Designing "lifestyle interventions" with the brain in mind.
The central hypothesis examined in this issue is that insulin resistance promotes maladaptive brain function and contributes to reduced neuronal plasticity, potentially accelerating brain aging. Therefore, if we were to prevent or treat insulin resistance, through weight loss and exercise, cognitive function would be improved. In this article, we argue that successful interventions influencing these outcomes depend upon overriding maladaptive neurobiology. This maladaptation may have developed over the course of the lifespan through interaction with modern environments. Furthermore, we emphasize the need to take this emergent neurobiology into account when designing interventions.


Actigraphic patterns, impulsivity and mood instability in bipolar disorder, borderline personality disorder and healthy controls.
To differentiate the relation between the structure and timing of rest-activity patterns and symptoms of impulsivity and mood instability in bipolar disorder (BD), borderline personality disorder (BPD) and healthy controls (HC). Eighty-seven participants (31 BD, 21 BPD and 35 HC) underwent actigraph monitoring for 28 days as part of the Automated Monitoring of Symptom Severity (AMoSS) study. Impulsivity was assessed at study entry using the BIS-11. Mood instability was subsequently longitudinally monitored using the digital Mood Zoom questionnaire. BPD participants show several robust and significant correlations between non-parametric circadian rest-activity variables and worsened symptoms. Impulsivity was associated with low interdaily stability (r = -0.663) and weak amplitude (r = -0.616). Mood instability was associated with low interdaily stability (r = -0.773), greater rhythm fragmentation (r = 0.662), weak amplitude (r = -0.694) and later onset of daily activity (r = 0.553). These associations were not present for BD or HCs. Classification analysis using actigraphic measures determined that later L5 onset reliably distinguished BPD from BD and HC but did not sufficiently discriminate between BD and HC. Rest-activity pattern disturbance indicative of perturbed sleep and circadian function is an important predictor of symptom severity in BPD. This appears to validate the greater subjective complaints of BPD individuals that are sometimes regarded as exaggerated by clinicians. We suggest that treatment strategies directed towards improving sleep and circadian entrainment may in the future be investigated in BPD.


Detecting reciprocity at a global scale.
Reciprocity stabilizes cooperation from the level of microbes all the way up to humans interacting in small groups, but does reciprocity also underlie stable cooperation between larger human agglomerations, such as nation states? Famously, evolutionary models show that reciprocity could emerge as a widespread strategy for achieving international cooperation. However, existing studies have only detected reciprocity-driven cooperation in a small number of country pairs. We apply a new method for detecting mutual influence in dynamical systems to a new large-scale data set that records state interactions with high temporal resolution. Doing so, we detect reciprocity between many country pairs in the international system and find that these reciprocating country pairs exhibit qualitatively different cooperative dynamics when compared to nonreciprocating pairs. Consistent with evolutionary theories of cooperation, reciprocating country pairs exhibit higher levels of stable cooperation and are more likely to punish instances of noncooperation. However, countries in reciprocity-based relationships are also quicker to forgive single acts of noncooperation by eventually returning to previous levels of mutual cooperation. By contrast, nonreciprocating pairs are more likely to exploit each other's cooperation via higher rates of defection. Together, these findings provide the strongest evidence to date that reciprocity is a widespread mechanism for achieving international cooperation.


Early variceal rebleeding after successful distal splenorenal shunt.
Of 77 patients with repeated variceal hemorrhage who underwent distal splenorenal shunt, five (6.5%) developed rebleeding despite a patent splenorenal anastomosis. Three of the five patients died. Early variceal rebleeding usually indicates shunt thrombosis but may occur with a patent anastomosis. Anatomic or functional left renal vein and/or splenic vein hypertension producing incomplete variceal decompression is generally the cause. Ineffective separation of the main portal vein from the gastrosplenic venous plexus may coexist and further intensify variceal congestion. Urgent angiographic studies and direct shunt catheterization with measurement of splenic vein, left renal vein, and inferior vena cava pressures should be performed to plan appropriate therapy. A significant gradient between the splenic and renal veins is evidence of an unsatisfactory anastomosis and should be managed by balloon angioplasty or reoperation. High splenic and left renal vein pressures with a gradient of more than 10 mm Hg between the renal vein and the inferior vena cava indicate renal vein hypertension. Initial therapy should include serial injection sclerotherapy, as renal vein hypertension will usually resolve over time as additional collaterals develop. However, persistent or recurrent variceal hemorrhage may require total portal decompression to bypass the restrictive left renal vein segment.


An unusual presentation of multiple myeloma in two cats.
Multiple myeloma is a rare neoplasm in cats. Common presenting signs (e.g., lethargy, anorexia, weight loss) usually are nonspecific. Two cats with multiple myeloma were presented with primary complaints of lameness; one had femoral osteolytic lesions, and the other likely had hyperviscosity syndrome. The cat with osteolytic lesions was treated with chemotherapy; the primary lesion responded, but the neoplasia metastasized.


Overcoming birth weight: can physical activity mitigate birth weight-related differences in adiposity?
Individuals born at low or high birth weight (BW) have elevated adiposity. The extent to which physical activity can mitigate this risk is unknown. The aim of this study was to determine if associations between BW and adiposity vary by self-reported moderate-to-vigorous physical activity (MVPA) among adolescents. We used data on adolescents in the National Health and Nutrition Examination Survey (1999-2006; 12-15 years; n = 4064). Using gender-stratified linear regression, we modelled body mass index (BMI) and waist circumference (WC) z-scores as a function of low, normal and high BW, MVPA (weekly Metabolic Equivalent of Task hours) and MVPA*BW cross-product terms, adjusting for sociodemographics, diet and, in WC models, BMI. Among girls with low MVPA, those born with high BW had greater BMI than normal BW; this difference diminished with greater MVPA (coefficient [95% confidence interval]: low MVPA: 0.72 [0.29, 1.14]; high MVPA: -0.04 [-0.48, 0.39]; P for interaction = 0.05). Among boys, MVPA did not modify the associations between BW and BMI. WC was unrelated to BW, regardless of MVPA. Findings suggest that effects of high BW in total adiposity can be more easily modified with MVPA in adolescent girls than in boys.


Laceration injuries and infections among workers in the poultry processing and pork meatpacking industries.
Workers in poultry processing and pork meatpacking have high rates of acute injuries and chronic disease among. The presence of zoonotic pathogens in these workplaces may interact with injury. We investigated incidence of worker injuries, lacerations, and infections reported by 10 companies from 2004 to 2009 and calculated annual incidence rates by industry and company along with temporal trends and job-related risk factors. Average annual mean total injury rates were 6.4 per 100 workers (poultry) and 13.2 per 100 workers (pork). Average annual mean rates for lacerations were 1.8 per 100 workers (poultry) and 1.9 per 100 (pork). Sharp tools and animal products were most frequently reported as sources for lacerations. Animal products were most frequently reported as sources of infected lacerations. The results indicate that these industries continue to have high injury rates. The results also suggest that zoonotic pathogens may be preventable health and safety risks.


Executive semantic processing is underpinned by a large-scale neural network: revealing the contribution of left prefrontal, posterior temporal, and parietal cortex to controlled retrieval and selection using TMS.
To understand the meanings of words and objects, we need to have knowledge about these items themselves plus executive mechanisms that compute and manipulate semantic information in a task-appropriate way. The neural basis for semantic control remains controversial. Neuroimaging studies have focused on the role of the left inferior frontal gyrus (LIFG), whereas neuropsychological research suggests that damage to a widely distributed network elicits impairments of semantic control. There is also debate about the relationship between semantic and executive control more widely. We used TMS in healthy human volunteers to create "virtual lesions" in structures typically damaged in patients with semantic control deficits: LIFG, left posterior middle temporal gyrus (pMTG), and intraparietal sulcus (IPS). The influence of TMS on tasks varying in semantic and nonsemantic control demands was examined for each region within this hypothesized network to gain insights into (i) their functional specialization (i.e., involvement in semantic representation, controlled retrieval, or selection) and (ii) their domain dependence (i.e., semantic or cognitive control). The results revealed that LIFG and pMTG jointly support both the controlled retrieval and selection of semantic knowledge. IPS specifically participates in semantic selection and responds to manipulations of nonsemantic control demands. These observations are consistent with a large-scale semantic control network, as predicted by lesion data, that draws on semantic-specific (LIFG and pMTG) and domain-independent executive components (IPS).


Plasticity of calcitonin gene related peptide neurotransmission in the spinal cord during peripheral inflammation.
Injection of complete Freund's adjuvant (CFA; 75 microL) into the plantar surface of the hind paw of the rat results in a mild inflammation that lasts for several days and is accompanied by hyperalgesia. Multiple components of calcitonin gene related peptide (CGRP) neurotransmission in the spinal cord are altered during the course of this peripheral inflammation. The content of immunoreactive (i) CGRP in the dorsal horn of the spinal cord, where primary afferent neurons terminate, is significantly decreased within 2 days after injection of CFA but increases to a level greater than that of the control at 8 days. The early decrease in iCGRP in the spinal cord suggests that the release of CGRP from primary afferent neurons is increased during the period of maximal hyperalgesia that accompanies peripheral inflammation. Changes in the mRNA for CGRP suggest that the increase in spinal content of iCGRP is due to an increase in synthesis of the peptide as the level of mRNA for CGRP is increased from 2 to 8 days after injection of CFA. Despite the decrease in the content of iCGRP in the spinal cord, there is no apparent decrease in the amount of iCGRP that can be released from the dorsal spinal cord by capsaicin; in fact, capsaicin-evoked release is increased at 4 days. Measurements of the binding of 125I-labelled CGRP in the dorsal spinal cord indicate that high affinity binding sites for CGRP are downregulated at 4 days after injection of CFA. In total, these data support the hypothesis that the activity of CGRP-containing primary afferent neurons is increased during peripheral inflammation. CGRP released from primary afferent neurons in the spinal cord may contribute to cellular changes that accompany peripheral inflammation.


