Multiple actuation microvalves in wax microfluidics.
Microvalves are an essential component of microfluidic devices. In this work, a low-consumption (<35 mJ), fast-response (<0.3 s), small footprint (<0.5 mm2) wax microvalve capable of multiple actuation is described. This phase-change microvalve is electrically controlled, simple to operate and can be easily fabricated as a fully integrated element of wax microfluidic devices through a special decal-transfer microlithographic process. The valve is inherently latched and leak-proof to at least 100 kPa. A minimum pressure of 3 kPa is required for valve opening. Maximum pressures for a successful closing in air and liquid are 90 and 40 kPa, respectively. The wax valve exhibits reversible open-close behaviour without failure for up to 10 actuation cycles in air (60 kPa) and 5 in water (30 kPa). To the best of our knowledge, this microvalve has the lowest energy consumption (two orders of magnitude lower) reported so far for a plug-type phase-change valve. Furthermore, its size, actuation mechanism and fabrication technology make it suitable for large-scale integration in microfluidic devices. Detailed characteristics in fabrication and actuation of the wax microfluidic valve as well as a test example of its performance for liquid dispensing are reported.


Nutrients and heavy metals distribution in thermally treated pig manure.
Ash from pig manure treated by combustion and thermal gasification was characterized and compared in terms of nutrient, i.e., potassium (K), phosphorus (P) and heavy metal, i.e., cadmium (Cd), chromium (Cr), copper (Cu), nickel (Ni) and zinc (Zn) contents. Total nutrient and metal concentrations were measured using energy dispersive X-ray fluorescence analysis. Acid (HNO3, H2SO4) and water-extractable concentrations were also measured both in non-classified ash and in selected ash particle size fractions using flame atomic absorption spectrometry and colorimetric spectrometry. Results indicate that ash from gasified manure contained more water-extractable K in comparison with combusted manure whereas the opposite was the case with respect to P. Heavy metals Ni, Cr and Cd were present in higher concentrations in the fine particle size fractions (< 30 microm of particle diameter), whereas K, P, Zn and Cu exhibited higher concentrations in the coarser particle size fractions (> 30 microm).


Gene expression profile of hepatopancreas from grass shrimp Palaemonetes pugio exposed to cyclic hypoxia.
Estuarine organisms often experience periods of cyclic hypoxia characterized by hypoxia in the early morning and normoxia in the afternoon. Here we examine the genomic responses of grass shrimp, Palaemonetes pugio, exposed to cyclic hypoxia in the laboratory. Differentially expressed genes in the hepatopancreas were determined in cyclic hypoxic vs. normoxic control groups after 1, 2, 5 and 10 days of exposure to cyclic hypoxia using microarrays printed with 661 annotated transcripts obtained from multiple EST (expressed sequence tag) libraries. Sampling on each day was conducted at two different time series, one in the morning (representing low concentration of dissolved oxygen (DO), designated C-AM) and one in the afternoon (representing high DO concentration, designated C-PM). Distinct differences were observed between the number and identity of specific genes that were significantly down- or up-regulated in shrimp collected at the low DO and high DO points of the cyclic DO cycle. However, cluster analysis showed that the overall response patterns of high (C-PM) and low DO (C-AM) exposures were in the same cluster at 1, 2, and 5 days. In contrast, the response patterns on different days were in different clusters. Day 1 was characterized by up-regulation of 17 unknown genes in the morning and a transient down-regulation of several hemocyanin genes, which returned to normoxic control levels in the afternoon. Days 2 and 5 showed significant down-regulation of 10 (C-AM) and 15 (C-PM) unknown genes, respectively. On day 10 the high DO samples showed a dramatic increase in the number of up-regulated genes, including several distinct hemocyanin genes, and this profile did not cluster with any of the other treatment groups. Vitellogenin, cathepsin L, cytochrome c oxidase subunit III, and fatty acid binding protein 10 were the signature down-regulated genes at day 10 (C-AM). According to GO annotation, the most abundant group of genes for both cyclic low (C-AM) and high (C-PM) DO exposure was associated with transport, defense response, and metabolic process. The differentially expressed genes were mapped to KEGG metabolic and regulatory pathways according to the gene distribution in Drosophila pathway database. Cyclic high (C-PM) DO affected a broad range of pathways compared to cyclic low (C-AM) DO.


Porphyromonas gingivalis attenuates ATP-mediated inflammasome activation and HMGB1 release through expression of a nucleoside-diphosphate kinase.
Many intracellular pathogens evade the innate immune response in order to survive and proliferate within infected cells. We show that Porphyromonas gingivalis, an intracellular opportunistic pathogen, uses a nucleoside-diphosphate kinase (NDK) homolog to inhibit innate immune responses due to stimulation by extracellular ATP, which acts as a danger signal that binds to P2X7 receptors and induces activation of an inflammasome and caspase-1. Thus, infection of gingival epithelial cells (GECs) with wild-type P. gingivalis results in inhibition of ATP-induced caspase-1 activation. However, ndk-deficient P. gingivalis is less effective than wild-type P. gingivalis in reducing ATP-mediated caspase-1 activation and secretion of the pro-inflammatory cytokine, IL-1β, from infected GECs. Furthermore, P. gingivalis NDK modulates release of high-mobility group protein B1 (HMGB1), a pro-inflammatory danger signal, which remains associated with chromatin in healthy cells. Unexpectedly, infection with either wild-type or ndk-deficient P. gingivalis causes release of HMGB1 from the nucleus to the cytosol. But HMGB1 is released to the extracellular space when uninfected GECs are further stimulated with ATP, and there is more HMGB1 released from the cells when ATP-treated cells are infected with ndk-deficient mutant than wild-type P. gingivalis. Our results reveal that NDK plays a significant role in inhibiting P2X7-dependent inflammasome activation and HMGB1 release from infected GECs.


Iterative evaluation of the Health Level 7--Logical Observation Identifiers Names and Codes Clinical Document Ontology for representing clinical document names: a case report.
The authors summarize their experience in iteratively testing the adequacy of three versions of the Health Level Seven (HL7) Logical Observation Identifiers Names and Codes (LOINC) Clinical Document Ontology (CDO) to represent document names at Columbia University Medical Center. The percentage of documents fully represented increased from 23.4% (Version 1) to 98.5% (Version 3). The proportion of unique representations increased from 7.9% (Analysis 1) to 39.4% (Analysis 4); the proportion reflects the level of specificity in the document names as well as the completeness and level of granularity of the CDO. The authors shared the findings of each analysis with the Clinical LOINC committee and participated in the decision-making regarding changes to the CDO on the basis of those analyses and those conducted by the Department of Veterans Affairs. The authors encourage other institutions to actively engage in testing healthcare standards and participating in standards development activities to increase the likelihood that the evolving standards will meet institutional needs.


Increased calvaria cell differentiation and bone matrix formation induced by fibroblast growth factor receptor 2 mutations in Apert syndrome.
Apert syndrome, associated with fibroblast growth factor receptor (FGFR) 2 mutations, is characterized by premature fusion of cranial sutures. We analyzed proliferation and differentiation of calvaria cells derived from Apert infants and fetuses with FGFR-2 mutations. Histological analysis revealed premature ossification, increased extent of subperiosteal bone formation, and alkaline phosphatase- positive preosteoblastic cells in Apert fetal calvaria compared with age-matched controls. Preosteoblastic calvaria cells isolated from Apert infants and fetuses showed normal cell growth in basal conditions or in response to exogenous FGF-2. In contrast, the number of alkaline phosphatase- positive calvaria cells was fourfold higher than normal in mutant fetal calvaria cells with the most frequent Apert FGFR-2 mutation (Ser252Trp), suggesting increased maturation rate of cells in the osteoblastic lineage. Biochemical and Northern blot analyses also showed that the expression of alkaline phosphatase and type 1 collagen were 2-10-fold greater than normal in mutant fetal calvaria cells. The in vitro production of mineralized matrix formed by immortalized mutant fetal calvaria cells cultured in aggregates was also increased markedly compared with control immortalized fetal calvaria cells. The results show that Apert FGFR-2 mutations lead to an increase in the number of precursor cells that enter the osteogenic pathway, leading ultimately to increased subperiosteal bone matrix formation and premature calvaria ossification during fetal development, which establishes a connection between the altered genotype and cellular phenotype in Apert syndromic craniosynostosis.


Should enteral nutrition be started in the first week of critical illness?
To review the mechanistic evidence for early enteral nutrition in critically ill patients within the first week of ICU admission. There is a lack of recent large randomized controlled trials showing clinically important outcome benefits related to early enteral nutrition. Most supporting studies are based on the animal models, and explore mechanisms of benefit related to gut immunity and oxidative stress. In addition, the impact of nutrient deprivation on the microbiome recently shown in a human trial is compelling. Large randomized controlled clinical trials have emerged in the last 2 years, however, comparing minimal enteral nutrition therapy and enteral nutrition versus parenteral nutrition. They call into question the low quality of clinical evidence and the widespread support for early enteral nutrition as a primary recommendation. As a result, the questions of whether or not enteral nutrition should be initiated in the first week versus standard of care or parenteral nutrition and how clinicians justify this recommendation are raised. Despite the wide range of quality in the current clinical outcomes evidence, early enteral nutrition within the first week of ICU admission, delivered to the appropriate patient, promotes gut-mediated immunity, lowers metabolic response to stress, maintains microbial diversity, and improves clinical outcomes versus standard of care or parenteral nutrition therapy.


