Culture adaptation of malaria parasites selects for convergent loss-of-function mutants.
Cultured human pathogens may differ significantly from source populations. To investigate the genetic basis of laboratory adaptation in malaria parasites, clinical Plasmodium falciparum isolates were sampled from patients and cultured in vitro for up to three months. Genome sequence analysis was performed on multiple culture time point samples from six monoclonal isolates, and single nucleotide polymorphism (SNP) variants emerging over time were detected. Out of a total of five positively selected SNPs, four represented nonsense mutations resulting in stop codons, three of these in a single ApiAP2 transcription factor gene, and one in SRPK1. To survey further for nonsense mutants associated with culture, genome sequences of eleven long-term laboratory-adapted parasite strains were examined, revealing four independently acquired nonsense mutations in two other ApiAP2 genes, and five in Epac. No mutants of these genes exist in a large database of parasite sequences from uncultured clinical samples. This implicates putative master regulator genes in which multiple independent stop codon mutations have convergently led to culture adaptation, affecting most laboratory lines of P. falciparum. Understanding the adaptive processes should guide development of experimental models, which could include targeted gene disruption to adapt fastidious malaria parasite species to culture.


The face inversion effect in infants is driven by high, and not low, spatial frequencies.
To investigate the mechanisms underlying development of upright face preferences in infants, the current study measured inversion effects for faces that were spatial frequency (SF) filtered, into low SF and high SF, with the notion that different SFs are analyzed by different visual mechanisms. For comparison to faces, we used object stimuli that consisted of pictures of strollers. In 4 month olds, 8 month olds, and adults, we measured the strength of the selective face inversion effect (sFIE), operationally defined as an upright over inverted looking preference that is greater for faces than objects. In Study 1, we employed unfiltered stimuli, and found a clear sFIE in both infants and adults. To determine what drove this sFIE, in Study 2, the sFIE was measured for low-SF and high-SF stimuli, with all stimuli being equated for visibility. For adults, the sFIE was equally strong for low-SF and high-SF stimuli. A different pattern was seen for infants. Infants exhibited a significantly greater sFIE for high-SF, than for low-SF, stimuli (and only for high SF was the sFIE significant). In fact, the strength of infants' upright face preference for high-SF stimuli was indistinguishable from that observed for unfiltered faces, indicating that in natural (unfiltered) stimuli, high SFs are sufficient to account for infants' upright face preferences.


Enterococcus faecalis internalization in human umbilical vein endothelial cells (HUVEC).
Initial Enterococcus faecalis-endothelial cell molecular interactions which lead to enterococci associating in the host endothelial tissue, colonizing it and proliferating there can be assessed using in vitro models. Cultured human umbilical vein endothelial cells (HUVEC) have been used to study other Gram-positive bacteria-cell interactions; however, few studies have been aimed at establishing the relationship of E. faecalis with endothelial cells. The aggregation substance (AS) family of adhesins represents an E. faecalis virulence factor which has been implicated in endocarditis severity and bacterial persistence. The Asc10 protein (a member of this family) promotes bacterium-bacterium aggregation and bacterium-host cell binding. Evaluating Asc10 role in bacterial internalization by cultured enterocytes has shown that this adhesin facilitates E. faecalis endocytosis by HT-29 cells. A few eukaryotic cell structural components, such as cytoskeletal proteins, have been involved in E. faecalis entry into cell-lines; it is thus relevant to determine whether Asc10, as well as microtubules and actin microfilaments, play a role in E. faecalis internalization by cultured endothelial cells. The role of Asc10 and cytoskeleton proteins in E. faecalis ability to enter HUVEC was assessed in the present study, as well as cell apoptosis induction by enterococcal internalization by HUVEC; the data indicated increased cell apoptosis and that cytoskeleton components were partially involved in E. faecalis entry to endothelial cells, thereby suggesting that E. faecalis Asc10 protein would not be a critical factor for bacterial entry to cultured HUVEC.


Frequent detection and characterization of hepatitis E virus variants in wild rats (Rattus rattus) in Indonesia.
One hundred sixteen rats (Rattus rattus) captured in Indonesia from 2011 to 2012 were investigated for the prevalence of hepatitis E virus (HEV)-specific antibodies and HEV RNA. Using an ELISA based on HEV genotype 4 with an ad hoc cutoff value of 0.500, 18.1 % of the rats tested positive for anti-HEV IgG. By nested RT-PCR, 14.7 % of the rats had rat HEV RNA, and none were positive for HEV genotype 1-4. A high HEV prevalence among rats was associated with lower sanitary conditions in areas with a high population density. Sixteen of the 17 HEV isolates obtained from infected rats showed >93.0 % nucleotide sequence identity within the 840-nucleotide ORF1-ORF2 sequence and were most closely related to a Vietnamese strain (85.9-87.9 % identity), while the remaining isolate differed from known rat HEV strains by 18.8-23.3 % and may belong to a novel lineage of rat HEV. These results suggest a wide distribution of rat HEV with divergent genomes.


Delta-shaped anastomosis in totally robotic Billroth I gastrectomy: technical aspects and short-term outcomes.
Delta-shaped anastomosis has been recognized as a method of intracorporeal Billroth I anastomosis in totally laparoscopic distal gastrectomy. However, the technical aspects and outcomes of the delta-shaped anastomosis in totally robotic distal gastrectomy have never been reported. A single-institutional, non-randomized, retrospective study was performed between 2009 and 2013. During the study period, 47 patients underwent robotic distal gastrectomy followed by robotic delta-shaped Billroth I reconstruction, and 165 patients underwent conventional laparoscopic distal gastrectomy followed by laparoscopic delta-shaped Billroth I reconstruction. After 64 were excluded because of insufficient intraoperative video, 43 patients in the robotic group and 105 patients in the laparoscopic group were enrolled in the study. Short-term outcomes were determined from medical records and full-length operative videos. There were no significant differences between the robotic and laparoscopic groups in terms of morbidity (4.7% vs 3.8%), anastomosis-related complications (0% vs 1.0%), non-anastomosis-related complications (2.3% vs 0%), or systemic complications (2.3% vs 0%). Time for reconstruction did not vary between the robotic group (16.6 min [8.8-42.9 min]) and the laparoscopic group (15.8 min [7.2-41.0 min]). There was no mortality in this series. In the conventional group, the morbidity rate was 3.8%. The anastomosis-related complication rate was 1.0% in the conventional group. Given the excellent short-term outcomes related to anastomosis, delta-shaped anastomosis after robotic distal gastrectomy was at least as feasible and safe as delta-shaped anastomosis after laparoscopic distal gastrectomy.


Early activation of wheat polyamine biosynthesis during Fusarium head blight implicates putrescine as an inducer of trichothecene mycotoxin production.
The fungal pathogen Fusarium graminearum causes Fusarium Head Blight (FHB) disease on wheat which can lead to trichothecene mycotoxin (e.g. deoxynivalenol, DON) contamination of grain, harmful to mammalian health. DON is produced at low levels under standard culture conditions when compared to plant infection but specific polyamines (e.g. putrescine and agmatine) and amino acids (e.g. arginine and ornithine) are potent inducers of DON by F. graminearum in axenic culture. Currently, host factors that promote mycotoxin synthesis during FHB are unknown, but plant derived polyamines could contribute to DON induction in infected heads. However, the temporal and spatial accumulation of polyamines and amino acids in relation to that of DON has not been studied. Following inoculation of susceptible wheat heads by F. graminearum, DON accumulation was detected at two days after inoculation. The accumulation of putrescine was detected as early as one day following inoculation while arginine and cadaverine were also produced at three and four days post-inoculation. Transcripts of ornithine decarboxylase (ODC) and arginine decarboxylase (ADC), two key biosynthetic enzymes for putrescine biosynthesis, were also strongly induced in heads at two days after inoculation. These results indicated that elicitation of the polyamine biosynthetic pathway is an early response to FHB. Transcripts for genes encoding enzymes acting upstream in the polyamine biosynthetic pathway as well as those of ODC and ADC, and putrescine levels were also induced in the rachis, a flower organ supporting DON production and an important route for pathogen colonisation during FHB. A survey of 24 wheat genotypes with varying responses to FHB showed putrescine induction is a general response to inoculation and no correlation was observed between the accumulation of putrescine and infection or DON accumulation. The activation of the polyamine biosynthetic pathway and putrescine in infected heads prior to detectable DON accumulation is consistent with a model where the pathogen exploits the generic host stress response of polyamine synthesis as a cue for production of trichothecene mycotoxins during FHB disease. However, it is likely that this mechanism is complicated by other factors contributing to resistance and susceptibility in diverse wheat genetic backgrounds.