The impact of self-incompatibility systems on the prevention of biparental inbreeding.
Inbreeding in hermaphroditic plants can occur through two different mechanisms: biparental inbreeding, when a plant mates with a related individual, or self-fertilization, when a plant mates with itself. To avoid inbreeding, many hermaphroditic plants have evolved self-incompatibility (SI) systems which prevent or limit self-fertilization. One particular SI system-homomorphic SI-can also reduce biparental inbreeding. Homomorphic SI is found in many angiosperm species, and it is often assumed that the additional benefit of reduced biparental inbreeding may be a factor in the success of this SI system. To test this assumption, we developed a spatially-explicit, individual-based simulation of plant populations that displayed three different types of homomorphic SI. We measured the total level of inbreeding avoidance by comparing each population to a self-compatible population (NSI), and we measured biparental inbreeding avoidance by comparing to a population of self-incompatible plants that were free to mate with any other individual (PSI). Because biparental inbreeding is more common when offspring dispersal is limited, we examined the levels of biparental inbreeding over a range of dispersal distances. We also tested whether the introduction of inbreeding depression affected the level of biparental inbreeding avoidance. We found that there was a statistically significant decrease in autozygosity in each of the homomorphic SI populations compared to the PSI population and, as expected, this was more pronounced when seed and pollen dispersal was limited. However, levels of homozygosity and inbreeding depression were not reduced. At low dispersal, homomorphic SI populations also suffered reduced female fecundity and had smaller census population sizes. Overall, our simulations showed that the homomorphic SI systems had little impact on the amount of biparental inbreeding in the population especially when compared to the overall reduction in inbreeding compared to the NSI population. With further study, this observation may have important consequences for research into the origin and evolution of homomorphic self-incompatibility systems.


The native cyclobutane pyrimidine dimer photolyase of rice is phosphorylated.
The cyclobutane pyrimidine dimer (CPD) is a major type of DNA damage induced by ultraviolet B (UVB) radiation. CPD photolyase, which absorbs blue/UVA light as an energy source to monomerize dimers, is a crucial factor for determining the sensitivity of rice (Oryza sativa) to UVB radiation. Here, we purified native class II CPD photolyase from rice leaves. As the final purification step, CPD photolyase was bound to CPD-containing DNA conjugated to magnetic beads and then released by blue-light irradiation. The final purified fraction contained 54- and 56-kD proteins, whereas rice CPD photolyase expressed from Escherichia coli was a single 55-kD protein. Western-blot analysis using anti-rice CPD photolyase antiserum suggested that both the 54- and 56-kD proteins were the CPD photolyase. Treatment with protein phosphatase revealed that the 56-kD native rice CPD photolyase was phosphorylated, whereas the E. coli-expressed rice CPD photolyase was not. The purified native rice CPD photolyase also had significantly higher CPD photorepair activity than the E. coli-expressed CPD photolyase. According to the absorption, emission, and excitation spectra, the purified native rice CPD photolyase possesses both a pterin-like chromophore and an FAD chromophore. The binding activity of the native rice CPD photolyase to thymine dimers was higher than that of the E. coli-expressed CPD photolyase. These results suggest that the structure of the native rice CPD photolyase differs significantly from that of the E. coli-expressed rice CPD photolyase, and the structural modification of the native CPD photolyase leads to higher activity in rice.


Temporal variability of visibility and its parameterizations in Ningbo, China.
Simultaneous and continuous measurements of visibility, meteorological parameters and air pollutants were carried out at a suburban site in Ningbo from June 1, 2013 to May 31, 2015. The characteristics of visibility and their relationships with air pollutants and meteorological factors were investigated using multiple statistical methods. Daily visibility ranged from 0.6 to 34.1 km, with a mean value of 11.8 km. During the 2-year experiment, 43.4% of daily visibility was found to be less than 10.0 km and only 9.2% was greater than 20.0 km. Visibility was lower in winter with a frequency of 53.4% in the range of 0.0-5.0 km. Annual visibility had an obvious diurnal variation, with the lowest and highest visibility being 7.5 km at approximately 06:00 local time and 15.6 km at approximately 14:00 local time, respectively. Multiple correspondence analysis (MCA) indicated that the different ranges of visibility were significantly affected by different levels of pollutants and meteorological conditions. Based on the analyses, visibility was found to be an exponential function of PM2.5 concentrations within a certain range of relative humidity. Thus, non-linear models combining multiple linear regressions with exponential regression were subsequently developed using the data collected from June 2014 to May 2015, and the data from June 2013 to May 2014 was used to evaluate the performance of the model. It was demonstrated that the derived models can quantitatively describe the relationships between visibility, air quality and meteorological parameters in Ningbo.


Impact of study oximeter masking algorithm on titration of oxygen therapy in the Canadian oxygen trial.
To compare oxygen saturations as displayed to caregivers on offset pulse oximeters in the 2 groups of the Canadian Oxygen Trial. In 5 double-blind randomized trials of oxygen saturation targeting, displayed saturations between 88% and 92% were offset by 3% above or below the true values but returned to true values below 84% and above 96%. During the transition, displayed values remained static at 96% in the lower and at 84% in the higher target group during a 3% change in true saturations. In contrast, displayed values changed rapidly from 88% to 84% in the lower and from 92% to 96% in the higher target group during a 1% change in true saturations. We plotted the distributions of median displayed saturations on days with >12 hours of supplemental oxygen in 1075 Canadian Oxygen Trial participants to reconstruct what caregivers observed at the bedside. The oximeter masking algorithm was associated with an increase in both stability and instability of displayed saturations that occurred during the transition between offset and true displayed values at opposite ends of the 2 target ranges. Caregivers maintained saturations at lower displayed values in the higher than in the lower target group. This differential management reduced the separation between the median true saturations in the 2 groups by approximately 3.5%. The design of the oximeter masking algorithm may have contributed to the smaller-than-expected separation between true saturations in the 2 study groups of recent saturation targeting trials in extremely preterm infants.


Human Auditory and Adjacent Nonauditory Cerebral Cortices Are Hypermetabolic in Tinnitus as Measured by Functional Near-Infrared Spectroscopy (fNIRS).
Tinnitus is the phantom perception of sound in the absence of an acoustic stimulus. To date, the purported neural correlates of tinnitus from animal models have not been adequately characterized with translational technology in the human brain. The aim of the present study was to measure changes in oxy-hemoglobin concentration from regions of interest (ROI; auditory cortex) and non-ROI (adjacent nonauditory cortices) during auditory stimulation and silence in participants with subjective tinnitus appreciated equally in both ears and in nontinnitus controls using functional near-infrared spectroscopy (fNIRS). Control and tinnitus participants with normal/near-normal hearing were tested during a passive auditory task. Hemodynamic activity was monitored over ROI and non-ROI under episodic periods of auditory stimulation with 750 or 8000 Hz tones, broadband noise, and silence. During periods of silence, tinnitus participants maintained increased hemodynamic responses in ROI, while a significant deactivation was seen in controls. Interestingly, non-ROI activity was also increased in the tinnitus group as compared to controls during silence. The present results demonstrate that both auditory and select nonauditory cortices have elevated hemodynamic activity in participants with tinnitus in the absence of an external auditory stimulus, a finding that may reflect basic science neural correlates of tinnitus that ultimately contribute to phantom sound perception.


Plant genome editing made easy: targeted mutagenesis in model and crop plants using the CRISPR/Cas system.
Targeted genome engineering (also known as genome editing) has emerged as an alternative to classical plant breeding and transgenic (GMO) methods to improve crop plants. Until recently, available tools for introducing site-specific double strand DNA breaks were restricted to zinc finger nucleases (ZFNs) and TAL effector nucleases (TALENs). However, these technologies have not been widely adopted by the plant research community due to complicated design and laborious assembly of specific DNA binding proteins for each target gene. Recently, an easier method has emerged based on the bacterial type II CRISPR (clustered regularly interspaced short palindromic repeats)/Cas (CRISPR-associated) immune system. The CRISPR/Cas system allows targeted cleavage of genomic DNA guided by a customizable small noncoding RNA, resulting in gene modifications by both non-homologous end joining (NHEJ) and homology-directed repair (HDR) mechanisms. In this review we summarize and discuss recent applications of the CRISPR/Cas technology in plants.


Maintenance of G2 arrest in the Xenopus oocyte: a role for 14-3-3-mediated inhibition of Cdc25 nuclear import.
Cdc2-cyclin B1 in the G2-arrested Xenopus oocyte is held inactive by phosphorylation of Cdc2 at two negative regulatory sites, Thr14 and Tyr15. Upon treatment with progesterone, these sites are dephosphorylated by the dual specificity phosphatase, Cdc25, leading to Cdc2-cyclin B1 activation. Whereas maintenance of the G2 arrest depends upon preventing Cdc25-induced Cdc2 dephosphorylation, the mechanisms responsible for keeping Cdc25 in check in these cells have not yet been described. Here we report that Cdc25 in the G2-arrested oocyte is bound to 14-3-3 proteins and that progesterone treatment abrogates this binding. We demonstrate that Cdc25, apparently statically localized in the cytoplasm, is actually capable of shuttling in and out of the oocyte nucleus. Binding of 14-3-3 protein markedly reduces the nuclear import rate of Cdc25, allowing nuclear export mediated by a nuclear export sequence present in the N-terminus of Cdc25 to predominate. If 14-3-3 binding to Cdc25 is prevented while nuclear export is inhibited, the coordinate nuclear accumulation of Cdc25 and Cdc2-cyclin B1 facilitates their mutual activation, thereby promoting oocyte maturation.


Carotid-cavernous fistula after functional endoscopic sinus surgery.
Carotid-cavernous fistulas (CCFs) are anomalous communications between the carotid arterial system and the venous cavernous sinus. They can arise because of spontaneous or trauma causes. Most caroticocavernous fistulas are of spontaneous origin and unknown etiology. Spontaneous CCF may also be associated with cavernous sinus pathology such as arteriosclerotic changes of the arterial wall, fibromuscular dysplasia, or Ehler-Danlos syndrome. Traumatic CCFs may occur after either blunt or penetrating head trauma. Their clinical presentation is related to their size and to the type of venous drainage, which can lead to a variety of symptoms, such as visual loss, proptosis, bruit, chemosis, cranial nerve impairment, intracranial hemorrhage (rare), and so on. Treatment by endovascular transarterial embolization with electrolytically detachable coils is a very effective method for CCF with good outcomes. Carotid-cavernous fistulas have been rarely reported after craniofacial surgery and are uncommon pathologies in otolaryngology practice. In this study, we report a 40-year-old woman with CCF secondary to blunt trauma of functional endoscopic sinus surgery.