Time to abandon the hygiene hypothesis: new perspectives on allergic disease, the human microbiome, infectious disease prevention and the role of targeted hygiene.
To review the burden of allergic and infectious diseases and the evidence for a link to microbial exposure, the human microbiome and immune system, and to assess whether we could develop lifestyles which reconnect us with exposures which could reduce the risk of allergic disease while also protecting against infectious disease. Using methodology based on the Delphi technique, six experts in infectious and allergic disease were surveyed to allow for elicitation of group judgement and consensus view on issues pertinent to the aim. Key themes emerged where evidence shows that interaction with microbes that inhabit the natural environment and human microbiome plays an essential role in immune regulation. Changes in lifestyle and environmental exposure, rapid urbanisation, altered diet and antibiotic use have had profound effects on the human microbiome, leading to failure of immunotolerance and increased risk of allergic disease. Although evidence supports the concept of immune regulation driven by microbe-host interactions, the term 'hygiene hypothesis' is a misleading misnomer. There is no good evidence that hygiene, as the public understands, is responsible for the clinically relevant changes to microbial exposures. Evidence suggests a combination of strategies, including natural childbirth, breast feeding, increased social exposure through sport, other outdoor activities, less time spent indoors, diet and appropriate antibiotic use, may help restore the microbiome and perhaps reduce risks of allergic disease. Preventive efforts must focus on early life. The term 'hygiene hypothesis' must be abandoned. Promotion of a risk assessment approach (targeted hygiene) provides a framework for maximising protection against pathogen exposure while allowing spread of essential microbes between family members. To build on these findings, we must change public, public health and professional perceptions about the microbiome and about hygiene. We need to restore public understanding of hygiene as a means to prevent infectious disease.


Nanostructural adsorption of vanadium oxide on functionalized graphene: a DFT study.
Rutile-monoclinic phase transitions of vanadium oxide (VO2) nanocrystals adsorbed on graphene-based substrates are of current scientific interest, although their adsorption and growth mechanisms have not been investigated theoretically. In this study, we use density functional theory (DFT) calculations for determining the binding energies and predicting the corresponding directions of growth of VO2 nanostructures (rutile and M1-monoclinic) interacting with both pure graphene and functionalized graphene nanoribbons. Several adsorption sites of pure graphene including the top, bridge, and hollow sites are considered, while additional adsorption sites of functionalized graphene nanoribbons, epoxy, alcohol and carboxylate are investigated. Vanadium oxide nanostructures are found to favor physical adsorption on the hollow sites of pure graphene, while chemical adsorption is favored on the carboxylate sites of functionalized graphene nanoribbons (FGNRs). Charge density maps showed the electron distribution originating from the interaction between VO2 and graphene substrates, helping to understand the mechanism of charge transfer. Electronic local potentials showed vertical growth tendencies for rutile VO2, while M1-monoclinic VO2 showed horizontal growth tendencies. Partial density of states (PDOS) helped examine the electronic structure of metallic rutile VO2 binding to hollow and carboxylate sites of functionalized graphene. These results provide an improved understanding of the controlled and oriented growth of VO2 nanocrystals on graphene-based substrates which can enable various properties such as the metal-insulator transition (MIT) of VO2 in light regulation applications.


Wildland Fires Worsened Population Exposure to PM2.5 Pollution in the Contiguous United States.
As wildland fires become more frequent and intense, fire smoke has significantly worsened the ambient air quality, posing greater health risks. To better understand the impact of wildfire smoke on air quality, we developed a modeling system to estimate daily PM2.5 concentrations attributed to both fire smoke and nonsmoke sources across the contiguous U.S. We found that wildfire smoke has the most significant impact on air quality in the West Coast, followed by the Southeastern U.S. Between 2007 and 2018, fire smoke contributed over 25% of daily PM2.5 concentrations at ∼40% of all regulatory air monitors in the EPA's air quality system (AQS) for more than one month per year. People residing outside the vicinity of an EPA AQS monitor (defined by a 5 km radius) were subject to 36% more smoke impact days compared with those residing nearby. Lowering the national ambient air quality standard (NAAQS) for annual mean PM2.5 concentrations to between 9 and 10 μg/m3 would result in approximately 35-49% of the AQS monitors falling in nonattainment areas, taking into account the impact of fire smoke. If fire smoke contribution is excluded, this percentage would be reduced by 6 and 9%, demonstrating the significant negative impact of wildland fires on air quality.


[Factors determining patients' choice of hospital].
To identify the best way to inform patients about centers and services so that they can make a responsible choice, and to identify which factors determine patients' decisions. Data on three illnesses (breast cancer, benign prostatic hypertrophy, and cataracts) in three fictitious hospitals were shown to patients. The data were presented to 1,259 subjects through four different means: a brochure including a table, a brochure with a bar graph, three brochures with tables, and three brochures with bar graphs. Data on each of the hospitals included: professional profiles, medical and surgical outcomes, side effects, patients' and relatives' satisfaction, and hospital complaints. The subjects completed a questionnaire evaluating the information received. Most patients (88%) preferred to receive full information and to share decision-making with their physician. Sixty-four percent of the patients chose the hospital with the best medical or surgical outcomes. Patients preferred the data presented through a brochure with a table (p < 0.001). Three factors were identified with severe illness (satisfaction, outcomes and complaints) and two factors with non-severe illness (satisfaction and outcomes). Men paid more attention to medical and surgical outcomes (p < 0.001), relapses (p < 0.001), and side effects (p < 0.001). When performing comparisons, health professionals paid greater attention to medical and surgical outcomes than to patients' and relatives' satisfaction (t range between 0.40 and 0.90, p < 0.01). Differences were found in the need for information and in comprehension depending on the way information was presented and on the type of disease. Gender and health knowledge seem to be predictors of patients' information needs when choosing a hospital.


Accuracy of urea breath test in Helicobacter pylori infection: meta-analysis.
To quantitatively summarize and appraise the available evidence of urea breath test (UBT) use to diagnose Helicobacter pylori (H. pylori) infection in patients with dyspepsia and provide pooled diagnostic accuracy measures. We searched MEDLINE, EMBASE, Cochrane library and other databases for studies addressing the value of UBT in the diagnosis of H. pylori infection. We included cross-sectional studies that evaluated the diagnostic accuracy of UBT in adult patients with dyspeptic symptoms. Risk of bias was assessed using QUADAS (Quality Assessment of Diagnostic Accuracy Studies)-2 tool. Diagnostic accuracy measures were pooled using the random-effects model. Subgroup analysis was conducted by UBT type (13C vs 14C) and by measurement technique (Infrared spectrometry vs Isotope Ratio Mass Spectrometry). Out of 1380 studies identified, only 23 met the eligibility criteria. Fourteen studies (61%) evaluated 13C UBT and 9 studies (39%) evaluated 14C UBT. There was significant variation in the type of reference standard tests used across studies.Pooled sensitivity was 0.96 (95%CI: 0.95-0.97) andpooled specificity was 0.93 (95%CI: 0.91-0.94). Likelihood ratio for a positive test was 12 and for a negative test was 0.05 with an area under thecurve of 0.985. Meta-analyses were associated with a significant statistical heterogeneity that remained unexplained after subgroup analysis. The included studies had a moderate risk of bias. UBT has high diagnostic accuracy for detecting H. pylori infection in patients with dyspepsia. The reliability of diagnostic meta-analytic estimates however is limited by significant heterogeneity.


The kinesin family member BimC contains a second microtubule binding region attached to the N terminus of the motor domain.
The kinesin family member BimC has a highly positively charged domain of approximately 70 amino acids at the N terminus of the motor domain. Motor domain constructs of BimC were prepared with and without this extra domain to determine its influence. The level of microtubules needed for half saturation of the ATPase of BimC motor domain constructs is reduced by approximately 7000-fold at low ionic strength upon addition of this extra N-terminal extension. Although the change in microtubule affinity is less at higher salt, addition of the N-terminal domain still produces a 20-fold increase in affinity for microtubules in 200 mm potassium acetate. A fusion protein of the N-terminal domain and thioredoxin binds tightly to MTs at low salt, consistent with the increased affinity of motor domain constructs (which contain the N-terminal domain) being due to the additional binding of the N-terminal domain to the microtubule. Hydrodynamic analysis indicates that the N-terminal extension is in a highly extended conformation, suggesting that it may be intrinsically disordered. Fusion of the N-terminal extension of BimC onto the motor domain of conventional kinesin produces a similar large increase in microtubule affinity without significant reduction in kcat or velocity in an in vitro motility assay, suggesting that the N-terminal extension can act in a modular manner to increase the microtubule affinity of kinesin motor domains without a decrease in velocity.


Biochemical markers of bone turnover in Paget's disease.
The two most commonly used biochemical markers of bone turnover are the serum alkaline phosphatase and the urinary excretion of peptide-bound hydroxyproline, both of which are increased in Paget's disease. Serum alkaline phosphatase is assumed to be derived from osteoblasts during the process of bone formation, whereas small peptides containing hydroxyproline are excreted in the urine following the degradation of bone collagen. The alkaline phosphatase is probably the more useful measurement for diagnosis and for following response to treatment, whereas hydroxyproline, although very sensitive, presents technical difficulties in collection and measurement. Several other biochemical changes in Paget's disease indicate abnormal bone metabolism. These include increased urinary excretion of hydroxylysine and its glycosides derived from collagen, as well as the release into the circulation and subsequent urinary excretion of fragments of pro-collagen indicative of increased collagen formation. Proteins specific to bone, such as osteocalcin, are increased in serum, bone, such as osteocalcin, are increased in serum, as are various enzymes possibly derived from bone cells, including acid phosphatase and proline imino-peptidase. Treatment of Paget's disease results in a fall in urinary hydroxyproline before alkaline phosphatase. This indicates that drug treatment, whether with diphosphonates, calcitonin or mithramycin, has a primary action to inhibit bone resorption, with a subsequent adaptive reduction in bone formation rate.


Designing payment for Collaborative Care for Depression in primary care.
To design a bundled case rate for Collaborative Care for Depression (CCD) that aligns incentives with evidence-based depression care in primary care. A clinical information system used by all care managers in a randomized controlled trial of CCD for older primary care patients. We conducted an empirical investigation of factors accounting for variation in CCD resource use over time and across patients. CCD resource use at the patient-episode and patient-month levels was measured by number of care manager contacts and direct patient contact time and analyzed with count data (Poisson or negative binomial) models. Episode-level resource use varies substantially with patient's time in the program. Monthly use declines sharply in the first 6 months regardless of treatment response or remission status, but it remains stable afterwards. An adjusted episode or monthly case rate design better matches payment with variation in resource use compared with a fixed design. Our findings lend support to an episode payment adjusted by number of months receiving CCD and a monthly payment adjusted by the ordinal month. Nonpayment tools including program certification and performance evaluation and reward systems are needed to fully align incentives.