Protective Effect of Hydrogen Gas Inhalation on Muscular Damage Using a Mouse Hindlimb Ischemia-Reperfusion Injury Model.
Ischemia-reperfusion injury is one of the leading causes of tissue damage and dysfunction, in particular, free tissue transfer, traumatically amputated extremity, and prolonged tourniquet application during extremity surgery. In this study, the authors investigated the therapeutic effects of hydrogen gas on skeletal muscle ischemia-reperfusion injury. The authors compared the concentration of hydrogen in a muscle on intraperitoneal administration of hydrogen-rich saline and on inhalation of hydrogen gas. Animals were subjected to ischemia-reperfusion. Mice were treated with inhalation of hydrogen gas, and the hind gastrocnemius muscle was collected. Muscle morphology and inflammatory change were evaluated after ischemia-reperfusion. Moreover, a footprint test was performed to assess the functional effect of hydrogen. Hydrogen concentration of tissue was significantly higher, and the elevated level was maintained longer by hydrogen gas inhalation than by intraperitoneal administration of hydrogen-rich saline. Infarct zone and area with loss of tissue structure and marked cellular infiltration were significantly decreased in groups treated by hydrogen gas inhalation during ischemia-reperfusion; however, these effects were not observed by posttreatment of hydrogen. One week after ischemia-reperfusion, mice that had been pretreated with hydrogen gas recovered faster and achieved smoother walking in appearance compared with mice in the other groups as assessed by the footprint test. Inhalation of hydrogen gas attenuates muscle damage, inhibits inflammatory response, and enhances functional recovery. These findings suggest that the optimal route for hydrogen delivery is continuous inhalation of hydrogen gas, which could be a novel clinical mode of treatment in ischemia-reperfusion injury.


[Distribution and trends of tuberculosis by age groups and municipalities in La Habana City, Cuba (1986-1998)].
The province of Havana City, the capital and most densely populated city in Cuba, totals around 27% of the new cases of tuberculosis. This article is for the purpose of describing the distribution and trend of the tuberculosis cases rates from 1986 to 1998, by municipalities and age groups in this province. A calculation was made of the rates and of the total percentage of deviation of these rates, in addition to the annual average deviation both throughout two time periods and between the two (1986-1993 and 1994-1998). The trends were calculated by simple linear regression. Throughout the 1986-1993 period, the case rate for this province underwent a 6% increase. Throughout the 1994-1998 period, it dropped by 7.3%. The three most densely-populated cities showed a higher rate increase. Of the 15 municipalities, only La Lisa showed a drop in the rates throughout the entire period in question. The rates by ages were similar throughout all of the municipalities, being higher for individuals > 65 years of age. Six tenths (0.6%) of the cases reported involved people under 15 years of age. The infantile tuberculosis rates were very low for the time period under analysis. The differences among the municipalities in question make it necessary for different measures to be taken to achieve a greater impact and repercussion in Cuba. The drop in the rates detected over the last three years is indicative of the epidemic being brought under control and the national tuberculosis control program being reinstated.


An evaluation of a morphine public health programme for cancer and AIDS pain relief in Sub-Saharan Africa.
Despite growing HIV and cancer prevalence in Sub-Saharan Africa, and WHO advocacy for a public health approach to palliative care provision, opioid availability is severely limited. Uganda has achieved a morphine roll-out programme in partnership with the Ministry of Health. This study aimed to evaluate that programme by identifying challenges to implementation that may inform replication. A multi-methods protocol appraised morphine regulation, storage, prescribing, and consumption in three phases: key informant interviews throughout the opioid supply chain, and direct observation and audit of clinical practice. Regulation had achieved its goal of preventing misuse and leakage from the supply chain. However, the Government felt that relaxation of regulation was now appropriate. Confusion and complexity in storage and authorization rules led to discontinuation of opioid pain management at the patient level and also wasted service time in trying to obtain supplies to which they were entitled. Continued neglect to prescribe among clinicians and public fear of opioids led to under prescribing, and clinical skills showed some evidence of need for improvement with respect to physical assessment and follow-up. The Ugandan programme offers a successful model for both advocacy and Governmental support in achieving opioid roll-out across health districts. Despite initial concerns, abuse of opioids has not been evident. Further work is required to ensure that available supplies of opioids are prescribed to those in need, and that clinical standards are met. However, the programme for roll-out has proved a useful model to expand opioid availability as the first step in improving patient care, and may prove a useful template for other Sub-Saharan African countries.


Structural analogues of roscovitine rescue the intracellular traffic and the function of ER-retained ABCB4 variants in cell models.
Adenosine triphosphate binding cassette transporter, subfamily B member 4 (ABCB4) is the transporter of phosphatidylcholine at the canalicular membrane of hepatocytes. ABCB4 deficiency, due to genetic variations, is responsible for progressive familial intrahepatic cholestasis type 3 (PFIC3) and other rare biliary diseases. Roscovitine is a molecule in clinical trial that was shown to correct the F508del variant of cystic fibrosis transmembrane conductance regulator (CFTR), another ABC transporter. In the present study, we hypothesized that roscovitine could act as a corrector of ABCB4 traffic-defective variants. Using HEK and HepG2 cells, we showed that roscovitine corrected the traffic and localisation at the plasma membrane of ABCB4-I541F, a prototypical intracellularly retained variant. However, roscovitine caused cytotoxicity, which urged us to synthesize non-toxic structural analogues. Roscovitine analogues were able to correct the intracellular traffic of ABCB4-I541F in HepG2 cells. Importantly, the phospholipid secretion activity of this variant was substantially rescued by three analogues (MRT2-235, MRT2-237 and MRT2-243) in HEK cells. We showed that these analogues also triggered the rescue of intracellular traffic and function of two other intracellularly retained ABCB4 variants, i.e. I490T and L556R. Our results indicate that structural analogues of roscovitine can rescue genetic variations altering the intracellular traffic of ABCB4 and should be considered as therapeutic means for severe biliary diseases caused by this class of variations.


Membrane proteins with immunoglobulin-like domains--a master superfamily of interaction molecules.
Recent genomic analysis has shown that proteins with immunoglobulin superfamily (IgSF) domains are extremely abundant and their number has increased markedly in evolution correlating with the development of the adaptive immune system. The IgSF domain is particularly good at being recognised and is involved in many different kinds of interactions. Thus, analysis of the properties of these domains can act as a paradigm for thinking about the roles of newly identified gene products. This review summarises the identification, function and properties of IgSF domains including, their size, variety of interactions, their strength of binding, role of glycosylation and organisation with other proteins.


Collaborative coupling between polymerase and helicase for leading-strand synthesis.
Rapid and processive leading-strand DNA synthesis in the bacteriophage T4 system requires functional coupling between the helicase and the holoenzyme, consisting of the polymerase and trimeric clamp loaded by the clamp loader. We investigated the mechanism of this coupling on a DNA hairpin substrate manipulated by a magnetic trap. In stark contrast to the isolated enzymes, the coupled system synthesized DNA at the maximum rate without exhibiting fork regression or pauses. DNA synthesis and unwinding activities were coupled at low forces, but became uncoupled displaying separate activities at high forces or low dNTP concentration. We propose a collaborative model in which the helicase releases the fork regression pressure on the holoenzyme allowing it to adopt a processive polymerization conformation and the holoenzyme destabilizes the first few base pairs of the fork thereby increasing the efficiency of helicase unwinding. The model implies that both enzymes are localized at the fork, but does not require a specific interaction between them. The model quantitatively reproduces homologous and heterologous coupling results under various experimental conditions.


