A resolution criterion for electron tomography based on cross-validation.
Despite much progress in electron tomography, quantitative assessment of resolution has remained a problematic issue. The criteria that are used in single particle analysis, based on gauging the consistency between density maps calculated from half data sets, are not directly applicable because of the uniqueness of a tomographic volume. Here, we propose two criteria based on a cross-validation approach. One, called FSC(e/o), is based on a Fourier shell correlation comparison between tomograms calculated from the even and odd members of a tilt series. The other, called noise-compensated leave-one-out (NLOO), is based on Fourier ring correlation comparisons between an original projection and the corresponding reprojection of the tomogram calculated from all the other projections, taking into account the differing noise statistics. Plotted as a function of tilt angle, they allow assessment of the angular dependence of resolution and quality control over the series of projections. Integrated over all projections, the results give a global figure for resolution. Tests on simulated tomograms established consistency between these criteria and the FSC(ref), a correlation coefficient calculated between a known reference structure and the corresponding portion of a tomogram containing that structure. The two criteria-FSC(e/o) and NLOO-are mutually consistent when residual noise is the major resolution-limiting factor. When the size of the tilt increment becomes a significant factor, NLOO provides a more reliable criterion, as expected, although it is computationally intensive. Applicable to entire tomograms or selected structures, NLOO has also been tested on experimental tomographic data.


Screening of Bauhinia purpurea Linn. for analgesic and anti-inflammatory activities.
Ethanol extract of the stem of Bauhinia purpurea Linn. was subjected to analgesic and anti-inflammatory activities in animal models. Albino Wistar rats and mice were the experimental animals respectively. Different CNS depressant paradigms like analgesic activity (determined by Eddy's hot plate method and acetic acid writhing method) and anti-inflammatory activity determined by carrageenan induced paw edema using plethysmometer in albino rats) were carried out, following the intra-peritoneal administration of ethanol extract of Bauhinia purpurea Linn. (BP) at the dose level of 50 mg/kg and 100 mg/kg. The analgesic and anti-inflammatory activities of ethanol extracts of BP were significant (P < 0.001). The maximum analgesic effect was observed at 120 min at the dose of 100 mg/kg (i.p.) and was comparable to that of standard analgin (150 mg/kg) and the percentage of edema inhibition effect was 46.4% and 77% for 50 mg/kg and 100 mg/kg (i.p) respectively. Anti-inflammatory activity was compared with standard Diclofenac sodium (5 mg/kg). Ethanol extract of Bauhinia purpurea has shown significant analgesic and anti-inflammatory activities at the dose of 100 mg/kg and was comparable with corresponding standard drugs. The activity was attributed to the presence of phytoconstituents in the tested extract.


Relevance of carbon stocks of marine sediments for national greenhouse gas inventories of maritime nations.
Determining national carbon stocks is essential in the framework of ongoing climate change mitigation actions. Presently, assessment of carbon stocks in the context of greenhouse gas (GHG)-reporting on a nation-by-nation basis focuses on the terrestrial realm, i.e., carbon held in living plant biomass and soils, and on potential changes in these stocks in response to anthropogenic activities. However, while the ocean and underlying sediments store substantial quantities of carbon, this pool is presently not considered in the context of national inventories. The ongoing disturbances to both terrestrial and marine ecosystems as a consequence of food production, pollution, climate change and other factors, as well as alteration of linkages and C-exchange between continental and oceanic realms, highlight the need for a better understanding of the quantity and vulnerability of carbon stocks in both systems. We present a preliminary comparison of the stocks of organic carbon held in continental margin sediments within the Exclusive Economic Zone of maritime nations with those in their soils. Our study focuses on Namibia, where there is a wealth of marine sediment data, and draws comparisons with sediment data from two other countries with different characteristics, which are Pakistan and the United Kingdom. Results indicate that marine sediment carbon stocks in maritime nations can be similar in magnitude to those of soils. Therefore, if human activities in these areas are managed, carbon stocks in the oceanic realm-particularly over continental margins-could be considered as part of national GHG inventories. This study shows that marine sediment organic carbon stocks can be equal in size or exceed terrestrial carbon stocks of maritime nations. This provides motivation both for improved assessment of sedimentary carbon inventories and for reevaluation of the way that carbon stocks are assessed and valued. The latter carries potential implications for the management of human activities on coastal environments and for their GHG inventories.


Enhanced wetting of Cu on ZnO by migration of subsurface oxygen vacancies.
Metal adhesion on metal oxides is strongly controlled by the oxide surface structure and composition, but lack of control over the surface conditions often limits the possibilities to exploit this in opto- and micro-electronics applications and heterogeneous catalysis where nanostructural control is of utmost importance. The Cu/ZnO system is among the most investigated of such systems in model studies, but the presence of subsurface ZnO defects and their important role for adhesion on ZnO have been unappreciated so far. Here we reveal that the surface-directed migration of subsurface defects affects the Cu adhesion on polar ZnO(0001) in the technologically interesting temperature range up to 550 K. This leads to enhanced adhesion and ultimately complete wetting of ZnO(0001) by a Cu overlayer. On the basis of our experimental and computational results we demonstrate a mechanism which implies that defect concentrations in the bulk are an important, and possibly controllable, parameter for the metal-on-oxide growth.


Ketamine effects on anxiety and fear-related behaviors: Current literature evidence and new findings.
Ketamine, a non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist, presents a rapid and sustained antidepressant effect in clinical and preclinical studies. Regarding ketamine effects on anxiety, there is a widespread discordance among pre-clinical studies. To address this issue, the present study reviewed the literature (electronic database MEDLINE) to summarize the profile of ketamine effects in animal tests of anxiety/fear. We found that ketamine anxiety/fear-related effects may depend on the anxiety paradigm, schedule of ketamine administration and tested species. Moreover, there was no report of ketamine effects in animal tests of fear related to panic disorder (PD). Based on that finding, we evaluated if treatment with ketamine and another NMDA antagonist, MK-801, would induce acute and sustained (24 hours later) anxiolytic and/or panicolytic-like effects in animals exposed to the elevated T-maze (ETM). The ETM evaluates, in the same animal, conflict-evoked and fear behaviors, which are related, respectively, to generalized anxiety disorder and PD. Male Wistar rats were systemically treated with racemic ketamine (10, 30 and 80 mg/kg) or MK-801 (0.05 and 0.1 mg/kg) and tested in the ETM in the same day or 24 hours after their administration. Ketamine did not affect the behavioral tasks performed in the ETM acutely or 24 h later. MK-801 impaired inhibitory avoidance in the ETM only at 45 min post-injection, suggesting a rapid but not sustained anxiolytic-like effect. Altogether our results suggest that ketamine might have mixed effects in anxiety tests while it does not affect panic-related behaviors.


TERT promoter mutations in familial and sporadic melanoma.
Cutaneous melanoma occurs in both familial and sporadic forms. We investigated a melanoma-prone family through linkage analysis and high-throughput sequencing and identified a disease-segregating germline mutation in the promoter of the telomerase reverse transcriptase (TERT) gene, which encodes the catalytic subunit of telomerase. The mutation creates a new binding motif for Ets transcription factors and ternary complex factors (TCFs) near the transcription start and, in reporter gene assays, caused up to twofold increase in transcription. We then screened the TERT promoter in sporadic melanoma and observed recurrent ultraviolet signature somatic mutations in 125 of 168 (74%) of human cell lines derived from metastatic melanomas, 45 of 53 corresponding metastatic tumor tissues (85%), and 25 of 77 (33%) primary melanomas. The majority of those mutations occurred at two positions in the TERT promoter and also generated binding motifs for Ets/TCF transcription factors.


A Novel Function of the Lysophosphatidic Acid Receptor 3 (LPAR3) Gene in Zebrafish on Modulating Anxiety, Circadian Rhythm Locomotor Activity, and Short-Term Memory.
Lysophosphatidic acid (LPA) is a small lysophospholipid molecule that activates multiple cellular functions through pathways with G-protein-coupled receptors. So far, six LPA receptors (LPAR1 to LPAR6) have been discovered and each one of them can connect to the downstream cell message-transmitting network. A previous study demonstrated that LPA receptors found in blood-producing stem cells can enhance erythropoietic processes through the activation of LPAR3. In the current study, newly discovered functions of LPAR3 were identified through extensive behavioral tests in lpar3 knockout (KO) zebrafish. It was found that the adult lpar3 KO zebrafish display an abnormal movement orientation and altered exploratory behavior compared to that of the control group in the three-dimensional locomotor and novel tank tests, respectively. Furthermore, consistent with those results, in the circadian rhythm locomotor activity test, the lpar3 KO zebrafish showed a lower level of angular velocity and average speed during the light cycles, indicating an hyperactivity-like behavior. In addition, the mutant fish also exhibited considerably higher locomotor activity during the dark cycle. Supporting those findings, this phenomenon was also displayed in the lpar3 KO zebrafish larvae. Furthermore, several important behavior alterations were also observed in the adult lpar3 KO fish, including a lower degree of aggression, less interest in conspecific social interaction, and looser shoal formation. However, there was no significant difference regarding the predator avoidance behavior between the mutant and the control fish. In addition, lpar3 KO zebrafish displayed memory deficiency in the passive avoidance test. These in vivo results support for the first time that the lpar3 gene plays a novel role in modulating behaviors of anxiety, aggression, social interaction, circadian rhythm locomotor activity, and memory retention in zebrafish.


Tutorial in biostatistics: competing risks and multi-state models.
Standard survival data measure the time span from some time origin until the occurrence of one type of event. If several types of events occur, a model describing progression to each of these competing risks is needed. Multi-state models generalize competing risks models by also describing transitions to intermediate events. Methods to analyze such models have been developed over the last two decades. Fortunately, most of the analyzes can be performed within the standard statistical packages, but may require some extra effort with respect to data preparation and programming. This tutorial aims to review statistical methods for the analysis of competing risks and multi-state models. Although some conceptual issues are covered, the emphasis is on practical issues like data preparation, estimation of the effect of covariates, and estimation of cumulative incidence functions and state and transition probabilities. Examples of analysis with standard software are shown.