Contact Mixing Patterns and Population Movement among Migrant Workers in an Urban Setting in Thailand.
Data relating to contact mixing patterns among humans are essential for the accurate modeling of infectious disease transmission dynamics. Here, we describe contact mixing patterns among migrant workers in urban settings in Thailand, based on a survey of 369 migrant workers of three nationalities. Respondents recorded their demographic data, including age, sex, nationality, workplace, income, and education. Each respondent chose a single day to record their contacts; this resulted in a total of more than 8300 contacts. The characteristics of contacts were recorded, including their age, sex, nationality, location of contact, and occurrence of physical contact. More than 75% of all contacts occurred among migrants aged 15 to 39 years. The contacts were highly clustered in this age group among migrant workers of all three nationalities. There were far fewer contacts between migrant workers with younger and older age groups. The pattern varied slightly among different nationalities, which was mostly dependent upon the types of jobs taken. Half of migrant workers always returned to their home country at most once a year and on a seasonal basis. The present study has helped us gain a better understanding of contact mixing patterns among migrant workers in urban settings. This information is useful both when simulating disease epidemics and for guiding optimal disease control strategies among this vulnerable section of the population.


Evaluation of the effects of penicillin G potassium and potassium chloride on the motility of the large intestine in horses.
To evaluate effects of IV administration of penicillin G potassium (KPEN) or potassium chloride (KCl) on defecation and myoelectric activity of the cecum and pelvic flexure of horses. 5 healthy horses. Horses with 12 bipolar electrodes on the cecum and pelvic flexure received KPEN or KCl solution by IV bolus 4 hours apart. Each horse received the following: 2 X 10(7) U of KPEN (high-dose KPEN) followed by 34 mEq of KCl (high-dose KCl), 1 X 10(7) U of KPEN (low-dose KPEN) followed by 17 mEq of KCl (low-dose KCl), high-dose KCl followed by high-dose KPEN, and low-dose KCl followed by low-dose KPEN. Number of defecations and myoelectric activity were recorded for 60 minutes. The first three 5-minute segments and first four 15-minute segments of myoelectric activity were analyzed. Number of defecations during the first 15-minute segment was greater after high-dose KPEN treatment than after high-dose or low-dose KCl treatment. Compared with reference indexes, myoelectric activity was greater in the pelvic flexure for the first 5-minute segment after high-dose KCl treatment, in the cecum and pelvic flexure for the first 5-minute segment and in the pelvic flexure for the first 15-minute segment after low-dose KPEN treatment, and in the pelvic flexure for the first and second 5-minute segments and the first three 15-minute segments after high-dose KPEN treatment. IV administration of KPEN stimulates defecation and myoelectric activity of the cecum and pelvic flexure in horses. Effects of KPEN may be beneficial during episodes of ileus.


Epidemiology and prevention of osteoporosis in urbanized Asian populations.
The incidence rate of hip fracture in Hong Kong in 1989 was calculated from hospital admission data and the census population. The incidence rate has increased 3-fold since 1966 to reach 9 per 1000 in men and 13 per 1000 in women 80 years and over. The age-adjusted rate was comparable to that in Singapore but higher than that in Japan. A case-control study was conducted on 280 women and 120 men with hip fracture and 800 controls. The relative risk for hip fracture increased with a low calcium intake, being 2.9 (95% confidence interval 1.9 to 4.6) in the lowest quartile. The relative risk for hip fracture was also significantly increased in subjects who did not perform load-bearing activity every day. Osteoporosis may be prevented by an adequate calcium intake and load-bearing activity in urbanized Asian populations.


Cyclic AMP-dependent protein kinase opens chloride channels in normal but not cystic fibrosis airway epithelium.
Chloride (Cl-) secretion by the airway epithelium regulates, in part, the quantity and composition of the respiratory tract fluid, thereby facilitating mucociliary clearance. The rate of Cl- secretion is controlled by apical membrane Cl- channels. Apical Cl- channels are opened and Cl- secretion is stimulated by a variety of hormones and neurotransmitters that increase intracellular levels of cyclic AMP (cAMP). In cystic fibrosis (CF), a common lethal genetic disease of Caucasians, airway, sweat-gland duct, secretory-coil and possibly other epithelia are anion impermeable. This abnormality may explain several of the clinical manifestations of the disease. The Cl- impermeability in CF-airway epithelia has been localized to the apical cell membrane, where regulation of Cl- channels is abnormal: hormonal secretagogues stimulate cAMP accumulation appropriately but Cl- channels fail to open. Here we report that the purified catalytic subunit of cAMP-dependent protein kinase plus ATP opens Cl- channels in excised, cell-free patches of membrane from normal cells, but fails to open Cl- channels in CF cells. These results indicate that in normal cells, the cAMP-dependent protein kinase phosphorylates the Cl- channel or an associated regulatory protein, causing the channel to open. The failure of CF Cl- channels to open suggests a defect either in the channel or in such an associated regulatory protein.


Origins of neurogenesis, a cnidarian view.
New perspectives on the origin of neurogenesis emerged with the identification of genes encoding post-synaptic proteins as well as many "neurogenic" regulators as the NK, Six, Pax, bHLH proteins in the Demosponge genome, a species that might differentiate sensory cells but no neurons. However, poriferans seem to miss some key regulators of the neurogenic circuitry as the Hox/paraHox and Otx-like gene families. Moreover as a general feature, many gene families encoding evolutionarily-conserved signaling proteins and transcription factors were submitted to a wave of gene duplication in the last common eumetazoan ancestor, after Porifera divergence. In contrast gene duplications in the last common bilaterian ancestor, Urbilateria, are limited, except for the bHLH Atonal-class. Hence Cnidaria share with Bilateria a large number of genetic tools. The expression and functional analyses currently available suggest a neurogenic function for numerous orthologs in developing or adult cnidarians where neurogenesis takes place continuously. As an example, in the Hydra polyp, the Clytia medusa and the Acropora coral, the Gsx/cnox2/Anthox-2 ParaHox gene likely supports neurogenesis. Also neurons and nematocytes (mechanosensory cells) share in hydrozoans a common stem cell and several regulatory genes indicating that they can be considered as sister cells. Performed in anthozoan and medusozoan species, these studies should tell us more about the way(s) evolution hazards achieved the transition from epithelial to neuronal cell fate, and about the robustness of the genetic circuitry that allowed neuromuscular transmission to arise and be maintained across evolution.


Current scientific understanding of the environmental biosafety of transgenic fish and shellfish.
A fluorescent zebrafish was the first genetically engineered animal to be marketed, and biotechnologists are developing many transgenic fish and shellfish. Biosafety science is not sufficiently advanced to be able to draw scientifically reliable and broadly trusted conclusions about the environmental effects of these animals. The science is best developed for identifying hazards posed by environmental spread of a transgenic fish or shellfish and least developed for assessing potential ecological harms of spread. Environmental spread of certain transgenic fish or shellfish could be an indirect route of entry into the human food supply. The management of predicted environmental risks is in its infancy and has thus far focused on the first step of the risk management process, i.e. risk reduction, via a few confinement methods. There is a critical need to improve scientific methods of environmental safety assessment and management and to gather empirical data needed to substantiate biosafety conclusions and to effectively manage transgenic fish and shellfish. Scientists and potentially affected parties should participate in prioritising the knowledge gaps to be addressed.


Genetic measurement of theory of epistatic effects.
Epistasis is defined as the influence of the genotype at one locus on the effect of a mutation at another locus. As such it plays a crucial role in a variety of evolutionary phenomena such as speciation, population bottle necks, and the evolution of genetic architecture (i.e., the evolution of dominance, canalization, and genetic correlations). In mathematical population genetics, however, epistasis is often represented as a mere noise term in an additive model of gene effects. In this paper it is argued that epistasis needs to be scaled in a way that is more directly related to the mechanisms of evolutionary change. A review of general measurement theory shows that the scaling of a quantitative concept has to reflect the empirical relationships among the objects. To apply these ideas to epistatic mutation effects, it is proposed to scale A x A epistatic effects as the change in the magnitude of the additive effect of a mutation at one locus due to a mutation at a second locus. It is shown that the absolute change in the additive effect at locus A due to a substitution at locus B is always identical to the absolute change in B due to the substitution at A. The absolute A x A epistatic effects of A on B and of B on A are identical, even if the relative effects can be different. The proposed scaling of A x A epistasis leads to particularly simple equations for the decomposition of genotypic variance. The Kacser Burns model of metabolic flux is analyzed for the presence of epistatic effects on flux. It is shown that the non-linearity of the Kacser Burns model is not sufficient to cause A x A epistasis among the genes coding for enzymes. It is concluded that non-linearity of the genotype-phenotype map is not sufficient to cause epistasis. Finally, it is shown that there exist correlations among the additive and epistatic effects among pairs of loci, caused by the inherent symmetries of Mendelian genetic systems. For instance, it is shown that a mutation that has a larger than average additive effect will tend to decrease the additive effect of a second mutation, i.e., it will tend to have a negative (canalizing) interaction with a subsequent gene substitution. This is confirmed in a preliminary analysis of QTL-data for adult body weight in mice.


A Comparison of Personality, Life Events, Comorbidity, and Health in Monozygotic Twins Discordant for Anorexia Nervosa.
Genetic and environmental factors contribute to the etiology of anorexia nervosa (AN). The co-twin control design is one of the most powerful methods available to evaluate environmental factors that could contribute to differences between monozygotic (MZ) twins who are discordant for AN. Using available data from a unique and rare sample of 22 Swedish female MZ pairs discordant for AN, we compared personality, life events, comorbidity, and health factors. Twins with AN had significantly higher perfectionism scores than unaffected co-twins and reported younger ages at first diet than unaffected co-twins who had dieted. Consistent with previous literature, more twins with AN reported gastrointestinal problems than unaffected co-twins. Although not significant due to low statistical power, more unaffected co-twins reported experiencing emotional neglect than twins with AN. Early dieting may be a harbinger of the development of AN or an early symptom. Higher perfectionism may represent a risk factor, sequela, or both. Sibling perception of neglect is noteworthy given the impact of an ill child with AN on family function and wellbeing. The health and wellbeing of siblings should be addressed clinically when one child in the family suffers from AN.


