Deoxysphingoid bases as plasma markers in diabetes mellitus.
Sphingoid bases are formed from the precursors L-serine and palmitoyl-CoA-a reaction which is catalyzed by the serine-palmitoyltransferase (SPT). SPT metabolizes, besides palmitoyl-CoA also other acyl-CoAs but shows also variability towards the use of other amino acid substrates. The enzyme is also able to metabolize alanine, which results in the formation of an atypical deoxy-sphingoid base (DSB). This promiscuous activity is greatly increased in the case of the sensory neuropathy HSAN1, and pathologically elevated DSB levels have been identified as the cause of this disease. Clinically, HSAN1 shows a pronounced similarity to the diabetic sensory neuropathy (DSN), which is the most common chronic complication of diabetes mellitus. Since serine and alanine metabolism is functionally linked to carbohydrate metabolism by their precursors 3-phosphoglycerate and pyruvate, we were interested to see whether the levels of certain sphingoid base metabolites are altered in patients with diabetes. In a case-control study we compared plasma sphingoid base levels between healthy and diabetic individuals. DSB levels were higher in the diabetic group whereas C16 and C18 sphingoid bases were not significantly different. Plasma serine, but not alanine levels were lower in the diabetic group. A subsequent lipoprotein fractionation showed that the DSBs are primarily present in the LDL and VLDL fraction. Our results suggest that DSBs are a novel category of plasma biomarkers in diabetes which reflect functional impairments of carbohydrate metabolism. Furthermore, elevated DSB levels as we see them in diabetic patients might also contribute to the progression of the diabetic sensory neuropathy, the most frequent complication of diabetes.


Proxies for healthcare need among populations: validation of alternatives--a study in Quebec.
To compare the use of a non-mortality based proxy for relative needs for healthcare among regional populations with a mortality based proxy for population relative needs and to evaluate the additional value of a proxy based on a combination of non-mortality and mortality based proxies. Analysis of cross sectional data on mortality, socioeconomic status, and self assessments of health taken from registrar general records, a population census, and a population health survey. The province of Quebec, Canada. COVERAGE: The populations of the 15 health regions in Quebec. The levels of correlation of indicators based on mortality data, socioeconomic data, and combined data with a standardised indicator of self assessed health. Variations in scores of a proxy based on socioeconomic data among regions explain 37% of the observed variation in self assessed health, 4% more than the level of variation explained by the standardised mortality rate scores. A weighted combination of both mortality and socioeconomic based proxies explains 56% of variation in self assessed health. Justification of "deprivation weights" reflecting variations in socioeconomic status among populations should be based on empirical support concerning the performance of such weights as proxies for relative levels of need among populations. The socioeconomic proxy developed in this study provides a closer correlation to the self assessed health of the populations under study than the mortality based proxy. The superior performance of the combined indicator suggests that the development of social deprivation indicators should be viewed as a complement to, as opposed to a substitute for, mortality based measures in needs based resource allocation exercises.


Experimental demonstration of 10 Gb/s multi-level carrier-less amplitude and phase modulation for short range optical communication systems.
Carrier-less amplitude and phase (CAP) modulation can be a good candidate for short range optical communications for considerable computational complexity reduction and simple system structure. In this paper, a detailed investigation on the digital filters in CAP modulation system is presented. An adaptive equalizer based on cascaded multi-modulus algorithm (CMMA) is used for the demodulation at the receiver. The impact of digital filter taps on system performance is investigated through comprehensive simulations and a 10 Gb/s CAP16 modulation system is demonstrated experimentally. The BER performance for different length of fiber link is measured. Compared with back-to-back (BTB) transmissions, 2 dB and 3.5 dB receiver power penalty are observed at BER of 10(-3) for 20 km and 40 km fiber link respectively. It clearly demonstrates the feasibility of the CAP16 modulation for the short range transmission systems.


Different pattern of chromosomal allele loss in multiple hepatocellular carcinomas as evidence of their multifocal origin.
One of the most problematic aspects of surgery for hepatocellular carcinoma (HCC) is the frequent development of multiple tumors. Determination of the origin of multiple tumors, i.e., multifocal or metastatic, is important for predicting the clinical course of the disease after surgery. In order to clarify the origin of multiple tumors of HCC genetically, we examined patterns of loss of heterozygosity (LOH) on chromosome 16 for DNA isolated from 43 HCCs resected from 19 patients by analysis of restriction fragment length polymorphism. The cases were classified macro- and microscopically into 3 groups: multifocal origin; metastatic origin; and undetermined. Classification based on morphological features was shown to be well correlated with patterns of LOH in multiple tumors of HCC. Different patterns of LOH on chromosome 16 were detected in 8 of 11 patients with tumors of morphologically multifocal origin, whereas they were detected in none of 5 patients with tumors of morphologically metastatic origin. Among five patients with tumors of morphologically undetermined origin, a difference of LOH pattern among the tumors was detected in two, whereas in the other three, the pattern was identical between the tumors. A different pattern of LOH among HCCs arising in situ showed that they were composed of different clones, strongly suggesting their independent clonal origin and multifocal development. These results show that not only appropriate morphological observation but also examination of the LOH pattern on a particular chromosome is useful in diagnosis of multifocal HCC.


Effect of very high-intensity statin therapy on regression of coronary atherosclerosis: the ASTEROID trial.
Prior intravascular ultrasound (IVUS) trials have demonstrated slowing or halting of atherosclerosis progression with statin therapy but have not shown convincing evidence of regression using percent atheroma volume (PAV), the most rigorous IVUS measure of disease progression and regression. To assess whether very intensive statin therapy could regress coronary atherosclerosis as determined by IVUS imaging. Prospective, open-label blinded end-points trial (A Study to Evaluate the Effect of Rosuvastatin on Intravascular Ultrasound-Derived Coronary Atheroma Burden [ASTEROID]) was performed at 53 community and tertiary care centers in the United States, Canada, Europe, and Australia. A motorized IVUS pullback was used to assess coronary atheroma burden at baseline and after 24 months of treatment. Each pair of baseline and follow-up IVUS assessments was analyzed in a blinded fashion. Between November 2002 and October 2003, 507 patients had a baseline IVUS examination and received at least 1 dose of study drug. After 24 months, 349 patients had evaluable serial IVUS examinations. All patients received intensive statin therapy with rosuvastatin, 40 mg/d. Two primary efficacy parameters were prespecified: the change in PAV and the change in nominal atheroma volume in the 10-mm subsegment with the greatest disease severity at baseline. A secondary efficacy variable, change in normalized total atheroma volume for the entire artery, was also prespecified. The mean (SD) baseline low-density lipoprotein cholesterol (LDL-C) level of 130.4 (34.3) mg/dL declined to 60.8 (20.0) mg/dL, a mean reduction of 53.2% (P<.001). Mean (SD) high-density lipoprotein cholesterol (HDL-C) level at baseline was 43.1 (11.1) mg/dL, increasing to 49.0 (12.6) mg/dL, an increase of 14.7% (P<.001). The mean (SD) change in PAV for the entire vessel was -0.98% (3.15%), with a median of -0.79% (97.5% CI, -1.21% to -0.53%) (P<.001 vs baseline). The mean (SD) change in atheroma volume in the most diseased 10-mm subsegment was -6.1 (10.1) mm3, with a median of -5.6 mm3 (97.5% CI, -6.8 to -4.0 mm3) (P<.001 vs baseline). Change in total atheroma volume showed a 6.8% median reduction; with a mean (SD) reduction of -14.7 (25.7) mm3, with a median of -12.5 mm3 (95% CI, -15.1 to -10.5 mm3) (P<.001 vs baseline). Adverse events were infrequent and similar to other statin trials. Very high-intensity statin therapy using rosuvastatin 40 mg/d achieved an average LDL-C of 60.8 mg/dL and increased HDL-C by 14.7%, resulting in significant regression of atherosclerosis for all 3 prespecified IVUS measures of disease burden. Treatment to LDL-C levels below currently accepted guidelines, when accompanied by significant HDL-C increases, can regress atherosclerosis in coronary disease patients. Further studies are needed to determine the effect of the observed changes on clinical outcome. ClinicalTrials.gov Identifier: NCT00240318.


Aspects of silane coupling agents and surface conditioning in dentistry: an overview.
To give an overview of aspects of silane coupling agents and surface conditioning in dentistry. Currently, silane coupling agents are used as adhesion promoters. Silanes are effective in enhancing adhesion between resin composite and silica-based ceramics. They do not bond effectively to non-silica based dental restorative materials. Surface conditioning of non-silica based ceramics with silica coating improves the bonding. This current overview will focus on the silane coupling agents: their properties, limitations in adhesion promotion and the clinical problems with the use of silanes. It will also focus on the current surface conditioning methods as well as new surface conditioning techniques to enhance the bonding through conventional silanization approaches. Several surface conditioning methods are being used clinically to enhance the adhesion of resin composites to non-silica based restorative materials. Other approaches are under investigation. The clinical problem of using silanes in adhesion promotion is the bond degradation over time in oral environment. The current silane coupling agents are not ideal. The current silane coupling agents can fulfill the minimum requirements in clinical practice to enhance the bonding of resin composite to dental restorative materials. Developments of novel surface conditioning methods and silane coupling agents are required to address the bond durability problem.


CHIT1-positive microglia drive motor neuron ageing in the primate spinal cord.
Ageing is a critical factor in spinal-cord-associated disorders1, yet the ageing-specific mechanisms underlying this relationship remain poorly understood. Here, to address this knowledge gap, we combined single-nucleus RNA-sequencing analysis with behavioural and neurophysiological analysis in non-human primates (NHPs). We identified motor neuron senescence and neuroinflammation with microglial hyperactivation as intertwined hallmarks of spinal cord ageing. As an underlying mechanism, we identified a neurotoxic microglial state demarcated by elevated expression of CHIT1 (a secreted mammalian chitinase) specific to the aged spinal cords in NHP and human biopsies. In the aged spinal cord, CHIT1-positive microglia preferentially localize around motor neurons, and they have the ability to trigger senescence, partly by activating SMAD signalling. We further validated the driving role of secreted CHIT1 on MN senescence using multimodal experiments both in vivo, using the NHP spinal cord as a model, and in vitro, using a sophisticated system modelling the human motor-neuron-microenvironment interplay. Moreover, we demonstrated that ascorbic acid, a geroprotective compound, counteracted the pro-senescent effect of CHIT1 and mitigated motor neuron senescence in aged monkeys. Our findings provide the single-cell resolution cellular and molecular landscape of the aged primate spinal cord and identify a new biomarker and intervention target for spinal cord degeneration.