Balance billing: the patients' perspective.
We study the effects of 'balance billing', i.e., allowing physicians to charge a fee from patients in addition to the fee paid by Medicare. First, we show that on pure efficiency grounds the optimal Medicare fee under balance billing is zero. An active Medicare policy thus can only be justified when distributional concerns are accounted for. Extending the analysis by Glazer and McGuire, we therefore analyze the optimal policy from the patients' point of view. We demonstrate that, from the patients' perspective, a positive fee can be superior under balance billing. Furthermore, patient welfare can be lower if balance billing is prohibited. In particular, this is the case if the administrative costs of Medicare are large. However, we cannot rule out that prohibiting balance billing may be superior. Finally, we show that payer fee discrimination increases patient welfare if Medicare's administrative costs are high or if Medicare's optimal fee under balance billing implies lower quality for fee-only patients.JEL-classification: I11, I18, H51.


Steroids and risk factors for airway compromise in multilevel cervical corpectomy patients: a prospective, randomized, double-blind study.
Prospective, randomized, double-blind study. To evaluate intravenous corticosteroids in preventing delayed extubation after multilevel corpectomy and strut graft reconstructive procedures and to identify risk factors for delayed extubation in these patients. We performed a prospective, randomized double-blind study in patients undergoing multilevel cervical corpectomy procedures. Our hypothesis was that high-dose perioperative steroids would decrease edema and thus decrease the incidence of delayed extubation. We studied patients undergoing 2- or 3-level anterior cervical corpectomy procedures with anterior strut graft reconstruction. Sixty-six patients were randomized to receive 3 doses of either intravenous dexamethasone (n = 35) or saline (n = 31). The first dose was given before the incision, with subsequent doses given 8 and 16 hours later. Patients remained intubated until postoperative day 1, at which time a cuff leak test was performed by the anesthesiology attending. If a leak was present, the patient was extubated. If not, the test was repeated each postoperative day until a leak was present, indicating a patent airway. Five of 35 (14%) in the steroid group and 6 of 31 (19%) in the saline group required delayed extubation (P = 0.22). There were no statistical differences in preoperative parameters of age, gender, diagnosis, smoking history, BMI, number of operative levels, or preoperative American Society of Anesthesiologists rating between the 2 groups. Similarly there were no differences between the groups for duration of anesthesia, intraoperative colloids or crystalloids, intraoperative blood loss, or intraoperative urine output. The data for both groups were pooled to evaluate risk factors for delayed extubation. The only statistically significant risk factor for delayed extubation in this study was female gender (P = 0.0001). Based on our data, we cannot recommend intravenous dexamethasone for prevention of delayed extubation after multilevel anterior cervical corpectomy and strut grafting procedures.


Current smoking predicts increased operative mortality and morbidity after cardiac surgery in the elderly.
The effect of preoperative smoking status on the outcome of cardiac surgery remains unclear. Preoperative cessation may be associated with reduced postoperative pulmonary complications and in older patients preoperative smoking status appears to have a greater impact on outcome. This study was designed to assess the relationship between age, preoperative smoking status and outcomes from cardiac surgery. We performed a single-centre, retrospective cohort study to compare in-patient cardiac surgical mortality and morbidity in current smokers and never-smokers. We analysed the cardiac surgical population in its entirety and in age-stratified subsets using univariate and logistic regression analyses. During a five-year period ending March 2007, 10.8% of all patients (n=554) undergoing cardiac surgery were current smokers. Five hundred and fifty-four never-smokers undergoing cardiac surgery during the same period were identified. Overall, the smokers had a tendency towards higher in-patient mortality (4.3 vs. 2.3%, P=0.067) and increased rates of morbidity. Amongst over 70-year-olds, the current smokers had significantly higher rates of pulmonary complications (24.7 vs. 8.2%, P<0.0002), new renal replacement therapy (17.3 vs. 3.1%, P<0.0001) and infections (44.4 vs. 23.8%, P<0.0007). They had longer intensive care stay (6.2 vs. 2.8 days, P=0.002) with more intensive care unit readmissions (19.8 vs. 5.2%, P<0.0002) and significantly increased in-patient mortality (14.8 vs. 2.1%, P<0.0001). In the elderly smokers, mortality was significantly associated with the rate of pulmonary complications (P=0.03). Preoperative smoking status remained a predictor of pulmonary complications after logistic regression. The current data strengthen the observation that preoperative smoking status is predictive of adverse outcomes of cardiac surgery in the elderly. Further study into the effect of preoperative smoking cessation in the elderly may inform cessation counselling and the timing of surgery.


Long-Term Outcomes From Repeated Smoking Cessation Assistance in Routine Primary Care.
To test the association between repeated clinical smoking cessation support and long-term cessation. Retrospective, observational cohort study using structured and free-text data from electronic health records. Six diverse health systems in the United States. Patients aged ≥18 years who were smokers in 2007 and had ≥1 primary care visit in each of the following 4 years (N = 33 691). Primary exposure was a composite categorical variable (comprised of documentation of smoking cessation medication, counseling, or referral) classifying the proportions of visits for which patients received any cessation assistance (<25% (reference), 25%-49%, 50%-74%, and ≥75% of visits). The dependent variable was long-term quit (LTQ; yes/no), defined as no indication of being a current smoker for ≥365 days following a visit where nonsmoker or former smoker was indicated. Mixed effects logistic regression analysis adjusted for age, sex, race, and comorbidities, with robust standard error estimation to account for within site correlation. Overall, 20% of the cohort achieved LTQ status. Patients with ≥75% of visits with any assistance had almost 3 times the odds of achieving LTQ status compared to those with <25% visits with assistance (odds ratio = 2.84; 95% confidence interval: 1.50-5.37). Results were similar for specific assistance types. These findings provide support for the importance of repeated assistance at primary care visits to increase long-term smoking cessation.


An innovative ovipositor for niche exploitation impacts genital coevolution between sexes in a fruit-damaging Drosophila.
Limited attention has been given to ecological factors influencing the coevolution of male and female genitalia. The innovative ovipositor of Drosophila suzukii, an invading fruit pest, represents an appealing case to document this phenomenon. The serrated saw-like ovipositor is used to pierce the hard skin of ripening fruits that are not used by other fruit flies that prefer soft decaying fruits. Here, we highlight another function of the ovipositor related to its involvement in genital coupling during copulation. We compared the morphology and coupling of male and female genitalia in this species to its sibling species, Drosophila subpulchrella, and to an outgroup species, Drosophila biarmipes These comparisons and a surgical manipulation indicated that the shape of male genitalia in D. suzukii has had to be adjusted to ensure tight coupling, despite having to abandon the use of a hook-like structure, paramere, because of the more linearly elongated ovipositor. This phenomenon demonstrates that ecological niche exploitation can directly affect the mechanics of genital coupling and potentially cause incompatibility among divergent forms. This model case provides new insights towards elucidating the importance of the dual functions of ovipositors in other insect species that potentially induce genital coevolution and ecological speciation.


Circulating C-reactive protein concentrations and risks of colon and rectal cancer: a nested case-control study within the European Prospective Investigation into Cancer and Nutrition.
The authors investigated associations between serum C-reactive protein (CRP) concentrations and colon and rectal cancer risk in a nested case-control study within the European Prospective Investigation into Cancer and Nutrition (1992-2003) among 1,096 incident cases and 1,096 controls selected using risk-set sampling and matched on study center, age, sex, time of blood collection, fasting status, menopausal status, menstrual cycle phase, and hormone replacement therapy. In conditional logistic regression with adjustment for education, smoking, nutritional factors, body mass index, and waist circumference, CRP showed a significant nonlinear association with colon cancer risk but not rectal cancer risk. Multivariable-adjusted relative risks for CRP concentrations of > or = 3.0 mg/L versus <1.0 mg/L were 1.36 (95% confidence interval (CI): 1.00, 1.85; P-trend = 0.01) for colon cancer and 1.02 (95% CI: 0.67, 1.57; P-trend = 0.65) for rectal cancer. Colon cancer risk was significantly increased in men (relative risk = 1.74, 95% CI: 1.11, 2.73; P-trend = 0.01) but not in women (relative risk = 1.06, 95% CI: 0.67, 1.68; P-trend = 0.13). Additional adjustment for C-peptide, glycated hemoglobin, and high density lipoprotein cholesterol did not attenuate these results. These data provide evidence that elevated CRP concentrations are related to a higher risk of colon cancer but not rectal cancer, predominantly among men and independently of obesity, insulin resistance, and dyslipidemia.


Deladenus posteroporus n. sp. (Nematoda: Neotylenchidae) Isolated from Packaging Wood from Canada and White Pine (Pinus monticola) Lumber from the United States and Intercepted in Ningbo, China.
Deladenus posteroporus n. sp. isolated from packaging wood originated from Canada and from white pine lumber from the United States, both intercepted in the port of Ningbo, China, is described and illustrated. Both mycetophagous and infective forms were recovered from the Canadian sample, whereas only the mycetophagous form was found in the U.S. The new species is characterized by the posteriorly positioned excretory pore relative to the hemizonid in both mycetophagous and infective forms, by a broadly rounded tail end in mycetophagous females and lateral fields with 11 to 12 lines midbody in both mycetophagous females and males. The partial 18S, complete internal transcribed spacer, and partial 28S D2/D3 rRNA genes were amplified and sequenced. Phylogenetic analyses of the genes grouped the new species with previously sequenced species of Deladenus in a fully supported clade. This is the first report of Deladenus species with a known infective stage to have the excretory pore positioned posterior to the hemizonid.


Dental and skeletal components of Class II open bite treatment with a modified Thurow appliance.
Due to the lack of studies that distinguish between dentoalveolar and basal changes caused by the Thurow appliance, this clinical study, carried out by the School of Dentistry--State University of São Paulo/Araraquara, aimed at assessing the dental and skeletal changes induced by modified Thurow appliance. The sample included an experimental group comprising 13 subjects aged between 7 and 10 years old, with Class II malocclusion and anterior open bite, and a control group comprising 22 subjects similar in age, sex and mandibular plane angle. Maxillary/mandibular, horizontal/vertical, dental/skeletal movements (ANS, PNS, U1, U6, Co, Go, Pog, L1, L6) were assessed, based on 14 landmarks, 8 angles (S-N-ANS, SNA, PPA, S-N-Pog, SNB, MPA, PP/MPA, ANB) and 3 linear measures (N-Me, ANS-Me, S-Go). Treatment caused significantly greater angle decrease between the palatal and the mandibular plane on the experimental group, primarily due to an increase in the palatal plane angle. ANB, SNA and S-N-ANS angles significantly decreased more in patients from the experimental group. PNS was superiorly remodeled. Lower face height (ANS-Me) decreased in the experimental group and increased in the control group. The modified Thurow appliance controlled vertical and horizontal displacements of the maxilla, rotated the maxilla and improved open bite malocclusion, decreasing lower facial height.