Side Benefits of Mass Drug Administration for Lymphatic Filariasis on Strongyloides stercoralis Prevalence on Pemba Island, Tanzania.
Strongyloides stercoralis, although endemic in many countries, is not included in helminth control programs. Few data are available on the prevalence and morbidity linked to this infection. We compared data from two studies conducted in 1998 and 2013 on Pemba Island, Tanzania, involving 525 and 509 schoolchildren, respectively. In 1998, the diagnostic method used was Harada Mori, whereas in 2013 diagnosis was made by both Koga agar plate and Baermann methods. The prevalence registered was 41% in 1998 and 7% in 2013. This data suggest that the prevalence of S. stercoralis on Pemba was significantly reduced 7 years after the last ivermectin administration for preventive chemotherapy and underlines the importance and impact of large-scale preventive chemotherapy, which often goes beyond its actual target. Preventive chemotherapy with ivermectin should be recommended in areas where S. stercoralis is endemic.


Immunostaining of pulpal nerve fibre bundle/arteriole associations in ground serial sections of whole human teeth embedded in technovit® 9100.
A technique for embedding human undecalcified tooth specimens in Technovit® 9100 was developed, which permits immunohistological evaluation of pulp tissue in serial ground sections. Human molars were divided into 14-18 sections of about 23 µm thickness. Immunohistological double staining for S-100 and CD34 revealed unique associations of myelinated nerve fibre bundles with arterioles, which continued through the entire tooth pulp. These arterioles were not only accompanied by, but partially or totally enveloped in longitudinally orientated myelinated nerve fibre bundles. We speculate that this unique arrangement may mechanically support the arterioles and alleviate detection or regulation of their contraction state by sensory nerve cells.


Heterobimetallic N-Heterocyclic Carbene Complexes: A Synthetic, Spectroscopic, and Theoretical Study.
A new synthetic methodology has been developed for the preparation of heterobimetallic group 11 and group 12 complexes of a symmetrical bis-NHC "pincer" ligand. The synthetic route involved the initial preparation of a mononuclear [Au(NHC)2](+) complex with pendent imidazole moieties on the NHC ligands. Subsequent alkylation of the imidazole groups with Et3OBF4 and metalation with a second metal ion (Ag(I) or Hg(II)) provided two heterobimetallic complexes. Four homobimetallic (Cu(I)2, Ag(I)2, Au(I)2, and Hg(II)2) complexes of the same bis-NHC "pincer" ligand were also prepared. The homobimetallic Cu(I)2, Au(I)2, and Hg(II)2 complexes and heterobimetallic Au(I)-Ag(I) and Au(I)-Hg(II) complexes and the synthetic intermediates for the heterobimetallic complexes were characterized by X-ray crystallography. These X-ray structures show that the bimetallic complexes adopt "twisted" conformations in the solid state, supporting short M···M interactions. Crystalline samples of the homobimetallic Ag(I)2 and Au(I)2 and heterobimetallic Au(I)-Ag(I) and Au(I)-Hg(II) complexes were emissive at room temperature and at 77 K. The geometries of the synthesized complexes were optimized at the M06-L/def2-SVP level of theory, and the electronic nature of the M···M interactions for all synthesized complexes was investigated using natural bond orbital (NBO) calculations.


A mnemonical or negative-co-operativity model for the activation of adenylate cyclase by a common G-protein-coupled calcitonin-gene-related neuropeptide (CGRP)/amylin receptor.
Both amylin and calcitonin-gene-related neuropeptide (CGRP) activated adenylate cyclase activity in hepatocyte membranes around 5-fold in a dose-dependent fashion, with EC50 values of 120 +/- 14 and 0.3 +/- 0.14 nM respectively. Whereas amylin exhibited normal activation kinetics (Hill coefficient, h approximately 1), CGRP showed kinetics indicative of either multiple sites/receptor species having different affinities for this ligand or a single receptor species exhibiting apparent negative co-operativity (h approximately 0.21). The CGRP antagonist CGRP-(8-37)-peptide inhibited adenylate cyclase stimulated by EC50 concentrations of either amylin or CGRP. Inhibition by CGRP-(8-37) was selective in that markedly lower concentrations were required to block the action of amylin (IC50 = 3 +/- 1 nM) compared with that of CGRP itself (IC50 = 120 +/- 11 nM). Dose-effect data for inhibition of CGRP action by CGRP-(8-37) showed normal saturation kinetics (h approximately 1), whereas CGRP-(8-37) inhibited amylin-stimulated adenylate cyclase activity in a fashion which was indicative of either multiple sites or apparent negative co-operativity (h approximately 0.24). Observed changes in the kinetics of inhibition by CGRP-(8-37) of CGRP, but not amylin-stimulated adenylate cyclase, at concentrations of agonists below their EC50 values militated against a model of two distinct populations of non-interacting receptors each able to bind both amylin and CGRP. A kinetic model is proposed whereby a single receptor, capable of being activated by both CGRP and amylin, obeys either a mnemonical kinetic mechanism or one of negative co-operativity with respect to CGRP but not to amylin. The relative merits of these two models are discussed together with a proposal suggesting that the activation of adenylate cyclase by various G-protein-linked receptors may be described by a mnemonical model mechanism.


Tertiary structural and functional analyses of a viroid RNA motif by isostericity matrix and mutagenesis reveal its essential role in replication.
RNA-templated RNA replication is essential for viral or viroid infection, as well as for regulation of cellular gene expression. Specific RNA motifs likely regulate various aspects of this replication. Viroids of the Pospiviroidae family, as represented by the Potato spindle tuber viroid (PSTVd), replicate in the nucleus by utilizing DNA-dependent RNA polymerase II. We investigated the role of the loop E (sarcin/ricin) motif of the PSTVd genomic RNA in replication. A tertiary-structural model of this motif, inferred by comparative sequence analysis and comparison with nuclear magnetic resonance and X-ray crystal structures of loop E motifs in other RNAs, is presented in which core non-Watson-Crick base pairs are precisely specified. Isostericity matrix analysis of these base pairs showed that the model accounts for the reported natural sequence variations and viable experimental mutations in loop E motifs of PSTVd and other viroids. Furthermore, isostericity matrix analysis allowed us to design disruptive, as well as compensatory, mutations of PSTVd loop E. Functional analyses of such mutants by in vitro and in vivo experiments demonstrated that loop E structural integrity is crucial for replication, specifically during transcription. Our results suggest that the PSTVd loop E motif exists and functions in vivo and provide loss-of-function genetic evidence for the essential role of a viroid RNA three-dimensional motif in rolling-circle replication. The use of isostericity matrix analysis of non-Watson-Crick base pairing to rationalize mutagenesis of tertiary motifs and systematic in vitro and in vivo functional assays of mutants offers a novel, comprehensive approach to elucidate the tertiary-structure-function relationships for RNA motifs of general biological significance.


Reversible flocculation with hydroxypropyl guar-borate, a labile anionic polyelectrolyte.
Cationic polystyrene latex was flocculated with mixtures of hydroxypropyl guar (HPG) and boric acid, which form an anionic polyelectrolyte HPG-borate. Flocculation could be reversed by either lowering pH or by adding fructoseboth procedures remove labile borate ions weakly bound to the HPG chains. Mixtures of phenyl boronic acid (PBA) and HPG also flocculate latex although the range of HPG-PBA concentrations, giving flocculation was much narrower than HPG-borate. The differences were explained by the tendency of borate ion to cross-link HPG chains, whereas PBA is monofunctional and cannot cross-link HPG.


Calcineurin-dependent Protein Phosphorylation Changes During Egg Activation in Drosophila melanogaster.
In almost all animals studied to date, the crucial process of egg activation, by which an arrested mature oocyte transitions into an actively developing embryo, initiates with an increase in Ca2+ in the oocyte's cytoplasm. This Ca2+ rise sets off a series of downstream events, including the completion of meiosis and the dynamic remodeling of the oocyte transcriptome and proteome, which prepares the oocyte for embryogenesis. Calcineurin is a highly conserved phosphatase that is activated by Ca2+ upon egg activation and that is required for the resumption of meiosis in Xenopus,, ascidians, and Drosophila. The molecular mechanisms by which calcineurin transduces the calcium signal to regulate meiosis and other downstream events are still unclear. In this study, we investigate the regulatory role of calcineurin during egg activation in Drosophila melanogaster,. Using mass spectrometry, we quantify the phosphoproteomic and proteomic changes that occur during egg activation, and we examine how these events are affected when calcineurin function is perturbed in female germ cells. Our results show that calcineurin regulates hundreds of phosphosites and also influences the abundance of numerous proteins during egg activation. We find calcineurin-dependent changes in cell cycle regulators including Fizzy (Fzy), Greatwall (Gwl) and Endosulfine (Endos); in protein translation modulators including PNG, NAT, eIF4G, and eIF4B; and in important components of signaling pathways including GSK3β and Akt1. Our results help elucidate the events that occur during the transition from oocyte to embryo.


Landscape of Tumor Mutation Load, Mismatch Repair Deficiency, and PD-L1 Expression in a Large Patient Cohort of Gastrointestinal Cancers.
The efficacy of immunotherapy varies widely among different gastrointestinal cancers. Response to immune checkpoint inhibitors is shown to correlate with tumor mutation load (TML), mismatch repair deficiency (dMMR) status, and programmed cell death-ligand 1 (PD-L1) expression. Herein, we quantify TML, dMMR, and PD-L1 expression and determine their interrelationship in gastrointestinal cancers. Here, a total of 4,125 tumors from 14 different gastrointestinal cancer sites were studied using validated assays. Next-generation sequencing was performed on genomic DNA isolated from formalin-fixed paraffin-embedded tumor specimens using the NextSeq platform. TML was calculated using only somatic nonsynonymous missense mutations sequenced with a 592-gene panel. Microsatellite instability (MSI) was assessed using direct analysis of altered known MSI loci in the target regions of the sequenced genes. PD-L1 expression was analyzed by IHC. Interestingly, right-sided colon and small-bowel adenocarcinomas had the highest prevalence of TML-high tumors (14.6% and 10.2%, respectively). Pancreatic neuroendocrine tumors and gastrointestinal stromal tumors had the lowest rates of TML-high (1.3% and 0%, respectively). TML-high was strongly associated with MSI-H (P < 0.0001). However, all TML-high anal cancers (8.3%) were microsatellite stable (MSS). Higher PD-L1 expression was more likely to be seen in MSI compared with MSS tumors (20.6% vs. 7.8%, P < 0.0001).Implications: TML-high rate varied widely among gastrointestinal cancers. Although MSI is conceivably the main driver for TML-high, other factors may be involved. Future clinical trials are needed to evaluate whether the integration of TML, MSI, and PD-L1 could better identify potential responders to immunotherapy. Mol Cancer Res; 16(5); 805-12. ©2018 AACR.


