing proteins, only one has been characterized at the protein level: hepatocyte growth factor activator inhibitor-1 (HAI-1). HAI-1 is an essential protein, as knockout mice die in utero due to placental defects. HAI-1 is an inhibitor of matriptase, hepsin and hepatocyte growth factor (HGF) activator, all serine proteases with important roles in epithelial development, cell growth and homoeostasis. Dysregulation of these proteases has been causatively implicated in pathological conditions such as skin diseases and cancer. Detailed functional understanding of HAI-1 and other MANEC-containing proteins is hampered by the lack of structural information on MANEC. Although many MANEC sequences exist, sequence-based database searches fail to predict structural homology. In the present paper, we present the NMR solution structure of the MANEC domain from HAI-1, the first three-dimensional (3D) structure from the MANEC domain family. Unexpectedly, MANEC is a new subclass of the PAN/apple domain family, with its own unifying features, such as two additional disulfide bonds, two extended loop regions and additional α-helical elements. As shown for other PAN/apple domain-containing proteins, we propose a similar active role of the MANEC domain in intramolecular and intermolecular interactions. The structure provides a tool for the further elucidation of HAI-1 function as well as a reference for the study of other MANEC-containing proteins.


mmm: an R package for analyzing multivariate longitudinal data with multivariate marginal models.
Modeling multivariate longitudinal data has many challenges in terms of both statistical and computational aspects. Statistical challenges occur due to complex dependence structures. Computational challenges are due to the complex algorithms, the use of numerical methods, and potential convergence problems. Therefore, there is a lack of software for such data. This paper introduces an R package mmm prepared for marginal modeling of multivariate longitudinal data. Parameter estimations are achieved by generalized estimating equations approach. A real life data set is applied to illustrate the core features of the package, and sample R code snippets are provided. It is shown that the multivariate marginal models considered in this paper and mmm are valid for binary, continuous and count multivariate longitudinal responses.


[Excessive daytime sleepiness].
When it is not due to an extrinsic origin, somnolence may be the main symptom of various diseases. Among these causes of excessive daytime sleepiness, obstructive sleep apnea syndrome is noteworthy for its very important prevalence, estimated at 4% in adult males. Due to repeated upper airway obstructions during sleep, this disease is efficiently treated by continuous positive airway pressure applied through a nasal masks during sleep. Another syndrome, periodic limb movements during sleep may also lead to a sleep fragmentation at the origin of daytime sleepiness. Its treatment is principally based on dopaminergic agonists. Narcolepsy-cataplexy and idiopathic hypersomnia are two causes of excessive daytime sleepiness in young people. The first is as frequent as multiple sclerosis and the second is ten times less frequent. The treatment of these two diseases is now based on a new French drug: modafinil(Modiodal). Sleep pathology still has only a small place in medical training. Excessive daytime sleepiness is therefore often misdiagnosed. In addition to their major risk of work or road accidents, numerous untreated patients continue to suffer from this very unpleasant symptom, at the origin of a major social handicap.


Toward an Information Infrastructure for Global Health Improvement.
Profound global challenges to individual and population health, alongside the opportunities to benefit from digital technology, have spawned the concept of the Learning Health System. Learning Health Systems (LHSs)--which can function at organizational, network, regional, and national levels of scale--have the capability of continuous data-driven self-study that promotes change and improvement. The LHS concept, which originated in the U.S. in 2007, is rapidly gaining attention around the world. LHSs require, but also transcend, the secondary use of health data. This paper describes the key features of LHSs, argues that effective and sustainable LHSs must be supported by infrastructures that allow them to function with economies of scale and scope, and describes the services that such infrastructures must provide. While it is relatively straightforward to describe LHSs, achieving them at the high level of capability necessary to promote significant health benefits will require advancements in science and engineering, engaging the field of informatics among a wider range of disciplines. It also follows from this vision that LHSs cannot be built from an imposed blueprint; LHSs will more likely evolve from efforts at smaller scales that compose into larger systems.


Geographic profiling as a novel spatial tool for targeting infectious disease control.
Geographic profiling is a statistical tool originally developed in criminology to prioritise large lists of suspects in cases of serial crime. Here, we use two data sets--one historical and one modern--to show how it can be used to locate the sources of infectious disease. First, we re-analyse data from a classic epidemiological study, the 1854 London cholera outbreak. Using 321 disease sites as input, we evaluate the locations of 13 neighbourhood water pumps. The Broad Street pump--the outbreak's source--ranks first, situated in the top 0.2% of the geoprofile. We extend our study with an analysis of reported malaria cases in Cairo, Egypt, using 139 disease case locations to rank 59 mosquitogenic local water sources, seven of which tested positive for the vector Anopheles sergentii. Geographic profiling ranks six of these seven sites in positions 1-6, all in the top 2% of the geoprofile. In both analyses the method outperformed other measures of spatial central tendency. We suggest that geographic profiling could form a useful component of integrated control strategies relating to a wide variety of infectious diseases, since evidence-based targeting of interventions is more efficient, environmentally friendly and cost-effective than untargeted intervention.


Temporal trends in liver transplant centre volume in the USA.
Although prior studies have suggested an inverse association between liver transplant centre volume and postoperative patient mortality, more recent analyses have failed to confirm this association. To date, all studies of the relationship between centre volume and outcomes in liver transplantation have been cross-sectional in design. The objective of our study was to examine temporal trends in the volume-outcomes relationship for liver transplantation. We used information obtained from the Scientific Registry of Transplant Recipients (SRTR) programme-specific data reports to examine the outcomes of adult liver transplant recipients stratified by annual centre volume. This relationship between centre volume and patient outcomes was assessed over three consecutive time periods from 2000 through 2007. The overall 25% increase in adult liver transplant volume in the USA from 2000 to 2007 appeared to be distributed fairly equally among existing transplant centres. In the earliest time period of our analysis, high-volume centres achieved superior risk-adjusted 1-year patient outcomes compared with low-volume centres. By the third and most recent time period of the analysis, this discrepancy between the outcomes of high- and low-volume centres was no longer statistically apparent. The relationship between centre volume and patient outcomes for liver transplantation in the USA has become less pronounced over time, suggesting that the use of procedure volume as a marker of liver transplant centre quality cannot be justified. The performance-based review process currently utilized in the USA may have contributed to this diminishing influence of centre volume on liver transplant recipient outcomes. This type of review process should be considered as a potential alternative to the volume-based referral initiatives that have been developed for other non-transplant, complex surgical procedures.