Neuropsychiatric manifestations and their clinical associations in southern Chinese patients with systemic lupus erythematosus.
To study the neuropsychiatric (NP) manifestations in a large cohort of southern Chinese patients with systemic lupus erythematosus (SLE) according to the new 1999 American College of Rheumatology (ACR) case definitions and their clinical associations. Patients with SLE who were followed from 1984 to 2000 were retrospectively reviewed. Patients with NP manifestations were ascertained and classified by at least 2 rheumatologists, with the collaboration of neurologists and psychiatrists. The association of NP manifestations with other clinical features and autoantibodies was studied by statistical analysis. Five hundred eighteen patients with SLE were studied. The female to male ratio was 7.8 to 1 and the mean age of disease onset was 29.5 +/- 12.0 years (range 9-80). The mean duration of followup was 7.3 +/- 6.7 years (range 0.3-23.0). Ninety-six patients (19%) had 133 NP events and the mean number of events per patient-year of followup was 0.035. In decreasing order of frequency. these events were: seizure disorder (28%), cerebrovascular disease (19%), acute confusional state (14%), psychosis (11%), myelopathy (8%), mood disorder (6%), headache (4%), movement disorder (2%), cranial neuropathy (3%), demyelinating syndrome (1.5%), anxiety disorder (1.5%), mononeuritis multiplex/mononeuropathy (1.5%), aseptic meningitis (1%), and polyneuropathy (1%). Cognitive dysfunction was not classified because of the lack of standard neuropsychological testing for every patient. Univariate analysis revealed that NP-SLE was associated with a positive lupus anticoagulant (LAC) (p = 0.001), a strongly positive IgG anticardiolipin (aCL) (p = 0.01). leukopenia (p = 0.01), lymphopenia (p = 0.03), thrombocytopenia (p = 0.03), and pulmonary involvement (p = 0.03). Multivariate analysis showed that a strongly positive IgG aCL [RR 3.1 (1.3-7.7), p = 0.01] and a history of cyclophosphamide treatment [RR 4.3 (2.1-9.0), p < 0.001] were independently associated with NP manifestations in our cohort. Among the NP features, cerebrovascular disorder was particularly associated with the presence of LAC [OR 3.3 (1.4-8.0), p = 0.01] and a strongly positive IgG aCL [OR 3.1 (1.1-8.2), p = 0.031]. The point prevalence of overt NP manifestations in our cohort of patients with SLE was 19%. This percentage was likely higher if subtle cognitive dysfunction was included. Seizure and cerebrovascular disorders were the most common NP features. The presence of antiphospholipid antibodies was significantly associated with NP manifestations, especially cerebrovascular disorders.


Public's Views toward Return of Secondary Results in Genomic Sequencing: It's (Almost) All about the Choice.
The therapeutic use of genomic sequencing creates novel and unresolved questions about cost, clinical efficacy, access, and the disclosure of sequencing results. The disclosure of the secondary results of sequencing poses a particularly challenging ethical problem. Experts disagree about which results should be shared and public input - especially important for the creation of disclosure policies - is complicated by the complex nature of genetics. Recognizing the value of deliberative democratic methods for soliciting informed public opinion on matters like these, we recruited participants from a clinical research site for an all-day deliberative democracy (DD) session. Participants were introduced to the clinical and ethical issues associated with genomic sequencing, after which they discussed the tradeoffs and offered their opinions about policies for the return of secondary results. Participants (n = 66; mean age = 57 (SD = 15); 70% female; 76% white) were divided into 10 small groups (5 to 8 participants each) allowing interactive deliberation on policy options for the return of three categories of secondary results: 1) medically actionable results; 2) risks for adult-onset disorders identified in children; and 3) carrier status for autosomal recessive disorders. In our qualitative analysis of the session transcripts, we found that while participants favored choice and had a preference for making information available, they also acknowledged the risks (and benefits) of learning such information. Our research reveals the nuanced reasoning used by members of the public when weighing the pros and cons of receiving genomic information, enriching our understanding of the findings of surveys of attitudes regarding access to secondary results.


Short-interval severe fire erodes the resilience of subalpine lodgepole pine forests.
Subalpine forests in the northern Rocky Mountains have been resilient to stand-replacing fires that historically burned at 100- to 300-year intervals. Fire intervals are projected to decline drastically as climate warms, and forests that reburn before recovering from previous fire may lose their ability to rebound. We studied recent fires in Greater Yellowstone (Wyoming, United States) and asked whether short-interval (<30 years) stand-replacing fires can erode lodgepole pine (Pinus contorta var. latifolia) forest resilience via increased burn severity, reduced early postfire tree regeneration, reduced carbon stocks, and slower carbon recovery. During 2016, fires reburned young lodgepole pine forests that regenerated after wildfires in 1988 and 2000. During 2017, we sampled 0.25-ha plots in stand-replacing reburns (n = 18) and nearby young forests that did not reburn (n = 9). We also simulated stand development with and without reburns to assess carbon recovery trajectories. Nearly all prefire biomass was combusted ("crown fire plus") in some reburns in which prefire trees were dense and small (≤4-cm basal diameter). Postfire tree seedling density was reduced sixfold relative to the previous (long-interval) fire, and high-density stands (>40,000 stems ha-1) were converted to sparse stands (<1,000 stems ha-1). In reburns, coarse wood biomass and aboveground carbon stocks were reduced by 65 and 62%, respectively, relative to areas that did not reburn. Increased carbon loss plus sparse tree regeneration delayed simulated carbon recovery by >150 years. Forests did not transition to nonforest, but extreme burn severity and reduced tree recovery foreshadow an erosion of forest resilience.


Simple, economical heat-shock devices for zebrafish housing racks.
One reason for the popularity of the zebrafish (Danio rerio) as a model vertebrate is the ability to manipulate gene expression in this organism. A common method is to induce gene expression transiently under control of a heat-shock promoter (e.g., hsp70l). By making simple mechanical adjustments to small aquarium heaters (25-50W), we were able to produce consistent and reliable heat-shock conditions within a conventional zebrafish housing system. Up to two heat-shock intervals per day (>37°C) could be maintained under conditions of continuous flow (5-25 mL/min). Temperature logging every 30 s indicated rapid warm up times, consistent heat-shock lengths, and accurate and precise peak water temperatures (mean±SD=38°C±0.2°C). The biological effects of these heat-shock treatments were confirmed by observing inducible expression of enhanced green fluorescent protein (EGFP) and inhibition of caudal fin regeneration in a transgenic fish line expressing a dominant negative fibroblast growth factor receptor (Tg(hsp70l:dnfgfr1-EGFP)(pd1)). These devices are inexpensive, easily modified, and can be calibrated to accommodate a variety of experimental designs. After setup on a programmable timer, the heaters require no intervention to produce consistent daily heat shocks, and all other standard care protocols can be followed in the fish facility. The simplicity and stability of these devices make them suitable for long-term heat shocks at any stage of the zebrafish lifecycle (>7 days postfertilization), and useful for both laboratory and classroom experiments on transgenic zebrafish.


Echovirus type 6 transmission clusters and the role of environmental surveillance in early warning, the Netherlands, 2007 to 2016.
BackgroundIn the Netherlands, echovirus type 6 (E6) is identified through clinical and environmental enterovirus surveillance (CEVS and EEVS). AimWe aimed to identify E6 transmission clusters and to assess the role of EEVS in surveillance and early warning of E6. MethodsWe included all E6 strains from CEVS and EEVS from 2007 through 2016. CEVS samples were from patients with enterovirus illness. EEVS samples came from sewage water at pre-specified sampling points. E6 strains were defined by partial VP1 sequence, month and 4-digit postcode. Phylogenetic E6 clusters were detected using pairwise genetic distances. We identified transmission clusters using a combined pairwise distance in time, place and phylogeny dimensions. ResultsE6 was identified in 157 of 3,506 CEVS clinical episodes and 92 of 1,067 EEVS samples. Increased E6 circulation was observed in 2009 and from 2014 onwards. Eight phylogenetic clusters were identified; five included both CEVS and EEVS strains. Among these, identification in EEVS did not consistently precede CEVS. One phylogenetic cluster was dominant until 2014, but genetic diversity increased thereafter. Of 14 identified transmission clusters, six included both EEVS and CEVS; in two of them, EEVS identification preceded CEVS identification. Transmission clusters were consistent with phylogenetic clusters, and with previous outbreak reports. ConclusionAlgorithms using combined time-place-phylogeny data allowed identification of clusters not detected by any of these variables alone. EEVS identified strains circulating in the population, but EEVS samples did not systematically precede clinical case surveillance, limiting EEVS usefulness for early warning in a context where E6 is endemic.


Carbon Nanotube Membranes: Synthesis, Properties, and Future Filtration Applications.
Over the course of the past decade, there has been growing interest in the development of different types of membranes composed of carbon nanotubes (CNTs), including buckypapers and composite materials, for an ever-widening range of filtration applications. This article provides an overview of how different types of CNT membranes are prepared and the results obtained from investigations into their suitability for different applications. The latter involve the removal of small particles from air samples, the filtration of aqueous solutions containing organic compounds and/or bacteria, and the separation of individual liquids present in mixtures. A growing number of reports have demonstrated that the incorporation of CNTs into composite membranes confers an improved resistance to fouling caused by biomacromolecules and bacteria. These results are discussed, along with evidence that demonstrates it is possible to further reduce fouling by taking advantage of the inherent conductivity of composite membranes containing CNTs, as well as by using different types of electrochemical stimuli.


The ICN code of ethics for nurses: a time for revision.
Since its development in 1953, the International Council of Nursing's Code of Ethics for Nurses has been revised a number of times and adopted or adapted as a guide for nurses in many countries. The Code serves as a critical model for ethical standards in the nursing profession, stressing responsibility and advocacy for the human rights of patients, their families and communities, as well as social issues, the work of nurses within health care systems and working with other health professionals. The most recent edition of the Code (2012) is now in the process of revision by an international Steering Committee. This policy paper describes the meaning of the Code, as well as the processes in place to develop a revised Code that is better fitted for the current needs of nurses around the globe.