Outcome after surgical or conservative management of cerebral cavernous malformations.
There have been few comparative studies of microsurgical excision vs conservative management of cerebral cavernous malformations (CCM) and none of them has reliably demonstrated a statistically and clinically significant difference. We conducted a prospective, population-based study to identify and independently validate definite CCM diagnoses first made in 1999-2003 in Scottish adult residents. We used multiple sources of prospective follow-up to assess adults' dependence and to identify and independently validate outcome events. We used univariate and multivariable survival analyses to test the influence of CCM excision on outcome, adjusted for prognostic factors and baseline imbalances. Of 134 adults, 25 underwent CCM excision; these adults were younger (34 vs 43 years at diagnosis, p = 0.004) and more likely to present with symptomatic intracranial hemorrhage or focal neurologic deficit than adults managed conservatively (48% vs 26%; odds ratio 2.7, 95% confidence interval [CI] 1.1-6.5). During 5 years of follow-up, CCM excision was associated with a deterioration to an Oxford Handicap Scale score 2-6 sustained over at least 2 successive years (adjusted hazard ratio [HR] 2.2, 95% CI 1.1-4.3) and the occurrence of symptomatic intracranial hemorrhage or new focal neurologic deficit (adjusted HR 3.6, 95% CI 1.3-10.0). CCM excision was associated with worse outcomes over 5 years compared to conservative management. Long-term follow-up will determine whether this difference is sustained over patients' lifetimes. Meanwhile, a randomized controlled trial appears justified. This study provides Class III evidence that CCM excision worsens short-term disability scores and increases the risk of symptomatic intracranial hemorrhage and new focal neurologic deficits.


Further development of the talent development environment questionnaire for sport.
Given the significance of monitoring the critical environmental factors that facilitate athlete performance, this two-phase research aimed to validate and refine the revised talent development environment questionnaire (TDEQ). The TDEQ is a multidimensional self-report scale that assesses talented athletes' environmental experiences. Study 1 (the first phase) involved the examination of the revised TDEQ through an exploratory factor analysis (n = 363). This exploratory investigation identified a 28-item five-factor structure (i.e., TDEQ-5) with adequate internal consistency. Study 2 (the second phase) examined the factorial structure of the TDEQ-5, including convergent validity, discriminant validity, and group invariance (i.e., gender and sports type). The second phase was carried out with 496 talented athletes through the application of confirmatory factor analyses and multigroup invariance tests. The results supported the convergent validity, discriminant validity, and group invariance of the TDEQ-5. In conclusion, the TDEQ-5 with 25 items appears to be a reliable and valid scale for use in talent development environments.


Volunteering: a physical activity intervention for older adults--The Experience Corps program in Baltimore.
There is compelling evidence supporting the benefits of increased regular physical activity in older adults. The Experience Corps program in Baltimore MD was designed in part as a community based approach to increasing physical activity that would also appeal to older adults who have historically not utilized health promotion programs. The Baltimore Experience Corps program places older volunteers in public elementary schools for 15 h a week in roles designed to improve the academic outcomes of children and, simultaneously, increase the physical, cognitive and social activity of volunteers. This paper reports on the change in physical activity levels among older adults associated with participation in the Baltimore Experience Corps. In a pilot randomized controlled evaluation, older adults were randomly assigned to Experience Corps (EC participants) or a waiting list control group. Ages ranged from 59-86 years, 96% were African American, 94% were women, and 84% had annual incomes less than $15,000. EC participants were required to serve >/=15 h a week. At follow-up after 4-8 months, an analysis of 113 randomized volunteers revealed 53% of the EC participants were more active than the previous year by self-report, as compared to 23% of the controls (p<0.01). When adjusted for age, gender and education, there was a trend toward increased physical activity in the EC participants as calculated by a kilocalorie per week increase of 40%, versus a 16% decrease in the controls (p=0.49). EC participants who reported "low activity" at baseline experienced an average 110% increase in their physical activity at follow-up. Among the controls who were in the "low activity" group at baseline, there was, on average, only a 12% increase in physical activity (p=0.03). Among those who were previously active, there was no significant difference (p=0.30). The pilot results suggest that a high intensity volunteer program that is designed as a health promotion intervention can lead, in the short-term, to significant improvements in the level of physical activity of previously inactive older adult volunteers.


Efficacy of ciprofloxacin versus doxycycline as prophylaxis against experimental murine Brucella melitensis infection.
The prophylactic potential of ciprofloxacin was assessed in comparison with doxycycline, an established therapeutic antibiotic, to limit or control infection by Brucella melitensis in an experimental mouse model. Ciprofloxacin treatment reduced bacterial loads in the spleens of challenged mice when administered prior to or at the same time as the bacterial challenge. In comparison, doxycycline provided much greater reductions in bacterial counts, even when treatment was initiated after infection. Doxycycline was able to protect against B. melitensis when administered for 5 days from 24 h after infection and for at least 28 days after cessation of the antibiotic. Overall, these results confirm that ciprofloxacin is less effective than doxycycline but suggest that it may have some utility in providing protection against low-level infections. Combination studies are indicated.


Non-suicidal self-injury in United States adults: prevalence, sociodemographics, topography and functions.
Non-suicidal self-injury (NSSI) has received increased attention in the mental health literature and has been proposed as a diagnostic entity for DSM-5. However, data on NSSI in the United States adult population are lacking. The prevalence and nature of NSSI were examined in a random-digit dialing sample of 439 adults in the United States. Participants were recruited during July and August of 2008. Lifetime prevalence of NSSI was 5.9%, including 2.7% who had self-injured five or more times. The 12-month prevalence was 0.9%. Methods of NSSI reported included cutting/carving, burning, biting, scraping/scratching skin, hitting, interfering with wound healing and skin picking. Half of self-injurers reported multiple methods. The average age of onset was 16 years (median 14 years). Instances of NSSI infrequently co-occurred with suicidal thoughts and with use of alcohol or drugs and rarely required medical treatment. Most injurers reported that NSSI functioned to alleviate negative emotions. Fewer reported that they self-injured to punish themselves, to communicate with others/get attention or to escape a situation or responsibility. NSSI was associated with younger age, being unmarried and a history of mental health treatment, but not with gender, ethnicity, educational history or household income. Results are largely consistent with previous research in adolescent and young adult samples. Study limitations notwithstanding, this study provides the most definitive and detailed information to date regarding the prevalence and characteristics of NSSI in US adults. In the future, it will be important for large-scale epidemiological studies of psychopathology to include questions about NSSI.


Perceived spatial displacement of motion-defined contours in peripheral vision.
The perceived displacement of motion-defined contours in peripheral vision was examined in four experiments. In Experiment 1, in line with Ramachandran and Anstis' finding [Ramachandran, V. S., & Anstis, S. M. (1990). Illusory displacement of equiluminous kinetic edges. Perception, 19, 611-616], the border between a field of drifting dots and a static dot pattern was apparently displaced in the same direction as the movement of the dots. When a uniform dark area was substituted for the static dots, a similar displacement was found, but this was smaller and statistically insignificant. In Experiment 2, the border between two fields of dots moving in opposite directions was displaced in the direction of motion of the dots in the more eccentric field, so that the location of a boundary defined by a diverging pattern is perceived as more eccentric, and that defined by a converging as less eccentric. Two explanations for this effect (that the displacement reflects a greater weight given to the more eccentric motion, or that the region containing stronger centripetal motion components expands perceptually into that containing centrifugal motion) were tested in Experiment 3, by varying the velocity of the more eccentric region. The results favoured the explanation based on the expansion of an area in centripetal motion. Experiment 4 showed that the difference in perceived location was unlikely to be due to differences in the discriminability of contours in diverging and converging patterns, and confirmed that this effect is due to a difference between centripetal and centrifugal motion rather than motion components in other directions. Our result provides new evidence for a bias towards centripetal motion in human vision, and suggests that the direction of motion-induced displacement of edges is not always in the direction of an adjacent moving pattern.


A cytoplasmic protein stimulates normal N-ras p21 GTPase, but does not affect oncogenic mutants.
The role of guanine nucleotides in ras p21 function was determined by using the ability of p21 protein to induce maturation of Xenopus oocytes as a quantitative assay for biological activity. Two oncogenic mutant human N-ras p21 proteins, Asp12 and Val12, actively induced maturation, whereas normal Gly12 p21 was relatively inactive in this assay. Both mutant proteins were found to be associated with guanosine triphosphate (GTP) in vivo. In contrast, Gly12 p21 was predominantly guanosine diphosphate (GDP)-bound because of a dramatic stimulation of Gly12 p21-associated guanosine triphosphatase (GTPase) activity. A cytoplasmic protein was shown to be responsible for this increase in activity. This protein stimulated GTP hydrolysis by purified Gly12 p21 more than 200-fold in vitro, but had no effect on Asp12 or Val12 mutants. A similar factor could be detected in extracts from mammalian cells. It thus appears that, in Xenopus oocytes, this protein maintains normal p21 in a biologically inactive, GDP-bound state through its effect on GTPase activity. Furthermore, it appears that the major effect of position 12 mutations is to prevent this protein from stimulating p21 GTPase activity, thereby allowing these mutants to remain in the active GTP-bound state.