Structure of protein geranylgeranyltransferase-I from the human pathogen Candida albicans complexed with a lipid substrate.
Protein geranylgeranyltransferase-I (GGTase-I) catalyzes the transfer of a 20-carbon isoprenoid lipid to the sulfur of a cysteine residue located near the C terminus of numerous cellular proteins, including members of the Rho superfamily of small GTPases and other essential signal transduction proteins. In humans, GGTase-I and the homologous protein farnesyltransferase (FTase) are targets of anticancer therapeutics because of the role small GTPases play in oncogenesis. Protein prenyltransferases are also essential for many fungal and protozoan pathogens that infect humans, and have therefore become important targets for treating infectious diseases. Candida albicans, a causative agent of systemic fungal infections in immunocompromised individuals, is one pathogen for which protein prenylation is essential for survival. Here we present the crystal structure of GGTase-I from C. albicans (CaGGTase-I) in complex with its cognate lipid substrate, geranylgeranylpyrophosphate. This structure provides a high-resolution picture of a non-mammalian protein prenyltransferase. There are significant variations between species in critical areas of the active site, including the isoprenoid-binding pocket, as well as the putative product exit groove. These differences indicate the regions where specific protein prenyltransferase inhibitors with antifungal activity can be designed.


Community-based restaurant interventions to promote healthy eating: a systematic review.
Eating in restaurants is associated with high caloric intake. This review summarizes and evaluates the evidence supporting community-based restaurant interventions. We searched all years of PubMed and Web of Knowledge through January 2014 for original articles describing or evaluating community-based restaurant interventions to promote healthy eating. We extracted summary information and classified the interventions into 9 categories according to the strategies implemented. A scoring system was adapted to evaluate the evidence, assigning 0 to 3 points to each intervention for study design, public awareness, and effectiveness. The average values were summed and then multiplied by 1 to 3 points, according to the volume of research available for each category. These summary scores were used to determine the level of evidence (insufficient, sufficient, or strong) supporting the effectiveness of each category. This review included 27 interventions described in 25 studies published since 1979. Most interventions took place in exclusively urban areas of the United States, either in the West or the South. The most common intervention categories were the use of point-of-purchase information with promotion and communication (n = 6), and point-of-purchase information with increased availability of healthy choices (n = 6). Only the latter category had sufficient evidence. The remaining 8 categories had insufficient evidence because of interventions showing no, minimal, or mixed findings; limited reporting of awareness and effectiveness; low volume of research; or weak study designs. No intervention reported an average negative impact on outcomes. Evidence about effective community-based strategies to promote healthy eating in restaurants is limited, especially for interventions in rural areas. To expand the evidence base, more studies should be conducted using robust study designs, standardized evaluation methods, and measures of sales, behavior, and health outcomes.


Understanding of and attitudes toward epilepsy among the urban Chinese population in Malaysia.
People with epilepsy are socially discriminated against on the grounds of widespread negative public attitudes, misunderstandings and defensive behaviour. The primary purpose of this study was to evaluate the public understanding of and attitudes toward epilepsy among the Chinese population in Malaysia. A validated, self-administered questionnaire comprising 23 questions was utilised to evaluate the understanding of and attitudes toward epilepsy among randomly approached respondents from the Chinese population living in the urban areas of Penang, Klang Valley, Kuala Lumpur and Sibu in Malaysia. Out of 1,000 people approached, 697 (69.7 percent) respondents agreed to participate in the study. When asked whether people with epilepsy are slow learners and have intellectual functioning below normal, 448 (64.3 percent) respondents answered 'no'. This positive answer was mainly provided by female (35.6 percent) as compared to male (28.6 percent) respondents. Moreover, more than half responded positively to the following statements: people with epilepsy should not be isolated from the normal population; epileptics can perform daily activities; epileptics can receive academic education; and epileptics can become useful members of society. In addition, significant associations were discovered between the education level of the respondents and several statements, including whether epileptics are as intelligent as everyone else (p-value is 0.009), whether epilepsy can be treated with drugs (p-value is 0.037) and whether epileptics can be as successful as other people in their chosen career (p-value is 0.009). Positive responses were mainly acquired from those with secondary education and above. A large number of the respondents felt that people with epilepsy should not be employed as lorry drivers, firefighters, doctors and army personnel. The general Chinese population in the urban areas of Malaysia had, at the time of the investigation, a relatively high level of understanding and positive attitudes toward certain aspects of epilepsy, although a minority of the study population demonstrated prejudice and discriminatory behaviours toward people with epilepsy.


Detection of resistance mutations and CD4 slopes in individuals experiencing sustained virological failure.
Several resistance mutations have been shown to affect viral fitness, and the presence of certain mutations might result in clinical benefit for patients kept on a virologically failing regimen due to an exhaustion of drug options. We sought to quantify the effect of resistance mutations on CD4 slopes in patients undergoing episodes of viral failure. Patients from the EuroSIDA and UK CHIC cohorts undergoing at least one episode of virological failure (>3 consecutive RNA measurements >500 on ART) with at least three CD4 measurements and a resistance test during the episode were included. Mutations were identified using the IAS-US (2013) list, and were presumed to be present from detection until the end of an episode. Multivariable linear mixed models with a random intercept and slope adjusted for age, baseline CD4 count, hepatitis C, drug type, RNA (log-scale), risk group and subtype were used to estimate CD4 slopes. Individual mutations with a population prevalence of >10% were tested for their effect on the CD4 slope. A total of 2731 patients experiencing a median of 1 (range 1-4) episodes were included in this analysis. The prevalence of any resistance per episode was 88.4%; NNRTI resistance was most common (78.5%). Overall, CD4 counts declined by 17.1 (-19.7; -14.5) cells per year; this decline was less marked with partial viral suppression (current HIV RNA more than 1.5 log below the setpoint; p=0.01). In multivariable models adjusting for viral load, CD4 decline was slower during episodes with detected resistance compared to episodes without detected resistance (21.0 cells/year less, 95% CI 11.75-30.31, p<0.001). Among those with more than one resistance mutation, there was only weak evidence that class-specific mutations had any effect on the CD4 slope (Table 1). The effects of individual mutations (incl. M184V) were explored, but none were significantly associated with the CD4 slope; for these comparisons, a Bonferroni-corrected p-value level was 0.003. In our study population, detected resistance was associated with slightly less steep CD4 declines. This may be due to a biological effect of resistance on CD4 slopes, or other unmeasured factors such as poor adherence among individuals without resistance. Among individuals with detected drug resistance, we found no evidence suggesting that the presence of individual mutations was associated with beneficial CD4 slope changes.


No difference in survivorship after unicompartmental knee arthroplasty with or without an intact anterior cruciate ligament.
Anterior cruciate ligament deficiency (ACLD) has been considered a contraindication for Oxford unicompartmental knee arthroplasty (UKA) because of the reported higher incidence of failure when implanted in ACLD knees. However, given the potential advantages of UKA over total knee arthroplasty (TKA), we have performed UKA in a limited number of patients with ACL deficiency and end-stage medial compartment osteoarthritis (OA) over the past 11 years. The primary aim of this study was to establish the clinical outcome of this cohort; the secondary aim was to compare both clinical and radiographic data with a matched cohort of ACL-intact (ACLI) patients who have undergone UKA for anteromedial OA. This retrospective observational study describes the clinical and radiological outcome in 46 medial Oxford UKAs implanted in 42 consecutive patients with ACL deficiency and concomitant symptomatic medial compartment OA at mean follow-up of 5 years. It also compares the outcomes with a matched cohort of UKA patients with an intact ACL (ACLI group). At the time of last follow-up, there was no significant difference in clinical results or survivorship between the two groups in this study. The successful short-term results of the ACLD group suggest ACL deficiency may not always be a contraindication to Oxford UKA as previously thought. Until long-term data is available, however, we maintain our recommendation that ACLD be considered a contraindication.


[Acute hepatitis in heroin addicts: etiologic and follow-up study of 110 cases].
In order to know the etiology and natural history of acute viral hepatitis in intravenous drugs abusers, a panel of 110 heroin addicts, suffering acute hepatitis, is studied. The B virus, by itself or in conjunction with the delta virus was responsible for 63 cases. 35 patients suffered of NANB hepatitis and only two patients were diagnosed of hepatitis A. Delta infection was detected in 38 of the 77 cases of HBsAg-positive acute hepatitis. All the hepatitis B patients had serum conversion within the first 6 months of the acute episode. All the patients suffering acute delta hepatitis and 75% of those suffering non-A, non-B hepatitis, developed chronic hepatic disease. We conclude that in our environment, the B virus is the main responsible for acute hepatitis in heroin addicts. Delta infection is detected in almost half of the heroin addicts with HBsAg-positive acute hepatitis. The acute hepatitis caused by B virus, or by B and delta viruses simultaneously, have a satisfactory evolution, thus, almost all the cases are cured within 6 months. On the contrary, hepatitis caused by NANB or delta viruses usually progresses to chronic hepatic disease.


Single- or two-stage revision for infected total hip arthroplasty? A systematic review of the literature.
The best approach for surgical treatment of an infected THA remains controversial. Two-stage revision is believed to result in lower reinfection rates but may result in significant functional impairment. Some authors now suggest that single-stage revision may provide comparable results in terms of infection eradication while providing superior functional outcomes. We performed a systematic review to determine whether single- or two-stage revision for an infected THA provides lower reinfection rates and higher functional outcome scores. We conducted a comprehensive search of PubMed and Embase, using the search string [Infection AND ("total hip replacement" OR "total hip arthroplasty") AND revision]. All studies comparing reinfection rates or functional scores for single- and two-stage revision were retrieved and reviewed. A systematic review was performed according to the PRISMA checklist. The initial search retrieved 1128 studies. Following strict exclusion criteria, we identified nine comparative studies comparing reinfection rates (all nine studies) or functional scores (four studies) between single- and two-stage revisions. The overall quality of studies was poor with no randomized studies being identified. Groups often varied in their baseline characteristics. There was no consensus among the studies regarding the relative incidence of reinfection between the two procedures. There was a trend toward better functional outcomes in single-stage surgery, but this reached significance in only one study. In appropriate patients, single-stage revision appears to be associated with similar reinfection rates when compared with two-stage revision with superior functional outcomes. This concurs with earlier studies, but given the methodologic quality of the included studies, these findings should be treated with caution. High-quality randomized studies are needed to compare the two approaches to confirm these findings, and, if appropriate, to determine which patients are appropriate for single-stage revision.