Subtalar distraction arthrodesis through a posterior approach.
Subtalar arthrodesis is a common procedure for treating subtalar arthrosis. An isolated lateral incision can be used as an approach to subtalar fusion alone. However, it can be a difficult approach for correction of varus or valgus hindfoot deformities through the subtalar joint; especially if it involves distraction to correct calcaneal alignment. Less commonly, two-incision or single-incision medial approaches also have been described, but distraction and angular correction can still be difficult. We present an alternative method, using a posterior Achilles tendon-splitting procedure, that gives excellent subtalar exposure and the ability to obtain distraction with angular correction and bone grafting. Six consecutive patients underwent subtalar fusion with a posterior approach. Both varus and valgus deformities secondary to trauma, tendon insufficiency, or congenital deformities were corrected. Distraction was accomplished with laminar spreaders, and fixation was obtained with two 7.3-mm partially threaded cannulated screws. All patients were followed until fusion, and clinical and radiographic results were recorded. No other concomitant procedures were performed. Fusion was observed radiographically in all patients by 3 months postoperatively. No serious wound complications occurred. Correction to slightly valgus alignment (for varus deformity) or neutral alignment (for valgus deformity) was obtained without collapse of autologous bone graft. Talocalcaneal height and all angular measurements were significantly increased (all p < 0.05). The posterior approach for distraction arthrodesis is an alternative to the standard lateral approach technique. It allows safe exposure with the ability to distract while also correcting varus or valgus deformity of the subtalar joint.


Molecule confirmation and structure characterization of pentatriacontatrienyl mycolate in Mycobacterium smegmatis.
Mycobacterium smegmatis is often used to study the different components of mycobacterial cell wall. Mycolic acids are important components of mycobacterial cell wall that have been associated with virulence. Recently, a novel lipid containing mycolic acids has been described in M. smegmatis. However, some uncertainties regarding the structure of this molecule named mycolate ester wax have been reported. The objective of this work was to perform an in depth structural study of this molecule for its precise characterization. Using 1H and 13C NMR spectroscopy, the molecular structure of mycolate ester wax found in M. smegmatis has been elucidated. The characterization was complemented with MS analyses. This molecule is formed by a carbon chain with three methyl substituted olefinic units and a mycolate structure with trans double bonds and cis cyclopropane rings. The present molecular study will facilitate the detection and identification of pentatriacontatrienyl mycolate in future studies by the performance of a simple 1D 1H NMR experiment.


Assessing the environmental impacts of halving food loss and waste along the food supply chain.
Reducing food loss and waste (FLW) is widely recognized as an important lever for lowering the environmental impacts of food systems. The United Nations Sustainable Development Agenda includes a goal to reduce FLW by 50% by 2030. Given differences in resource inputs along the food supply chain (FSC), the environmental benefits of FLW reduction will vary by stage of the FSC. Here, we identify the points along the supply chain where a 50% FLW reduction could yield the largest potential environmental benefits, assuming that decreases in consumption propagate back up the supply chain to reduce production. We use an environmentally extended input-output (EEIO) model combined with data on rates of FLW to calculate the scale of the total environmental impacts of the U.S. food system resulting from lost or wasted food. We evaluate the maximum potential environmental benefit resulting from 50% FLW reduction at all possible combinations of six supply chain stages (agricultural production, food processing, distribution/retail, restaurant foodservice, institutional foodservice, and households). We find that FLW reduction efforts should target the foodservice (restaurant) sector, food processing sector, and household consumption. Halving FLW in the foodservice sector has the highest potential to reduce greenhouse gas output and energy use. Halving FLW in the food processing sector could reduce the most land use and eutrophication potential, and reducing household consumption waste could avert the most water consumption. In contrast, FLW reduction at the retail, institutional foodservice, and farm level averts less environmental impact. Our findings may help determine optimal investment in FLW reduction strategies.


The pharmacology and subacute toxicology of dopamine.
Preclinical studies with dopamine showed a unique spectrum of biological activities which suggested that it might be of therapeutic use in the clinical syndromes of shock and low cardiac output. Most prominent among these were its effects on cardiac output, renal perfusion, and vital organ flow. The unique effect on renal function and the subsequent studies by Goldberg and his colleagues led to the recognition of a previously unknown catecholamine receptor site, the 'dopaminergic receptor'. Studies on the toxicology of dopamine in our laboratories suggested that dopamine could be safely used in the clinic.


Growth of ZnO nanowires on nonwoven polyethylene fibers.
We report the growth of ZnO nanowires on nonwoven polyethylene fibers using a simple hydrothermal method at a temperature below the boiling point of water. The ZnO nanowires were grown from seed ZnO nanoparticles affixed onto the fibers. The seed ZnO nanoparticles, with diameters of about 6-7 nm, were synthesized in isopropanol by reducing zinc acetate hydrate with sodium hydroxide. The growth process was carried out in a sealed chemical bath containing an equimolar solution of zinc nitrate hexahydrate and hexamethylene tetramine at a temperature of 95 °C over a period of up to 20 h. The thickness and length of the nanowires can be controlled by using different concentrations of the starting reactants and growth durations. A 0.5 mM chemical bath yielded nanowires with an average diameter of around 50 nm, while a 25 mM bath resulted in wires with a thickness of up to about 1 μm. The length of the wires depends both on the concentration of the precursor solution as well as the growth duration, and in 20 h, nanowires as long as 10 μm can be grown. The nonwoven mesh of polyethylene fibers covered with ZnO nanowires can be used for novel applications such as water treatment by degrading pollutants by photocatalysis. Photocatalysis tests carried out on standard test contaminants revealed that the polyethylene fibers with ZnO nanowires grown on them could accelerate the photocatalytic degradation process by a factor of 3.


Low Tidal Volumes Are Associated With Slightly Improved Oxygenation in Patients Having Cardiac Surgery: A Cohort Analysis.
Mechanical ventilation with low tidal volumes appears to provide benefit in patients having noncardiac surgery; however, whether it is beneficial in patients having cardiac surgery is unclear. We retrospectively examined patients having elective cardiac surgery requiring cardiopulmonary bypass through a median sternotomy approach who received mechanical ventilation with a single lumen endotracheal tube from January 2010 to mid-August 2016. Time-weighted average tidal volume (milliliter per kilogram predicted body weight [PBW]) during the duration of surgery excluding cardiopulmonary bypass was analyzed. The association between tidal volumes and postoperative oxygenation (measured by arterial partial pressure of oxygen (PaO2)/fraction of inspired oxygen ratio [PaO2/FIO2]), impaired oxygenation (PaO2/FIO2 <300), and clinical outcomes were examined. Of 9359 cardiac surgical patients, larger tidal volumes were associated with slightly worse postoperative oxygenation. Postoperative PaO2/FIO2 decreased an estimated 1.05% per 1 mL/kg PBW increase in tidal volume (97.5% confidence interval [CI], -1.74 to -0.37; PBon = .0005). An increase in intraoperative tidal volumes was also associated with increased odds of impaired oxygenation (odds ratio [OR; 97.5% CI]: 1.08 [1.02-1.14] per 1 mL/kg PBW increase in tidal volume; PBon = .0029), slightly longer intubation time (5% per 1 mL/kg increase in tidal volume (hazard ratio [98.33% CI], 0.95 [0.93-0.98] per 1 mL/kg PBW; PBon < .0001), and increased mortality (OR [98.33% CI], 1.34 [1.06-1.70] per 1 mL/kg PBW increase in tidal volume; PHolm = .0144). An increase in intraoperative tidal volumes was also associated with acute postoperative respiratory failure (OR [98.33% CI], 1.16 [1.03-1.32] per 1 mL/kg PBW increase in tidal volume; PHolm = .0146), but not other pulmonary complications. Lower time-weighted average intraoperative tidal volumes were associated with a very modest improvement in postoperative oxygenation in patients having cardiac surgery.


Lens making for scientific instrumentation in the seventeenth century.
With the invention of the telescope and the microscope early in the 17th century, the production of optical lenses became an important factor in the development of these instruments for scientific observation and investigation. In spite of improvements in equipment and techniques, the obstacles to the production of suitable lenses were not surmounted until the 18th century because of lack of knowledge of the optical properties of lenses, and the difficulties in producing glass of suitable clarity due to primitive grinding and polishing techniques. The early astronomical lenses were produced by means of the primitive equipment of the mirror makers and polishers of pietre dure in Murano and Venice. The first professional apparatus for lens grinding and polishing was developed by Ippolito Francini of Florence, and subsequently improved by Eustachio Divini and Carlo Antonio Manzini. A major advance in the equipment and techniques was made by Giuseppe Campani of Rome in the second half of the century. Other important contributions were made by Christiaan Huygens in Holland and John Marshall in England. Toward the end of the 17th century, craftsmen in England and France made great strides in the improvement of apparatus and techniques for lens grinding and polishing. In spite of them, however, optical workshop practice improved extremely slowly, and it remained virtually unchanged into the 19th century.


Soil response to a 3-year increase in temperature and nitrogen deposition measured in a mature boreal forest using ion-exchange membranes.
The projected increase in atmospheric N deposition and air/soil temperature will likely affect soil nutrient dynamics in boreal ecosystems. The potential effects of these changes on soil ion fluxes were studied in a mature balsam fir stand (Abies balsamea [L.] Mill) in Quebec, Canada that was subjected to 3 years of experimentally increased soil temperature (+4 °C) and increased inorganic N concentration in artificial precipitation (three times the current N concentrations using NH4NO3). Soil element fluxes (NO3, NH4, PO4, K, Ca, Mg, SO4, Al, and Fe) in the organic and upper mineral horizons were monitored using buried ion-exchange membranes (PRS™ probes). While N additions did not affect soil element fluxes, 3 years of soil warming increased the cumulative fluxes of K, Mg, and SO4 in the forest floor by 43, 44, and 79 %, respectively, and Mg, SO4, and Al in the mineral horizon by 29, 66, and 23 %, respectively. We attribute these changes to increased rates of soil organic matter decomposition. Significant interactions of the heating treatment with time were observed for most elements although no clear seasonal patterns emerged. The increase in soil K and Mg in heated plots resulted in a significant but small K increase in balsam fir foliage while no change was observed for Mg. A 6-15 % decrease in foliar Ca content with soil warming could be related to the increase in soil-available Al in heated plots, as Al can interfere with the root uptake of Ca.