Endovascular stenting for rescue of a failed donor graft during superficial temporal artery to middle cerebral artery bypass surgery: case report.
Direct bypass has been used to salvage failed endovascular treatment; however, little is known of the reversed role of endovascular management for failed bypass.The authors report the case of a 7-year-old patient who underwent a superficial temporal artery to middle cerebral artery (STA-MCA) bypass for treatment of a giant MCA aneurysm and describe the role of endovascular rescue in this case. Post-bypass catheter angiogram showed occlusion of the proximal extracranial STA donor with patent anastomosis, possibly due to STA dissection. A self-expanding Neuroform Atlas stent was deployed across the dissection flap, and follow-up images showed revascularization of the STA with good MCA runoff.This case demonstrates that direct extracranial-intracranial bypass failure can infrequently originate from the STA donor vessel and that superselective angiogram can be useful for identification and treatment in such cases. With more advanced endovascular techniques the tide has turned in the treatment of complex cerebrovascular cases, with this case being an early example of successful rescue stenting for endovascular management of a failed donor after STA-MCA bypass.


Correlation of changes in gravity, elevation, and strain in southern california.
Measurements made once or twice a year from 1977 through 1982 show large correlated changes in gravity, elevation, and strain in several southern California networks. Precise gravity surveys indicate changes of as much as 25 microgals between surveys 6 months apart. Repeated surveys show that annual elevation changes as large as 100 millimeters occur along baselines 40 to 100 kilometers long. Laser-ranging surveys reveal coherent changes in areal strain of 1 to 2 parts per million occurred over much of southern California during 1978 and 1979. Although the precision of these measuring systems has been questioned, the rather good agreement among them suggests that the observed changes reflect true crustal deformation.


Multisampling suprathreshold perimetry: a comparison with conventional suprathreshold and full-threshold strategies by computer simulation.
To compare a multisampling suprathreshold strategy with conventional suprathreshold and full-threshold strategies in detecting localized visual field defects and in quantifying the area of loss. Probability theory was applied to examine various suprathreshold pass criteria (i.e., the number of stimuli that have to be seen for a test location to be classified as normal). A suprathreshold strategy that requires three seen or three missed stimuli per test location (multisampling suprathreshold) was selected for further investigation. Simulation was used to determine how the multisampling suprathreshold, conventional suprathreshold, and full-threshold strategies detect localized field loss. To determine the systematic error and variability in estimates of loss area, artificial fields were generated with clustered defects (0-25 field locations with 8- and 16-dB loss) and, for each condition, the number of test locations classified as defective (suprathreshold strategies) and with pattern deviation probability less than 5% (full-threshold strategy), was derived from 1000 simulated test results. The full-threshold and multisampling suprathreshold strategies had similar sensitivity to field loss. Both detected defects earlier than the conventional suprathreshold strategy. The pattern deviation probability analyses of full-threshold results underestimated the area of field loss. The conventional suprathreshold perimetry also underestimated the defect area. With multisampling suprathreshold perimetry, the estimates of defect area were less variable and exhibited lower systematic error. Multisampling suprathreshold paradigms may be a powerful alternative to other strategies of visual field testing. Clinical trials are needed to verify these findings.


Revisiting EmbodiedQA: A Simple Baseline and Beyond.
In Embodied Question Answering (EmbodiedQA), an agent interacts with an environment to gather necessary information for answering user questions. Existing works have laid a solid foundation towards solving this interesting problem. But the current performance, especially in navigation, suggests that EmbodiedQA might be too challenging for the contemporary approaches. In this paper, we empirically study this problem and introduce 1) a simple yet effective baseline that achieves promising performance; 2) an easier and practical setting for EmbodiedQA where an agent has a chance to adapt the trained model to a new environment before it actually answers users questions. In this new setting, we randomly place a few objects in new environments, and upgrade the agent policy by a distillation network to retain the generalization ability from the trained model. On the EmbodiedQA v1 benchmark, under the standard setting, our simple baseline achieves very competitive results to the-state-of-the-art; in the new setting, we found the introduced small change in settings yields a notable gain in navigation.


Bisphosphonate treatment of benign multifocal and unifocal osteolytic tumours of bone.
Growth of benign tumours and tumour-like lesions of bone results in osteolysis which may cause pathological fracture. Bisphosphonates are anti-osteolytic agents which have proved effective in the treatment of number of osteolytic conditions. In this study we report the results of treatment with the aminobisphosphonate, pamidronate, of three benign osteolytic tumours of bone, two cases of fibrous dysplasia and one of Langerhans cell histiocytosis. In all three cases there was clinical and radiological improvement following treatment. Radiologically, bone lesions did not exhibit progressive enlargement. Two cases of fibrous dysplasia also showed features suggestive of increased bone formation. These findings indicate that bisphosphonates are likely to be useful in controlling the osteolysis of benign tumours/tumour-like lesions of bone, particularly in those cases where surgical intervention is not possible or multifocal lesions are present.


Coherent Rayleigh-Brillouin scattering measurement of atmospheric atomic and molecular gas temperature.
Broadband coherent Rayleigh-Brillouin scattering (CRBS) was used to measure translational gas temperatures for nitrogen, argon, and methane at the ambient pressure of 0.8 atm. Temperatures derived from spectral analysis were compared with experimentally-measured temperatures, with a maximum 5.2% difference for all gases at all temperatures; and with nitrogen, argon, and methane exhibiting average differences over the temperature range tested of 0.8%, 1.4% and -0.5%, respectively. These values are consistent with the 2% estimated, experimental error of the experiment. Improving upon the efficiency of previous line shape acquisition methods, CRBS data were spectrally de-convolved using a cost effective, purpose-designed, Fabry-Perot etalon spectrometer. The resulting line shapes were compared to models obtained from approximations to the 1D Boltzmann equation. Although this study employed broadband CRBS for explicit gas temperature measurement, similar line shape acquisition techniques could be used with broadband coherent Rayleigh scattering (CRS) to experimentally-measure gas temperatures, pressures and other transport properties in both the kinetic (CRBS) and rarefied (CRS) regimes.


Degradation of p27(Kip) cdk inhibitor triggered by Kaposi's sarcoma virus cyclin-cdk6 complex.
The Kaposi's sarcoma-associated human herpesvirus 8 (KSHV/HHV8) encodes a protein similar to cellular cyclins. This cyclin is most closely related to cellular D-type cyclins, but biochemically it behaves atypically in various respects. Complexes formed between the viral cyclin and the cyclin-dependent kinase subunit, cdk6, can phosphorylate a wider range of substrates and are resistant to cdk inhibitory proteins. We show here that the KSHV-cyclin-cdk6 complex phosphorylates p27(Kip) on a C-terminal threonine that is implicated in destabilization of this cdk inhibitor. Expression of the viral cyclin in tissue culture cells overcomes a cell cycle block by p27(Kip). However, full cell-cycle transit of these cells appears to depend on C-terminal phosphorylation of p27(Kip) and seems to involve transactivation of other cellular cyclin-dependent kinases. A p27(Kip)-phosphorylating cdk6 complex exists in cell lines derived from primary effusion lymphoma and in Kaposi's sarcoma, this indicating that virally induced p27(Kip) degradation may occur in KSHV-associated tumours.