Effects of high-frequency harmonics on cardiac relaxation indices.
This study was designed to determine the degree to which the systematic distortions in the pressure curve due to recording and filtering systems influence the relaxation indices. The purposes are 1) to analyze the dependency on the high frequencies of the relaxation and contractility parameters, 2) to determine the degree to which the catheter recordings modify those parameters, and 3) to test the monoexponential hypothesis in which the T index is based (Weiss, Frederiksen, and Weisfeldt, J. Clin. Invest. 58: 751-760, 1976). In 17 open-chest dogs, the left ventricular pressure (LVP) was recorded in four different ways: 1) with a large-bored metal cannula, 2) with a catheter-tip manometer, 3) with a Cournand catheter, and 4) with a pigtail catheter. The ECG and LVP were digitized by a computer. A Fourier analysis was performed, and the LVP waves were resynthesized with different numbers of harmonics. We conclude that 1) the catheter recordings are not recommended to calculate relaxation indices, 2) relaxation parameters present more sources of errors than contractility parameters, 3) the monoexponential hypothesis of T is fulfilled if the isovolumic relaxation is considered to end at the time the LVP is 10 mmHg more than left ventricular end-diastolic pressure level, 4) the dependency on higher harmonics of T index and +dP/dtmax is similar, and 5) -dP/dtmax is much more high-frequency dependent than T index.


ALIFE@Work: a randomised controlled trial of a distance counselling lifestyle programme for weight control among an overweight working population [ISRCTN04265725].
The prevalence of overweight is increasing and its consequences will cause a major public health burden in the near future. Cost-effective interventions for weight control among the general population are therefore needed. The ALIFE@Work study is investigating a novel lifestyle intervention, aimed at the working population, with individual counselling through either phone or e-mail. This article describes the design of the study and the participant flow up to and including randomisation. ALIFE@Work is a controlled trial, with randomisation to three arms: a control group, a phone based intervention group and an internet based intervention group. The intervention takes six months and is based on a cognitive behavioural approach, addressing physical activity and diet. It consists of 10 lessons with feedback from a personal counsellor, either by phone or e-mail, between each lesson. Lessons contain educational content combined with behaviour change strategies. Assignments in each lesson teach the participant to apply these strategies to every day life. The study population consists of employees from seven Dutch companies. The most important inclusion criteria are having a body mass index (BMI) > or = 25 kg/m2 and being an employed adult. Primary outcomes of the study are body weight and BMI, diet and physical activity. Other outcomes are: perceived health; empowerment; stage of change and self-efficacy concerning weight control, physical activity and eating habits; work performance/productivity; waist circumference, sum of skin folds, blood pressure, total blood cholesterol level and aerobic fitness. A cost-utility- and a cost-effectiveness analysis will be performed as well. Physiological outcomes are measured at baseline and after six and 24 months. Other outcomes are measured by questionnaire at baseline and after six, 12, 18 and 24 months. Statistical analyses for short term (six month) results are performed with multiple linear regression. Analyses for long term (two year) results are performed with multiple longitudinal regression. Analyses for cost-effectiveness and cost-utility are done at one and two years, using bootstrapping techniques. ALIFE@Work will make a substantial contribution to the development of cost-effective weight control- and lifestyle interventions that are applicable to and attractive for the large population at risk.


Analysis and IbM simulation of the stages in bacterial lag phase: basis for an updated definition.
The lag phase is the initial phase of a culture that precedes exponential growth and occurs when the conditions of the culture medium differ from the pre-inoculation conditions. It is usually defined by means of cell density because the number of individuals remains approximately constant or slowly increases, and it is quantified with the lag parameter lambda. The lag phase has been studied through mathematical modelling and by means of specific experiments. In recent years, Individual-based Modelling (IbM) has provided helpful insights into lag phase studies. In this paper, the definition of lag phase is thoroughly examined. Evolution of the total biomass and the total number of bacteria during lag phase is tackled separately. The lag phase lasts until the culture reaches a maximum growth rate both in biomass and cell density. Once in the exponential phase, both rates are constant over time and equal to each other. Both evolutions are split into an initial phase and a transition phase, according to their growth rates. A population-level mathematical model is presented to describe the transitional phase in cell density. INDividual DIScrete SIMulation (INDISIM) is used to check the outcomes of this analysis. Simulations allow the separate study of the evolution of cell density and total biomass in a batch culture, they provide a depiction of different observed cases in lag evolution at the individual-cell level, and are used to test the population-level model. The results show that the geometrical lag parameter lambda is not appropriate as a universal definition for the lag phase. Moreover, the lag phase cannot be characterized by a single parameter. For the studied cases, the lag phases of both the total biomass and the population are required to fully characterize the evolution of bacterial cultures. The results presented prove once more that the lag phase is a complex process that requires a more complete definition. This will be possible only after the phenomena governing the population dynamics at an individual level of description, and occurring during the lag and exponential growth phases, are well understood.


The unique probe selector: a comprehensive web service for probe design and oligonucleotide arrays.
Nucleic acid hybridization, a fundamental technique in molecular biology, can be modified into very effective and sensitive methods for detecting particular targets mixed with millions of non-target sequences. Therefore, avoiding cross-hybridization is the most crucial issue for developing diagnostic methods based on hybridization. To develop a probe with a high discriminating power, this study constructed a web service, the Unique Probe Selector (UPS), for customized probe design. The UPS service integrates a probe design mechanism and a scoring system for evaluating the performance of probe annealing and the uniqueness of a probe in a user-defined genetic background. Starting from an intuitive web interface, the UPS accepts a query with single or multiple sequences in fasta format. The best probe(s) for each sequence can be downloaded from result pages in a fasta or .csv format with a summary of probe characteristics. The option "Unique probe within group" selects the most unique probe for each target sequence with low probability to hybridize to the other sequences in the same submitted query. The option "Unique probe in the specific organism" devises probes for each submitted sequence to identify its target among selected genetic backgrounds based on Unigene. The UPS evaluates probe-to-target hybridization under a user-defined condition in silico to ensure high-performance hybridization and minimizes the possibility of non-specific reactions. UPS has been applied to design human arrays for gene expression studies and to develop several small arrays of gene families that were inferred as molecular signatures of cancer typing/staging or pathogen signatures. Notably, UPS is freely accessible at http://array.iis.sinica.edu.tw/ups/.


Chemodiversity of Calophyllum inophyllum L. oil bioactive components related to their specific geographical distribution in the South Pacific region.
Different parts of the tree Calophyllum inophyllum L. (nuts, leaves, roots, bark, fruits, nut oil and resin) are used as traditional medicines and cosmetics in most of the Pacific Islands. The oil efficiency as a natural cure and in traditional cosmetics has been largely described throughout the South Pacific, which led us to investigate C. inophyllum's chemical and genetic diversity. A correlative study of the nut resin and leaf DNA from three distinct archipelagos in the South Pacific was carried out in order to identify diversity patterns in C. inophyllum across the South Pacific. Calophyllum inophyllum plants were sampled from French Polynesia, New Caledonia and Fiji. We extracted tamanu oil (nut oil) resin for chemo-diversity studies and sampled leaf tissues for genetic studies. We applied an analysis method designed for small quantities (at a microscale level), and used High Performance Liquid Chromatography (HPLC) to establish the chemo-diversity of tamanu oil resin. In-house standards were co-eluted for qualitative determination. Genetic diversity was assessed using chloroplast barcoding markers (the Acetyl-CoA carboxylase (accD) gene and the psaA-ycf3 intergenic spacer region). Our HPLC analysis revealed 11 previously known tamanu oil constituents, with variability among plant samples. We also isolated and characterized two new neoflavonoids from tamanu oil resin namely, tamanolide E1 and E2 which are diastereoisomers. Although genetic analysis revealed low genetic variation, our multivariate analysis (PCA) of the tamanu oil resin chemical profiles revealed differentiation among geographic regions. We showed here that chromatographic analysis using formalized in-house standards of oil resin compounds for co-elution studies against oil resin samples could identify patterns of variation among samples of C. inophyllum, and discriminate samples from different geographical origins.


Effects of nicorandil, a new antianginal agent, and nifedipine on collateral blood flow in a chronic coronary occlusion model.
The effects of nicorandil and nifedipine on collateral blood flow were compared in anesthetized dogs with a well-developed collateral circulation produced by Ameroid constriction (6-8 weeks) of the left anterior descending (LAD) coronary artery. The radioactive microsphere technique was used to determine myocardial perfusion in the normal left circumflex (LC) region and in the LAD region distal to the Ameroid constrictor. Low and high doses of nicorandil (25 and 50 micrograms/kg/min) or nifedipine (1 and 3 micrograms/kg/min) were infused i.v. to reduce mean arterial and left ventricular systolic pressure approximately 10 and 25 mm Hg, respectively. A low dose of nicorandil had no effect on myocardial perfusion whereas nifedipine increased subepicardial blood flow in both the LC and LAD regions. The high dose of nifedipine further increased both subepicardial and subendocardial perfusion to the LC region and subepicardial blood flow to the LAD region whereas nicorandil had no effect. When aortic blood pressure was returned to control by occluding a snare around the descending thoracic aorta during infusion of the high dose, nicorandil and nifedipine increased subepicardial and subendocardial blood flow to LAD and LC regions. Whereas nicorandil increased flow to both tissue layers equally, nifedipine increased subepicardial perfusion primarily. In summary, nifedipine increased collateral blood flow in a chronic coronary occlusion model despite the presence of systemic hypotension, whereas nicorandil only increased flow when aortic blood pressure was maintained. However, nicorandil increased myocardial blood flow equally across the left ventricular wall, whereas nifedipine primarily increased subepicardial blood flow.