Navigating breast cancer: axon guidance molecules as breast cancer tumor suppressors and oncogenes.
Slit, Netrin, Ephrin, and Semaphorin's roles in development have expanded greatly in the past decade from their original characterization as axon guidance molecules (AGMs) to include roles as regulators of tissue morphogenesis and development in diverse organs. In the mammary gland, AGMs are important for maintaining normal cell proliferation and adhesion during development. The frequent dysregulation of AGM expression during tumorigenesis and tumor progression suggests that AGMs also play a crucial role as tumor suppressors and oncogenes in breast cancer. Moreover, these findings suggest that AGMs may be excellent targets for new breast cancer prognostic tests and more effective therapeutic strategies.


Soil Organic Matter in Its Native State: Unravelling the Most Complex Biomaterial on Earth.
Since the isolation of soil organic matter in 1786, tens of thousands of publications have searched for its structure. Nuclear magnetic resonance (NMR) spectroscopy has played a critical role in defining soil organic matter but traditional approaches remove key information such as the distribution of components at the soil-water interface and conformational information. Here a novel form of NMR with capabilities to study all physical phases termed Comprehensive Multiphase NMR, is applied to analyze soil in its natural swollen-state. The key structural components in soil organic matter are identified to be largely composed of macromolecular inputs from degrading biomass. Polar lipid heads and carbohydrates dominate the soil-water interface while lignin and microbes are arranged in a more hydrophobic interior. Lignin domains cannot be penetrated by aqueous solvents even at extreme pH indicating they are the most hydrophobic environment in soil and are ideal for sequestering hydrophobic contaminants. Here, for the first time, a complete range of physical states of a whole soil can be studied. This provides a more detailed understanding of soil organic matter at the molecular level itself key to develop the most efficient soil remediation and agricultural techniques, and better predict carbon sequestration and climate change.


Cocoa procyanidins attenuate 4-hydroxynonenal-induced apoptosis of PC12 cells by directly inhibiting mitogen-activated protein kinase kinase 4 activity.
Neurodegenerative disorders such as Alzheimer's disease (AD) are associated with oxidative stress, and it has been suggested that apoptosis is a crucial pathway in neuronal cell death in AD patients. 4-Hydroxynonenal (HNE), one of the aldehydic products of membrane lipid peroxidation, is reported to be elevated in the brains of AD patients and mediates the induction of neuronal apoptosis in the presence of oxidative stress. In this study, we investigated the HNE-induced apoptosis mechanism and the protective effects of the cocoa procyanidin fraction (CPF) and its major antioxidant procyanidin B2 against the apoptosis induced by HNE in rat pheochromocytoma (PC12) cells. HNE-induced nuclear condensation and increased sub-G1 fraction, both of which are markers of apoptotic cell death, were inhibited by CPF and procyanidin B2. Intracellular reactive oxygen species (ROS) accumulation was attenuated by pretreatment with CPF and procyanidin B2. CPF and procyanidin B2 also prevented HNE-induced poly(ADP-ribose) polymerase cleavage, antiapoptotic protein (Bcl-2 and Bcl-X(L)) down-regulation, and caspase-3 activation. Activation of c-Jun N-terminal protein kinase (JNK) and mitogen-activated protein kinase kinase 4 (MKK4) was attenuated by CPF and procyanidin B2. Moreover, CPF and procyanidin B2 bound directly to MKK4 and inhibited its activity. Data obtained with SP600125, a selective inhibitor of JNK, revealed that JNK is involved in HNE-induced apoptosis through the inhibition of PARP cleavage and caspase-3 activation in PC12 cells. Collectively, these results indicate that CPF and procyanidin B2 protect PC12 cells against HNE-induced apoptosis by blocking MKK4 activity as well as ROS accumulation.


Spatial and temporal visualisation techniques for crash analysis.
Understanding the underlying structure of single vehicle crashes (SVCs) is essential for improving safety on the roads. Past research has found that SVCs tend to cluster both spatially and temporally. However, limited research has been conducted to investigate the interaction between the location of SVCs and the time they occur, especially at different levels of scales or spatial extents. This paper applied spatial, temporal and spatio-temporal techniques to investigate patterns of SVCs in Western Australia between 1999 and 2008, at different levels of scale. Spider graphs were adapted to identify temporal patterns of vehicle crashes at two different levels of scales: daily and weekly with respect to their causes. The spatial structures of vehicle crashes were analysed using Kernel Density Estimation analysis at three different scales: West Australia, Metropolitan area, and Perth Local Government Area (LGA). These are illustrated using spatial zooming theory. Comap was then used to demonstrate the spatio-temporal interaction effect on vehicle crashes. The results show significant differences in spatio-temporal patterns of SVCs for various crash causes. The techniques used here have the potential to help decision makers in developing effective road safety strategies.


Detection capacity, information gaps and the design of surveillance programs for invasive forest pests.
Integrated pest risk maps and their underlying assessments provide broad guidance for establishing surveillance programs for invasive species, but they rarely account for knowledge gaps regarding the pest of interest or how these can be reduced. In this study we demonstrate how the somewhat competing notions of robustness to uncertainty and potential knowledge gains could be used in prioritizing large-scale surveillance activities. We illustrate this approach with the example of an invasive pest recently detected in North America, Sirex noctilio Fabricius. First, we formulate existing knowledge about the pest into a stochastic model and use the model to estimate the expected utility of surveillance efforts across the landscape. The expected utility accounts for the distribution, abundance and susceptibility of the host resource as well as the value of timely S. noctilio detections. Next, we make use of the info-gap decision theory framework to explore two alternative pest surveillance strategies. The first strategy aims for timely, certain detections and attempts to maximize the robustness to uncertainty about S. noctilio behavior; the second strategy aims to maximize the potential knowledge gain about the pest via unanticipated (i.e., opportune) detections. The results include a set of spatial outputs for each strategy that can be used independently to prioritize surveillance efforts. However, we demonstrate an alternative approach in which these outputs are combined via the Pareto ranking technique into a single priority map that outlines the survey regions with the best trade-offs between both surveillance strategies.


Generation of calcium oscillations in fibroblasts by positive feedback between calcium and IP3.
A wide variety of nonexcitable cells generate repetitive transient increases in cytosolic calcium ion concentration ([Ca2+]i) when stimulated with agonists that engage the phosphoinositide signalling pathway. Current theories regarding the mechanisms of oscillation disagree on whether Ca2+ inhibits or stimulates its own release from internal stores and whether inositol 1,4,5-trisphosphate (IP3) and diacylglycerol (DG) also undergo oscillations linked to the Ca2+ spikes. In this study, Ca2+ was found to stimulate its own release in REF52 fibroblasts primed by mitogens plus depolarization. However, unlike Ca2+ release in muscle and nerve cells, this amplification was insensitive to caffeine or ryanodine and required hormone receptor occupancy and functional IP3 receptors. Oscillations in [Ca2+]i were accompanied by oscillations in IP3 concentration but did not require functional protein kinase C. Therefore, the dominant feedback mechanism in this cell type appears to be Ca2+ stimulation of phospholipase C once this enzyme has been activated by hormone receptors.


Tobacco use in the Myeloproliferative neoplasms: symptom burden, patient opinions, and care.
Patients with Philadelphia-negative Myeloproliferative Neoplasms (MPN) suffer from numerous symptoms and decreased quality of life. Smoking is associated with an increased symptom burden in several malignancies. The aim of this study was to analyze the association between smoking and MPN-related symptom burden and explore MPN patients' opinions on smoking. A total of 435 patients with MPN participated in a cross-sectional internet-based survey developed by the Mayo Clinic and the Myeloproliferative Neoplasm Quality of Life Group. Patients reported their demographics, disease characteristics, tobacco use, and opinions on tobacco use. In addition, MPN-related symptoms were reported via the validated 10-item version of the Myeloproliferative Neoplasms Symptom Assessment Form. Current/former smokers reported worse fatigue (mean severity 5.6 vs. 5.0, p = 0.02) and inactivity (mean severity 4.0 vs. 3.4, p = 0.03) than never smokers. Moreover, current/former smokers more frequently experienced early satiety (68.5% vs. 58.3%, p = 0.03), inactivity (79.9% vs. 71.1%, p = 0.04), and concentration difficulties (82.1% vs. 73.1%, p = 0.04). Although not significant, a higher total symptom burden was observed for current/former smokers (mean 30.4 vs. 27.0, p = 0.07). Accordingly, overall quality of life was significantly better among never smokers than current/former smokers (mean 3.5 vs. 3.9, p = 0.03). Only 43.2% of the current/former smokers reported having discussed tobacco use with their physician, and 17.5% did not believe smoking increased the risk of thrombosis. The current study suggests that smoking may be associated with increased prevalence and severity of MPN symptoms and underscores the need to enhance patient education and address tobacco use in the care of MPN patients.


A new species and subgenus of Elseya (Testudines: Pleurodira: Chelidae) from New Guinea.
The New Guinea freshwater turtle, Elseya novaeguineae (senu lato) is a long-term, widespread resident of New Guinea and has been subject to substantial vicariance in one of the most geologically dynamic regions on earth. Thus, it should come as no surprise that the taxonomy of this turtle is poorly resolved. Elseya novaeguineae has long been recognized as a species complex, though which elements of this taxon warrant recognition as separate species has been debated. In this paper, we restrict Elseya novaeguineae to the Birds Head region of New Guinea, west of the Langguru Thrust and Fold Belt; we resurrect from synonymy Elseya schultzei for the populations north of the Central Ranges; and we describe a new species for the populations to the south of the Central Ranges. We revisit the classification of the genus Elseya throughout its range in the light of our work and recently published reports, and erect three subgenera that recognize the three major clades within this genus. Subgenus Elseya is the nominate subgenus, to which we assign Elseya dentata, E. branderhorsti and undescribed E. sp. aff. dentata [Sth Alligator]. Elseya novaeguineae, E. schultzei and our new species are assigned to a new subgenus, Hanwarachelys. Species of northern and eastern Queensland, E. albagula, E. irwini, E. lavarackorum and the fossil taxa E. uberrima and E. nadibajagu, are assigned to subgenus Pelocomastes, resurrected from synonymy.