Control of excitatory and inhibitory synapse formation by neuroligins.
The normal function of neural networks depends on a delicate balance between excitatory and inhibitory synaptic inputs. Synapse formation is thought to be regulated by bidirectional signaling between pre- and postsynaptic cells. We demonstrate that members of the Neuroligin family promote postsynaptic differentiation in cultured rat hippocampal neurons. Down-regulation of neuroligin isoform expression by RNA interference results in a loss of excitatory and inhibitory synapses. Electrophysiological analysis revealed a predominant reduction of inhibitory synaptic function. Thus, neuroligins control the formation and functional balance of excitatory and inhibitory synapses in hippocampal neurons.


Reframing "participation" and "inclusion" in public health policy and practice to address health inequalities: Evidence from a major resident-led neighbourhood improvement initiative.
There is a need for greater conceptual clarity in place-based initiatives that seek to give residents of disadvantaged neighbourhoods more control over action to address the social determinants of health inequalities at a local level. In this article, we address this issue as it relates to the concepts of participation and inclusion. We draw on qualitative data generated during the first phase of the Communities in Control Study, a longitudinal multisite independent evaluation of the impact of Big Local on the social determinants of health and health inequalities. Big Local is a resident-led area improvement initiative in England, funded by the UK Big Lottery Fund. Initiatives focused on community empowerment are increasingly prominent in public health policy and practice globally. Approaches emphasise the promotion of greater control over decisions and action among individuals, groups, and communities, particularly those living in disadvantaged circumstances. However, when it comes to participation and inclusion in taking action and making decisions, the field is characterised by conceptual confusion. This risks undermining the impact of these initiatives. While participation and inclusion are necessary conditions for empowerment and collective control, they are not necessarily sufficient. Sufficiency requires attention to the breadth of participation (i.e., to inclusion) and to the depth of participation (i.e., the extent to which it is experienced as empowering and ultimately enables the exercise of collective control over decisions and actions). In observing how different Big Local resident-led partnerships across England are tackling the day-to-day challenges of engaging with their communities, we reveal the potential for policy and practice of reframing, and therefore clarifying (to highlight the different roles they have) the concepts of participation and inclusion in terms of depth and breadth.


In vitro and in vivo postmarketing surveillance of valsartan, alone or in combination with amlodipine or hydrochlorothiazide, among Palestinian hypertensive patients.
The objectives of this study were to evaluate the general quality of the most prescribed products of valsartan (VL; alone or in combination) and to evaluate their efficacy and safety among Palestinian population through in vivo postmarketing surveillance. The first part was pharmacopeial quality control assay, including dissolution, disintegration, friability, and weight uniformity for VL. The second part was a 3-month cardiology clinics, observational, postmarketing surveillance pilot study that included 103 hypertensive patients who were prescribed 80 mg or 160 mg of VL as monotherapy or combination therapy. The end points were reduction in blood pressure (BP) and the rate of incidence of adverse effects (AEs) at weeks 4 and 8. According to our quality control tests, all VL products showed high-quality standards according to the international guidelines. A reduction in BP was observed at weeks 4 and 8, and no significant difference was observed between the strengths of 80 mg and 160 mg. Higher BP reduction was observed after the use of combination therapy. Moreover, VL was well tolerated; most of the AEs were of mild-to-moderate intensity. In general, the most frequently reported AEs included headache (17.5%), dizziness (11.75%), and weakness (11.7%). No serious AEs or death cases were reported during the study period. High quality of VL tablet products was used; hence, the observed efficacy and safety results should be related to patient's factors and not due to any product defects or substandard quality. Moreover, VL is an effective treatment for essential hypertension.


Perceptions and Attitudes of Medical Students towards Two Methods of Assessing Practical Anatomy Knowledge.
Traditionally, summative practical examination in anatomy takes the form of 'spotters' consisting of a stream of prosections, radiological images and dissections with pins indicating specific structures. Recently, we have started to administer similar examinations online using the quiz facility in Moodle™ (a free, open-source web application for producing modular internet-based courses) in addition to the traditional format. This paper reports on an investigation into students' perceptions of each assessment environment. Over a 3-year period, practical assessment in anatomy was conducted either in traditional format or online via learning management software called Moodle™. All students exposed to the two examination formats at the College of Medicine & Health Sciences, Sultan Qaboos University, Oman, were divided into two categories: junior (Year 3) and senior (Year 4). An evaluation of their perception of both examination formats was conducted using a self-administered questionnaire consisting of restricted and free response items. More than half of all students expressed a clear preference for the online environment and believed it was more exam-friendly. This preference was higher amongst senior students. Compared to females, male students preferred the online environment. Senior students were less likely to study on cadavers when the examination was conducted online. Specimen quality, ability to manage time, and seating arrangements were major advantages identified by students who preferred the online format. Computer-based practical examinations in anatomy appeared to be generally popular with our students. The students adopted a different approach to study when the exam was conducted online as compared to the traditional 'steeplechase' format.


Fermented Garlic Ameliorates Hypercholesterolemia and Inhibits Platelet Activation.
Dietary cholesterol augments the lipid profile and induces the production and activation of platelets, leading to the development of atherosclerosis with detrimental effects on cardiovascular health. Ethnomedicine and Mediterranean diets are natural and cost-effective approaches against several ailments including cardiovascular diseases. In addition, fermented foods have attracted interest due to their increased nutrient profile and enhanced bioavailability and efficacy. Garlic is known to reduce cholesterol and inhibit platelet activation. Therefore, we examined whether fermented garlic could effectively ameliorate the effects of hypercholesterolemia and platelet functions in rats. Male Sprague-Dawley rats were fed a hypercholesterolemic diet and treated with spirulina and fermented and nonfermented preparations of garlic for one month. Platelet aggregation and granule secretion were assessed to evaluate platelet activation. Analysis of the liver and kidney weights and lipid and enzymatic profiles of the serum and whole blood analysis was performed. The expression levels of SREBP-2, ACAT-2, and HMG-CoA were assessed by RT-PCR, while ACAT-1 and ACAT-2 were assessed by real-time PCR, and histological changes in the liver and adipose tissues were analyzed. Both fermented and nonfermented garlic inhibited platelet aggregation and granule secretion; however, fermented garlic exhibited a greater inhibitory effect. In comparison with nonfermented garlic, fermented garlic significantly reduced liver weight and triglyceride concentrations. Fermented garlic also markedly abrogated the detrimental effects of steatosis on liver and adipose tissues. We conclude that fermented garlic significantly improved the lipid profile and modulated platelet functions, thereby inhibiting atherosclerosis- and platelet-related cardiovascular disorders.


Persistent and transient antibody responses to hepatitis E virus detected by western immunoblot using open reading frame 2 and 3 and glutathione S-transferase fusion proteins.
Recombinant proteins containing amino acid sequences from open reading frame (ORF) 2 and ORF3 of a Chinese strain of hepatitis E virus (HEV) were constructed as fusions with glutathione S-transferase (GST). Stable fusion proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the proteins were transferred to nitrocellulose membranes, and the immobilized proteins were probed with sera from hepatitis E patients from various regions or from rhesus monkeys (Macaca mulatta) experimentally infected with the Chinese strain of HEV. Immunoglobulin G-class antibodies were detected with chemiluminescence and anti-human immunoglobulin G conjugated to horseradish peroxidase. Anti-ORF3 antibodies were detected in most patients and monkeys within 17 days of exposure, but this humoral response declined with time and was usually undetectable by approximately 100 days. Anti-ORF2.1 antibody was usually detected as early as anti-ORF3 but persisted in all animals and many patients, whereas reactivity to the larger GST-ORF2.2 fusion protein was more transient, even though all sequences present in GST-ORF2.1 are present in GST-ORF2.2. Rechallenge of these monkeys with HEV suggested that immunity to reinfection was incomplete, as levels of anti-ORF2.1 (but not anti-ORF2.2) were boosted after each rechallenge. The results demonstrate that the carboxy-terminal region of HEV ORF2 contains epitopes which are recognized by convalescent-phase antibody and are likely to be associated with limited immunity to infection, but these epitopes may be masked when larger portions of ORF2 are expressed as recombinant proteins.


Early detection of sporadic pancreatic cancer: strategic map for innovation--a white paper.
Innovation leading to significant advances in research and subsequent translation to clinical practice is urgently necessary in early detection of sporadic pancreatic cancer. Addressing this need, the Early Detection of Sporadic Pancreatic Cancer Summit Conference was conducted by Kenner Family Research Fund in conjunction with the 2014 American Pancreatic Association and Japan Pancreas Society Meeting. International interdisciplinary scientific representatives engaged in strategic facilitated conversations based on distinct areas of inquiry: Case for Early Detection: Definitions, Detection, Survival, and Challenges; Biomarkers for Early Detection; Imaging; and Collaborative Studies. Ideas generated from the summit have led to the development of a Strategic Map for Innovation built upon 3 components: formation of an international collaborative effort, design of an actionable strategic plan, and implementation of operational standards, research priorities, and first-phase initiatives. Through invested and committed efforts of leading researchers and institutions, philanthropic partners, government agencies, and supportive business entities, this endeavor will change the future of the field and consequently the survival rate of those diagnosed with pancreatic cancer.


Effects of third-order susceptibility in sum frequency generation spectra: a molecular dynamics study in liquid water.
When sum frequency generation (SFG) spectroscopy is applied to charged solid/liquid interfaces, the observed SFG signals include both the second-order and third-order polarizations. The latter is called the χ(3) effect, which mainly includes induced molecular orientation by electric fields at charged interfaces. We theoretically evaluate the χ(3) effect on the SFG spectroscopy of liquid water using molecular dynamics (MD) simulations. The MD simulations enable us to definitely calculate the χ(3) susceptibility as a bulk property, and thereby separating it from the usual χ(2) effect shown in the SFG spectra. The calculated results of χ(3) for liquid water are fairly consistent with the experimental estimates. The present finding is utilized to analyze the spectral change of SFG at the air/water interface under electric fields and at the charged silica/water interface. The present analysis of the spectral changes allows for distinguishing the intrinsic change of the interface structure and the χ(3) effect from bulk liquid.