Repression of RNA Polymerase II Transcription by B2 RNA Depends on a Specific Pattern of Structural Regions in the RNA.
B2 RNA is a mouse non-coding RNA that binds directly to RNA polymerase II (Pol II) and represses transcription by disrupting critical interactions between the polymerase and promoter DNA. How the structural regions within B2 RNA work together to mediate transcriptional repression is not well understood. To address this question, we systematically deleted structural regions from B2 RNA and determined the effects on transcriptional repression using a highly purified Pol II in vitro transcription system. Deletions that compromised the ability of B2 RNA to function as a transcriptional repressor were also tested for their ability to bind directly to Pol II, which enabled us to distinguish regions uniquely important for repression from those important for binding. We found that transcriptional repression requires a pattern of RNA structural motifs consisting of an extended single-stranded region bordered by two stem-loops. Hence, there is modularity in the function of the stem-loops in B2 RNA-when one stem-loop is deleted, another can take its place to enable transcriptional repression.


Reaction of trimethylchlorosilane in spin-on Silicalite-1 zeolite film.
We present a study on the hydrophobization of spin-on Silicalite-1 zeolite films through silylation with trimethylchlorosilane. Microporous and micro-mesoporous Silicalite-1 films were synthesized by spin coating of suspensions of Silicalite-1 nanozeolite crystallized for different times. Ellipsometric porosimetry with toluene and water adsorbates reveals that silylation decreases the porosity and makes the films hydrophobic. The decrease in porosity depends on the exposed surface area in the pores. Water contact angle measurements confirm the hydrophobicity. Fourier transform infrared spectroscopy reveals that the trimethylsilyl groups are chemisorbed selectively on isolated silanols and less on geminal and vicinal silanols due to steric limitations. Time-of-flight secondary-ion mass spectroscopy and in situ ellipsometry analysis of the reaction kinetics show that the silylation is a bulk process occurring in the absence of diffusion limitation. Electrical current leakage on films decreases upon silylation. Silylation with trimethylchlorosilane is shown to be an effective hydrophobization method for spin-on Silicalite-1 zeolite films.


Controlled release systems for DNA delivery.
Adapting controlled release technologies to the delivery of DNA has the potential to overcome extracellular barriers that limit gene therapy. Controlled release systems can enhance gene delivery and increase the extent and duration of transgene expression relative to more traditional delivery methods (e.g., injection). These systems typically deliver vectors locally, which can avoid distribution to distant tissues, decrease toxicity to nontarget cells, and reduce the immune response to the vector. Delivery vehicles for controlled release are fabricated from natural and synthetic polymers, which function either by releasing the vector into the local tissue environment or by maintaining the vector at the polymer surface. Vector release or binding is regulated by the effective affinity of the vector for the polymer, which depends upon the strength of molecular interactions. These interactions occur through nonspecific binding based on vector and polymer composition or through the incorporation of complementary binding sites (e.g., biotin-avidin). This review examines the delivery of nonviral and viral vectors from natural and synthetic polymers and presents opportunities for continuing developments to increase their applicability.


A Useful Experimental Model of Short Bowel Syndrome.
The short bowel syndrome is a well-known human clinical entity that produces serious metabolic disorders. This syndrome arises after a massive resection of more than 50% of the small intestine, when the intestine attempts to minimize the consequent irregularities by means of compensatory mechanisms. Many reports suggest that an exocrine and endocrine pancreatic dysfunction is associated with enterohormones and an abnormal altered nutrient flow. In this report, we present an experimental model of short bowel syndrome in rats. A massive intestine resection was performed in rats, followed by a histological study of the small intestine. We report the histological changes related to the compensatory changes that occurred in the remaining intestine. The residual intestine produces a hyperplasic response, and hypertrophy was seen in the portion proximal to the anastomosis. We believe this experimental model of short bowel syndrome could be a very useful tool for studying the enterohormonal changes related to an abnormal blood flow of nutrients.


[Sec-butyl ester of dodecenoate: synthesis and attractive properties].
2-Butyl 2-dodecenoate has been synthesized from lauric acid and sec-butanol. The study of the biological activity of this substance has demonstrated its property as a sex attractant for males of Jordanita graeca, Jordanita globulariae and Theresimima ampellophaga (Lepidoptera: Zygaenidae, Procridinae). The latter species is the grape pest in southern Europe. The attractant obtained can be used for the detection of species, for monitoring their numbers in nature and for the elaboration of ecological methods of pest control.


Comparing the impact of artemisinin-based combination therapies on malaria transmission in sub-Saharan Africa.
Artemisinin-based combination therapies (ACTs) are currently considered the first-line treatments for uncomplicated Plasmodium falciparum malaria. Among these, artemether-lumefantrine (AL) has been the most widely prescribed ACT in sub-Saharan Africa. Recent clinical trials conducted in sub-Saharan Africa have shown that dihydroartemisinin-piperaquine (DP), a most recent ACT, may have a longer post-treatment prophylactic period and post-treatment infection period (duration of gametocyte carriage) than AL. Using epidemiological and clinical data on the efficacy of AL and DP, we developed and parameterized a mathematical transmission model that we used to compare the population-level impact of AL and DP for reducing P. falciparum malaria transmission in sub-Saharan Africa. Our results showed that DP is likely to more effectively reduce malaria incidence of clinical episodes than AL. However in low P. falciparum transmission areas, DP and AL are likely to be equally effective in reducing malaria prevalence. The predictions of our model were shown to be robust to the empirical uncertainty summarizing the epidemiological parameters. DP should be considered as a replacement for AL as first-line treatment of uncomplicated malaria in highly endemic P. falciparum communities. To optimize the effectiveness of ACTs, it is necessary to tailor treatment policies to the transmission intensity in different settings.


The hippocampus as the switchboard between perception and memory.
Adaptive memory recall requires a rapid and flexible switch from external perceptual reminders to internal mnemonic representations. However, owing to the limited temporal or spatial resolution of brain imaging modalities used in isolation, the hippocampal-cortical dynamics supporting this process remain unknown. We thus employed an object-scene cued recall paradigm across two studies, including intracranial electroencephalography (iEEG) and high-density scalp EEG. First, a sustained increase in hippocampal high gamma power (55 to 110 Hz) emerged 500 ms after cue onset and distinguished successful vs. unsuccessful recall. This increase in gamma power for successful recall was followed by a decrease in hippocampal alpha power (8 to 12 Hz). Intriguingly, the hippocampal gamma power increase marked the moment at which extrahippocampal activation patterns shifted from perceptual cue toward mnemonic target representations. In parallel, source-localized EEG alpha power revealed that the recall signal progresses from hippocampus to posterior parietal cortex and then to medial prefrontal cortex. Together, these results identify the hippocampus as the switchboard between perception and memory and elucidate the ensuing hippocampal-cortical dynamics supporting the recall process.


What is the value of the routine use of patient-reported outcome measures toward improvement of patient outcomes, processes of care, and health service outcomes in cancer care? A systematic review of controlled trials.
The systematic use of patient-reported outcome measures (PROMs) has been advocated as an effective way to standardize cancer practice. Yet, the question of whether PROMs can lead to actual improvements in the quality of patient care remains under debate. This review examined whether inclusion of PROM in routine clinical practice is associated with improvements in patient outcomes, processes of care, and health service outcomes during active anticancer treatment. A systematic review of five electronic databases (Medline, EMBASE, CINAHL [Cumulative Index to Nursing and Allied Health Literature], PsycINFO, and Psychology and Behavioral Sciences Collection [PBSC]) was conducted from database inception to May 2012 to locate randomized and nonrandomized controlled trials of patients receiving active anticancer treatment or supportive care irrespective of type of cancer. Based on prespecified eligibility criteria, we included 26 articles that reported on 24 unique controlled trials. Wide variability in the design and use of interventions delivered, outcomes evaluated, and cancer- and modality-specific context was apparent. Health service outcomes were only scarcely included as end points. Overall, the number of statistically significant findings were limited and PROMs' intervention effect sizes were predominantly small-to-moderate. The routine use of PROMs increases the frequency of discussion of patient outcomes during consultations. In some studies, PROMs are associated with improved symptom control, increased supportive care measures, and patient satisfaction. Additional effort is required to ensure patient adherence, as well as additional support to clinicians who will respond to patient concerns and issues, with clear system guidelines in place to guide their responses. More research is required to support PROM cost-benefit in terms of patient safety, clinician burden, and health services usage.


Medical support in the Tangshan earthquake: a review of the management of mass casualties and certain major injuries.
The Tangshan earthquake was probably the worst catastrophe in this century. It took a death toll of 242,769, with 164,851 injured in addition. This presentation describes the organization of disaster relief work after the earthquake, the rescue of buried victims, the organization of medical resources, and the sanitation work to forestall epidemics. It also presents the author's reflections on the management of three major injuries, namely, crush syndrome, fracture of pelvis, and traumatic paraplegia, by reviewing the available data pertaining to these injuries. The author concurs with the prevailing opinion that fasciotomy plays an important role in the successful management of crush injury. It not only prevented acute renal failure subsequent to intracompartmental increase of pressure, but also the occurrence of Volkmann's ischemic contracture as a late sequela. Herbs to induce catharsis and diuresis were used to alleviate intracompartmental pressure. For the management of pelvic fractures, two newly developed treatment techniques are described. On analysis of clinical data, it is the author's opinion that traumatic paraplegia should not be given the priority of early surgery in the circumstances of mass casualties. The primary concern should be the stability and restoration of normal curvature of the spine, especially in cases of complete paraplegia. Decompression of the spinal cord through an anterolateral approach gave promising results in hyperflexion type of spinal fracture.


Sedimentary phosphate method for estimating paleosalinities: a paleontological assumption.
Paleosalinity values in certain rocks determined by the sedimentary phosphate method differ from salinity estimates based upon contained fossil assemblages, geochemical methods, and existing stratigraphic controls. Some anomalous values are related to the abundance of fossil organisms known to be concentrators of calcium phosphate. Because of the abundance and diversity of organisms which might introduce significant errors into paleosalinity estimates, the sedimentary phosphate method seemingly is of limited applicability.