Energy and environmental assessment of a traction lithium-ion battery pack for plug-in hybrid electric vehicles.
Traction batteries are a key factor in the environmental sustainability of electric mobility and, therefore, it is necessary to evaluate their environmental performance to allow a comprehensive sustainability assessment of electric mobility. This article presents an environmental assessment of a lithium-ion traction battery for plug-in hybrid electric vehicles, characterized by a composite cathode material of lithium manganese oxide (LiMn2O4) and lithium nickel manganese cobalt oxide Li(NixCoyMn1-x-y)O2. Composite cathode material is an emerging technology that promises to combine the merits of several active materials into a hybrid electrode to optimize performance and reduce costs. In this study, the environmental assessment of one battery pack (with a nominal capacity of 11.4 kWh able to be used for about 140,000 km of driving) is carried out by using the Life Cycle Assessment methodology consistent with ISO 14040. The system boundaries are the battery production, the operation phase and recycling at the end of life, including the recovery of various material fractions. The composite cathode technology examined besides a good compromise between the higher and the lower performance of NMC and LMO cathodes, can present good environmental performances. The results of the analysis show that the manufacturing phase is relevant to all assessed impact categories (contribution higher than 60%). With regard to electricity losses due to battery efficiency and battery transport, the contribution to the use phase impact of battery efficiency is larger than that of battery transport. Recycling the battery pack contributes less than 11% to all of the assessed impact categories, with the exception of freshwater ecotoxicity (60% of the life cycle impact). The environmental credits related to the recovery of valuable materials (e.g. cobalt and nickel sulphates) and other metal fractions (e.g. aluminium and steel) are particularly relevant to impact categories such as marine eutrophication, human toxicity and abiotic resource depletion. The main innovations of this article are that (1) it presents the first bill of materials of a lithium-ion battery cell for plug-in hybrid electric vehicles with a composite cathode active material; (2) it describes one of the first applications of the life cycle assessment to a lithium-ion battery pack for plug-in hybrid electric vehicles with a composite cathode active material with the aim of identifying the "hot spots" of this technology and providing useful information to battery manufacturers on potentially improving its environmental sustainability; (3) it evaluates the impacts associated with the use phase based on primary data about the battery pack's lifetime, in terms of kilometres driven; and (4) it models the end-of-life phase of the battery components through processes specifically created for or adapted to the case study.


Comparison of ocular straylight after implantation of multifocal intraocular lenses.
A comprehensive review of the effect of multifocal intraocular lens (IOL) designs on postoperative ocular straylight was performed. Studies reporting straylight values obtained with the natural pupil using the C-Quant device after uneventful multifocal IOL implantation were included. The IOLs were categorized based on their material characteristics; that is, hydrophobicity and presence of colored chromophores. Age adjustment was achieved using the straylight age-dependency norm for pseudophakic eyes. This norm also served as a reference for comparing mean straylight levels of the various IOLs. The literature review identified 10 studies reporting 9 multifocal IOL designs. The hydrophilic IOLs showed less straylight than the hydrophobic IOLs by 0.08 log(s) (P = .001). Blue violet light-filtering IOLs showed less straylight than standard IOLs by 0.04 log(s), which was not statistically significant (P = .32). Hydrophobicity was a factor that significantly affected straylight in multifocal IOLs. The Netherlands Academy of Arts and Sciences owns a patent on straylight measurement, with Dr. van den Berg as the inventor, and licenses it to Oculus Optikgeräte GmbH for the C-Quant instrument. No other author has a financial or proprietary interest in any material or method mentioned.


Analysis of peanut leaf proteome.
Peanut (Arachis hypogaea) is one of the most important sources of plant protein. Current selection of genotypes requires molecular characterization of available populations. Peanut genome database has several EST cDNAs which can be used to analyze gene expression. Analysis of proteins is a direct approach to define function of their associated genes. Proteome analysis linked to genome sequence information is critical for functional genomics. However, the available protein expression data is extremely inadequate. Proteome analysis of peanut leaf was conducted using two-dimensional gel electrophoresis in combination with sequence identification using MALDI/TOF to determine their identity and function related to growth, development and responses to stresses. Peanut leaf proteins were resolved into 300 polypeptides with pI values between 3.5 and 8.0 and relative molecular masses from 12 to 100 kDa. A master leaf polypeptide profile was generated based on the consistently expressed protein pattern. Proteins present in 205 spots were identified using GPS software and Viridiplantae database (NCBI). Identity of some of these proteins included RuBisCO, glutamine synthetase, glyoxisomal malate dehydrogenase, oxygen evolving enhancer protein and tubulin. Bioinformatical analyses showed that there are 133 unique protein identities. They were categorized into 10 and 8 groups according to their cellular compartmentalization and biological functionality, respectively. Enzymes necessary for carbohydrate metabolism and photosynthesis dominated in the set of identified proteins. The reference map derived from a drought-tolerant cv.Vemana should serve as the basis for further investigations of peanut physiology such as detection of expressed changes due to biotic and abiotic stresses, plant development. Furthermore, the leaf proteome map will lead to development of protein markers for cultivar identification at seedling stage of the plant. Overall, this study will contribute to improve our understanding of plant genetics and metabolism, and overall assist in the selection and breeding programs geared toward crop improvement.


Cr(vi) uptake and reduction by biogenic iron (oxyhydr)oxides.
The mobility and toxicity of chromium (Cr) in soil and water systems are largely controlled by its oxidation state and interactions with solid phases. Relative to abiotic minerals, biogenic iron (Fe) (oxyhydr)oxides (BIOS) may enhance Cr(vi) adsorption and reduction due to their poorly ordered structures, large surface areas, and incorporation of cell derived organic matter. To determine the extent and mechanisms of the reaction between Cr(vi) and BIOS, sorption isotherm and kinetic studies were conducted using two-line ferrihydrite, BIOS, and BIOS amended with 0.135 M ferrozine (an Fe(ii) chelator). X-ray absorption near edge structure (XANES) spectroscopy of BIOS reacted with Cr(vi) showed approximately 50% reduction of the total sorbed Cr from Cr(vi) to Cr(iii) after 14 days of exposure. Sorbed Cr(iii) was best fit with an organic carboxylate complex after 1 d of reaction, but after 7 d mineral-associated Cr(iii) was the predominant form. In the presence of ferrozine, Cr(vi) reduction by BIOS was inhibited, confirming a key role for Fe(ii) as the Cr(vi) reductant. However, the lack of a 3 : 1 reaction stoichiometry between Fe(ii) and Cr(iii) produced suggests roles for reaction with organic matter and Cr(v) autoreduction in Cr(iii) production. This study thus elucidates an unrecognized mechanism of Cr sequestration by ubiquitous natural Fe (oxyhydr)oxide deposits. Furthermore, the redox transformation of mobile Cr(vi) to less soluble Cr(iii) species observed in our study implies that biogenic Fe (oxyhydr)oxides in soils and natural waters may naturally attenuate Cr(vi) concentrations through sorption and reduction processes, thus limiting its transport to downstream environments.


Effects of long-term cranberry supplementation on endocrine pancreas in aging rats.
The effects of long-term cranberry consumption on age-related changes in endocrine pancreas are not fully understood. Here we treated male Fischer 344 rats with either 2% whole cranberry powder supplemented or normal rodent chow from 6 to 22 month old. Both groups displayed an age-related decline in basal plasma insulin concentrations, but this age-related decline was delayed by cranberry. Cranberry supplementation led to increased β-cell glucose responsiveness during the oral glucose tolerance test. Portal insulin concentration was 7.6-fold higher in rats fed cranberry, coupled with improved β-cell function. However, insulin resistance values were similar in both groups. Total β-cell mass and expression of pancreatic and duodenal homeobox 1 and insulin within islets were significantly enhanced in rats fed cranberry relative to controls. Furthermore, cranberry increased insulin release of an insulin-producing β-cell line, revealing its insulinotropic effect. These findings suggest that cranberry is of particular benefit to β-cell function in normal aging rats.