Health service use by patients with heart failure living in a community setting: a cross-sectional analysis in North West London.
The complex nature of heart failure (HF) management, often involving multidimensional care, is widely recognised, but overall health service utilisation by patients with HF has not previously been described. To describe overall health service use by adults with HF living in a community setting. Cross-sectional analysis of prevalent HF cases from January 2015 to December 2018 using an administrative dataset covering primary and secondary care, and 'other' (community, mental health, social care) services in North West London. Healthcare use of each service was described overall and by individual components of secondary care (such as, outpatient appointments), and 'other' services (such as, nursing contacts). Usage patterns were identified using k-means cluster analysis, using all distinct contacts for the whole study period, and visualised with a heatmap. A total of 39 301 patients with a prevalent diagnosis of HF between 1 January 2015 and 31 December 2018 were found. Of those, approximately 90% used health services during the study period, most commonly outpatient services, GP consultations, unplanned accident and emergency visits, and community services. Use of cardiology-specific services ranged from around 3% (cardiology-related community care) to around 20% (outpatient cardiology visits). GP consultations decreased by 11% over the study period. Five clusters of patients were identified, each with statistically significantly different care usage patterns and patient characteristics. Patients with HF make heavy but heterogeneous use of services. Relatively low and falling use of GP consultations, and the apparently low uptake of community rehabilitation services by patients with HF, is concerning and suggests challenges in primary care access and integration of care.


Getting Grounded Gracefully: effectiveness and acceptability of Feldenkrais in improving balance.
The Getting Grounded Gracefully program, based on the Awareness Through Movement lessons of the Feldenkrais method, was designed to improve balance and function in older people. Fifty-five participants (mean age 75, 85% women) were randomized to an intervention (twice-weekly group classes over 8 wk) or a control group (continued with their usual activity) after being assessed at baseline and then reassessed 8 wk later. Significant improvement was identified for the intervention group relative to the control group using ANOVA between-groups repeated-measures analysis for the Modified Falls Efficacy Scale score (p = .003) and gait speed (p = .028), and a strong trend was evident in the timed up-and-go (p = .056). High class attendance (88%) and survey feedback indicate that the program was viewed positively by participants and might therefore be acceptable to other older people. Further investigation of the Getting Grounded Gracefully program is warranted.


Traumatic Brain Injury and Posttraumatic Stress Disorder: Comorbid Consequences of War.
Scientific literature is reviewed supporting a "consequence of war syndrome (CWS)" in Operation Enduring Freedom/Operation Iraqi Freedom/Operation New Dawn soldiers. CWS constituents include chronic pain and insomnia, other physical complaints, posttraumatic stress disorder (PTSD), anxiety, depression, and neuropsychological deficits. The foundation of CWS lies with the chronic stressors inherent to deployment and the cascade of biological events mediated and maintained by hypothalamic-pituitary-adrenal (HPA) axis dysregulation. Such dysregulation is modified by the individual's specific experiences at war, difficulty reintegrating to post-deployment life, and the onset or exacerbation of the chronic and comorbid physical, emotional, and cognitive disorders. The circuit network between the prefrontal cortex (PFC), amygdala, and hippocampus is particularly sensitive to the consequences of war. The review's specific conclusions are as follows: HPA axis dysregulation contributes to the chronic insomnia and hyperarousal seen in soldiers. There is considerable symptom overlap between PTSD and blast-related head injury, and it is difficult to determine the relative contributions of the two disorders to abnormal imaging studies. In some cases, traumatic brain injury (TBI) may directly precipitate PTSD symptoms. While not intuitive, the relationship between TBI and postconcussion syndrome appears indirect and mediated through PTSD. Blast-related or conventional head injury may have little long-term impact on neuropsychological functioning; contrarily, PTSD particularly accounts for current cognitive deficits. The psychological experience of CWS includes a "war-within" where soldiers continue to battle an internalized enemy. Successful treatment of CWS entails transdisciplinary care that addresses each of the constituent disorders.


Annual summary of vital statistics: 2010-2011.
The number of births in the United States declined by 1% between 2010 and 2011, to a total of 3 953 593. The general fertility rate also declined by 1% to 63.2 births per 1000 women, the lowest rate ever reported. The total fertility rate was down by 2% in 2011 (to 1894.5 births per 1000 women). The teenage birth rate fell to another historic low in 2011, 31.3 births per 1000 women. Birth rates also declined for women aged 20 to 29 years, but the rates increased for women aged 35 to 39 and 40 to 44 years. The percentage of all births to unmarried women declined slightly to 40.7% in 2011, from 40.8% in 2010. In 2011, the cesarean delivery rate was unchanged from 2010 at 32.8%. The preterm birth rate declined for the fifth straight year in 2011 to 11.72%; the low birth weight rate declined slightly to 8.10%. The infant mortality rate was 6.05 infant deaths per 1000 live births in 2011, which was not significantly lower than the rate of 6.15 deaths in 2010. Life expectancy at birth was 78.7 years in 2011, which was unchanged from 2010. Crude death rates for children aged 1 to 19 years did not change significantly between 2010 and 2011. Unintentional injuries and homicide were the first and second leading causes of death, respectively, in this age group. These 2 causes of death jointly accounted for 47.0% of all deaths of children and adolescents in 2011.


Mitochondrial matrix localization of a protein altered in mutants resistant to the microtubule inhibitor podophyllotoxin.
Specific antibodies to a protein designated P1 (Mr approximately equal to 63,000), which is specifically altered in mutants resistant to the microtubule inhibitor podophyllotoxin, bind to mitochondria in cells of various vertebrate and invertebrate species (Eur. J. Cell Biol. 44, 278-285 (1987); Can. J. Biochem. Cell Biol. 63, 489-502 (1985)). To investigate the relationship of this protein to mitochondria, rat liver mitochondria have been purified and immunoblot analysis with these provide evidence that the P1 protein is a major component of mitochondria. Two-dimensional gel electrophoretic analysis of mitochondrial proteins from Chinese hamster ovary (CHO) cells also show the P1 protein to be a major mitochondrial component. Subfractionation of rat liver mitochondria into various compartments indicates that the P1 protein is mainly associated with the matrix fraction. Effect of treatment of CHO cells with mitochondrial inhibitors on the synthesis of P1 protein was also investigated. Treatment with the K+ ionophores nonactin and valinomycin, which abolish mitochondrial membrane potential, inhibited synthesis of the mature forms of the P1 protein as well as a number of other mitochondrial proteins, as seen by two-dimensional gel electrophoresis of labeled polypeptides. Treatment of the podophyllotoxin-resistant mutant of CHO cells with the above inhibitors affected both the wild-type and the mutant forms of the P1 protein in a similar manner. Concomitant with the disappearance of the above proteins, new basic proteins of higher molecular masses, related to the P1 and other proteins by peptide analysis, were observed in the drug-treated cells.(ABSTRACT TRUNCATED AT 250 WORDS)


Global haplotype partitioning for maximal associated SNP pairs.
Global partitioning based on pairwise associations of SNPs has not previously been used to define haplotype blocks within genomes. Here, we define an association index based on LD between SNP pairs. We use the Fisher's exact test to assess the statistical significance of the LD estimator. By this test, each SNP pair is characterized as associated, independent, or not-statistically-significant. We set limits on the maximum acceptable proportion of independent pairs within all blocks and search for the partitioning with maximal proportion of associated SNP pairs. Essentially, this model is reduced to a constrained optimization problem, the solution of which is obtained by iterating a dynamic programming algorithm. In comparison with other methods, our algorithm reports blocks of larger average size. Nevertheless, the haplotype diversity within the blocks is captured by a small number of tagSNPs. Resampling HapMap haplotypes under a block-based model of recombination showed that our algorithm is robust in reproducing the same partitioning for recombinant samples. Our algorithm performed better than previously reported models in a case-control association study aimed at mapping a single locus trait, based on simulation results that were evaluated by a block-based statistical test. Compared to methods of haplotype block partitioning, we performed best on detection of recombination hotspots. Our proposed method divides chromosomes into the regions within which allelic associations of SNP pairs are maximized. This approach presents a native design for dimension reduction in genome-wide association studies. Our results show that the pairwise allelic association of SNPs can describe various features of genomic variation, in particular recombination hotspots.


Evolution of the AT-rich mitochondrial DNA of the root knot nematode, Meloidogyne hapla.
Mitochondrial DNA of the root knot nematode Meloidogyne hapla was investigated for intraspecific diversity and divergence from other parthenogenetic root knot nematodes. A 1,900-bp fragment containing COII, tRNAHis, 16S rRNA, ND3 and Cyt b genes has been cloned and sequenced from one individual and an 1,188-bp region within this region was sequenced from four other Australian isolates. M. hapla mtDNA is more than 80% AT-rich, like other Meloidogyne spp. Nucleotide diversity within M. hapla is some 10-fold higher than across three other parthenogenetic species of root-knot nematode (M. arenaria, M. javanica, and M. incognita), implying an earlier origin for M. hapla. Nucleotide divergence between M. hapla and its congener M. javanica is as great as that between Ascaris suum and Caenorhabditis elegans, members of different nematode subclasses, while amino acid sequence difference between Meloidogyne is more than twice as great. This is interpreted as an AT-bias-induced acceleration of the amino acid substitution rate, over and above saturation of nucleotide divergence in the strongly AT-biased DNA, on three lines of evidence: (1) in conserved blocks in 16S rDNA congeneric Meloidogyne have no more differences than between A. suum and C. elegans; (2) the Meloidogyne lineage has more amino acid changes relative to the Ascaris/Caenorhabditis lineage with respect to four of five outgroups, the exceptional outgroup being the only species (Apis) as AT-rich as Meloidogyne; and (3) between the two Meloidogyne there are more first and second but fewer third codon position changes than between the other nematode species. M. hapla is also found to contain a 102-bp tandem repeat of at least 40 copies; a size, arrangement, and position the same as in M. javanica, but sequence comparisons did not demonstrate homology between the two repeats.


Expression of skeletal but not cardiac Na+ channel isoform preserves normal conduction in a depolarized cardiac syncytium.
Reentrant arrhythmias often develop in the setting of myocardial infarction and ensuing slow propagation. Increased Na(+) channel expression could prevent or disrupt reentrant circuits by speeding conduction if channel availability is not limited by membrane depolarization within the diseased myocardium. We therefore asked if, in the setting of membrane depolarization, action potential (AP) upstroke and normal conduction can be better preserved by the expression of a Na(+) channel isoform with altered biophysical properties compared to the native cardiac Na(+) channel isoform, namely having a positively shifted, voltage-dependent inactivation. The skeletal Na(+) channel isoform (SkM1) and the cardiac Na(+) channel isoform (Nav1.5) were expressed in newborn rat ventricular myocyte cultures with a point mutation introduced in Nav1.5 to increase tetrodotoxin (TTX) sensitivity so native and expressed currents could be distinguished. External K(+) was increased from 5.4 to 10 mmol/L to induce membrane depolarization. APs, Na(+) currents, and conduction velocity (CV) were measured. In control cultures, elevated K(+) significantly reduced AP upstroke ( approximately 75%) and CV ( approximately 25%). Expression of Nav1.5 did not protect AP upstroke from K(+) depolarization. In contrast, in SkM1 expressing cultures, high K(+) reduced AP upstroke <50% and conduction was not significantly reduced. In a simulated anatomical reentry setting (using a void), the angular velocity (AV) of induced reentry was faster and the excitable gap shorter in SkM1 cultures compared to control for both normal and high K(+). Expression of SkM1 but not Nav1.5 preserves AP upstroke and CV in a K(+)-depolarized syncytium. The higher AV and shorter excitable gap observed during reentry excitation around a void in SkM1 cultures would be expected to facilitate reentry self-termination. SkM1 Na(+) channel expression represents a novel gene therapy for the treatment of reentrant arrhythmias.