The PF4/PPBP/CXCL5 Gene Cluster Is Associated with Periodontitis.
Periodontitis is a common dysbiotic inflammatory disease with an estimated heritability of 50%. Due to the limited sample size of available periodontitis cohorts and the underlying trait heterogeneity, genome-wide association studies (GWAS) of chronic periodontitis (CP) have been unsuccessful in discovering susceptibility factors. A strategy that combines agnostic GWAS with a well-powered candidate-gene approach has the potential to discover novel loci. We combined RNA-seq data from gingival tissues with quantitative trait loci (QTLs) that were identified in a F2-cross of mice resistant and susceptible to infection with oral bacterial pathogens. Four genes, which were located within the mapped QTLs, showed differential expression. The chromosomal regions across the human orthologous were interrogated for putative periodontitis-associated variants using existing GWAS data from a German case-control sample of aggressive periodontitis (AgP; 651 cases, 4,001 controls), the most severe and early onset form of periodontitis. Two haplotype blocks, one upstream to the coding region of UGT2A1 (rs146712414, P = 9.1 × 10-5; odds ratio [OR], 1.34; 95% confidence interval [CI], 1.16-1.56) and one downstream of the genes PF4/PPBP/CXCL5 (rs1595009, P = 1.3 × 10-4; OR, 1.32; 95% CI, 1.15-1.52), were associated with AgP. The association of rs1595009 was validated in an independent cohort of CP of European Americans (1,961 cases and 1,864 controls; P = 0.03; OR, 1.45; 95% CI, 1.01-1.29). This association was further replicated in another sample of 399 German CP cases (disease onset <60 y of age) and 1,633 controls ( P = 0.03; OR, 1.75; 95% CI, 1.06-2.90). The combined estimates of association from all samples were P = 2.9 × 10-5 (OR, 1.2; 95% CI, 1.1-1.3). This study shows the strength of combining QTL mapping and RNA-Seq data from a mouse model with association studies in human case-control samples to identify genetic risk variants of periodontitis.


Extensive remodeling of a cyanobacterial photosynthetic apparatus in far-red light.
Cyanobacteria are unique among bacteria in performing oxygenic photosynthesis, often together with nitrogen fixation and, thus, are major primary producers in many ecosystems. The cyanobacterium, Leptolyngbya sp. strain JSC-1, exhibits an extensive photoacclimative response to growth in far-red light that includes the synthesis of chlorophylls d and f. During far-red acclimation, transcript levels increase more than twofold for ~900 genes and decrease by more than half for ~2000 genes. Core subunits of photosystem I, photosystem II, and phycobilisomes are replaced by proteins encoded in a 21-gene cluster that includes a knotless red/far-red phytochrome and two response regulators. This acclimative response enhances light harvesting for wavelengths complementary to the growth light (λ = 700 to 750 nanometers) and enhances oxygen evolution in far-red light.


Evaluation of the potential of Raman microspectroscopy for prediction of chemotherapeutic response to cisplatin in lung adenocarcinoma.
The study of the interaction of anticancer drugs with mammalian cells in vitro is important to elucidate the mechanisms of action of the drug on its biological targets. In this context, Raman spectroscopy is a potential candidate for high throughput, non-invasive analysis. To explore this potential, the interaction of cis-diamminedichloroplatinum(II) (cisplatin) with a human lung adenocarcinoma cell line (A549) was investigated using Raman microspectroscopy. The results were correlated with parallel measurements from the MTT cytotoxicity assay, which yielded an IC(50) value of 1.2 ± 0.2 µM. To further confirm the spectral results, Raman spectra were also acquired from DNA extracted from A549 cells exposed to cisplatin and from unexposed controls. Partial least squares (PLS) multivariate regression and PLS Jackknifing were employed to highlight spectral regions which varied in a statistically significant manner with exposure to cisplatin and with the resultant changes in cellular physiology measured by the MTT assay. The results demonstrate the potential of the cellular Raman spectrum to non-invasively elucidate spectral changes that have their origin either in the biochemical interaction of external agents with the cell or its physiological response, allowing the prediction of the cellular response and the identification of the origin of the chemotherapeutic response at a molecular level in the cell.


Use of social network sites and instant messaging does not lead to increased offline social network size, or to emotionally closer relationships with offline network members.
The effect of Internet use on social relationships is still a matter of intense debate. This study examined the relationships between use of social media (instant messaging and social network sites), network size, and emotional closeness in a sample of 117 individuals aged 18 to 63 years old. Time spent using social media was associated with a larger number of online social network "friends." However, time spent using social media was not associated with larger offline networks, or feeling emotionally closer to offline network members. Further, those that used social media, as compared to non-users of social media, did not have larger offline networks, and were not emotionally closer to offline network members. These results highlight the importance of considering potential time and cognitive constraints on offline social networks when examining the impact of social media use on social relationships.


Rab18 dynamics in adipocytes in relation to lipogenesis, lipolysis and obesity.
Lipid droplets (LDs) are organelles that coordinate lipid storage and mobilization, both processes being especially important in cells specialized in managing fat, the adipocytes. Proteomic analyses of LDs have consistently identified the small GTPase Rab18 as a component of the LD coat. However, the specific contribution of Rab18 to adipocyte function remains to be elucidated. Herein, we have analyzed Rab18 expression, intracellular localization and function in relation to the metabolic status of adipocytes. We show that Rab18 production increases during adipogenic differentiation of 3T3-L1 cells. In addition, our data show that insulin induces, via phosphatidylinositol 3-kinase (PI3K), the recruitment of Rab18 to the surface of LDs. Furthermore, Rab18 overexpression increased basal lipogenesis and Rab18 silencing impaired the lipogenic response to insulin, thereby suggesting that this GTPase promotes fat accumulation in adipocytes. On the other hand, studies of the β-adrenergic receptor agonist isoproterenol confirmed and extended previous evidence for the participation of Rab18 in lipolysis. Together, our data support the view that Rab18 is a common mediator of lipolysis and lipogenesis and suggests that the endoplasmic reticulum (ER) is the link that enables Rab18 action on these two processes. Finally, we describe, for the first time, the presence of Rab18 in human adipose tissue, wherein the expression of this GTPase exhibits sex- and depot-specific differences and is correlated to obesity. Taken together, these findings indicate that Rab18 is involved in insulin-mediated lipogenesis, as well as in β-adrenergic-induced lipolysis, likely facilitating interaction of LDs with ER membranes and the exchange of lipids between these compartments. A role for Rab18 in the regulation of adipocyte biology under both normal and pathological conditions is proposed.


Production of an antibiotic substance by Bacillus licheniformis within the digestive tract of gnotobiotic mice.
In monoxenic mice, vegetative cells and spores of Bacillus licheniformis were enumerated and in vivo antibiotic production was measured at various levels of the digestive tract and in the feces. The apparent independence of vegetative cell and spore populations in the cecum and feces, as well as the marked fluctuations of these two populations in the stomach and small intestine, suggested that sporulation of B. licheniformis and production of antibiotic occur only in the upper levels of the digestive tract. Study of dixenic models wherein B. licheniformis was inoculated after Clostridium perfringens or Lactobacillus sp. or before Lactobacillus sp. revealed the simultaneous disappearance of B. licheniformis from the stomach and antibiotic from the feces. This strain persisted in the cecum, but only in the form of vegetative cells. These models demonstrate that the activity of the microflora in the upper segments can affect the equilibrium of the microflora throughout the digestive tract.


Effectiveness of High-Fidelity Simulation in Nursing Education for End-of-Life Care: A Quasi-experimental Design.
Providing end of life (EOL) care is a component of palliative care but dealing with dying patients and their family members is stressful for the healthcare providers. To prepare them for providing EOL care, the high-fidelity simulation could be used as a pedagogy in which real-life scenarios are used on the computerized manikins mimicking the real patients. The aim of this study was to measure the effectiveness of high-fidelity simulation to teach EOL care in the palliative nursing course in the undergraduate nursing education program at the School of Nursing and Midwifery at Aga Khan University which is private university in Karachi, Pakistan. This study was approved by the ethics review committee of Aga Khan University. It was hypothesized that exposure to high-fidelity simulation will lead to an increased positive attitude in participants towards the care of dying. A quasi-experimental design was used. In line with the design, there was no control group. The same group of students (n = 42) were assessed through Frommelt Attitudes Toward Care of the Dying (FATCOD) Part B assessment tool. Permission for using this tool was obtained from Dr. Katherine Frommelt, the author of this tool. Research participants filled this tool before and after the intervention, i.e., providing EOL care to a patient in a high-fidelity simulation lab. Out of 30-FATCOD items, significant attitude change was detected on 11-items of which 8 were positively worded statements and 3 were negatively worded statements. As per the hypothesis, it was expected for the positively worded statements that the mean score for the posttest would be significantly greater than the pretest mean score (pretest score < posttest score). The hypothesis was proved for items 1, 4, 10, 18, 22, 25, 27, and 30 as their t-value was significant at 0.05 alpha value (one-tailed). For the negatively worded statements, it was expected that the mean score for the posttest would be significantly lower than the pretest (pretest score > posttest score). The hypothesis was proved for items 5, 6, and 11 as their t-value was significant at 0.05 alpha value (one-tailed). In this research teaching, EOL care through high-fidelity simulation had improved the attitudes of students toward providing care. This pedagogy also provided the participants with a learning opportunity to deal with their own emotions. These findings provide a way forward for teaching EOL and other complex skills of clinical practice.


Does a TV Public Service Advertisement Campaign for Suicide Prevention Really Work?
One of the critical measures in suicide prevention is promoting public awareness of crisis hotline numbers so that individuals can more readily seek help in a time of crisis. Although public service advertisements (PSA) may be effective in raising the rates of both awareness and use of a suicide hotline, few investigations have been performed regarding their effectiveness in South Korea, where the suicide rate is the highest among OECD countries. The goal of this study was to evaluate the effectiveness of a television PSA campaign. We analyzed a database of crisis phone calls compiled by the Korean Ministry of Health and Welfare to track changes in call volume to a crisis hotline that was promoted in a TV campaign. We compared daily call counts for three periods of equal length: before, during, and after the campaign. The number of crisis calls during the campaign was about 1.6 times greater than the number before or after the campaign. Relative to the number of suicide-related calls in the previous year, the number of calls during the campaign period surged, displaying a noticeable increase. The findings confirmed that this campaign had a positive impact on call volume to the suicide hotline.