Decreased 6-mercaptopurine retention by two resistant variants of mouse neuroblastoma with normal hypoxanthine-guanine-phospho-ribosyltransferase activities.
In an effort to propose more effective chemotherapeutic regimens for the treatment of neuroblastoma, we have characterized mouse, neuroblastoma variants whose growth in tisue culture are resistant to antimetabolites. We report the partial characterization of two lines resistant to 6-mercaptopurine (6-MP). Concentrations of drug required to inhibit their growth rates 50% are 110- and 575-fold higher, respectively, than that inhibiting the sensitive parental clone. Unlike most 6-MP-resistant cell lines described previously, both neuroblastoma populations display normal activities of hypoxanthine-guanine phosphoribosyltransferase but greatly reduced accumulation of 14-C-labeled 6-MP. Drug accumulation was inhibited by adenine, blocked by dinitrophenol but not ouabain and strongly temperature dependent suggesting a need for cytoplasmic phosphoribosylation. Possible mechanisms for this reduction in 6-MP retention are discussed. Importantly, eight clones isolated in 6-MP-free media from the 110-fold resistant population of cells demonstrated quantitatively identical growth inhibition at all drug concentrations tested suggesting that the original 110-fold resistant neuroblastoma population was homogenous with respect to its mechanisms of resistance.


Telomerase activity in human breast cancer and benign breast lesions: diagnostic applications in clinical specimens, including fine needle aspirates.
We analysed telomerase activity in normal, benign and malignant breast tissues and in fine needle aspirates by a PCR-based assay. The tissue samples we used in this assay consisted of 20 cryostat sections, 10 microns thick, from each breast biopsy. This method was used to obtain effective extraction from small samples and to confirm the histological identity of the specimen by microscopical examination of serial sections. Fifty-two of 71 breast carcinomas were positive for telomerase activity, and the intensity of this was strong in most cases, whereas all 6 samples of normal breast tissue and 17 of fibrocystic disease were negative and only 1 of 15 fibroadenomas was positive. Invasive ductal carcinomas were more frequently positive than invasive lobular carcinomas. There was no correlation of telomerase activity with tumour size or the occurrence of lymph node metastasis. Evaluation of our assay system showed that a signal of telomerase activity was detectable in extracts from single cryostat sections (< 1 mm2) of a cancer specimen and from as few as 4 cells of a human breast cancer cell line. On the basis of the above data, we applied this assay to fine needle aspirates of breast lesions. Ten of 15 aspirates which had been cytopathologically diagnosed as cancer were strongly positive, while 26 of 29 benign aspirates were totally negative and the remaining 3 showed only borderline activity. In 3 cases, the telomerase result could have helped establish a diagnosis when the cytological observations were inconclusive. Our results indicate that this sensitive assay could become a useful new modality for supplementing microscopic cytopathology in the detection of cancer cells in small tissue biopsies and fine needle aspirates.


The effect of immunomodulatory drugs on aortic stenosis: a Mendelian randomisation analysis.
There are currently no approved pharmacological treatment options for aortic stenosis (AS), and there are limited identified drug targets for this chronic condition. It remains unclear whether inflammation plays a role in AS pathogenesis and whether immunomodulation could become a therapeutic target. We evaluated the potentially causal association between inflammation and AS by investigating the genetically proxied effects of tocilizumab (IL6 receptor, IL6R, inhibitor), canakinumab (IL1β inhibitor) and colchicine (β-tubulin inhibitor) through a Mendelian randomisation (MR) approach. Genetic proxies for these drugs were identified as single nucleotide polymorphisms (SNPs) in the gene, enhancer or promoter regions of IL6R, IL1β or β-tubulin gene isoforms, respectively, that were significantly associated with serum C-reactive protein (CRP) in a large European genome-wide association study (GWAS; 575,531 participants). These were paired with summary statistics from a large GWAS of AS in European patients (653,867 participants) to then perform primary inverse-variance weighted random effect and sensitivity MR analyses for each exposure. This analysis showed that genetically proxied tocilizumab was associated with reduced risk of AS (OR 0.56, 95% CI 0.45-0.70 per unit decrease in genetically predicted log-transformed CRP). Genetically proxied canakinumab was not associated with risk of AS (OR 0.80, 95% CI 0.51-1.26), and only one suitable SNP was identified to proxy the effect of colchicine (OR 34.37, 95% CI 1.99-592.89). The finding that genetically proxied tocilizumab was associated with reduced risk of AS is concordant with an inflammatory hypothesis of AS pathogenesis. Inhibition of IL6R may be a promising therapeutic target for AS management.


Effects of cisplatin on mitochondrial function in Jurkat cells.
In this work, we measured the effects of pharmacological concentrations of cisplatin (cis-diaminedichloroplatinum II) on mitochondrial function, cell viability, and DNA fragmentation in Jurkat cells. The exposure of cells to 0-25 microM cisplatin for 3 h had no immediate effect on cellular mitochondrial oxygen consumption, measured using a palladium-porphyrin oxygen sensing phosphor. Similarly, the cell viability as measured by trypan blue staining was unchanged immediately following exposure to the drug, and no small DNA fragments, characteristic of drug-induced apoptosis, appeared. At 24 h after exposure to cisplatin, cellular respiration and viability decreased relative to controls and the amount of small DNA fragments, measured using quantitative agarose gel electrophoresis, was proportional to the concentration of cisplatin present during the drug exposure period. The small DNA fragments showed the banding pattern (with a spacing of approximately 300 bp) characteristic of drug-induced cell death by apoptosis. The changes in respiration and DNA fragmentation correlated linearly with the amount of platinum bound to DNA, determined by atomic absorption spectroscopy immediately following drug exposure. The oxygen consumption by beef heart mitochondria was not affected 0-24 h after exposure to 25 microM cisplatin or to solutions containing the monoaquated form of the drug, suggesting that the drug does not attack the mitochondrial respiratory chain directly. Cells exposed to the peptide benzyloxycarbonyl-val-ala-asp-fluoromethyl ketone, which blocks apoptosis by the caspase pathway, showed a decrease in cisplatin-induced DNA fragmentation but not in the impairment of cellular respiration. Thus, although apoptosis is caspase-dependent, the impairment of cellular respiration is independent of the caspase system. Collectively, these results suggest that alteration in mitochondrial function is a secondary effect of cisplatin cytotoxicity in Jurkat cells.


Direct identification of ligand-receptor interactions on living cells and tissues.
Many cellular responses are triggered by proteins, drugs or pathogens binding to cell-surface receptors, but it can be challenging to identify which receptors are bound by a given ligand. Here we describe TRICEPS, a chemoproteomic reagent with three moieties--one that binds ligands containing an amino group, a second that binds glycosylated receptors on living cells and a biotin tag for purifying the receptor peptides for identification by quantitative mass spectrometry. We validated this ligand-based, receptor-capture (LRC) technology using insulin, transferrin, apelin, epidermal growth factor, the therapeutic antibody trastuzumab and two DARPins targeting ErbB2. In some cases, we could also determine the approximate ligand-binding sites on the receptors. Using TRICEPS to label intact mature vaccinia viruses, we identified the cell surface proteins AXL, M6PR, DAG1, CSPG4 and CDH13 as binding factors on human cells. This technology enables the identification of receptors for many types of ligands under near-physiological conditions and without the need for genetic manipulations.


Sequence- and base-specific delivery of nitric oxide to cytidine and 5-methylcytidine leading to efficient deamination.
Nitric oxide (NO) is an important endogenous regulatory molecule, and S-nitrosothiols are believed to play a significant role in NO storage, transport, and delivery. On the basis of their ability to generate NO in vivo, S-nitrosothiols can be used as therapeutic drugs. In this study, we have developed an innovative method for sequence- and base-specific delivery of NO to a specific site of DNA followed by specific deamination. We designed a NO transfer reaction from S-nitroso thioguanine to an imino tautomer of cytosine. Nitrosation of the thioguanosine-containing ODN 1 was carried out with S-nitroso-N-acetylpenicillamine (SNAP) to produce ODN 2. An interstrand NO transfer reaction was performed using ODN 2 and its complementary ODN 3 having dC or dmC at the target site, and a rapid NO transfer reaction was observed. In contrast, a transfer reaction was not observed either with ODN 3 having dT, dA, or dG at the target site or with ODN 5-7 having dC at a nontarget site. In the analysis of deaminated products of the NO-transferred ODN 4, it was found that the transformation ratio from dmC to dT was as high as 42% together with the dmC-diazoate (13%). In conclusion, we have demonstrated the innovative method of sequence- and base-specific delivery of nitric oxide to cytidine and 5-methylcytidine. The selectivity and efficiency of NO transfer followed by deamination exhibited in this study are extremely high compared to those of the conventional methods.