Vanin-1(-/-) mice show decreased NSAID- and Schistosoma-induced intestinal inflammation associated with higher glutathione stores.
Vanin-1 is a membrane-anchored pantetheinase highly expressed in the gut and liver. It hydrolyzes pantetheine to pantothenic acid (vitamin B5) and the low-molecular-weight thiol cysteamine. The latter is believed to be a key regulating factor of several essential metabolic pathways, acting through sulfhydryl-disulfide exchange reactions between sulfhydryl groups of the enzymes and the oxidized form, cystamine. Its physiological importance remains to be elucidated, however. To explore this point, we developed Vanin-1-deficient mice that lack free cysteamine. We examined the susceptibility of deficient mice to intestinal inflammation, either acute (NSAID administration) or chronic (Schistosoma infection). We found that Vanin-1(-/-) mice better controlled inflammatory reaction and intestinal injury in both experiments. This protection was associated with increased gamma-glutamylcysteine synthetase activity and increased stores of reduced glutathione, as well as reduced inflammatory cell activation in inflamed tissues. Oral administration of cystamine reversed all aspects of the deficient phenotype. These findings suggest that one cysteamine function is to upregulate inflammation. Consequently, the pantetheinase activity of Vanin-1 molecule could be a target for a new anti-inflammatory strategy.


The First Step of Neurospora crassa Molybdenum Cofactor Biosynthesis: Regulatory Aspects under N-Derepressing and Nitrate-Inducing Conditions.
Molybdenum cofactor (Moco) is the active site prosthetic group found in all Moco dependent enzymes, except for nitrogenase. Mo-enzymes are crucial for viability throughout all kingdoms of life as they catalyze a diverse set of two electron transfer reactions. The highly conserved Moco biosynthesis pathway consists of four different steps in which guanosine triphosphate is converted into cyclic pyranopterin monophosphate, molybdopterin (MPT), and subsequently adenylated MPT and Moco. Although the enzymes and mechanisms involved in these steps are well characterized, the regulation of eukaryotic Moco biosynthesis is not. Within this work, we described the regulation of Moco biosynthesis in the filamentous fungus Neurospora crassa, which revealed the first step of the multi-step pathway to be under transcriptional control. We found, that upon the induction of high cellular Moco demand a single transcript variant of the nit-7 gene is increasingly formed pointing towards, that essentially the encoded enzyme NIT7-A is the key player for Moco biosynthesis activity in Neurospora.


Associations between an obesity related genetic variant (FTO rs9939609) and prostate cancer risk.
Observational studies suggest that obese men have a lower risk of incident prostate cancer, but an increased risk of advanced and fatal cancers. These observations could be due to confounding, detection bias, or a biological effect of obesity. Genetic studies are less susceptible to confounding than observational epidemiology and can suggest how associations between phenotypes (such as obesity) and diseases arise. To determine whether the associations between obesity and prostate cancer are causal, we conducted a genetic association study of the relationship between a single nucleotide polymorphism known to be associated with obesity (FTO rs9939609) and prostate cancer. Data are from a population-based sample of 1550 screen-detected prostate cancers, 1815 age- and general practice matched controls with unrestricted prostate specific antigen (PSA) values and 1175 low-PSA controls (PSA <0.5 ng/ml). The rs9939609 A allele, which was associated with higher BMI in the sample, was inversely associated with overall (odds ratio (OR) versus all controls = 0.93; 95% confidence interval (CI): 0.85-1.02 p = 0.12 per allele) and low-grade (OR = 0.90; 0.81-0.99 p = 0.03 per allele) prostate cancer risk, but positively associated with high-grade cancer among cases (OR high- versus low-grade cancer = 1.16; 0.99-1.37 p = 0.07 per allele). Although evidence for these effects was weak, they are consistent with observational data based on BMI phenotypes and suggest that the observed association between obesity and prostate cancer is not due to confounding. Further research should confirm these findings, extend them to other BMI-related genetic variants and determine whether they are due to detection bias or obesity-related hormonal changes. Controlled-Trials.com ISRCTN20141297.


Draft Genome Sequence of Bifidobacterium longum ZJ1, Isolated from a Centenarian in Anhui, China.
Here, we present the complete genome sequence of a Bifidobacterium longum isolate, that of strain ZJ1, and this strain showed a cholesterol degradation ability that is greater than that of five strains we chose for comparison (Bifidobacterium longum 536, B. infantis 1912, B. longum 1941, B. breve ATCC 15698, B. infantis ATCC 17930). The draft genome of strain ZJ1 consists of 2,414,672 bp, with 2,042 protein-coding genes, 69 noncoding RNA genes, and 60.16% G+C content.


Management of child and adolescent obesity: attitudes, barriers, skills, and training needs among health care professionals.
The primary aim of this study was to evaluate among health care professionals their attitudes, perceived barriers, perceived skill level, and training needs in the management of child and adolescent obesity. A national needs assessment consisting of a mailed questionnaire was conducted among a random sample of health care professionals. The survey was completed by 202 pediatricians, 293 pediatric nurse practitioners, and 444 registered dietitians. The majority of all respondents felt that childhood obesity was a condition that needs treatment (75%-93%), and affects chronic disease risk (76%-89%) and future quality of life (83%-93%). The most frequent barriers were lack of parent involvement, lack of patient motivation, and lack of support services. Registered dietitians were less likely to identify barriers to treatment compared with pediatricians or pediatric nurse practitioners. The most common areas of self-perceived low proficiency were in the use of behavioral management strategies, guidance in parenting techniques, and addressing family conflicts. All 3 groups expressed high interest in additional training on obesity management of children and adolescents, especially in the area of behavioral management strategies and parenting techniques. Those practitioners with >10 years of practice reported the greatest interest in training. Pediatric practitioners view child and adolescent obesity with concern and feel that intervention is important. However, several important barriers interfere with treatment efforts and will need to be addressed. There is also a need for increased training opportunities related to obesity prevention and treatment. The results of this study provide directions and priorities for training, education, and advocacy efforts.


Influence of Eysenckian Personality Traits in Choice of Specialization by Young Omani Doctors.
The role of personality in occupational specialty choices has been explored in many parts of the world. To our knowledge, there is a dearth of such studies in the Arab/Islamic population and Oman is no exception. This study aimed to explore the relationship between personality traits and specialty choice among residents of Oman Medical Specialty Board (OMSB). A cross-sectional study was carried out among Omani resident physicians working under OMSB. The Eysenck Personality Questionnaire-Revised was employed to quantify personality subtypes (e.g., psychoticism, extraversion, and neuroticism). Specialties were categorized as surgical, medical, and diagnostics as per standard of North American medical specialties. A total of 255 residents in 17 medical specialties participated in the study (m = 40.4%; f = 59.6%) of 300 eligible subjects giving a response rate of 85.0%. Respondents who had chosen surgical specialties scored significantly higher on the psychoticism subscale than those who had opted for medical and diagnostic specialties. As for individual specialties, orthopedic respondents had statistically significant higher mean scores on psychoticism and neuroticism compared to radiologists and psychiatrists who scored the lowest in the two personality traits, respectively. This study found statistically significant associations between personality traits and choices of specialty by young Omani doctors. We recommend more detailed studies that examine further psychological and cultural variables that are likely to affect the choices of specializations by young Omani professionals in both medical and non-medical fields.


Managing the symptoms of neuropathic pain: an exploration of patients' experiences.
The debilitating effects of chronic neuropathic pain on everyday life are considerable, but little is known about how individual sufferers manage these effects. Virtually nothing is known about what patients prefer, what measures they take themselves and when, or in what combinations. The aim of this study was to explore patients' reports of how they managed their neuropathic pain symptoms. Three focus groups (10 participants in total) were used to generate qualitative data on both individual and shared experiences of managing symptoms of neuropathic pain. Discussions were recorded and transcribed verbatim. Data were analyzed using thematic analysis, identifying categories and broader themes of importance to patients. The most common management strategy was the use of conventional medications, often associated with poor effectiveness and unpleasant side effects. Complementary and alternative medicine was ineffective, but many found resting or retreating helpful. They exhibited a repeated cycle of seeking help to manage the pain, with each unsuccessful attempt followed by new attempts. Some had tried to accept their pain, but there was insufficient psychological, social, emotional, and practical support to allow them to do this successfully. This exploratory study provides a basis from which to develop a larger study to validate and extend the findings. Other issues meriting research are the effectiveness of cognitive-behavioral therapies for those with neuropathic pain, and an exploration and subsequent evaluation of different types of social, practical, and emotional support needed to help live with neuropathic pain.


Application of Psychological Theories in Agent-Based Modeling: The Case of the Theory of Planned Behavior.
It is likely that computer simulations will assume a greater role in the next future to investigate and understand reality (Rand & Rust, 2011). Particularly, agent-based models (ABMs) represent a method of investigation of social phenomena that blend the knowledge of social sciences with the advantages of virtual simulations. Within this context, the development of algorithms able to recreate the reasoning engine of autonomous virtual agents represents one of the most fragile aspects and it is indeed crucial to establish such models on well-supported psychological theoretical frameworks. For this reason, the present work discusses the application case of the theory of planned behavior (TPB; Ajzen, 1991) in the context of agent-based modeling: It is argued that this framework might be helpful more than others to develop a valid representation of human behavior in computer simulations. Accordingly, the current contribution considers issues related with the application of the model proposed by the TPB inside computer simulations and suggests potential solutions with the hope to contribute to shorten the distance between the fields of psychology and computer science.