Improvement in sensitivity of screening mammography with computer-aided detection: a multiinstitutional trial.
Our study evaluated radiologist detection of breast cancer using a computer-aided detection system. Three radiologists reviewed 377 screening mammograms interpreted as showing normal or benign findings 9-24 months before cancer diagnosis from 17 of the 18 participating centers. In 313 cases, study radiologists recommended additional mammographic evaluation. In 177 cases, the area warranting additional workup precisely correlated with the subsequently diagnosed cancer. These 177 missed cancers were evaluated with computer-aided detection. The proportion of radiologists identifying the missed cancers was used to determine radiologist sensitivity without computer-aided detection. The study radiologists determined that 123 of the 377 missed cancer cases warranted workup. Therefore, 123 additional cancers cases could have been found. The calculated radiologist sensitivity without computer-aided detection was therefore 75.4% (377 / [377 + 123]). Similarly, using the performance of the system on the missed cancers, we estimated that 80 (65.0%) of these 123 missed cancer cases would have been identified with the use of computer-aided detection. Consequently, the estimated sensitivity of radiologists using computer-aided detection was 91.4% ([377 + 80] / [377 + 123])-resulting in a 21.2% ([91.4% / 75.4%] - 1) increase in radiologist sensitivity with computer-aided detection. Use of the computer-aided detection system significantly improved the detection of breast cancer by increasing radiologist sensitivity by 21.2%. Therefore, for every 100,000 women with breast cancer identified without the use of computer-aided detection, an estimated additional 21,200 cancers would be found with the use of computer-aided detection.


Asymmetric histone modifications between the original and derived loci of human segmental duplications.
Sequencing and annotation of several mammalian genomes have revealed that segmental duplications are a common architectural feature of primate genomes; in fact, about 5% of the human genome is composed of large blocks of interspersed segmental duplications. These segmental duplications have been implicated in genomic copy-number variation, gene novelty, and various genomic disorders. However, the molecular processes involved in the evolution and regulation of duplicated sequences remain largely unexplored. In this study, the profile of about 20 histone modifications within human segmental duplications was characterized using high-resolution, genome-wide data derived from a ChIP-Seq study. The analysis demonstrates that derivative loci of segmental duplications often differ significantly from the original with respect to many histone methylations. Further investigation showed that genes are present three times more frequently in the original than in the derivative, whereas pseudogenes exhibit the opposite trend. These asymmetries tend to increase with the age of segmental duplications. The uneven distribution of genes and pseudogenes does not, however, fully account for the asymmetry in the profile of histone modifications. The first systematic analysis of histone modifications between segmental duplications demonstrates that two seemingly 'identical' genomic copies are distinct in their epigenomic properties. Results here suggest that local chromatin environments may be implicated in the discrimination of derived copies of segmental duplications from their originals, leading to a biased pseudogenization of the new duplicates. The data also indicate that further exploration of the interactions between histone modification and sequence degeneration is necessary in order to understand the divergence of duplicated sequences.


Stress-induced DNA methylation changes and their heritability in asexual dandelions.
*DNA methylation can cause heritable phenotypic modifications in the absence of changes in DNA sequence. Environmental stresses can trigger methylation changes and this may have evolutionary consequences, even in the absence of sequence variation. However, it remains largely unknown to what extent environmentally induced methylation changes are transmitted to offspring, and whether observed methylation variation is truly independent or a downstream consequence of genetic variation between individuals. *Genetically identical apomictic dandelion (Taraxacum officinale) plants were exposed to different ecological stresses, and apomictic offspring were raised in a common unstressed environment. We used methylation-sensitive amplified fragment length polymorphism markers to screen genome-wide methylation alterations triggered by stress treatments and to assess the heritability of induced changes. *Various stresses, most notably chemical induction of herbivore and pathogen defenses, triggered considerable methylation variation throughout the genome. Many modifications were faithfully transmitted to offspring. Stresses caused some epigenetic divergence between treatment and controls, but also increased epigenetic variation among plants within treatments. *These results show the following. First, stress-induced methylation changes are common and are mostly heritable. Second, sequence-independent, autonomous methylation variation is readily generated. This highlights the potential of epigenetic inheritance to play an independent role in evolutionary processes, which is superimposed on the system of genetic inheritance.


Acceptability of reminder letters for Papanicolaou tests: a survey of women from 23 Family Health Networks in Ontario.
To explore women's perspectives on the acceptability and content of reminder letters from the family physician for Papanicolaou (Pap) test screening and the effect of reminder letters on compliance with screening recommendations. A population-based survey was conducted in 23 Family Health Networks and Primary Care Networks participating in a demonstration project to increase the delivery of preventive services in Ontario. Questionnaires were mailed to randomly selected women aged 35 to 69 years who had received a reminder letter for a Pap test from their family physician within the previous six months. Two focus groups were conducted with a volunteer sample of respondents. The usable response rate was 54.3% (406/748). Two-thirds (65.8%, 267/406) of women who completed the survey recalled receiving the reminder letter. Overall, 52.3% (212/405) reported having a Pap test in the past six months. Among women who recalled the reminder letter and scheduled or had a Pap test, 71.4% (125/175) reported that the letter influenced their decision to be screened. The majority of respondents (80.8%, 328/406) wanted to continue to receive reminder letters for Pap tests from their physician, and 34.5% (140/406) wanted to receive additional information about cervical screening. Focus group interviews indicated that women who have had a Pap test may still be unsure about screening recommendations, what the test detects, and the rationale for follow-up procedures. Reminder letters in family practice were viewed as useful and influenced women's decisions to undergo Pap test screening. Women who have had a Pap test may still need additional information about the test.


Addressing Research Bottlenecks to Crop Productivity.
Asymmetry of investment in crop research leads to knowledge gaps and lost opportunities to accelerate genetic gain through identifying new sources and combinations of traits and alleles. On the basis of consultation with scientists from most major seed companies, we identified several research areas with three common features: (i) relatively underrepresented in the literature; (ii) high probability of boosting productivity in a wide range of crops and environments; and (iii) could be researched in 'precompetitive' space, leveraging previous knowledge, and thereby improving models that guide crop breeding and management decisions. Areas identified included research into hormones, recombination, respiration, roots, and source-sink, which, along with new opportunities in phenomics, genomics, and bioinformatics, make it more feasible to explore crop genetic resources and improve breeding strategies.


Hypomethylation of CYP11B2 in Aldosterone-Producing Adenoma.
The purpose of this study was to evaluate the DNA methylation levels of steroidogenic enzyme genes in aldosterone-producing adenoma (APA) and the effects of gene mutations in APA on the DNA methylation levels. DNA methylation array analysis was conducted using nonfunctioning adrenocortical adenoma (n=12) and APA (n=35) samples, including some with a KCNJ5 mutation (n=21), an ATP1A1 mutation (n=5), and without the known mutations (n=9). The quantitative polymerase chain reaction assay was performed for the detection of CYP11B2 and CYP11B1 expression levels in nonfunctioning adrenocortical adenoma and APA. We introduced the KCNJ5 T158A mutation using lentivirus delivery in the human adrenocortical 15 cell line, and analyzed the effects of the mutation on DNA methylation levels. We analyzed the 83 presumed DNA methylation sites of steroidogenic enzymes. In APA, we found 7 hypomethylated sites in CYP11B2 and 1 hypomethylated and 6 hypermethylated sites in CYP11B1 There were no differences in the steroidogenic enzymes gene DNA methylation of peripheral leukocytes between nonfunctioning adrenocortical adenoma and APA. No CYP11B2 methylation level was associated with CYP11B2 transcription levels in APA. All methylation sites, except for a CYP11B2 region, showed no difference among APAs with or without gene mutations. Human adrenocortical 15 cells with the KCNJ5 mutation showed no changes in CYP11B2 or CYP11B1 methylation levels compared with control cells. We demonstrated that CYP11B2 in APA was extensively hypomethylated, and CYP11B2 methylation in the region with hypomethylation was not induced by KCNJ5 or ATP1A1 mutations that cause aldosterone overproduction in APA and a KCNJ5 mutation human adrenocortical 15 cells.


mtDNA affinities of the peoples of North-Central Mexico.
mtDNA haplotypes of representatives of the cosmopolitan peoples of north-central Mexico were studied. Two hundred twenty-three samples from individuals residing in vicinities of two localities in north-central Mexico were analyzed. A combination of strategies was employed to identify the origin of each haplotype, including length variation analysis of the COII and tRNALYS intergenic region, nucleotide sequence analysis of control region hypervariable segment 1, and RFLP analysis of PCR products spanning diagnostic sites. Analysis of these data revealed that the majority of the mtDNA haplotypes were of Native American origin, belonging to one of four primary Native American haplogroups. Others were of European or African origin, and the frequency of African haplotypes was equivalent to that of haplotypes of European derivation. These results provide diagnostic, discrete character, molecular genetic evidence that, together with results of previous studies of classical genetic systems, is informative with regard to both the magnitude of African admixture and the relative maternal contribution of African, European, and Native American peoples to the genetic heritage of Mexico. Phylogenetic analysis revealed that African sequences formed a basal, paraphyletic group.


Experimental radiotherapy of murine lymphoma with 131I-labeled anti-Thy 1.1 monoclonal antibody.
Monoclonal antibodies against the Thy 1.1 differentiation antigen are ineffective in the treatment of transplanted AKR T-cell lymphoma once a palpable tumor nodule is present, due to the inability of the host to eliminate antibody-coated tumor cells. To overcome this limitation, we have evaluated the use of 131I-labeled anti-Thy 1.1 antibodies for the therapy of established AKR/J SL2 lymphoma (Thy 1.1+) nodules growing in congeneic AKR/Cu mice (Thy 1.2+). In these experiments, 131I-anti-Thy 1.1 antibody specifically localized to a s.c. tumor with a mean of 6.5% of the infused dose per g of tumor at 24 h after infusion. The proportion of infused anti-Thy 1.1 antibody localizing to tumor was constant following antibody doses of up to 400 micrograms/animal. Antibody iodinated with up to 2 atoms of iodine per antibody of molecule maintained binding activity and localization to tumor equivalent to antibody labeled with less iodine. The concentrations of 131I-anti-Thy 1.1 in tumor would result in delivery of a mean of 1600 cGy to tumor following infusion of 500 muCi of 131I-labeled anti-Thy 1.1 antibody. In comparison, 500 muCi 131I-labeled irrelevant antibody would deliver a mean of 380 cGy to tumor. Treatment of animals with palpable tumor nodules with 500 muCi 131I-anti-Thy 1.1 led to regression of the tumor nodule in 44% of animals, significantly prolonged survival, and cured two of five of the animals treated prior to the development of metastatic disease. In contrast, unlabeled anti-Thy 1.1 led to tumor response in 6% of animals, and up to 1000 muCi 131I-labeled irrelevant antibody had no effect on tumor growth. Therapy was limited by the emergence of variant tumor cells lacking the target antigen and by bone marrow toxicity following 131I-labeled antibody doses of greater than or equal to 1000 muCi/animal. These studies demonstrate that 131I-labeled monoclonal antibodies can have a significant antitumor effect in a situation where unmodified antibody is ineffective.