Sedimentation in parasitological coproscopy.
A sedimentation technique is described, in which a fecal suspension is placed on top of an aqueous sucrose solution of specific gravity 1.015 g/cm3. Using 100 by 15 mm test tubes, duplicate gravity sedimentation experiments were made using homogenized fecal suspensions (single-columns) and fecal suspensions placed on top of clear columns (double-columns). Egg and cyst counts, and turbidity determinations were made in the sediments obtained after definite time intervals. Most Ascaris lumbricoides, Trichuris trichiura and Ancylostomidae eggs sedimented within 20 minutes in single-and between 30 and 60 minutes in double-columns. Giardia duodenalis cysts required longer periods to sediment in double-than in single-columns; after 180 minutes (the maximum period of observation), double-column sediments produced 60.0% of the counts of single-columns. Double-column sediments were consistently less turbid than single-column ones.


Gluten exacerbates IgA nephropathy in humanized mice through gliadin-CD89 interaction.
IgA1 complexes containing deglycosylated IgA1, IgG autoantibodies, and a soluble form of the IgA receptor (sCD89), are hallmarks of IgA nephropathy (IgAN). Food antigens, notably gluten, are associated with increased mucosal response and IgAN onset, but their implication in the pathology remains unknown. Here, an IgAN mouse model expressing human IgA1 and CD89 was used to examine the role of gluten in IgAN. Mice were given a gluten-free diet for three generations to produce gluten sensitivity, and then challenged for 30 days with a gluten diet. A gluten-free diet resulted in a decrease of mesangial IgA1 deposits, transferrin 1 receptor, and transglutaminase 2 expression, as well as hematuria. Mice on a gluten-free diet lacked IgA1-sCD89 complexes in serum and kidney eluates. Disease severity depended on gluten and CD89, as shown by reappearance of IgAN features in mice on a gluten diet and by direct binding of the gluten-subcomponent gliadin to sCD89. A gluten diet exacerbated intestinal IgA1 secretion, inflammation, and villous atrophy, and increased serum IgA1 anti-gliadin antibodies, which correlated with proteinuria in mice and patients. Moreover, early treatment of humanized mice with a gluten-free diet prevented mesangial IgA1 deposits and hematuria. Thus, gliadin-CD89 interaction may aggravate IgAN development through induction of IgA1-sCD89 complex formation and a mucosal immune response. Hence, early-stage treatment with a gluten-free diet could be beneficial to prevent disease.


Simultaneous storage and utilization of polyhydroxyalkanoates and glycogen under aerobic conditions.
This study aims to investigate the storage response of an activated sludge system in the presence of two different substrates which are stored as two different polymers. The objective of the study was to determine the changes in the response of an activated sludge system when two different storage mechanisms could occur simultaneously. Acetic acid (HAc) and soluble starch (SolS) were selected as model substrates and three different feeding conditions, namely (i) when both HAc and SolS were initially present in the reactor, (ii) only HAc was present, and (iii) only SolS was present in the substrate solution. The batch reactors were monitored for acetic acid, polyhydroxyalkanoates (PHA), poly-glucose (glycogen like substances) and oxygen uptake rate (OUR). The experiments have shown that, while the main portion of hydrolyzed starch was stored as poly-glucose, which was further used for heterotrophic growth, the rest was utilized for direct growth. However, acetic acid was totally stored as PHA and the stored PHA was used for biomass growth under the presented experimental conditions. When the system was fed with the substrate mixture, the storage mechanism was not significantly affected. Both PHA and poly-glucose storage took place simultaneously with the same stoichiometry and kinetics defined for single substrate utilization.


Adalimumab effectiveness in Behçet's disease: short and long-term data from a multicenter retrospective observational study.
Our aim was to evaluate the effectiveness of adalimumab (ADA) during a 24-month study period in patients affected with Behçet's disease (BD). Clinical and therapeutic data from 100 consecutive BD patients treated with ADA were retrospectively collected and statistically analyzed. At 12-week follow-up, ADA induced clinical efficacy in 81 patients, with a mean time to response of 7.63 ± 3.43 weeks; 25 (30.9 %) patients underwent a disease relapse after 22.17 ± 1.57 months, but treatment adjustments allowed a recovery of efficacy in 11 cases. At 24-month follow-up, 67/100 patients were still on ADA therapy despite concomitant treatments. No differences were identified between ADA monotherapy and co-treatment with DMARDs about efficacy (p = 0.09), time to response (p = 0.61), relapses (p = 0.36), and ADA discontinuation (p = 0.40). No differences existed in patients switched from other tumor necrosis factor (TNF)-α inhibitors about efficacy at 12 weeks (p = 0.13) and rapidity of response (p = 0.93) while relapses (p = 0.01) and ADA discontinuation at 24 months (p = 0.001) were significantly more common. Adverse events occurred in 10 patients. ADA confirmed its effectiveness in BD. Combination therapy with DMARDs seems not significantly superior to monotherapy. Frequency and time to response for ADA was not conditioned by a previous lack or loss of efficacy to other TNF-α inhibitors, but long-term loss of efficacy seemed more likely in patients switched from other anti-TNF agents.


Traffic Accidents Involving Cyclists Identifying Causal Factors Using Questionnaire Survey, Traffic Accident Data, and Real-World Observation.
The purpose of this study is to clarify the mechanism of traffic accidents involving cyclists. The focus is on the characteristics of cyclist accidents and scenarios, because the number of traffic accidents involving cyclists in Tokyo is the highest in Japan. First, dangerous situations in traffic incidents were investigated by collecting data from 304 cyclists in one city in Tokyo using a questionnaire survey. The survey indicated that cyclists used their bicycles generally while commuting to work or school in the morning. Second, the study investigated the characteristics of 250 accident situations involving cyclists that happened in the city using real-world bicycle accident data. The results revealed that the traffic accidents occurred at intersections of local streets, where cyclists collided most often with vehicles during commute time in the morning. Third, cyclists' behavior was observed at a local street intersection in the morning in the city using video pictures. In one hour during the morning commute period, 250 bicycles passed through the intersection. The results indicated that one of the reasons for traffic accidents involving cyclists might be the combined effect of low visibility, caused by the presence of box-like building structures close to the intersections, and the cyclists' behavior in terms of their velocity and no confirming safety. It was observed that, on average, bicycle velocity was 3.1 m/s at the initial line of an intersection. The findings from this study could be useful in developing new technologies to improve cyclist safety, such as alert devices for cyclists and vehicle drivers, wireless communication systems between cyclists and vehicle drivers, or advanced vehicles with bicycle detection and collision mitigation systems.


Modulation change detection in human auditory cortex: Evidence for asymmetric, non-linear edge detection.
Changes in modulation rate are important cues for parsing acoustic signals, such as speech. We parametrically controlled modulation rate via the correlation coefficient (r) of amplitude spectra across fixed frequency channels between adjacent time frames: broadband modulation spectra are biased toward slow modulate rates with increasing r, and vice versa. By concatenating segments with different r, acoustic changes of various directions (e.g., changes from low to high correlation coefficients, that is, random-to-correlated or vice versa) and sizes (e.g., changes from low to high or from medium to high correlation coefficients) can be obtained. Participants listened to sound blocks and detected changes in correlation while MEG was recorded. Evoked responses to changes in correlation demonstrated (a) an asymmetric representation of change direction: random-to-correlated changes produced a prominent evoked field around 180 ms, while correlated-to-random changes evoked an earlier response with peaks at around 70 and 120 ms, whose topographies resemble those of the canonical P50m and N100m responses, respectively, and (b) a highly non-linear representation of correlation structure, whereby even small changes involving segments with a high correlation coefficient were much more salient than relatively large changes that did not involve segments with high correlation coefficients. Induced responses revealed phase tracking in the delta and theta frequency bands for the high correlation stimuli. The results confirm a high sensitivity for low modulation rates in human auditory cortex, both in terms of their representation and their segregation from other modulation rates.