Btk regulates macrophage polarization in response to lipopolysaccharide.
Bacterial Lipopolysaccharide (LPS) is a strong inducer of inflammation and does so by inducing polarization of macrophages to the classic inflammatory M1 population. Given the role of Btk as a critical signal transducer downstream of TLR4, we investigated its role in M1/M2 induction. In Btk deficient (Btk (-\-)) mice we observed markedly reduced recruitment of M1 macrophages following intraperitoneal administration of LPS. Ex vivo analysis demonstrated an impaired ability of Btk(-/-) macrophages to polarize into M1 macrophages, instead showing enhanced induction of immunosuppressive M2-associated markers in response to M1 polarizing stimuli, a finding accompanied by reduced phosphorylation of STAT1 and enhanced STAT6 phosphorylation. In addition to STAT activation, M1 and M2 polarizing signals modulate the expression of inflammatory genes via differential activation of transcription factors and regulatory proteins, including NF-κB and SHIP1. In keeping with a critical role for Btk in macrophage polarization, we observed reduced levels of NF-κB p65 and Akt phosphorylation, as well as reduced induction of the M1 associated marker iNOS in Btk(-/-) macrophages in response to M1 polarizing stimuli. Additionally enhanced expression of SHIP1, a key negative regulator of macrophage polarisation, was observed in Btk(-/-) macrophages in response to M2 polarizing stimuli. Employing classic models of allergic M2 inflammation, treatment of Btk (-/-) mice with either Schistosoma mansoni eggs or chitin resulted in increased recruitment of M2 macrophages and induction of M2-associated genes. This demonstrates an enhanced M2 skew in the absence of Btk, thus promoting the development of allergic inflammation.


Influence of quercetin-rich food intake on microRNA expression in lung cancer tissues.
Epidemiologic studies have reported that frequent consumption of quercetin-rich foods is inversely associated with lung cancer incidence. A quercetin-rich diet might modulate microRNA (miR) expression; however, this mechanism has not been fully examined. miR expression data were measured by a custom-made array in formalin-fixed paraffin-embedded tissue samples from 264 lung cancer cases (144 adenocarcinomas and 120 squamous cell carcinomas). Intake of quercetin-rich foods was derived from a food-frequency questionnaire. In individual-miR-based analyses, we compared the expression of miRs (n = 198) between lung cancer cases consuming high versus low quercetin-rich food intake using multivariate ANOVA tests. In family-miR-based analyses, we used Functional Class Scoring (FCS) to assess differential effect on biologically functional miR families. We accounted for multiple testing using 10,000 global permutations (significance at P(global) < 0.10). All multivariate analyses were conducted separately by histology and by smoking status (former and current smokers). Family-based analyses showed that a quercetin-rich diet differentiated miR expression profiles of the tumor suppressor let-7 family among adenocarcinomas (P(FCS) < 0.001). Other significantly differentiated miR families included carcinogenesis-related miR-146, miR-26, and miR-17 (P (FCS) < 0.05). In individual-based analyses, we found that among former and current smokers with adenocarcinoma, 33 miRs were observed to be differentiated between highest and lowest quercetin-rich food consumers (23 expected by chance; P(global) = 0.047). We observed differential expression of key biologically functional miRs between high versus low consumers of quercetin-rich foods in adenocarcinoma cases. Our findings provide preliminary evidence on the mechanism underlying quercetin-related lung carcinogenesis.


Fatal or harmless: extreme bistability induced by sterilizing, sexually transmitted pathogens.
Models of sexually transmitted infections have become a fixture of mathematical epidemiology. A common attribute of all these models is treating reproduction and mating, and hence pathogen transmission, as uncoupled events. This is fine for humans, for example, where only a tiny fraction of sexual intercourses ends up with having a baby. But it can be a deficiency for animals in which mating and giving birth are tightly coupled, and mating thus mediates both reproduction and pathogen transmission. Here, we model dynamics of sterilizing, sexually transmitted infections in such animals, assuming structural consistency between the processes of reproduction and pathogen transmission. We show that highly sterilizing, sexually transmitted pathogens trigger bistability in the host population. In particular, the host population can end up in two extreme alternative states, disease-free persistence and pathogen-driven extinction, depending on its initial state. Given that sterilizing, sexually transmitted infections that affect animals are abundant, our results might implicate an effective pest control tactic that consists of releasing the corresponding pathogens, possibly after genetically enhancing their sterilization power.


Epigallocatechin-3-gallate promotes apoptosis and expression of the caspase 9a splice variant in PC3 prostate cancer cells.
Growing evidence suggests that the flavonoid epigallocatechin-3-gallate (EGCG), notably abundant in green tea, has health-promoting properties. We examined the effect of EGCG on cell survival and apoptosis in the prostate cancer cell line PC3. Cell survival was reduced and apoptosis increased significantly with a low dose of 1 µM EGCG. The ability of the anticancer drug cisplatin to promote apoptosis was enhanced by EGCG. Furthermore, EGCG, both alone and in combination with cisplatin, promoted the expression of the pro-apoptotic splice isoform of caspase 9.


Assessment of patient safety culture in Palestinian public hospitals.
To assess the prevalent patient safety culture in Palestinian public hospitals. A cross-sectional design, Arabic translated version of the Hospital Survey on Patient Safety Culture was used. All the 11 general public hospitals in the West Bank. A total of 1460 clinical and non-clinical hospital staff. No. Twelve patient safety culture composites and 2 outcome variables (patient safety grade and events reported in the past year) were measured. Most of the participants were nurses and physicians (69.2%) with direct contact with patients (92%), mainly employed in medical/surgical units (55.1%). The patient safety composites with the highest positive scores were teamwork within units (71%), organizational learning and continuous improvement (62%) and supervisor/manager expectations and actions promoting patient safety (56%). The composites with the lowest scores were non-punitive response to error (17%), frequency of events reported (35%), communication openness (36%), hospital management support for patient safety (37%) and staffing (38%). Although 53.2% of the respondents did not report any event in the past year, 63.5% rated patient safety level as 'excellent/very good'. Significant differences in patient safety scores and outcome variables were found between hospitals of different size and in relation to staff positions and work hours. This study highlights the existence of a punitive and blame culture, under-reporting of events, lack of communication openness and inadequate management support that are key challenges for patient safe hospital care. The baseline survey results are valuable for designing and implementing the patient safety program and for measuring future progress.


Are carbon nanotubes a natural solution? Applications in biology and medicine.
Carbon nanotubes and materials based on carbon nanotubes have many perceived applications in the field of biomedicine. Several highly promising examples have been highlighted in the literature, ranging from their use as growth substrates or tissue scaffolds to acting as intracellular transporters for various therapeutic and diagnostic agents. In addition, carbon nanotubes have a strong optical absorption in the near-infrared region (in which tissue is transparent), which enables their use for biological imaging applications and photothermal ablation of tumors. Although these advances are potentially game-changing, excitement must be tempered somewhat as several bottlenecks exist. Carbon nanotube-based technologies ultimately have to compete with and out-perform existing technologies in terms of performance and price. Moreover, issues have been highlighted relating to toxicity, which presents an obstacle for the transition from preclinical to clinical use. Although many studies have suggested that well-functionalized carbon nanotubes appear to be safe to the treated animals, mainly rodents, long-term toxicity issues remains to be elucidated. In this report, we systematically highlight some of the most promising biomedical application areas of carbon nanotubes and review the interaction of carbon nanotubes with cultured cells and living organisms with a particular focus on in vivo biodistribution and potential adverse health effects. To conclude, future challenges and prospects of carbon nanotubes for biomedical applications will be addressed.


Effects of self-monitoring of glucose in non-insulin treated patients with type 2 diabetes: design of the IN CONTROL-trial.
Diabetes specific emotional problems interfere with the demanding daily management of living with type 2 diabetes mellitus (T2DM). Possibly, offering direct feedback on diabetes management may diminish the presence of diabetes specific emotional problems and might enhance the patients' belief they are able to manage their illness. It is hypothesized that self-monitoring of glucose in combination with an algorithm how and when to act will motivate T2DM patients to become more active participants in their own care leading to a decrease in diabetes related distress and an increased self-efficacy. Six hundred patients with T2DM (45 < or = 75 years) who receive care in a structured diabetes care system, HbA1c > or = 7.0%, and not using insulin will be recruited and randomized into 3 groups; Self-monitoring of Blood Glucose (SMBG), Self-monitoring of Urine Glucose (SMUG) and usual care (n = 200 per group). Participants are eligible if they have a known disease duration of over 1 year and have used SMBG or SMUG less than 3 times in the previous year. All 3 groups will receive standardized diabetes care. The intervention groups will receive additional instructions on how to perform self-monitoring of glucose and how to interpret the results. Main outcome measures are changes in diabetes specific emotional distress and self-efficacy. Secondary outcome measures include difference in HbA1c, patient satisfaction, occurrence of hypoglycaemia, physical activity, costs of direct and indirect healthcare and changes in illness beliefs. The IN CONTROL-trial is designed to explore whether feedback from self-monitoring of glucose in T2DM patients who do not require insulin can affect diabetes specific emotional distress and increase self-efficacy. Based on the self-regulation model it is hypothesized that glucose self-monitoring feedback changes illness perceptions, guiding the patient to reduce emotional responses to experienced threats, and influences the patients ability to perform and maintain self-management skills.


Efficient Identification of Causal Mutations through Sequencing of Bulked F 2 from Two Allelic Bloomless Mutants of Sorghum bicolor.
Sorghum (Sorghum bicolor Moench, L.) plant accumulates copious layers of epi-cuticular wax (EW) on its aerial surfaces, to a greater extent than most other crops. EW provides a vapor barrier that reduces water loss, and is therefore considered to be a major determinant of sorghum's drought tolerance. However, little is known about the genes responsible for wax accumulation in sorghum. We isolated two allelic mutants, bloomless40-1 (bm40-1) and bm40-2, from a mutant library constructed from ethyl methane sulfonate (EMS) treated seeds of an inbred, BTx623. Both mutants were nearly devoid of the EW layer. Each bm mutant was crossed to the un-mutated BTx623 to generated F2 populations that segregated for the bm phenotype. Genomic DNA from 20 bm F2 plants from each population was bulked for whole genome sequencing. A single gene, Sobic.001G228100, encoding a GDSL-like lipase/acylhydrolase, had unique homozygous mutations in each bulked F2 population. Mutant bm40-1 harbored a missense mutation in the gene, whereas bm40-2 had a splice donor site mutation. Our findings thus provide strong evidence that mutation in this GDSL-like lipase gene causes the bm phenotype, and further demonstrate that this approach of sequencing two independent allelic mutant populations is an efficient method for identifying causal mutations. Combined with allelic mutants, MutMap provides powerful method to identify all causal genes for the large collection of bm mutants in sorghum, which will provide insight into how sorghum plants accumulate such abundant EW on their aerial surface. This knowledge may facilitate the development of tools for engineering drought-tolerant crops with reduced water loss.