CO2 adsorption on gas-phase Cu4-xPtx (x = 0-4) clusters: a DFT study.
Transition and noble metal clusters have proven to be critical novel materials, potentially offering major advantages over conventional catalysts in a range of value-added catalytic processess such as carbon dioxide transformation to methanol. In this work, a systematic computational study of CO2 adsorption on gas-phase Cu4-xPtx (x = 0-4) clusters is performed. An exhaustive potential energy surface exploration is initially performed using our recent density functional theory basin-hopping global optimization implementation. Ground-state and low-lying energy isomers are identified for Cu4-xPtx clusters. Secondly, a CO2 molecule adsorption process is analyzed on the ground-state Cu4-xPtx configurations, as a function of cluster composition. Our results show that the gas-phase linear CO2 molecule is deformed upon adsorption, with its bend angle varying from about 132° to 139°. Cu4-xPtx cluster geometries remain unchanged after CO2 adsorption, with the exception of Cu3Pt1 and Pt4 clusters. For these particular cases, a structural conversion between the ground-state geometry and the corresponding first isomer configurations is found to be assisted by the CO2 adsorption. For all clusters, the energy barriers between the ground-state and first isomer structures are explored. Our calculated CO2 adsorption energies are found to be larger for Pt-rich clusters, exhibiting a volcano-type plot. The overall effect of a hybrid functional including dispersion forces is also discussed.


Near-infrared spectral image analysis of pork marbling based on Gabor filter and wide line detector techniques.
Marbling is an important quality attribute of pork. Detection of pork marbling usually involves subjective scoring, which raises the efficiency costs to the processor. In this study, the ability to predict pork marbling using near-infrared (NIR) hyperspectral imaging (900-1700 nm) and the proper image processing techniques were studied. Near-infrared images were collected from pork after marbling evaluation according to current standard chart from the National Pork Producers Council. Image analysis techniques-Gabor filter, wide line detector, and spectral averaging-were applied to extract texture, line, and spectral features, respectively, from NIR images of pork. Samples were grouped into calibration and validation sets. Wavelength selection was performed on calibration set by stepwise regression procedure. Prediction models of pork marbling scores were built using multiple linear regressions based on derivatives of mean spectra and line features at key wavelengths. The results showed that the derivatives of both texture and spectral features produced good results, with correlation coefficients of validation of 0.90 and 0.86, respectively, using wavelengths of 961, 1186, and 1220 nm. The results revealed the great potential of the Gabor filter for analyzing NIR images of pork for the effective and efficient objective evaluation of pork marbling.


Review of neural rehabilitation programs for dyslexia: how can an allophonic system be changed into a phonemic one?
Neural investigations suggest that there are three possible core deficits in dyslexia: phonemic, grapho-phonemic, and graphemic. These investigations also suggest that the phonemic deficit resides in a different mode of speech perception which is based on allophonic (subphonemic) units rather than phonemic units. Here we review the results of remediation methods that tap into each of these core deficits, and examine how the methods that tap into the phonemic deficit might contribute to the remediation of allophonic perception. Remediation of grapho-phonemic deficiencies with a new computerized phonics training program (GraphoGame) might be able to surpass the limits of classical phonics training programs, particularly with regard to reading fluency. Remediation of visuo-graphemic deficiencies through exposure to enhanced letter spacing is also promising, although children with dyslexia continued to read more slowly than typical readers after this type of training. Remediation of phonemic deficiencies in dyslexia with programs based solely on phonemic awareness has a limited impact on reading. This might be due to the persistence of a covert deficit in phonemic perception. Methods based on slowed speech enhance the perception not only of phonemic features but also of allophonic features, and this is probably why they have not been found to be effective in meta-analyses. Training of phonemic perception with a perceptual fading paradigm, a method that improves precision in identification and discrimination around phonemic boundaries, has yielded promising results. However, studies with children at risk for dyslexia and dyslexic adults have found that even when behavioral data do not reflect allophonic perception, it can nevertheless be present in neural recordings. Further investigations should seek to confirm that the perceptual fading paradigm is beneficial for reading, and that it renders perception truly phonemic.


Is the learning value of workplace-based assessment being realised? A qualitative study of trainer and trainee perceptions and experiences.
Workplace-based assessments (WBAs) were originally intended to inform learning and development by structuring effective observation-based feedback. The success of this innovation has not yet been established due in part to the widely varied tools, implementation strategies and research approaches. Using a conceptual framework of experience, trajectories and reifications in workplace learning, we aimed to explore trainer and trainee experiences and perceptions of the learning value of WBAs. Trainers and trainees who had used at least one WBA in the previous year were invited to participate in semistructured interviews for this phenomenological study. We used a template analysis method to explore and compare the experiences of the two groups, using the emergent themes to develop an understanding of the impact of these experiences on perceptions of learning value. Nine trainers and eight trainees participated in the study. Common themes emerged among the two groups around issues of responsibility and engagement along with (mis)understandings of the purpose of the individual tools. Trainer-specific themes emerged related to the concurrent implementation of a new e-portfolio and perceptions of increased workload. Trainees associated WBA with a training structure support value but could not translate experiences into learning values. WBAs provide trainees with a justified reason to approach trainers for feedback. WBAs, however, are not being reified as the formative assessments originally intended. A culture change may be required to change the focus of WBA research and reconceptualise this set of tools and methods as a workplace learning practice.


Insect repellent jacket tests against biting midges (diptera: Culicoides) in Panama.
Light-weight net jackets treated with N,N-diethyl-m-toluamide (deet) were field-tested in Panama against five species of biting midges, principally Culicoides furens (Poey) and C. barbosai Wirth and Blanton. The deet-treated jacket provided 87-93% protection. Time of day as well as season appeared to influence the proportionate numbers of species collected. The mean coefficient of protection was slightly lower during morning tests when C. barbosai was most abundant and higher during evening tests when C. furens was most abundant.


Advance information and movement sequencing in Gilles de la Tourette's syndrome.
Tourette's syndrome is a chronic neurological disorder manifested by involuntary motor tics and vocalisations. Because the basal ganglia have been implicated in the pathology underlying Tourette's syndrome, the present two procedures, both involving sequential movements, sought to determine the extent to which patients with Tourette's syndrome were reliant on, and could utilise different levels of advance information. Patients with Tourette's syndrome were found to be more reliant than controls on external visual cues to execute rather than to initiate a motor programme. When there was a high level of reduction in advance information--that is, a visual pathway to be followed was extinguished well in advance of each successive movement--executions progressively slowed as the sequence was traversed. Similarly, if no advance information was provided before each move, movement execution was slower than that of controls. The movement initiation times of patients with Tourette's syndrome were, however, similar to those of controls, as were their movement execution times when advance visual information was available. It seems that patients with Tourette's syndrome, like parkinsonian patients who are known to have a basal ganglia disorder, require external sensory cues to sequence a motor programme effectively. The present study found evidence consistent with the hypothesis that patients with Tourette's syndrome, like patients with Parkinson's disease, may be dysfunctional in internal switching mechanisms. Alternatively, with limited visual guidance, patients with Tourette's syndrome, regardless of medication or depression state, may require more time to plan and programme each next submovement, and under such conditions may require external visual cues to direct attention effectively to given targets. Although the underlying pathogenesis is still speculative, it is concluded that there is much to support the notion that Tourette's syndrome may stem from abnormalities of the major pathways between the basal ganglia and the frontal lobes.


Vitamin d receptor gene variants and esophageal adenocarcinoma risk: a population-based case-control study.
Polymorphisms in the vitamin D receptor (VDR) gene may be of etiological importance in determining cancer risk. The aim of this study was to assess the association between common VDR gene polymorphisms and esophageal adenocarcinoma (EAC) risk in an all-Ireland population-based case-control study. EAC cases and frequency-matched controls by age and gender recruited between March 2002 and December 2004 throughout Ireland were included. Participants were interviewed, and a blood sample collected for DNA extraction. Twenty-seven single nucleotide polymorphisms in the VDR gene were genotyped using Sequenom or TaqMan assays while the poly(A) microsatellite was genotyped by fluorescent fragment analysis. Unconditional logistic regression was applied to assess the association between VDR polymorphisms and EAC risk. A total of 224 cases of EAC and 256 controls were involved in analyses. After adjustment for potential confounders, TT homozygotes at rs2238139 and rs2107301 had significantly reduced risks of EAC compared with CC homozygotes. In contrast, SS alleles of the poly(A) microsatellite had significantly elevated risks of EAC compared with SL/LL alleles. However, following permutation analyses to adjust for multiple comparisons, no significant associations were observed between any VDR gene polymorphism and EAC risk. VDR gene polymorphisms were not significantly associated with EAC development in this Irish population. Confirmation is required from larger studies.