CLOCK deubiquitylation by USP8 inhibits CLK/CYC transcription in Drosophila.
A conserved transcriptional feedback loop underlies animal circadian rhythms. In Drosophila, the transcription factors CLOCK (CLK) and CYCLE (CYC) activate the transcription of direct target genes like period (per) and timeless (tim). They encode the proteins PER and TIM, respectively, which repress CLK/CYC activity. Previous work indicates that repression is due to a direct PER-CLK/CYC interaction as well as CLK/CYC phosphorylation. We describe here the role of ubiquitin-specific protease 8 (USP8) in circadian transcriptional repression as well as the importance of CLK ubiquitylation in CLK/CYC transcription activity. usp8 loss of function (RNAi) or expression of a dominant-negative form of the protein (USP8-DN) enhances CLK/CYC transcriptional activity and alters fly locomotor activity rhythms. Clock protein and mRNA molecular oscillations are virtually absent within circadian neurons of USP8-DN flies. Furthermore, CLK ubiquitylation cycles robustly in wild-type flies and peaks coincident with maximal CLK/CYC transcription. As USP8 interacts with CLK and expression of USP8-DN increases CLK ubiquitylation, the data indicate that USP8 deubiquitylates CLK, which down-regulates CLK/CYC transcriptional activity. Taken together with the facts that usp8 mRNA cycles and that its transcription is activated directly by CLK/CYC, USP8, like PER and TIM, contributes to the transcriptional feedback loop cycle that underlies circadian rhythms.


Fragmentation of negative ions from N-linked carbohydrates: part 6. Glycans containing one N-acetylglucosamine in the core.
Negative ion collision-induced dissociation (CID) spectra of N-glycans contain many diagnostic ions that provide more structural information than positive ion spectra. EndoH or endoS release of glycans from glycoproteins, as used by many investigators, cleaves glycans between the GlcNAc residues of the chitobiose core leaving the glycan without the reducing-terminal GlcNAc residue. However, their negative ion CID spectra do not appear to have been studied in detail. This paper examines the CID and ion mobility properties of these endoH-released glycans to determine if the missing GlcNAc influences the production of diagnostic fragment ions. N-Glycans were released from ribonuclease B, ovalbumin and gp120 with endoH to give high-mannose and hybrid glycans, and from IgG with endoS to produce biantennary complex glycans, all missing the reducing-terminal GlcNAc residue. Negative ion CID and travelling wave ion mobility spectra were recorded with a Waters Synapt G2 mass spectrometer using nanospray sample introduction. The majority of glycans yielded CID spectra exhibiting the same diagnostic fragments, which were equivalently informative, as the fully released structures. However, the ability of ion mobility to separate isomers was generally found to be inferior to its use with the full glycans despite the smaller nature of the compounds. The exception was the partial resolution of a pair of biantennary monogalactosylated glycans from IgG where, as chloride adducts, slight separation of the isomers was observed. The results show that the CID spectra of endoH- and endoS-released glycans are as useful as the corresponding spectra of the intact glycans (as released by PNGase F) in providing structural information on N-glycans.


Health promoting behaviours and lifestyle characteristics of students at seven universities in the UK.
University students' wellbeing and health promoting and damaging behaviours are important and comprise many parameters. The purpose of this study was to assess a range of health behaviours and lifestyle characteristics of 3,706 undergraduate students from seven universities in England, Wales and Northern Ireland. We compared differences in these parameters between males and females, and across the participating universities. A self-administered questionnaire assessed socio-demographic information (e.g., gender, age), nutrition, dietary intake and food consumption patterns, as well as the importance of healthy eating, three levels of physical activity, restful sleep, tobacco smoking, use of illicit substance (recreational drugs), frequency of binge drinking and problem drinking. The data was collected in 2007-2008. While females generally reported lower use of tobacco, illicit substances and alcohol (binge drinking/problem drinking) and consumed more fruits and vegetables, male students had a higher level of physical activity, consumed less sweets and had more restful sleep. When lifestyle characteristics of students were compared between the different universities we observed some 'clustering' of the parameters under study, whereby favourable health practices would be exhibited at some universities; and conversely, the clustering of less favourable practices exhibited at other participating sites. We conclude that only a minority of students exhibited positive health practices above recommended levels and the level of binge drinking and problem drinking was high. This calls for increased awareness of university administrators, leaders and policy makers to the risky health habits of their students. The observed clustering effects also indicate the need for local (university-specific) health profiles as basis and guidance for relevant health promotion programmes at universities.


Automated turbidimetry of serum lipoprotein(a).
We describe a simple immunoturbidimetric method for quantifying lipoprotein(a) in serum based on latex-enhanced particle agglutination technology. Carboxylated latex particles (diameter 240 nm) covalently coated with F(ab')2 fragments of anti-lipoprotein(a) antibodies are incubated with the sample for 5 min at 37 degrees C, and the resulting agglutination is quantified by measuring the change of turbidity produced at 700 nm. The assay is rapid, precise and fully automated on the Hitachi 911 analyser. The assay range is about 0.03-0.9 g/l. Average analytical recovery was 97.8%. Precision (CV) ranged from 1.9 to 3.1% at different lipoprotein(a) values. There was no interference from bilirubin, Intralipid, haemoglobin, plasminogen or apolipoprotein B. Comparisons with a latex nephelometric assay carried out on the Behring nephelometer analyser, and with three commercially available methods, a radioimmunoassay and two ELISA assays, gave good correlations (r > 0.95), although a large among-method variation in lipoprotein(a) values was found. We conclude that the proposed latex turbidimetric immunoassay method is suitable for routine use in clinical laboratories.


Flat and complex temperate reefs provide similar support for fish: Evidence for a unimodal species-habitat relationship.
Structural complexity, a form of habitat heterogeneity, influences the structure and function of ecological communities, generally supporting increased species density, richness, and diversity. Recent research, however, suggests the most complex habitats may not harbor the highest density of individuals and number of species, especially in areas with elevated human influence. Understanding nuances in relationships between habitat heterogeneity and ecological communities is warranted to guide habitat-focused conservation and management efforts. We conducted fish and structural habitat surveys of thirty warm-temperate reefs on the southeastern US continental shelf to quantify how structural complexity influences fish communities. We found that intermediate complexity maximizes fish abundance on natural and artificial reefs, as well as species richness on natural reefs, challenging the current paradigm that abundance and other fish community metrics increase with increasing complexity. Naturally occurring rocky reefs of flat and complex morphologies supported equivalent abundance, biomass, species richness, and community composition of fishes. For flat and complex morphologies of rocky reefs to receive equal consideration as essential fish habitat (EFH), special attention should be given to detecting pavement type rocky reefs because their ephemeral nature makes them difficult to detect with typical seafloor mapping methods. Artificial reefs of intermediate complexity also maximized fish abundance, but human-made structures composed of low-lying concrete and metal ships differed in community types, with less complex, concrete structures supporting lower numbers of fishes classified largely as demersal species and metal ships protruding into the water column harboring higher numbers of fishes, including more pelagic species. Results of this study are essential to the process of evaluating habitat function provided by different types and shapes of reefs on the seafloor so that all EFH across a wide range of habitat complexity may be accurately identified and properly managed.


Write-once recording for multilayered optical waveguide-type holographic cards.
We propose a write-once recording technique for multilayered optical waveguide-type holographic cards. The card medium has a construction created by adding a recording layer and a holographic grating layer to the multilayered optical waveguide composed of core and cladding layers. Individual data for each medium were recorded as an arrangement of optically transparent holes formed in the recording layer. Holograms common to all media were designed in the holographic grating layer so that diffracted lights from the holograms could pass through the holes and focus on an image sensor. We succeeded in write-once recording with a memory capacity potential of more than 128 bits.


Quantum cascade laser master-oscillator power-amplifier with 1.5 W output power at 300 K.
We report quantum cascade laser (QCL) master-oscillator power-amplifiers (MOPAs) at 300 K reaching output power of 1.5 W for tapered devices and 0.9 W for untapered devices. The devices display single-longitudinal-mode emission at λ = 7.26 µm and single-transverse-mode emission at TM(00). The maximum amplification factor is 12 dB for the tapered devices.


Pectobacterium carotovorum elicits plant cell death with DspE/F but the P. carotovorum DspE does not suppress callose or induce expression of plant genes early in plant-microbe interactions.
The broad-host-range bacterial soft rot pathogen Pectobacterium carotovorum causes a DspE/F-dependent plant cell death on Nicotiana benthamiana within 24 h postinoculation (hpi) followed by leaf maceration within 48 hpi. P. carotovorum strains with mutations in type III secretion system (T3SS) regulatory and structural genes, including the dspE/F operon, did not cause hypersensitive response (HR)-like cell death and or leaf maceration. A strain with a mutation in the type II secretion system caused HR-like plant cell death but no maceration. P. carotovorum was unable to impede callose deposition in N. benthamiana leaves, suggesting that P. carotovorum does not suppress this basal immunity function. Within 24 hpi, there was callose deposition along leaf veins and examination showed that the pathogen cells were localized along the veins. To further examine HR-like plant cell death induced by P. carotovorum, gene expression profiles in N. benthamiana leaves inoculated with wild-type and mutant P. carotovorum and Pseudomonas syringae strains were compared. The N. benthamiana gene expression profile of leaves infiltrated with Pectobacterium carotovorum was similar to leaves infiltrated with a Pseudomonas syringae T3SS mutant. These data support a model where Pectobacterium carotovorum uses the T3SS to induce plant cell death in order to promote leaf maceration rather than to suppress plant immunity.


Small chromosomal regions position themselves autonomously according to their chromatin class.
The spatial arrangement of chromatin is linked to the regulation of nuclear processes. One striking aspect of nuclear organization is the spatial segregation of heterochromatic and euchromatic domains. The mechanisms of this chromatin segregation are still poorly understood. In this work, we investigated the link between the primary genomic sequence and chromatin domains. We analyzed the spatial intranuclear arrangement of a human artificial chromosome (HAC) in a xenospecific mouse background in comparison to an orthologous region of native mouse chromosome. The two orthologous regions include segments that can be assigned to three major chromatin classes according to their gene abundance and repeat repertoire: (1) gene-rich and SINE-rich euchromatin; (2) gene-poor and LINE/LTR-rich heterochromatin; and (3) gene-depleted and satellite DNA-containing constitutive heterochromatin. We show, using fluorescence in situ hybridization (FISH) and 4C-seq technologies, that chromatin segments ranging from 0.6 to 3 Mb cluster with segments of the same chromatin class. As a consequence, the chromatin segments acquire corresponding positions in the nucleus irrespective of their chromosomal context, thereby strongly suggesting that this is their autonomous property. Interactions with the nuclear lamina, although largely retained in the HAC, reveal less autonomy. Taken together, our results suggest that building of a functional nucleus is largely a self-organizing process based on mutual recognition of chromosome segments belonging to the major chromatin classes.