In vitro and in vivo stability and pharmacokinetic profile of unacylated ghrelin (UAG) analogues.
Ghrelin, an endocrine hormone predominantly produced by the stomach, exists in acylated and unacylated forms in the circulation. Unacylated ghrelin (UAG), the more abundant form in blood, possesses similar, independent or opposite physiological actions as acylated ghrelin (AG). AZP502, a linear 8-amino acid peptide from the central region of UAG (UAG(6-13)), and its full (AZP531) and partially (AZP533) cyclised derivatives, exhibit the same pharmacological profile as UAG both in vitro and in vivo, independently of AG receptor binding. We investigated the stability of these three fragments in vitro in human blood samples and in vivo after subcutaneous and intravenous injection in rats and dogs using liquid chromatography-mass spectrometry. In both species, AZP502 is rapidly degraded generating two major metabolites. Partial cyclisation of AZP502 and acylation at its N-terminus (AZP533 peptide) improves its stability in human plasma in vitro. Full cyclisation of AZP502 (AZP531 peptide) also completely protects the peptide from peptidase degradation in vitro in human blood samples. Moreover this cyclisation strongly improves the stability and the bioavailability of this peptide in vivo in both dogs and rats (mean bioavailability of 10-15% and 85-95% for AZP502 and AZP531 respectively). Taken together these results support the rationale for developing AZP531 as a long-acting UAG analogue for subcutaneous injection for the treatment of type 2 diabetes mellitus and other metabolic disorders.


Canadian economic evaluation of budesonide-formoterol as maintenance and reliever treatment in patients with moderate to severe asthma.
To compare the cost-effectiveness of budesonide-formoterol in a single inhaler used as both maintenance and reliever medication versus clinician-directed titration of salmeterol-fluticasone as maintenance medication, plus salbutamol taken as needed, in controlling asthma in adults and adolescents. A Canadian economic evaluation was conducted based on the results of a large (n=2143), open-label, randomized, controlled effectiveness trial in which health resource use was prospectively collected. The primary outcome measurement was the time to the first severe exacerbation. Costs included direct medical costs (physician and emergency room visits, hospitalizations, asthma drug costs, etc) and productivity (absenteeism). The time horizon was one year, which corresponded to the duration of the clinical trial. Prices were obtained from 2005 Canadian sources. Both health care and societal perspectives were considered, and deterministic univariate sensitivity analyses were conducted. In the clinical trial, budesonide-formoterol as maintenance and reliever treatment was superior to salmeterol-fluticasone with respect to the time to the first severe exacerbation, overall rate of exacerbations and use of as-needed reliever medication. The annualized rate of severe exacerbations was 0.24 events/patient in the budesonide-formoterol arm and 0.31 events/patient in the salmeterol-fluticasone arm (P=0.0025). From a health care perspective, the mean cost per patient-year was $1,315 in the budesonide-formoterol arm versus $1,541 in the salmeterol-fluticasone arm. From a societal perspective, the mean cost per patient-year was $1,538 in the budesonide-formoterol arm and $1,854 in the salmeterol-fluticasone arm. Budesonide-formoterol was dominant (more effective and less expensive) in the base case analysis from both perspectives. The results were robust under sensitivity testing. The strategy that allows budesonide-formoterol to be used in a single inhaler as both maintenance and reliever medication proved to be more effective and less expensive than a strategy of clinician-directed titration of salmeterol-fluticasone with salbutamol as reliever therapy.


Laboratory tests commonly used in complementary and alternative medicine: a review of the evidence.
It is increasingly easy for the general public to access a wide range of laboratory tests. Tests can be ordered online with little or no input from a health professional. The complementary and alternative medicine (CAM) community promote and sell a wide range of tests, many of which are of dubious clinical significance. Many have little or no clinical utility and have been widely discredited, whilst others are established tests that are used for unvalidated purposes. They range from the highly complex, employing state of the art technology, e.g. heavy metal analysis using inductively coupled plasma-mass spectrometry, to the rudimentary, e.g. live blood cell analysis. Results of 'CAM tests' are often accompanied by extensive clinical interpretations which may recommend, or be used to justify, unnecessary or harmful treatments. There are now a small number of laboratories across the globe that specialize in CAM testing. Some CAM laboratories operate completely outside of any accreditation programme whilst others are fully accredited to the standard of established clinical laboratories. In this review, we explore CAM testing in the United States, the United Kingdom and Australia with a focus on the common tests on offer, how they are reported, the evidence base for their clinical application and the regulations governing their use. We will also review proposed changed to in-vitro diagnostic device regulations and how these might impact on CAM testing.


CD2 sets quantitative thresholds in T cell activation.
It has been proposed that CD2, which is highly expressed on T cells, serves to enhance T cell-antigen presenting cell (APC) adhesion and costimulate T cell activation. Here we analyzed the role of CD2 using CD2-deficient mice crossed with transgenic mice expressing a T cell receptor specific for lymphocytic choriomeningitis virus (LCMV)-derived peptide p33. We found that absence of CD2 on T cells shifted the p33-specific dose-response curve in vitro by a factor of 3-10. In comparison, stimulation of T cells in the absence of lymphocyte function-associated antigen (LFA)-1-intercellular adhesion molecule (ICAM)-1 interaction shifted the dose-response curve by a factor of 10, whereas absence of both CD2-CD48 and LFA-1-ICAM-1 interactions shifted the response by a factor of approximately 100. This indicates that CD2 and LFA-1 facilitate T cell activation additively. T cell activation at low antigen density was blocked at its very first steps, as T cell APC conjugate formation, TCR triggering, and Ca(2+) fluxes were affected by the absence of CD2. In vivo, LCMV-specific, CD2-deficient T cells proliferated normally upon infection with live virus but responded in a reduced fashion upon cross-priming. Thus, CD2 sets quantitative thresholds and fine-tunes T cell activation both in vitro and in vivo.


Characterization of cell clones isolated from hypoxia-selected renal proximal tubular cells.
Under hypoxia, some cells survive and others are irreversibly injured and die. The factors that determine cell fate under stress remain largely unknown. We recently selected death-resistant cells via repeated episodes of hypoxia. In the present study, 80 clones were isolated from the selected cells and their response to apoptotic injury was characterized. Compared with the wild-type cells, the isolated clones showed a general resistance to apoptosis: 13 were extremely resistant to azide-induced apoptosis, 10 to staurosporine, and 9 to cisplatin. The cell clones that most consistently demonstrated resistance or sensitivity to injury were further studied for their response to azide treatment. Azide induced comparable ATP depletion in these clones and wild-type cells. Hypoxia inducible factor-1 (HIF-1) was upregulated in several clones, but the upregulation did not correlate with cell death resistance. The selected clones maintained an epithelial phenotype, showing typical epithelial morphology, forming "domes" at high density, and expressing E-cadherin. Azide-induced Bax translocation and cytochrome c release, two critical mitochondrial events of apoptosis, were abrogated in death-resistant clones. In addition, cell lysates isolated from these clones showed lower caspase activation on addition of exogenous cytochrome c. Bax, Bak, and Bid expression in these clones was similar to that in wild-type cells, whereas Bcl-2 expression was higher in all the selected clones and, interestingly, Bcl-xL was markedly upregulated in the most death-resistant clones. The results suggest that apoptotic resistance of the selected clones is not determined by a single factor or molecule but, rather, by various alterations at the core apoptotic pathway.