Zika Virus Infection as a Cause of Congenital Brain Abnormalities and Guillain-Barré Syndrome: Systematic Review.
The World Health Organization (WHO) stated in March 2016 that there was scientific consensus that the mosquito-borne Zika virus was a cause of the neurological disorder Guillain-Barré syndrome (GBS) and of microcephaly and other congenital brain abnormalities based on rapid evidence assessments. Decisions about causality require systematic assessment to guide public health actions. The objectives of this study were to update and reassess the evidence for causality through a rapid and systematic review about links between Zika virus infection and (a) congenital brain abnormalities, including microcephaly, in the foetuses and offspring of pregnant women and (b) GBS in any population, and to describe the process and outcomes of an expert assessment of the evidence about causality. The study had three linked components. First, in February 2016, we developed a causality framework that defined questions about the relationship between Zika virus infection and each of the two clinical outcomes in ten dimensions: temporality, biological plausibility, strength of association, alternative explanations, cessation, dose-response relationship, animal experiments, analogy, specificity, and consistency. Second, we did a systematic review (protocol number CRD42016036693). We searched multiple online sources up to May 30, 2016 to find studies that directly addressed either outcome and any causality dimension, used methods to expedite study selection, data extraction, and quality assessment, and summarised evidence descriptively. Third, WHO convened a multidisciplinary panel of experts who assessed the review findings and reached consensus statements to update the WHO position on causality. We found 1,091 unique items up to May 30, 2016. For congenital brain abnormalities, including microcephaly, we included 72 items; for eight of ten causality dimensions (all except dose-response relationship and specificity), we found that more than half the relevant studies supported a causal association with Zika virus infection. For GBS, we included 36 items, of which more than half the relevant studies supported a causal association in seven of ten dimensions (all except dose-response relationship, specificity, and animal experimental evidence). Articles identified nonsystematically from May 30 to July 29, 2016 strengthened the review findings. The expert panel concluded that (a) the most likely explanation of available evidence from outbreaks of Zika virus infection and clusters of microcephaly is that Zika virus infection during pregnancy is a cause of congenital brain abnormalities including microcephaly, and (b) the most likely explanation of available evidence from outbreaks of Zika virus infection and GBS is that Zika virus infection is a trigger of GBS. The expert panel recognised that Zika virus alone may not be sufficient to cause either congenital brain abnormalities or GBS but agreed that the evidence was sufficient to recommend increased public health measures. Weaknesses are the limited assessment of the role of dengue virus and other possible cofactors, the small number of comparative epidemiological studies, and the difficulty in keeping the review up to date with the pace of publication of new research. Rapid and systematic reviews with frequent updating and open dissemination are now needed both for appraisal of the evidence about Zika virus infection and for the next public health threats that will emerge. This systematic review found sufficient evidence to say that Zika virus is a cause of congenital abnormalities and is a trigger of GBS.


Cause of hoarseness in elderly patients.
Hoarseness is a common symptom in older individuals and may reflect a wide variety of pathologic, medical, physiologic, and/or functional causes. Although vocal fold atrophy is one of the more common reported findings in the elderly, inconclusive information is known about the differential diagnosis and cause of dysphonia in older individuals. The purpose of this investigation was to review the cause of hoarseness in all patients older than 65 years and to determine any correlation with advancing age and other demographic factors. Additionally, we wanted to determine the effect vocal pathology has on objective voice measures with advancing age. The two most common causes of hoarseness found in 393 patients older than 65 years were vocal fold bowing and unilateral vocal fold paralysis, followed by benign vocal fold lesions, voice tremor, and spasmodic dysphonia. Although objective measures of vocal function were abnormal compared with reported normative data, they did not increase in severity with advancing age. Apparently, the compounding effect of age on underlying vocal pathology does not increase the severity of the vocal disturbance, at least as represented by objective voice measures. The high incidence of medical illnesses seen in this population also needs to be kept in mind because it may further affect the underlying voice disturbance. It might be interesting to compare data on the patients' perceptions of their vocal disturbance for each disorder as a function of age. It would also be helpful to know whether patients responded to treatment differentially based on age.


Dietary patterns and mortality in a Chinese population.
Limited research has examined the association between dietary patterns and mortality, especially in non-Western populations. We examined the association of dietary patterns with all-cause mortality and cause-specific mortality in the Singapore Chinese Health Study, which included a unique ethnic population with strong Western and South Asian cultural influences. We conducted a prospective data analysis of the Singapore Chinese Health Study, which included 52,584 Chinese men and women (aged 45-74 y) who were free of diabetes, cardiovascular disease (CVD), and cancer at baseline (1993-1998) and followed through 2011 with 10,029 deaths. The following 2 major dietary patterns were identified by using a principal components analysis: a vegetable-, fruit-, and soy-rich (VFS) pattern and a dim sum- and meat-rich (DSM) dietary pattern. Pattern scores for each participant were calculated and examined with all-cause and cause-specific mortality risks by using a Cox proportional hazards regression. The VFS pattern was inversely associated with all-cause mortality and each cause-specific category (CVD, cancer, and respiratory) of mortality during the follow-up period. Compared with the lowest quintile of the VFS pattern, HRs for quintiles 2-5 for all-cause mortality were 0.90, 0.79, 0.80, and 0.75, respectively (P-trend < 0.0001). The DSM pattern was positively associated with CVD mortality in the whole population (HR for fifth quintile compared with first quintile: 1.23; 95% CI: 1.07, 1.40; P-trend = 0.001). Positive associations between the DSM pattern and cancer and all-cause mortality were only present in ever-smokers. In ever-smokers, relative to the first quintile, HRs for quintiles 2-5 of the DSM pattern for all-cause mortality were 1.04, 1.04, 1.13, and 1.24, respectively (P-trend < 0.0001). Similarly, HRs for quintiles 2-5 for cancer mortality were 1.08, 1.03, 1.25, and 1.34, respectively (P-trend < 0.0001). The DSM pattern was not associated with respiratory mortality. Dietary patterns are strongly associated with mortality in Chinese Singaporeans.


A new function of BMP4: dual role for BMP4 in regulation of Sonic hedgehog expression in the mouse tooth germ.
The murine tooth development is governed by sequential and reciprocal epithelial-mesenchymal interactions. Multiple signaling molecules are expressed in the developing tooth germ and interact each other to mediate the inductive tissue interactions. Among them are Sonic hedgehog (SHH), Bone Morphogenetic Protein-2 (BMP2) and Bone Morphogenetic Protein-4 (BMP4). We have investigated the interactions between these signaling molecules during early tooth development. We found that the expression of Shh and Bmp2 is downregulated at E12.5 and E13.5 in the dental epithelium of the Msx1 mutant tooth germ where Bmp4 expression is significantly reduced in the dental mesenchyme. Inhibition of BMP4 activity by noggin resulted in repression of Shh and Bmp2 in wild-type dental epithelium. When implanted into the dental mesenchyme of Msx1 mutants, beads soaked with BMP4 protein were able to restore the expression of both Shh and Bmp2 in the Msx1 mutant epithelium. These results demonstrated that mesenchymal BMP4 represents one component of the signal acting on the epithelium to maintain Shh and Bmp2 expression. In contrast, BMP4-soaked beads repressed Shh and Bmp2 expression in the wild-type dental epithelium. TUNEL assay indicated that this suppression of gene expression by exogenous BMP4 was not the result of an increase in programmed cell death in the tooth germ. Ectopic expression of human Bmp4 to the dental mesenchyme driven by the mouse Msx1 promoter restored Shh expression in the Msx1 mutant dental epithelium but repressed Shh in the wild-type tooth germ in vivo. We further demonstrated that this regulation of Shh expression by BMP4 is conserved in the mouse developing limb bud. In addition, Shh expression was unaffected in the developing limb buds of the transgenic mice in which a constitutively active Bmpr-IB is ectopically expressed in the forelimb posterior mesenchyme and throughout the hindlimb mesenchyme, suggesting that the repression of Shh expression by BMP4 may not be mediated by BMP receptor-IB. These results provide evidence for a new function of BMP4. BMP4 can act upstream to Shh by regulating Shh expression in mouse developing tooth germ and limb bud. Taken together, our data provide insight into a new regulatory mechanism for Shh expression, and suggest that this BMP4-mediated pathway in Shh regulation may have a general implication in vertebrate organogenesis.


Signaling factor interactions with polysaccharide aggregates of bacterial biofilms.
Biofilms are surface-attached colonies of bacteria embedded in an extracellular polymeric substance (EPS). Inside the eukaryotic hosts, bacterial biofilms interact with the host cells through signaling factors (SFs). These signaling processes play important roles in the interaction between bacteria and host cells and the outcome of infections and symbiosis. However, how host immune factors diffuse through biofilms is not well understood. Here, we describe synergistic molecular dynamics and experimental approaches for studying the translocation of signaling factors through polysaccharide chain aggregates present in the extracellular matrix of bacterial biofilms. The effect of polysaccharide chain degradation on the energetics of SF-EPS interactions was examined by simulating an EPS consisting of various polysaccharide chain lengths. It is shown that the SF stabilization energy, defined as the average potential of mean force difference between the environments outside and within the matrix, increases linearly with decreasing chain length. This effect has been explained based on the changes in the polysaccharide configurations around the SF. Specifically, shorter chains are packed tightly around the SF, promoting favorable SF-EPS interactions, while longer chains are packed loosely resulting in screening of interactions with neighboring chains. We further investigated the translocation of SFs through the host cell membrane using molecular dynamics simulations. Further, simulations predict the existence of energy barriers greater than 1000 kJ mol(-1) associated with the translocation of the signaling factors necrosis factor-alpha (TNF-α) and granulocyte macrophage colony stimulating factor (GM-CSF) across the lipid bilayer. The agreement of computational and experimental findings motivates future computational studies using a more detailed description of the EPS aimed at understanding the role of the extracellular matrix on biofilm drug resistance.