A wellness program for individuals with disabilities: Using a student wellness coach approach.
Individuals with disabilities are at higher risk of health conditions; thus, there is a need to provide hands-on opportunities for pre-healthcare professionals to interact with individual with disabilities as well as deliver wellness services to this population. Examine the feasibility and effectiveness of a student-led wellness program for individuals with disabilities. Thirty-two undergraduate student wellness coaches between the ages of 19-23 years, and fifteen participants with disabilities, ranging in ages from 28 to 74 years were included in this study. Every participant was assigned to at least 1 student wellness coach with the purpose of establishing an individualized wellness plan. After 3 months (fall 2013 academic semester), all wellness coaches demonstrated improved clinical interaction and confidence toward working with the participants. The participants had an average weight loss of 2.0 ± 2.9 kg, ranging from 0.0 to 9.0 kg. All participants had improved functionality and fitness and reported high satisfaction toward the program. This study demonstrated the impact of a unique program on the education of pre-healthcare professionals and the overall wellness of participants with disabilities. The program model has the potential to provide clinical health education among pre-healthcare professionals through interacting with individuals with disabilities.


Wound healing in pre-tibial injuries--an observation study.
Pre-tibial lacerations are complex wounds affecting a primarily aged population, with poor healing and a potentially significant impact on social well-being. Management of these wounds has changed little in 20 years, despite significant advances in wound care. A retrospective observational study was undertaken to observe current wound care practice and to assess the effect of various medical factors on wound healing time on 24 elderly patients throughout their wound journey. Wound length was found to be substantively and significantly associated with wound healing time, with a reduction in instantaneous healing rate of about 30% for every increase of 1 cm in wound length. Hence, longer wounds are associated with longer wound healing times. Prescription of several categories of drugs, including those for ischaemic heart disease (IHD), hypertension, respiratory disease or asthma; and the age of the patient were not significantly associated with wound healing times, although substantive significance could be inferred in the case of prescription for IHD and asthma. Despite the small sample size, this study identified a clear association between healing and length of wound. Neither the comorbidities nor prescriptions explored showed any significant association although some seem to be more prevalent in this patient group. The study also highlighted other issues that require further exploration including the social and economic impact of these wounds.


The relationship between temperament and character and psychotic-like experiences in healthy children and adolescents.
Prior work by our group identified personality profiles associated with psychotic-like experiences (PLE's) in healthy adults that were strikingly similar to those found in schizophrenia patients, with the exception of two key differences. Specifically, higher levels of PLE's were linked to higher persistence and cooperativeness, suggesting that these characteristics might represent personality-based resilience factors. Notably, age and personality were significantly correlated in these data, raising questions about whether healthy children and adolescents would show similar results. To date, no study has examined personality profiles associated with both positive and negative PLE's in healthy children and adolescents using Cloninger's Temperament and Character Inventory (TCI). Thus, this study examined the relationship between TCI dimensions and PLE's in healthy children and adolescents. The TCI and the Community Assessment of Psychic Experiences (CAPE) were administered to 123 healthy children and adolescents aged 8-18. Multiple regression models were used to examine personality dimensions associated with overall severity of PLE's as well as severity of positive and negative PLE's separately. Positive, negative, and overall PLE severity were all associated with a personality pattern of higher harm avoidance and lower self-directedness. Negative PLE severity was also associated with lower persistence. Personality correlates of PLE's in healthy children and adolescents were largely consistent with our past work on PLE's in healthy adults. However, our previously identified resilience factors were notably absent in this sample. These findings may suggest that these personality characteristics have not yet crystallized or emerged to aid in coping with PLE's.


The association of serum vitamin D concentration with serious complications after noncardiac surgery.
Vitamin D deficiency is a global health problem. Epidemiological studies demonstrate that vitamin D is both cardioprotective and neuroprotective. Vitamin D also plays a substantial role in innate and acquired immunity. Our goal was to evaluate the association of serum vitamin D concentration on serious postoperative complications and death in noncardiac surgical patients. We retrospectively analyzed the data of 3509 patients who had noncardiac surgery at the Cleveland Clinic Main Campus and had a serum vitamin D measurement. The relationship between serum vitamin D concentration and all-cause in-hospital mortality, in-hospital cardiovascular morbidity, and serious in-hospital infections was assessed as a common effect odds ratio (OR) by using a multivariate generalized estimating equation model with adjustment for demographic, medical history variables, and type and duration of surgery. Higher vitamin D concentrations were associated with decreased odds of in-hospital mortality/morbidity (P = 0.003). There was a linear reduction of the corresponding common effect odds ratio (OR 0.93, 95% confidence interval, 0.88-0.97) for severe in-hospital outcomes for each 5 ng/mL increase in vitamin D concentration over the range from 4 to 44 ng/mL. In addition, we found that the odds versus patients with vitamin D <13 ng/mL (i.e., 1st quintile) were significantly lower in patients with vitamin D 13-20, 20-27, 27-36, and > 36 ng/mL (i.e., 2nd-5th quintiles); the corresponding estimated ORs were 0.65 (99% confidence interval, 0.43-0.98), 0.53 (0.35-0.80), 0.44 (0.28-0.70), and 0.49 (0.31-0.78), respectively. However, there was no statistically significant difference among individual quintiles >13 ng/mL. Vitamin D concentrations were associated with a composite of in-hospital death, serious infections, and serious cardiovascular events in patients recovering from noncardiac surgery. While causality cannot be determined from our retrospective analysis, the association suggests that a large randomized trial of preoperative vitamin D supplementation and postoperative outcomes is warranted.


mtDNA from hair and nail clarifies the genetic relationship of the 15th century Qilakitsoq Inuit mummies.
The 15th century Inuit mummies excavated at Qilakitsoq in Greenland in 1978 were exceptionally well preserved and represent the largest find of naturally mummified specimens from the Arctic. The estimated ages of the individuals, their distribution between two adjacent graves, the results of tissue typing, and incomplete STR results led researchers to conclude that the eight mummies formed two distinct family groups: A grandmother (I/5), two daughters (I/3, I/4), and their two children (I/1, I/2) in one grave, and two sisters (II/6, II/8) and a daughter (II/7) of one of them in the other. Using mtDNA from hair and nail, we have reanalyzed the mummies. The results allowed the unambiguous assignment of each of the mummies to one of three mtDNA haplogroups: A2b (I/5); A2a (I/2, I/3, II/6, II/8); A2a-311 (I/1, I/4, II/7), excluded some of the previous relations, and pointed to new ones. I/5 is not the grandmother/mother of the individuals in Grave I, and she is not maternally related to any of the seven other mummies; I/3 and I/4 are not sisters and II/7 is neither the daughter of II/6 nor of II/8. However, I/1 may be the child of either I/4 or II/7 and these two may be sisters. I/2 may be the son of I/3, who may be the daughter of either II/6 or II/8, and these two may be sisters. The observation of haplogroups A2a and A2b amongst the 550-year-old Inuit puts a lower limit on the age of the two lineages in Greenland.


Gradient-index filters: conversion into a two-index solution by taking into account dispersion.
The possibility of converting the continuous refractive-index profile into a two-index solution is shown. The technique of conversion that is developed makes it possible to take dispersion into account. This ensures that it is possible to achieve a good agreement between theory and practice in a broad spectral range. Several gradient-index filters have been developed and produced, and measuring results are presented.


Randomised controlled trial of GM-CSF in critically ill patients with impaired neutrophil phagocytosis.
Critically ill patients with impaired neutrophil phagocytosis have significantly increased risk of nosocomial infection. Granulocyte-macrophage colony-stimulating factor (GM-CSF) improves phagocytosis by neutrophils ex vivo. This study tested the hypothesis that GM-CSF improves neutrophil phagocytosis in critically ill patients in whom phagocytosis is known to be impaired. This was a multicentre, phase IIa randomised, placebo-controlled clinical trial. Using a personalised medicine approach, only critically ill patients with impaired neutrophil phagocytosis were included. Patients were randomised 1:1 to subcutaneous GM-CSF (3 μg/kg/day) or placebo, once daily for 4 days. The primary outcome measure was neutrophil phagocytosis 2 days after initiation of GM-CSF. Secondary outcomes included neutrophil phagocytosis over time, neutrophil functions other than phagocytosis, monocyte HLA-DR expression and safety. Thirty-eight patients were recruited from five intensive care units (17 randomised to GM-CSF). Mean neutrophil phagocytosis at day 2 was 57.2% (SD 13.2%) in the GM-CSF group and 49.8% (13.4%) in the placebo group, p=0.73. The proportion of patients with neutrophil phagocytosis≥50% at day 2, and monocyte HLA-DR, appeared significantly higher in the GM-CSF group. Neutrophil functions other than phagocytosis did not appear significantly different between the groups. The most common adverse event associated with GM-CSF was fever. GM-CSF did not improve mean neutrophil phagocytosis at day 2, but was safe and appeared to increase the proportion of patients with adequate phagocytosis. The study suggests proof of principle for a pharmacological effect on neutrophil function in a subset of critically ill patients.