[Age dependence of the level of copia retrotransposon expression in the testes of Drosophila melanogaster].
Expression of the lacZ reporter gene controlled by various deletion derivatives of the regulatory region of the copia retrotransposon was studied in the testes of adult transgenic males of the Drosophila melanogaster y1W67c23(2) strain at the age of 3, 6-9, 12-15, 18-21, and 24-27 days. When the construct contained the full-length regulatory region, which included the 5'-long terminal repeat (LTR) and the 5'-untranslated region (UTR), expression was the lowest in males aged 12-15 days and the highest in males aged 3 or 24-27 days. A similar V-shaped age dependence was previously observed for the copia transposition rate and RNA content in a strain with a high rate of copia transposition. Thus, the V-shaped age dependence of expression, which is unusual for Drosophila, proved to be characteristic of copia regardless of its transposition rate. Deletion of the 5' or 3' end of the LTR, but not of the UTR, changed the age dependence of the level of reporter gene expression. In this case, expression was the highest in 3-day-old males and gradually decreased with age, as typical for many Drosophila genes. It was assumed that the 5'- and 3'-terminal regions of the copia LTR contain regulatory elements responsible for the V-shaped age dependence of expression, while the expression level depends to a greater extent on the regulatory elements of UTR.


Synthetic Cannabis Analogues and Suicidal Behavior: Case Report.
Despite growing legal control, a wide range of synthetic cannabis analogues is currently used for recreational purposes, notwithstanding their well adverse outcomes, which appear to be more frequent and more serious than those associated with cannabis use. We present the case report of a patient with paranoid schizophrenia, who attempted suicide by serious bodily harm after a single use of "Shiva Ultra Strong," a compound of several synthetic cannabis analogues. A 32-year-old male patient with paranoid schizophrenia was brought to the emergency department presenting with a severe self-inflicted wound to the neck which lacerated the right jugular vein and ipsilateral airway, and narrowly missed the carotid bifurcation. On examination, the patient exhibited psychomotor agitation and anxiety. Laboratory tests, which included routine substance use screening, proved unremarkable. The patient was admitted to the ENT Department for surgical treatment, after which he was transferred to our Psychiatry Department, exhibiting consistent improvement with his usual antipsychotic regimen, to which he had good previous adherence. Later, after discharge, he admitted to having used a smartshop drug, so-called "Shiva Ultra Strong," shortly before the suicide attempt. Although current data on the suicide risk of synthetic cannabis analogues are limited, there is growing evidence of relevant psychiatric effects after their use. Patients with serious mental disorders could prove particularly vulnerable to these drugs, resulting in severe behavioral changes and self-harm.


Microsatellite testing in colon cancer
Significant progress has been made in understanding the molecular genetic basis of colorectal cancer (CRC). That information is paving the way to understanding the genetic basis of other tumors, as well. Oncology nurses should anticipate the routine integration of this information and testing of CRC tumors to understand the molecular basis of the disease in clinical practice. Molecular testing can lead to the identification of families at risk for hereditary cancer syndromes, particularly Lynch syndrome, which sometimes is referred to as hereditary nonpolyposis colorectal cancer. Knowledge of the genetic basis of CRC also contributes valuable information aimed at selecting appropriate and effective targeted therapy.


Bioinformatics Analysis of Estrogen-Responsive Genes.
Estrogen is a steroid hormone that plays critical roles in a myriad of intracellular pathways. The expression of many genes is regulated through the steroid hormone receptors ESR1 and ESR2. These bind to DNA and modulate the expression of target genes. Identification of estrogen target genes is greatly facilitated by the use of transcriptomic methods, such as RNA-seq and expression microarrays, and chromatin immunoprecipitation with massively parallel sequencing (ChIP-seq). Combining transcriptomic and ChIP-seq data enables a distinction to be drawn between direct and indirect estrogen target genes. This chapter discusses some methods of identifying estrogen target genes that do not require any expertise in programming languages or complex bioinformatics.


The experience among patients with multiple dental loss as a consequence of treatment for head and neck cancer: A qualitative study.
To explore the experience among patients with multiple dental loss as a consequence of treatment for head and neck cancer. Semi-structured qualitative interviews were conducted with a purposive sample of fifteen people with head and neck cancer, who had multiple teeth removed as part of their tumour resection or extracted pre-radiotherapy. The interviews were digitally recorded and transcribed for analysis by two researchers, independently. The interview data were categorised into four themes: 1. Pretreatment experience of being informed that teeth had to be removed, 2. Impact of dental loss post-treatment, 3. Coping with dental loss, and 4. Getting dentures and implants. Patients receiving primary radiotherapy felt the time between being informed of requirement for dental extractions and actual extractions was short. Dental loss was detrimental to all patients in terms of eating, speaking, socially and their intimate lives. Patients whose primary treatment was surgery, appeared more accepting of dental loss. Whereas, those having primary radiotherapy, there seemed to be a focus on prevention of radiotherapy-induced complications, with limited choices and recognition on post-treatment dental functionality. This study showed the negative impact of dental loss on patients' quality of life. Dentists should be aware of the post-treatment implications of dental loss on patients' lives; presenting this, as well as the clinical advantages for extractions, to aid decision-making. Patients should be informed of the lack of conclusive research evidence regarding pre-radiotherapy dental extraction. There should also be clear pathways regarding post-treatment dental-related rehabilitation.


Environmental history impacts gene expression during diapause development in the alfalfa leafcutting bee, Megachile rotundata.
Our understanding of the mechanisms controlling insect diapause has increased dramatically with the introduction of global gene expression techniques, such as RNA sequencing (RNA-seq). However, little attention has been given to how ecologically relevant field conditions may affect gene expression during diapause development because previous studies have focused on laboratory-reared and -maintained insects. To determine whether gene expression differs between laboratory and field conditions, prepupae of the alfalfa leafcutting bee, Megachile rotundata, entering diapause early or late in the growing season were collected. These two groups were further subdivided in early autumn into laboratory- and field-maintained groups, resulting in four experimental treatments of diapausing prepupae: early and late field, and early and late laboratory. RNA-seq and differential expression analyses were performed on bees from the four treatment groups in November, January, March and May. The number of treatment-specific differentially expressed genes (97 to 1249) outnumbered the number of differentially regulated genes common to all four treatments (14 to 229), indicating that exposure to laboratory or field conditions had a major impact on gene expression during diapause development. Principle component analysis and hierarchical cluster analysis yielded similar grouping of treatments, confirming that the treatments form distinct clusters. Our results support the conclusion that gene expression during the course of diapause development is not a simple ordered sequence, but rather a highly plastic response determined primarily by the environmental history of the individual insect.


Open-book tests: search behaviour, time used and test scores.
Because of the increasing medical knowledge and the focus of medical education on acquiring competencies, the use of open-book tests seems inevitable. Dealing with a large body of information, indicating which kind of information is needed to solve a problem, and finding and understanding that knowledge at the right moment are behaviours that cannot be assessed during closed-book tests. To examine whether there is a relationship between students' search behaviour - using references or not when answering a question - during open-book tests and their test scores. Second- (N = 491) and third-year medical students (N = 325) participated in this study. Search behaviour was operationalized as the number of questions for which students consulted their references. Furthermore, we collected data on the time students spent on answering all open-book questions and their test scores. To determine the relations, we calculated Spearman's and Pearson's correlations. Second- and third-year students consulted their references for 87% and 73% of the questions and spent 5.0 and 4.3 min on answering an open-book question, respectively. We did not find significant correlations between search behaviour and test scores. Both 'well' and 'poorer performing' students often consulted their references. Spending almost 5 min per open-book question in multiple choice format seems to be too much. More research is needed to establish optimal open-book test time and to explore how 'well performing' students use their references during open-book tests.


The GPIIIA PlA2 polymorphism is associated with an increased risk of cardiovascular adverse events.
The clinical impact of PlA2 polymorphism has been investigated in several diseases, but the definition of its specific role on thrombotic cardiovascular complications has been challenging. We aimed to explore the effect of PlA2 polymorphism on outcome in patients with atherosclerosis. We studied 400 consecutive patients with coronary artery disease (CAD) undergoing percutaneous coronary intervention. A replication study was conducted in 74 hypertensive patients with cerebrovascular events while a group of 100 healthy subjects was included as control population. PlA genotype was determined by PCR-RFLP on genomic DNA from peripheral blood cells. Major adverse cardiac events (MACE), were considered as end points, and recorded at a mean follow up of 24 ± 4.3 months. The frequencies of PlA2 polymorphism was similar between groups and genotype distribution was in Hardy-Weinberg equilibrium. In patients with CAD, the presence of PlA2 allele was associated with higher incidence of cardiac death (13.1% vs. 1.5%, p = 0.0001), myocardial infarction (10.7% vs. 2.6%, p = 0.004) and needs of new revascularization (34.8% vs. 17.7%, p = 0.010). Accordingly, the Kaplan-Meier analysis for event free survival in patients harboring the PlA2 allele showed worse long-term outcome for these patients (p = 0.015). Cox regression analysis identified the presence of PlA2 as an independent predictor of cardiac death (OR: 9.594, 95% CI: 2.6 to 35.3, p = 0.002) and overall MACE (OR: 1.829, 95% CI: 1.054 to 3.176, p = 0.032). In the replication study, the PlA2 polymorphism increased the risk of stroke (OR: 4.1, 95% CI: 1.63-12.4, p = 0.02) over TIA and was identified as an independent risk factor for stroke (B:-1.39; Wald: 7.15; p = 0.001). Our study demonstrates that in patients with severe atherosclerosis the presence of PlA2 allele is associated with thrombotic cardiovascular complications.