Standard versus biofilm antimicrobial susceptibility testing to guide antibiotic therapy in cystic fibrosis.
The antibiotics used to treat pulmonary infections in people with cystic fibrosis are typically chosen based on the results of antimicrobial susceptibility testing performed on bacteria traditionally grown in a planktonic mode (grown in a liquid). However, there is considerable evidence to suggest that Pseudomonas aeruginosa actually grows in a biofilm (or slime layer) in the airways of people with cystic fibrosis with chronic pulmonary infections. Therefore, choosing antibiotics based on biofilm rather than conventional antimicrobial susceptibility testing could potentially improve response to treatment of Pseudomonas aeruginosa in people with cystic fibrosis. This is an update of a previously published Cochrane Review. To compare biofilm antimicrobial susceptibility testing-driven therapy to conventional antimicrobial susceptibility testing-driven therapy in the treatment of Pseudomonas aeruginosa infection in people with cystic fibrosis. We searched the Cochrane Cystic Fibrosis Trials Register, compiled from electronic database searches and handsearching of journals and conference abstract books. We also searched a registry of ongoing trials and the reference lists of relevant articles and reviews.Most recent search: 19 November 2014. Randomized controlled trials of antibiotic therapy based on biofilm antimicrobial susceptibility testing compared to antibiotic therapy based on conventional antimicrobial susceptibility testing in the treatment of Pseudomonas aeruginosa pulmonary infection in people with cystic fibrosis. Both authors independently selected trials, assessed their risk of bias and extracted data from eligible trials. Additionally, the review authors contacted the trial investigators to obtain further information. The searches identified two multicentre, randomized, double-blind controlled clinical trials eligible for inclusion in the review with a total of 78 participants; one trial was done in people who were clinically stable, the other in people experiencing pulmonary exacerbations. These trials prospectively assessed whether the use of biofilm antimicrobial susceptibility testing improved microbiological and clinical outcomes in participants with cystic fibrosis who were infected with Pseudomonas aeruginosa. The primary outcome was the change in sputum Pseudomonas aeruginosa density from the beginning to the end of antibiotic therapy.Although the intervention was shown to be safe, the data from these two trials did not provide evidence that biofilm susceptibility testing was superior to conventional susceptibility testing either in terms of microbiological or lung function outcomes. One of the trials also measured risk and time to subsequent exacerbation as well as quality of life measures and did not demonstrate any difference between groups in these outcomes. Both trials had an overall low risk of bias. The current evidence is insufficient to recommend choosing antibiotics based on biofilm antimicrobial susceptibility testing rather than conventional antimicrobial susceptibility testing in the treatment of Pseudomonas aeruginosa pulmonary infections in people with cystic fibrosis. Biofilm antimicrobial susceptibility testing may be more appropriate in the development of newer, more effective formulations of drugs which can then be tested in clinical trials.


[Non-steroidal anti-inflammatory drugs and risk of stroke in older patients: current controversies and research directions].
Recent professional statements from established bodies, i.e. American Heart Association, European Medicines Agency, and Medicines and Healthcare products Regulatory Agency, have cautioned on the use of non-steroidal anti-inflammatory drugs (NSAIDs) given the potential increase in atherothrombotic risk associated with their long-term use. However, pharmacoepidemiological studies on the association between NSAID use and the risk of stroke, one of the leading causes of death, disability, and institutionalisation in old age, have shown contrasting results. Notably, very few such studies have addressed the risk in the older population, in particular patients >75-80 years, perhaps the biggest consumer group of these drugs. This article reviews the current evidence on the association between NSAIDs and risk of stroke in the older population. It also discusses the potential clinical, demographic, and pathophysiological factors potentially accounting for the discrepancies in the results of the pharmacoepidemiological studies and provides suggestions for future research directions.


Self-reported radiation safety behaviors among veterinary specialists and residents performing fluoroscopic procedures on small animals.
To describe the radiation safety behaviors of veterinary specialists performing small animal fluoroscopic procedures and examine potential risk factors for these behaviors, including knowledge of radiation risk and training regarding machine operating parameters. 197 veterinary specialists and residents in training. An electronic questionnaire was distributed to members of the American Colleges of Veterinary Internal Medicine (subspecialties of cardiology and small animal internal medicine), Veterinary Radiology, and Veterinary Surgery. The overall survey response rate was 6% (240/4,274 email recipients). Of the 240 respondents, 197 (82%) had operated an x-ray unit for a small animal fluoroscopic procedure in the preceding year and fully completed the questionnaire. More than 95% of respondents believed that radiation causes cancer, yet approximately 60% of respondents never wore hand or eye protection during fluoroscopic procedures, and 28% never adjusted the fluoroscopy machine operating parameters for the purpose of reducing their radiation dose. The most common reasons for not wearing eye shielding included no requirement to wear eyeglasses, poor fit, discomfort, and interference of eyeglasses with task performance. Respondents who had received training regarding machine operating parameters adjusted those parameters to reduce their radiation dose during procedures significantly more frequently than did respondents who had not received training. On the basis of the self-reported suboptimal radiation safety practices among veterinary fluoroscopy users, we recommend formal incorporation of radiation safety education into residency training programs. All fluoroscopy machine operators should be trained regarding the machine operating parameters that can be adjusted to reduce occupational radiation exposure.


Conserved repeats in diverged ice nucleation structural genes from two species of Pseudomonas.
Sequence analysis shows that an ice nucleation gene (inaW) from Pseudomonas fluorescens is related to the inaZ gene of Pseudomonas syringae. The two genes have diverged by many amino acid substitutions, and have effectively randomized the third bases of homologous codons. By reference to their potential for change, it is shown that certain conserved features must have been maintained by selection pressure. In particular, their conservation of internal sequence repetition, with three orders of repeat periodicity in each gene, suggests that the pattern of repetition is significant to the gene products' function. We propose models for the structure of the gene products in which each order of periodicity would be required for the nucleation function.


Multipoint approximations of identity-by-descent probabilities for accurate linkage analysis of distantly related individuals.
We propose an analytical approximation method for the estimation of multipoint identity by descent (IBD) probabilities in pedigrees containing a moderate number of distantly related individuals. We show that in large pedigrees where cases are related through untyped ancestors only, it is possible to formulate the hidden Markov model of the Lander-Green algorithm in terms of the IBD configurations of the cases. We use a first-order Markov approximation to model the changes in this IBD-configuration variable along the chromosome. In simulated and real data sets, we demonstrate that estimates of parametric and nonparametric linkage statistics based on the first-order Markov approximation are accurate. The computation time is exponential in the number of cases instead of in the number of meioses separating the cases. We have implemented our approach in the computer program ALADIN (accurate linkage analysis of distantly related individuals). ALADIN can be applied to general pedigrees and marker types and has the ability to model marker-marker linkage disequilibrium with a clustered-markers approach. Using ALADIN is straightforward: It requires no parameters to be specified and accepts standard input files.


Choppers to optimise the repetition rate multiplication technique on a direct geometry neutron chopper spectrometer.
In recent years the use of repetition rate multiplication (RRM) on direct geometry neutron spectrometers has been established and is the common mode of operation on a growing number of instruments. However, the chopper configurations are not ideally optimised for RRM with a resultant 100 fold flux difference across a broad wavelength band. This paper presents chopper configurations that will produce a relative constant (RC) energy resolution and a relative variable (RV) energy resolution for optimised use of RRM. The RC configuration provides an almost uniform ΔE/E for all incident wavelengths and enables an efficient use of time as the entire dynamic range is probed with equivalent statistics, ideal for single shot measurements of transient phenomena. The RV energy configuration provides an almost uniform opening time at the sample for all incident wavelengths with three orders of magnitude in time resolution probed for a single European Spallation Source (ESS) period, which is ideal to probe complex relaxational behaviour. These two chopper configurations have been simulated for the Versatile Optimal Resolution direct geometry spectrometer, VOR, that will be built at ESS.


The effect of selective phosphodiesterase inhibitors on plasma insulin concentrations and insulin secretion in vitro in the rat.
We have examined in rats the effects of Org 9935 (4,5-dihydro-6-(5,6-dimethoxy-benzo[b]-thien-2-yl)-methyl-1-(2H)-p yridazinone), a selective inhibitor of type 3 phosphodiesterase (phosphodiesterase 3) and Org 30029 (N-hydroxy-5,6-dimethoxy-benzo[b]-thiophene-2-carboximidamide HCl), an inhibitor of phosphodiesterase 3/4 on rat plasma insulin and glucose concentrations in pentobarbitone-anaesthetised rats and on insulin secretion by rat isolated islets. We have also compared their effects on islet phosphodiesterase activity. Org 9935 (0.1 and 1.0 mg kg(-1) i.v. 15 min previously) dose dependently elevated fasting and post-glucose (0.25 g kg(-1) i.v.) plasma insulin concentrations. Org 30029 in a dose of 10 mg kg(-1), but not 1 mg kg(-1), also increased plasma insulin concentrations. Neither drug modified either fasting or post-glucose plasma glucose concentrations. Each drug augmented glucose-induced insulin release by rat isolated islets in a static incubation system, with approximate EC50 values of 1.5 microM for Org 9935 and 20 microM for Org 30029. Phosphodiesterase activity, in both supernatant and pellet fractions of islet homogenates, was inhibited concentration dependently by each drug. Although the shape of the concentration-inhibition curve for Org 30029 precluded estimation of an IC50 value, this drug was clearly much less potent than Org 9935 (IC50 about 50 nM) in inhibiting islet phosphodiesterase activity. We conclude that the increase in plasma insulin produced by each drug is a consequence of augmented insulin secretion, probably secondary to inhibition of phosphodiesterase 3 in the islet beta cell, with a resultant elevation in cAMP. The failure of the drugs to modify plasma glucose may be due to concomitant inhibition of cAMP phosphodiesterase in liver and adipose tissue.


Recovery of dominant, autosomal flightless mutants of Drosophila melanogaster and identification of a new gene required for normal muscle structure and function.
To identify further mutations affecting muscle function and development in Drosophila melanogaster we recovered 22 autosomal dominant flightless mutations. From these we have isolated eight viable and lethal alleles of the muscle myosin heavy chain gene, and seven viable alleles of the indirect flight muscle (IFM)-specific Act88F actin gene. The Mhc mutations display a variety of phenotypic effects, ranging from reductions in myosin heavy chain content in the indirect flight muscles only, to reductions in the levels of this protein in other muscles. The Act88F mutations range from those which produce no stable actin and have severely abnormal myofibrillar structure, to those which accumulate apparently normal levels of actin in the flight muscles but which still have abnormal myofibrils and fly very poorly. We also recovered two recessive flightless mutants on the third chromosome. The remaining five dominant flightless mutations are all lethal alleles of a gene named lethal(3)Laker. The Laker alleles have been characterized and the gene located in polytene bands 62A10,B1-62B2,4. Laker is a previously unidentified locus which is haplo-insufficient for flight. In addition, adult wild-type heterozygotes and the lethal larval trans-heterozygotes show abnormalities of muscle structure indicating that the Laker gene product is an important component of muscle.