Chewing gum alleviates negative mood and reduces cortisol during acute laboratory psychological stress.
The notion that chewing gum may relieve stress was investigated in a controlled setting. A multi-tasking framework which reliably evokes stress and also includes performance measures was used to induce acute stress in the laboratory. Using a randomised crossover design forty participants (mean age 21.98 years) performed on the multi-tasking framework at two intensities (on separate days) both while chewing and not chewing. Order of workload intensity and chewing conditions were counterbalanced. Before and after undergoing the platform participants completed the state portion of the State-Trait Anxiety Inventory, Bond-Lader visual analogue mood scales, a single Stress Visual Analogue Scale and provided saliva samples for cortisol measurement. Baseline measures showed that both levels of the multi-tasking framework were effective in significantly reducing self-rated alertness, calmness and contentment while increasing self-rated stress and state anxiety. Cortisol levels fell during both levels of the stressor during the morning, reflecting the predominance of a.m. diurnal changes, but this effect was reversed in the afternoon which may reflect a measurable stress response. Pre-post stressor changes (Delta) for each measure at baseline were subtracted from Delta scores under chewing and no chewing conditions. During both levels of stress the chewing gum condition was associated with significantly better alertness and reduced state anxiety, stress and salivary cortisol. Overall performance on the framework was also significantly better in the chewing condition. The mechanisms underlying these effects are unknown but may involve improved cerebral blood flow and/or effects secondary to performance improvement during gum chewing.


Association of schistosome infection with adiposity in Tanzania.
Observational studies in humans have reported a link between schistosome infection and lower adiposity, but this may be explained by socioeconomic and demographic factors, intensity of infection, or common co-infections such as HIV. This was a cross-sectional study that investigated the relationship between schistosome infection and adiposity in a large, well-described cohort of Tanzanian adults living with and without HIV. Cross-sectional data were collected among adults living in Mwanza, Tanzania who were enrolled in the Chronic Infections, Co-morbidities and Diabetes in Africa (CICADA) cohort study. Schistosome circulating anodic antigen, secreted by both Schistosoma mansoni and haematobium which are endemic to Tanzania, was quantified from stored samples. Schistosome infection diagnosed by serum circulating anodic antigen levels. The primary outcome was fat mass measured by bioimpedance analysis. Secondary outcomes included fat-free mass, waist circumference, mid-upper arm circumference, and body mass index. The study enrolled 1,947 adults, of whom 1,923 (98.8%) had serum available for schistosome testing. Of these, 873 (45.4%) had a serum circulating anodic antigen ≥30 pg/mL, indicating schistosome infection. Compared to uninfected individuals, those with schistosome infections had -1.1 kg [95% CI -1.9 to -0.3] lower fat mass after adjusting for age, sex, physical activity, tobacco use, education level, and socioeconomic status. Infected participants also had lower waist circumference, mid-upper arm circumference, and body mass index. Fat-free mass was not different between the two groups. Neither being HIV-infected, nor receiving antiretroviral therapy, modified associations between schistosome infection and adiposity. These associations were also not affected by Schistosoma worm burden. Schistosome infection was associated with lower fat mass and less central adiposity without a difference in muscle mass, irrespective of confounders, HIV status, or the intensity of schistosome infection. Future studies should adjust for socioeconomic and demographic factors that are associated with schistosome infection and adiposity. Identifying mechanistic pathways by which schistosome infection reduces adiposity while preserving muscle mass could yield new strategies for obesity control and cardiovascular disease prevention.


Spatiotemporal mathematical modelling of mutations of the dhps gene in African Plasmodium falciparum.
Plasmodium falciparum has repeatedly evolved resistance to first-line anti-malarial drugs, thwarting efforts to control and eliminate the disease and in some period of time this contributed largely to an increase in mortality. Here a mathematical model was developed to map the spatiotemporal trends in the distribution of mutations in the P. falciparum dihydropteroate synthetase (dhps) gene that confer resistance to the anti-malarial sulphadoxine, and are a useful marker for the combination of alleles in dhfr and dhps that is highly correlated with resistance to sulphadoxine-pyrimethamine (SP). The aim of this study was to present a proof of concept for spatiotemporal modelling of trends in anti-malarial drug resistance that can be applied to monitor trends in resistance to components of artemisinin combination therapy (ACT) or other anti-malarials, as they emerge or spread. Prevalence measurements of single nucleotide polymorphisms in three codon positions of the dihydropteroate synthetase (dhps) gene from published studies of dhps mutations across Africa were used. A model-based geostatistics approach was adopted to create predictive surfaces of the dhps540E mutation over the spatial domain of sub-Saharan Africa from 1990-2010. The statistical model was implemented within a Bayesian framework and hence quantified the associated uncertainty of the prediction of the prevalence of the dhps540E mutation in sub-Saharan Africa. The maps presented visualize the changing prevalence of the dhps540E mutation in sub-Saharan Africa. These allow prediction of space-time trends in the parasite resistance to SP, and provide probability distributions of resistance prevalence in places where no data are available as well as insight on the spread of resistance in a way that the data alone do not allow. The results of this work will be extended to design optimal sampling strategies for the future molecular surveillance of resistance, providing a proof of concept for similar techniques to design optimal strategies to monitor resistance to ACT.


Comparative transcriptome analyses of genes involved in sulforaphane metabolism at different treatment in Chinese kale using full-length transcriptome sequencing.
Sulforaphane is a natural isothiocyanate available from cruciferous vegetables with multiple characteristics including antioxidant, antitumor and anti-inflammatory effect. Single-molecule real-time (SMRT) sequencing has been used for long-read de novo assembly of plant genome. Here, we investigated the molecular mechanism related to glucosinolates biosynthesis in Chinese kale using combined NGS and SMRT sequencing. SMRT sequencing produced 185,134 unigenes, higher than 129,325 in next-generation sequencing (NGS). NaCl (75 mM), methyl jasmonate (MeJA, 40 μM), selenate (Se, sodium selenite 100 μM), and brassinolide (BR, 1.5 μM) treatment induced 6893, 13,287, 13,659 and 11,041 differentially expressed genes (DEGs) in Chinese kale seedlings comparing with control. These genes were associated with pathways of glucosinolates biosynthesis, including phenylalanine, tyrosine and tryptophan biosynthesis, cysteine and methionine metabolism, and glucosinolate biosynthesis. We found NaCl decreased sulforaphane and glucosinolates (indolic and aliphatic) contents and downregulated expression of cytochrome P45083b1 (CYP83b1), S-alkyl-thiohydroximatelyase or carbon-sulfur lyase (SUR1) and UDP-glycosyltransferase 74B1 (UGT74b1). MeJA increased sulforaphane and glucosinolates contents and upregulated the expression of CYP83b1, SUR1 and UGT74b1; Se increased sulforaphane; BR increased expression of CYP83b1, SUR1 and UGT74b1, and increased glucosinolates contents. The desulfoglucosinolate sulfotransferases ST5a_b_c were decreased by all treatments. We confirmed that NaCl inhibited the biosynthesis of both indolic and aliphatic glucosinolates, while MeJA and BR increased them. MeJA and BR treatments, conferred the biosynthesis of glucosinolates, and Se and MeJA contributed to sulforaphane in Chinese kale via regulating the expression of CYP83b1, SUR1 and UGT74b1.


KIDFamMap: a database of kinase-inhibitor-disease family maps for kinase inhibitor selectivity and binding mechanisms.
Kinases play central roles in signaling pathways and are promising therapeutic targets for many diseases. Designing selective kinase inhibitors is an emergent and challenging task, because kinases share an evolutionary conserved ATP-binding site. KIDFamMap (http://gemdock.life.nctu.edu.tw/KIDFamMap/) is the first database to explore kinase-inhibitor families (KIFs) and kinase-inhibitor-disease (KID) relationships for kinase inhibitor selectivity and mechanisms. This database includes 1208 KIFs, 962 KIDs, 55 603 kinase-inhibitor interactions (KIIs), 35 788 kinase inhibitors, 399 human protein kinases, 339 diseases and 638 disease allelic variants. Here, a KIF can be defined as follows: (i) the kinases in the KIF with significant sequence similarity, (ii) the inhibitors in the KIF with significant topology similarity and (iii) the KIIs in the KIF with significant interaction similarity. The KIIs within a KIF are often conserved on some consensus KIDFamMap anchors, which represent conserved interactions between the kinase subsites and consensus moieties of their inhibitors. Our experimental results reveal that the members of a KIF often possess similar inhibition profiles. The KIDFamMap anchors can reflect kinase conformations types, kinase functions and kinase inhibitor selectivity. We believe that KIDFamMap provides biological insights into kinase inhibitor selectivity and binding mechanisms.


Guinea pig immune response-related histocompatibility antigens. Partial characterization and distribution.
We have previously demonstrated that guinea pig alloantisera directed at strain 2 and strain 13 membrane antigens block specific lymphocyte activation in immune response gene-controlled systems. In this communication we describe the partial characterization of the antigens against which these antisera are directed (the 2 and 13 antigens) and, in addition, that of the B antigen which by distribution resembles the human HL-A and mouse H-2 major histocompatibility antigens. Lymphoid cells from strain 2 and strain 13 guinea pigs were surface labeled with 125I by the lactoperoxidase technique. Nonidet P-40 extracts of these labeled cells were precipitated by sandwiches of strain 2 antistrain 13, strain 13 antistrain 2, or outbred anti-B antisera, followed by rabbit antiguinea pig immunoglobulin antisera. Precipitates were dissolved in sodium dodecyl sulfate (SDS) and electrophoresed on SDS polyacrylamide gels. Radioactive peaks representing the 2 and B-cell membrane antigens were obtained from strain 2 lymph node cells, as well as from a B-lymphoid cell population (L2C leukemia cells) and a T-lymphocyte population (STRAIN 2 PERITONEAL EXUDATE LYMPHOCYTES [PELs]). Radioactive peaks representing the 13 and B-cell membrane antigens were obtained from strain 13 lymph node cells and strain 13 PELs. All anti-B precipitates produced two peaks when electrophoresed on SDS polyacrylamide gels; one representing an antigen with a mol wt of approximately 45,000, and one representing an antigen with a mol wt of about 12,000. Both may be components of a single protein. All anti-2 and anti-13 precipitates produced a single peak when electrophoresed on SDS polyacrylamide gels. Both the 2 and 13 antigens were found by this technique to have mol wt of approximately 25,000. By molecular weight criteria, as well as by previously investigated distributional criteria, the B antigen is similar to the human LA and Four antigens, and to the mouse D and K antigens, and the 2 and 13 antigens are similar to the mouse Ia antigens.