Anthropometric and immunological success of antiretroviral therapy and prediction of virological success in west African adults.
The 6 month assessment of the response to antiretroviral therapy (ART) is a critical step. In sub-Saharan Africa, few people have access to plasma viral-load measurement. We assessed the gain or loss in body mass index (BMI), alone or in combination with the gain or loss in CD4+ T-cell count (CD4), as a tool for predicting the response to ART. In a cohort of 622 adults in Abidjan, Côte d'Ivoire, we calculated the sensitivity, specificity and predictive values of BMI and CD4 for treatment success defined as viral-load undetectability (< 300 copies/ml) as gold standard. After 6 months of ART, the median change in BMI was an increase of 1.0 kg/m(2) (interquartile range, IQR: 0.0-2.1), the median change in CD4 an increase of 148/ml (IQR: 54-230) and 84% of patients reached viral-load undetectability. The distribution of change in BMI was similar among patients who reached undetectability and those who did not (increases of 1.06 kg/m(2) versus 0.99 kg/m(2), P = 0.51). With larger changes in BMI, the specificity for treatment success increased but its sensitivity decreased and its positive predictive value was stable around 85%. All results remained similar when combining changes in BMI with those in CD4 and when stratifying by groups of baseline BMI or CD4. In settings where viral-load measurement is not available, a high BMI gain does not reflect virological success, even when combined with a high CD4 gain. In our population, most patients with detectable viral-load had probably adhered to the drug regimen sufficiently to reach significant gains in body mass and CD4 count but had adhered insufficiently to reach viral suppression.


Structure and function of RGD peptides derived from disintegrin proteins.
The Arg-Gly-Asp (RGD) sequence serves as the primary recognition site in extracellular matrix proteins, and peptides containing this sequence can mimic the biological activities of matrix proteins. We have initiated structure-function studies of two RGD containing peptides, RGD-5(AGGDD) and cyclic RGD-6(CARGDDC). Assays have shown that cyclic RGD-peptides inhibit platelet aggregation more efficiently than linear ones. NMR data revealed that RGD-5 and RGD-6 have entirely different conformation. RGD-5 has a linear extended structure and RGD-6 has a stable loop conformation. In RGD-5 the guanidinium group of Arg2 and the carboxyl group of Asp4 lie in parallel, whereas the side-chains of Arg3 and Asp5 of RGD-6 are located in different planes, supporting the idea that the stability of the cyclic form derives from the packing of the side chain of the Arg and Asp residues. The structural features of these peptides could provide a basis for designing new drugs against diseases related to platelet aggregation and as cancer antagonists.


Biochemical and endocrine responses to impact and collision during elite Rugby League match play.
The purpose of this study was to investigate the relationship between the prematch and short-term postmatch biochemical and endocrine responses to the intensity, number, and distribution of impacts associated with collisions during elite Rugby League match play. Seventeen elite male Rugby League players each provided blood and saliva samples 24 hours prematch, 30 minutes prematch, 30 minutes postmatch, and then at 24-hour intervals for a period of 5 days postmatch to determine plasma creatine kinase concentration ([CK]) and salivary cortisol concentration ([sCort]). The intensity, number, and distribution of impact forces experienced by players during match play were recorded using portable global positioning systems (GPSs). The change in the dependent variables at each sample collection time was compared to 24 hours prematch and 30-minute prematch measures. The [CK] and [sCort] increased significantly (p < 0.05) during match play. Significant correlations (p < 0.05) were observed between the number of hit-ups and peak [CK] 24 hours postmatch, 48 hours postmatch, and 72 hours postmatch (p < 0.05). The number of impacts recorded in zone 5 (8.1-10.0G) and zone 6 (>10.1G) during match play was significantly correlated (p < 0.05) to [CK] 30 minutes postmatch, 24 hours post, 48 hours post, and 72 hours postmatch. The GPS was able to provide data on the intensity, number, and distribution of impacts resulting from collisions during match play. Elite Rugby League match play resulted in significant skeletal muscle damage and was highly dependent on the number of heavy collisions >8.1G. [CK] remained elevated 120 hours postmatch identifying that at least 5 days modified activity is required to achieve full recovery after elite Rugby League match play.


Supporting employees' work-family needs improves health care quality: Longitudinal evidence from long-term care.
We analyzed qualitative and quantitative data from U.S.-based employees in 30 long-term care facilities. Analysis of semi-structured interviews from 154 managers informed quantitative analyses. Quantitative data include 1214 employees' scoring of their supervisors and their organizations on family supportiveness (individual scores and aggregated to facility level), and three outcomes: (1), care quality indicators assessed at facility level (n = 30) and collected monthly for six months after employees' data collection; (2), employees' dichotomous survey response on having additional off-site jobs; and (3), proportion of employees with additional jobs at each facility. Thematic analyses revealed that managers operate within the constraints of an industry that simultaneously: (a) employs low-wage employees with multiple work-family challenges, and (b) has firmly institutionalized goals of prioritizing quality of care and minimizing labor costs. Managers universally described providing work-family support and prioritizing care quality as antithetical to each other. Concerns surfaced that family-supportiveness encouraged employees to work additional jobs off-site, compromising care quality. Multivariable linear regression analysis of facility-level data revealed that higher family-supportive supervision was associated with significant decreases in residents' incidence of all pressure ulcers (-2.62%) and other injuries (-9.79%). Higher family-supportive organizational climate was associated with significant decreases in all falls (-17.94%) and falls with injuries (-7.57%). Managers' concerns about additional jobs were not entirely unwarranted: multivariable logistic regression of employee-level data revealed that among employees with children, having family-supportive supervision was associated with significantly higher likelihood of additional off-site jobs (RR 1.46, 95%CI 1.08-1.99), but family-supportive organizational climate was associated with lower likelihood (RR 0.76, 95%CI 0.59-0.99). However, proportion of workers with additional off-site jobs did not significantly predict care quality at facility levels. Although managers perceived providing work-family support and ensuring high care quality as conflicting goals, results suggest that family-supportiveness is associated with better care quality.


Roles of alpha-toxin and beta-toxin in virulence of Staphylococcus aureus for the mouse mammary gland.
Mutants of Staphylococcus aureus which fail to express alpha-toxin (Hly), beta-toxin (Hlb), or both have been constructed by site-specific mutagenesis. The virulence of the mutants was compared with that of wild-type toxigenic strains by intramammary inoculation of lactating mice. A bovine strain, M60, and a laboratory strain, 8325-4, caused acute mastitis and death within 48 h for 60% of the mice inoculated. Animals inoculated with Hly mutants also developed acute mastitis, but no deaths occurred. Comparisons of Hly- or Hlb-positive strains with the double mutation Hly Hlb showed that both toxins led to a significantly higher recovery of S. aureus from the gland 48 h postinfection. Histopathological examination of mammary glands showed that phagocytosis of bacteria occurred irrespective of toxigenicity, but toxigenic strains, particularly those which were Hly+, continued to multiply, invaded the interalveolar tissues, and produced severe lesions. Stimulation of an inflammatory response by inoculation of the mammary gland with endotoxin prior to challenge with S. aureus reduced recovery of the bacteria 10- to 100-fold and, under these conditions, neither alpha-toxin nor beta-toxin contributed significantly to growth and survival.


Qualitative grading of aortic regurgitation: a pilot study comparing CMR 4D flow and echocardiography.
Over the past 10 years there has been intense research in the development of volumetric visualization of intracardiac flow by cardiac magnetic resonance (CMR).This volumetric time resolved technique called CMR 4D flow imaging has several advantages over standard CMR. It offers anatomical, functional and flow information in a single free-breathing, ten-minute acquisition. However, the data obtained is large and its processing requires dedicated software. We evaluated a cloud-based application package that combines volumetric data correction and visualization of CMR 4D flow data, and assessed its accuracy for the detection and grading of aortic valve regurgitation using transthoracic echocardiography as reference. Between June 2014 and January 2015, patients planned for clinical CMR were consecutively approached to undergo the supplementary CMR 4D flow acquisition. Fifty four patients(median age 39 years, 32 males) were included. Detection and grading of the aortic valve regurgitation using CMR4D flow imaging were evaluated against transthoracic echocardiography. The agreement between 4D flow CMR and transthoracic echocardiography for grading of aortic valve regurgitation was good (j = 0.73). To identify relevant,more than mild aortic valve regurgitation, CMR 4D flow imaging had a sensitivity of 100 % and specificity of 98 %. Aortic regurgitation can be well visualized, in a similar manner as transthoracic echocardiography, when using CMR 4D flow imaging.


Extensive sympatry, cryptic diversity and introgression throughout the geographic distribution of two coral species complexes.
The identification of species is one of the most basic, and yet critically important, issues in biology with far-reaching potential implications for fields such as biodiversity conservation, population ecology and epidemiology. Morphology has long been the primary tool biologists have used to categorize life. However, we now know that a significant portion of natural diversity is morphologically hidden, and therefore, we must integrate nonmorphological tools into the description of biodiversity. Here, we demonstrate the utility of multilocus population genetic data for identifying and characterizing cryptic species complexes, even when species share large amounts of genetic variability. Specifically, we have used DNA sequence data from 12 genomic regions to characterize two widespread species complexes in the coral genus Acropora: A. cytherea and A. hyacinthus. These two morphospecies have each been sampled from 5 to 7 locations throughout their Indo-Pacific distributions, and with the use of structure and hierarchical clustering, we demonstrate the presence of at least six widespread cryptic species within these two morphospecies complexes. After identifying cryptic lineages, we then utilize the genetic data to examine the history of introgressive hybridization within and between these morphospecies complexes. Our data indicate that these two complexes form a global syngameon with consistent patterns of introgression between species across large geographic distributions.


Late Holocene environmental change and anthropogenic: Ecosystem interaction on the Laikipia Plateau, Kenya.
East African ecosystems have been shaped by long-term socio-ecological-environmental interactions. Although much previous work on human-environment interrelationships have emphasised the negative impacts of human interventions, a growing body of work shows that there have also often been strong beneficial connections between people and ecosystems, especially in savanna environments. However, limited information and understanding of past interactions between humans and ecosystems of periods longer than a century hampers effective management of contemporary environments. Here, we present a late Holocene study of pollen, fern spore, fungal spore, and charcoal analyses from radiocarbon-dated sediment sequences and assess this record against archaeological and historical data to describe socio-ecological changes on the Laikipia Plateau in Rift Valley Province, Kenya. The results suggest a landscape characterised by closed forests between 2268 years before present (cal year BP) and 1615 cal year BP when there was a significant change to a more open woodland/grassland mosaic that continues to prevail across the study area. Increased amounts of charcoal in the sediment are observed for this same period, becoming particularly common from around 900 cal year BP associated with fungal spores commonly linked to the presence of herbivores. It is likely these trends reflect changes in land use management as pastoral populations improved and extended pasture, using fire to eradicate disease-prone habitats. Implications for contemporary land use management are discussed in the light of these findings.


Etelcalcetide in Patients on Hemodialysis with Severe Secondary Hyperparathyroidism. Multicenter Study in "Real Life".
Etelcalcetide is a new calcimimetic indicated for the treatment of secondary hyperparathyroidism (SHPT) in dialysis patients. Etelcalcetide efficacy in SHPT has been ascertained only in randomized controlled trials. This multicenter study was carried out in "real world" setting that is different from randomized controlled trials (RCTs) to (1) evaluate the effectiveness of etelcalcetide in SHPT, (2) to assess calcium, phosphorus, alkaline phosphatase changes, (3) to register gastrointestinal side effects. Data were collected from twenty-three dialysis units with n = 1190 patients on the charge. From this cohort, n = 168 (14%) patients were on treatment with etelcalcetide, and they were evaluated for statistics. A median weekly dose of etelcalcetide was 15 mg (7.5-45 mg). Patients were either naïve (33%) or switched from cinacalcet to obtain better control of SHPT with reduced side effects or pills burden. Serum parathyroid hormone (PTH) declined over time from a median value of 636 pg/mL to 357 pg/mL. The median time for responders (intact PTH (iPTH) range: two to nine times the upper normal limit) was 53 days; the percentage of responders increased (from baseline 27% to 63%) being similar in switched-patients and naïve-patients. Few patients had symptomatic hypocalcemia requiring etelcalcetide withdrawal (four cases (3%) at 30-day control, two cases (2%) at 60-day, one case (1%) at 90-day control). Side effects with etelcalcetide were lower (3-4%) than that registered during cinacalcet treatment (53%). Etelcalcetide is a new therapeutic option for SHPT with low side effects and pills burden. Etelcalcetide may improve adherence to therapy, avoiding unremitting SHP. It remains to be assessed whether etelcalcetide may reduce parathyroidectomy, vascular calcification, or mortality. Being etelcalcetide very potent in suppressing PTH levels, even in severe SHPT, future studies should evaluate the potential risk of more adynamic bone disease during long-term therapy.


The 68-kDa telomeric repeat binding factor 1 (TRF1)-associated protein (TAP68) interacts with and recruits TRF1 to the spindle pole during mitosis.
The telomere capping protein TRF1 is a component of the multiprotein complex "shelterin," which organizes the telomere into a high order structure. Besides telomere maintenance, telomere-associated proteins also have nontelomeric functions. For example, tankyrase 1 and TRF1 are required for the maintenance of faithful mitotic progression. However, the functional relevance of their centrosomal localization has not been established. Here, we report the identification of a TRF1-binding protein, TAP68, that interacts with TRF1 in mitotic cells. TAP68 contains two coiled-coil domains and a structural maintenance of chromosome motifs and co-localizes with TRF1 to telomeres during interphase. Immediately after nuclear envelope breakdown, TAP68 translocates toward the spindle poles followed by TRF1. Dissociation of TAP68 from the telomere is concurrent with the Nek2A-dependent phosphorylation at Thr-221. Biochemical characterization demonstrated that the first coiled-coil domain of TAP68 binds and recruits TRF1 to the centrosome. Inhibition of TAP68 expression by siRNA blocked the localization of TRF1 and tankyrase 1 to the centrosome. Furthermore, siRNA-mediated depletion of TAP68 perturbed faithful chromosome segregation and genomic stability. These findings suggest that TAP68 functions in mediating TRF1-tankyrase 1 localization to the centrosome and in mitotic regulation.


16S pan-bacterial PCR can accurately identify patients with ventilator-associated pneumonia.
Ventilator-associated pneumonia (VAP) remains a challenge to intensive care units, with secure diagnosis relying on microbiological cultures that take up to 72 hours to provide a result. We sought to derive and validate a novel, real-time 16S rRNA gene PCR for rapid exclusion of VAP. Bronchoalveolar lavage (BAL) was obtained from two independent cohorts of patients with suspected VAP. Patients were recruited in a 2-centre derivation cohort and a 12-centre confirmation cohort. Confirmed VAP was defined as growth of >104 colony forming units/ml on semiquantitative culture and compared with a 16S PCR assay. Samples were tested from 67 patients in the derivation cohort, 10 (15%) of whom had confirmed VAP. Using cycles to cross threshold (Ct) values as the result of the 16S PCR test, the area under the receiver operating characteristic (ROC) curve (AUROC) was 0.94 (95% CI 0.86 to 1.0, p<0.0001). Samples from 92 patients were available from the confirmation cohort, 26 (28%) of whom had confirmed VAP. The AUROC for Ct in this cohort was 0.89 (95% CI 0.83 to 0.95, p<0.0001). This study has derived and assessed the diagnostic accuracy of a novel application for 16S PCR. This suggests that 16S PCR in BAL could be used as a rapid test in suspected VAP and may allow better stewardship of antibiotics. VAPRAPID trial ref NCT01972425.


Radiation damage to nucleoprotein complexes in macromolecular crystallography.
Significant progress has been made in macromolecular crystallography over recent years in both the understanding and mitigation of X-ray induced radiation damage when collecting diffraction data from crystalline proteins. In contrast, despite the large field that is productively engaged in the study of radiation chemistry of nucleic acids, particularly of DNA, there are currently very few X-ray crystallographic studies on radiation damage mechanisms in nucleic acids. Quantitative comparison of damage to protein and DNA crystals separately is challenging, but many of the issues are circumvented by studying pre-formed biological nucleoprotein complexes where direct comparison of each component can be made under the same controlled conditions. Here a model protein-DNA complex C.Esp1396I is employed to investigate specific damage mechanisms for protein and DNA in a biologically relevant complex over a large dose range (2.07-44.63 MGy). In order to allow a quantitative analysis of radiation damage sites from a complex series of macromolecular diffraction data, a computational method has been developed that is generally applicable to the field. Typical specific damage was observed for both the protein on particular amino acids and for the DNA on, for example, the cleavage of base-sugar N1-C and sugar-phosphate C-O bonds. Strikingly the DNA component was determined to be far more resistant to specific damage than the protein for the investigated dose range. At low doses the protein was observed to be susceptible to radiation damage while the DNA was far more resistant, damage only being observed at significantly higher doses.


Arabidopsis PUB22 and PUB23 are homologous U-Box E3 ubiquitin ligases that play combinatory roles in response to drought stress.
Ubiquitination is involved in diverse cellular processes in higher plants. In this report, we describe Arabidopsis thaliana PUB22 and PUB23, two homologous U-box-containing E3 ubiquitin (Ub) ligases. The PUB22 and PUB23 genes were rapidly and coordinately induced by abiotic stresses but not by abscisic acid. PUB22- and PUB23-overexpressing transgenic plants were hypersensitive to drought stress. By contrast, loss-of-function pub22 and pub23 mutant plants were significantly more drought-tolerant, and a pub22 pub23 double mutant displayed even greater drought tolerance. These results indicate that PUB22 and PUB23 function as negative regulators in the water stress response. Yeast two-hybrid, in vitro pull-down, and in vivo coimmunoprecipitation experiments revealed that PUB22 and PUB23 physically interacted with RPN12a, a subunit of the 19S regulatory particle (RP) in the 26S proteasome. Bacterially expressed RPN12a was effectively ubiquitinated in a PUB-dependent fashion. RPN12a was highly ubiquitinated in 35S:PUB22 plants, but not in pub22 pub23 double mutant plants, consistent with RPN12a being a substrate of PUB22 and PUB23 in vivo. In water-stressed wild-type and PUB-overexpressing plants, a significant amount of RPN12a was dissociated from the 19S RP and appeared to be associated with small-molecular-mass protein complexes in cytosolic fractions, where PUB22 and PUB23 are localized. Overall, our results suggest that PUB22 and PUB23 coordinately control a drought signaling pathway by ubiquitinating cytosolic RPN12a in Arabidopsis.


Autoregulation of Sinorhizobium meliloti exoR gene expression.
The successful nitrogen-fixing symbiosis between the gram-negative soil bacterium Sinorhizobium meliloti and its leguminous plant host alfalfa (Medicago sativa) requires the bacterial exopolysaccharide succinoglycan. Succinoglycan and flagellum production, along with the ability to metabolize more than 20 different carbon sources and control the expression of a large number of S. meliloti genes, is regulated by the ExoR-ExoS/ChvI signalling pathway. The ExoR protein interacts with and suppresses the sensing activities of ExoS, the membrane-bound sensor of the ExoS/ChvI two-component regulatory system. Here we show that exoR expression is clearly upregulated in the absence of any functional ExoR protein. This upregulation was suppressed by the presence of the wild-type ExoR protein but not by a mutated ExoR protein lacking signal peptide. The levels of exoR expression could be directly modified in real time by changing the levels of total ExoR protein. The expression of exoR was also upregulated by the constitutively active sensor mutation exoS96, and blocked by two single mutations, exoS* and exoS(supA), in the ExoS sensing domain. Presence of the wild-type ExoS protein further elevated the levels of exoR expression in the absence of functional ExoR protein, and reversed the effects of exoS96, exoS* and exoS(supA) mutations. Altogether, these data suggest that ExoR protein autoregulates exoR expression through the ExoS/ChvI system, allowing S. meliloti cells to maintain the levels of exoR expression based on the amount of total ExoR protein.


A synthetic oscillatory network of transcriptional regulators.
Networks of interacting biomolecules carry out many essential functions in living cells, but the 'design principles' underlying the functioning of such intracellular networks remain poorly understood, despite intensive efforts including quantitative analysis of relatively simple systems. Here we present a complementary approach to this problem: the design and construction of a synthetic network to implement a particular function. We used three transcriptional repressor systems that are not part of any natural biological clock to build an oscillating network, termed the repressilator, in Escherichia coli. The network periodically induces the synthesis of green fluorescent protein as a readout of its state in individual cells. The resulting oscillations, with typical periods of hours, are slower than the cell-division cycle, so the state of the oscillator has to be transmitted from generation to generation. This artificial clock displays noisy behaviour, possibly because of stochastic fluctuations of its components. Such 'rational network design may lead both to the engineering of new cellular behaviours and to an improved understanding of naturally occurring networks.