Temporal control of neuronal diversity: common regulatory principles in insects and vertebrates?
It is well established in species as diverse as insects and mammals that different neuronal and glial subtypes are born at distinct times during central nervous system development. In Drosophila, there is now compelling evidence that individual multipotent neuroblasts express a sequence of progenitor transcription factors which, in turn, regulates the postmitotic transcription factors that specify neuronal/glial temporal identities. Here, we examine the hypothesis that the regulatory principles underlying this mode of temporal specification are shared between insects and mammals, even if some of the factors themselves are not. We also propose a general model for birth-order-dependent neural specification and suggest some experiments to test its validity.


Presurgical cleft lip management: nasal alveolar molding.
Continued advancement in the comprehensive care of children with cleft lip and palate has resulted in numerous innovative treatment protocols. The cooperation of numerous specialties has continued to expand the options for difficult cases. A high standard for aesthetic and functional results has been established. Presurgical treatment of a cleft alveolus and its associated soft tissues with presurgical orthopedics is just one of these techniques. Differing opinions of the effectiveness, growth disturbances, and additional costs of presurgical molding are often debated, but the concepts should be understood by any cleft surgeon or orthodontist involved in cleft care.


Whole Genome Sequencing Defines the Genetic Heterogeneity of Familial Pancreatic Cancer.
Pancreatic cancer is projected to become the second leading cause of cancer-related death in the United States by 2020. A familial aggregation of pancreatic cancer has been established, but the cause of this aggregation in most families is unknown. To determine the genetic basis of susceptibility in these families, we sequenced the germline genomes of 638 patients with familial pancreatic cancer and the tumor exomes of 39 familial pancreatic adenocarcinomas. Our analyses support the role of previously identified familial pancreatic cancer susceptibility genes such as BRCA2, CDKN2A, and ATM, and identify novel candidate genes harboring rare, deleterious germline variants for further characterization. We also show how somatic point mutations that occur during hematopoiesis can affect the interpretation of genome-wide studies of hereditary traits. Our observations have important implications for the etiology of pancreatic cancer and for the identification of susceptibility genes in other common cancer types. The genetic basis of disease susceptibility in the majority of patients with familial pancreatic cancer is unknown. We whole genome sequenced 638 patients with familial pancreatic cancer and demonstrate that the genetic underpinning of inherited pancreatic cancer is highly heterogeneous. This has significant implications for the management of patients with familial pancreatic cancer.


Evaluation of Sclerotinia Stem Rot Resistance in Oilseed Brassica napus Using a Petiole Inoculation Technique Under Greenhouse Conditions.
A petiole inoculation technique was adapted for evaluating resistance of oilseed Brassica napus seedlings to Sclerotinia sclerotiorum. In the first of four experiments, four isolates of S. sclerotiorum were tested, two originating from soybean and two from B. napus. In all, 10 to 47 B. napus accessions were inoculated in the seedling stage and responses to isolates were evaluated using days to wilt (DW) and a lesion phenotype index (LP). There were no significant differences in virulence among the four isolates for DW and only slight differences for LP. However, significant differences (P < 0.0001) were observed among the B. napus accessions for DW and LP in this experiment and in subsequent experiments using one isolate. The responses of accessions were consistent among experiments and among evaluation criteria. Higher levels of resistance were found among winter-type than spring-type accessions, and among rapeseed-quality compared with canola-quality accessions. The most resistant accessions identified also were the most resistant when inoculated at the flowering stage. Terminal stems were inoculated immediately below the lowest flower and stem lesion length (SLL) was used to characterize the interaction phenotype of each accession. The petiole inoculation technique can be used successfully to differentiate oilseed B. napus germ plasm for response to S. sclerotiorum. This inoculation technique and the sources of resistance identified in this study may be used to determine inheritance resistance to S. sclerotiorum and for improving oilseed B. napus cultivars for resistance to this important pathogen.


A recurrent glycine substitution mutation, G2043R, in the type VII collagen gene (COL7A1) in dominant dystrophic epidermolysis bullosa.
Dystrophic epidermolysis bullosa (DEB) is caused by mutations in the type VII collagen gene (COL7A1). Nearly all cases of dominant DEB are caused by glycine substitution mutations occurring within the triple helical region of type VII collagen, and most of the mutations are unique to individual families. In this study, we identified a patient of Hispanic-Mexican origin with a mild form of DEB, which resulted from a de novo dominant glycine substitution, G2043R, in exon 73 of COL7A1. We also investigated a Scottish family with a three-generation pedigree of dominant DEB, in whom the same glycine to arginine substitution mutation was demonstrated. This particular mutation has also been detected previously in three other families with dominant DEB: one Italian, one Hungarian and one Norwegian. Given the widespread geographical distribution of this mutation and the demonstration of its occurrence as a de novo event, G2043R therefore represents the first example of a mutational hotspot in dominant DEB. Interestingly, although both the Mexican and Scottish families we studied had some clinical features in keeping with the Pasini form of the disorder, there was considerable interfamilial variability as well as intrafamilial diversity in the affected individuals.


Sensitization of nanocrystalline TiO2 anchored with pendant catechol functionality using a new tetracyanato ruthenium(II) polypyridyl complex.
We have synthesized a new photoactive ruthenium(II) complex having a pendant catechol functionality (K(2)[Ru(CN)(4)(L)] (1) (L is 4-[2-(4'-methyl-2,2'-bipyridinyl-4-yl)vinyl]benzene-1,2-diol) for studying the dynamics of the interfacial electron transfer between nanoparticulate TiO(2) and the photoexcited states of this Ru(II) complex using femtosecond transient absorption spectroscopy. Steady-state absorption and emission studies revealed that the complex 1 showed a strong solvatochromic behavior in solvents or solvent mixtures of varying polarity. Our steady-state absorption studies further revealed that 1 is bound to TiO(2) surfaces through the catechol functionality, though 1 has two different types of functionalities (catecholate and cyanato) for binding to TiO(2) surfaces. The longer wavelength absorption band tail for 1, bound to TiO(2) through the proposed catecholate functionality, could also be explained on the basis of the DFT calculations. Dynamics of the interfacial electron transfer between 1 and TiO(2) nanoparticles was investigated by studying kinetics at various wavelengths in the visible and near-infrared region. Electron injection to the conduction band of the nanoparticulate TiO(2) was confirmed by detection of the conduction band electron in TiO(2) ([e(-)](TiO(2))(CB)) and cation radical of the adsorbed dye (1(*+)) in real time as monitored by transient absorption spectroscopy. A single exponential and pulse-width limited (<100 fs) electron injection was observed. Back electron transfer dynamics was determined by monitoring the decay kinetics of 1(*+) and [e(-)](TiO(2))(CB). This is the first report on ultrafast ET dynamics on TiO(2) nanoparticle surface using a solvatochromic sensitizer molecule.


AKT1 moderation of cannabis-induced cognitive alterations in psychotic disorder.
Genetic variation in AKT1 may be associated with sensitivity to the psychotomimetic effects of cannabis as well as with increased risk for psychotic disorder following cannabis use. Investigation of the effect of this interaction on relevant intermediate phenotypes for psychosis, such as cognition, may help to clarify the underlying mechanism. Thus, verbal memory (visually presented Word Learning Task), sustained attention (Continuous Performance Test, CPT), AKT1 rs2494732 genotype, and cannabis use were examined in a large cohort of patients with psychotic disorder. No evidence was found for AKT1 × cannabis interaction on verbal memory. Cannabis use preceding onset of psychotic disorder did interact significantly with AKT1 rs2494732 genotype to affect CPT reaction time (β=8.0, SE 3.9, p=0.037) and CPT accuracy (β=-1.2, SE 0.4, p=0.003). Cannabis-using patients with the a priori vulnerability C/C genotype were slower and less accurate on the CPT, whereas cannabis-using patients with the T/T genotype had similar or better performance than non-using patients with psychotic disorder. The interaction was also apparent in patients with psychotic disorder who had not used cannabis in the 12 months preceding assessment, but was absent in the unaffected siblings of these patients and in healthy controls. In conclusion, cannabis use before onset of psychosis may have long-lasting effects on measures of sustained attention, even in the absence of current use, contingent on AKT1 rs2494732 genotype. The results suggest that long-term changes in cognition may mediate the risk-increasing effect of the AKT1 × cannabis interaction on psychotic disorder.


Increased production of reactive oxygen species in hyperglycemic conditions requires dynamic change of mitochondrial morphology.
Increased production of mitochondrial reactive oxygen species (ROS) by hyperglycemia is recognized as a major cause of the clinical complications associated with diabetes and obesity [Brownlee, M. (2001) Nature 414, 813-820]. We observed that dynamic changes in mitochondrial morphology are associated with high glucose-induced overproduction of ROS. Mitochondria undergo rapid fragmentation with a concomitant increase in ROS formation after exposure to high glucose concentrations. Neither ROS increase nor mitochondrial fragmentation was observed after incubation of cells with the nonmetabolizable stereoisomer L-glucose. However, inhibition of mitochondrial pyruvate uptake that blocked ROS increase did not prevent mitochondrial fragmentation in high glucose conditions. Importantly, we found that mitochondrial fragmentation mediated by the fission process is a necessary component for high glucose-induced respiration increase and ROS overproduction. Extended exposure to high glucose conditions, which may mimic untreated diabetic conditions, provoked a periodic and prolonged increase in ROS production concomitant with mitochondrial morphology change. Inhibition of mitochondrial fission prevented periodic fluctuation of ROS production during high glucose exposure. These results indicate that the dynamic change of mitochondrial morphology in high glucose conditions contributes to ROS overproduction and that mitochondrial fission/fusion machinery can be a previously unrecognized target to control acute and chronic production of ROS in hyperglycemia-associated disorders.