Considerations for using distributed research networks to conduct aspects of randomized trials.
Stakeholders in the clinical research enterprise are aligned around the need to make clinical research in general, and randomized controlled trials in particular, more meaningful and efficient. To that end, we have built distributed research networks (DRNs) for the Sentinel System, the National Institutes of Health (NIH) Collaboratory, and the National Patient-Centered Clinical Research Network (PCORnet). DRNs reuse electronic health record (EHR) and claims data for research. The design and use of health data DRNs is complicated by lack of uniformity in data collection, a fragmented healthcare system, and the imperative to protect research participants. We describe the key elements of successful DRNs, as well as methods, challenges, and solutions we have encountered in using DRNs to support different phases of randomized, multi-site, clinical research. This work supports "real-world" efforts to build a learning health system and will enable others to conduct randomized clinical trial research using a DRN.


Urinary proteome signature of Renal Cysts and Diabetes syndrome in children.
Renal Cysts and Diabetes Syndrome (RCAD) is an autosomal dominant disorder caused by mutations in the HNF1B gene encoding for the transcriptional factor hepatocyte nuclear factor-1B. RCAD is characterized as a multi-organ disease, with a broad spectrum of symptoms including kidney abnormalities (renal cysts, renal hypodysplasia, single kidney, horseshoe kidneys, hydronephrosis), early-onset diabetes mellitus, abnormal liver function, pancreatic hypoplasia and genital tract malformations. In the present study, using capillary electrophoresis coupled to mass spectrometry (CE-MS), we investigated the urinary proteome of a pediatric cohort of RCAD patients and different controls to identify peptide biomarkers and obtain further insights into the pathophysiology of this disorder. As a result, 146 peptides were found to be associated with RCAD in 22 pediatric patients when compared to 22 healthy age-matched controls. A classifier based on these peptides was generated and further tested on an independent cohort, clearly discriminating RCAD patients from different groups of controls. This study demonstrates that the urinary proteome of pediatric RCAD patients differs from autosomal dominant polycystic kidney disease (PKD1, PKD2), congenital nephrotic syndrome (NPHS1, NPHS2, NPHS4, NPHS9) as well as from chronic kidney disease conditions, suggesting differences between the pathophysiology behind these disorders.


Phosphorus-nitrogen compounds. 21. Syntheses, structural investigations, biological activities, and DNA interactions of new N/O spirocyclic phosphazene derivatives. The NMR behaviors of chiral phosphazenes with stereogenic centers upon the addition of chiral solvating agents.
The reactions of hexachlorocyclotriphosphazatriene, N(3)P(3)Cl(6), with N/O-donor-type N-alkyl (or aryl)-o-hydroxybenzylamines (1a-1e) produce mono- (2a-2e), di- (3a-3d), and tri- (4a and 4b) spirocyclic phosphazenes. The tetrapyrrolidino monospirocyclic phosphazenes (2f-2i) are prepared from the reactions of partly substituted compounds (2a-2d) with excess pyrrolidine. The dispirodipyrrolidinophosphazenes (3e-3h) and trispirophosphazenes (3i-3k) are obtained from the reactions of trans-dispirophosphazenes with excess pyrrolidine and sodium (3-amino-1-propanoxide), respectively. Compounds 3a-3d have cis and trans geometric isomers. Only the trans isomers of these compounds are isolated. Compounds 3a-3h have two stereogenic P atoms. They are expected to be in cis (meso) and trans (racemic) geometric isomers. In the trans trispiro compounds (3i-3k), there are three stereogenic P atoms. They are expected to be in racemic mixtures. The stereogenic properties of 3a-3k are confirmed by (31)P NMR spectroscopy upon the addition of the chiral solvating agent; (S)-(+)-2,2,2-trifluoro-1-(9'-anthryl)ethanol. The molecular structures of 3i-3k, 4a, and 4b look similar to a propeller, where the chemical environment of one P atom is different from that of others. Additionally, 4a and 4b are also expected to exist as cis-trans-trans and cis-cis-cis geometric isomers, but both of them are found to be in cis-trans-trans geometries. The solid-state structures of 2a, 2e, 2f, 3e, and 3f are determined by X-ray crystallography. The compounds 2f-2i, 3e-3i, and 3k are screened for antibacterial activity against gram-positive and gram-negative bacteria and for antifungal activity against yeast strains. These compounds (except 3f) have shown a strong affinity against most of the bacteria. Minimum inhibitory concentrations (MIC) are determined for 2f-2i, 3e-3i, and 3k. DNA binding and the nature of interaction with pUC18 plasmid DNA are studied. The compounds 2f-2i, 3e-3i, and 3k induce changes on the DNA mobility. The prevention of BamHI and HindIII digestion (except 2g) with compounds indicates that the compounds bind with nucleotides in DNA.


Reliability of vertebral fracture assessment using multidetector CT lateral scout views: the Framingham Osteoporosis Study.
Two radiologists evaluated images of the spine from computed tomography (CT) scans on two occasions to diagnose vertebral fracture in 100 individuals. Agreement was fair to good for mild fractures, and agreement was good to excellent for more severe fractures. CT scout views are useful to assess vertebral fracture. We investigated inter-reader agreement between two radiologists and intra-reader agreement between duplicate readings for each radiologist, in assessment of vertebral fracture using a semi-quantitative method from lateral scout views obtained by CT. Participants included 50 women and 50 men (age 50-87 years, mean 70 years) in the Framingham Study. T4-L4 vertebrae were assessed independently by two radiologists on two occasions using a semi-quantitative scale as normal, mild, moderate, or severe fracture. Vertebra-specific prevalence of grade ≥ 1 (mild) fracture ranged from 3% to 5%. We found fair (κ = 56-59%) inter-reader agreement for grade ≥ 1 vertebral fractures and good (κ = 68-72%) inter-reader agreement for grade ≥ 2 fractures. Intra-reader agreement for grade ≥ 1 vertebral fracture was fair (κ = 55%) for one reader and excellent for another reader (κ = 77%), whereas intra-reader agreement for grade ≥ 2 vertebral fracture was excellent for both readers (κ = 76% and 98%). Thoracic vertebrae were more difficult to evaluate than the lumbar region, and agreement was lowest (inter-reader κ = 43%) for fracture at the upper (T4-T9) thoracic levels and highest (inter-reader κ = 76-78%) for the lumbar spine (L1-L4). Based on a semi-quantitative method to classify vertebral fractures using CT scout views, agreement within and between readers was fair to good, with the greatest source of variation occurring for fractures of mild severity and for the upper thoracic region. Agreement was good to excellent for fractures of at least moderate severity. Lateral CT scout views can be useful in clinical research settings to assess vertebral fracture.


A Common Mechanism Links Activities of Butyrate in the Colon.
Two biological activities of butyrate in the colon (suppression of proliferation of colonic epithelial stem cells and inflammation) correlate with inhibition of the activity of histone deacetylases. Cellular and biochemical studies of molecules similar in structure to butyrate, but different in molecular details (functional groups, chain-length, deuteration, oxidation level, fluorination, or degree of unsaturation), demonstrated that these activities were sensitive to molecular structure, and were compatible with the hypothesis that butyrate acts by binding to the Zn2+ in the catalytic site of histone deacetylases. Structure-activity relationships drawn from a set of 36 compounds offer a starting point for the design of new compounds targeting the inhibition of histone deacetylases. The observation that butyrate was more potent than other short-chain fatty acids is compatible with the hypothesis that crypts evolved (at least in part), to separate stem cells at the base of crypts from butyrate produced by commensal bacteria.


Meditation training modulates brain electric microstates and felt states of awareness.
Meditation practice is believed to foster states of mindful awareness and mental quiescence in everyday life. If so, then the cultivation of these qualities with training ought to leave its imprint on the activity of intrinsic functional brain networks. In an intensive longitudinal study, we investigated associations between meditation practitioners' experiences of felt mindful awareness and changes in the spontaneous electrophysiological dynamics of functional brain networks. Experienced meditators were randomly assigned to complete 3 months of full-time training in focused-attention meditation (during an initial intervention) or to serve as waiting-list controls and receive training second (during a later intervention). We collected broadband electroencephalogram (EEG) during rest at the beginning, middle, and end of the two training periods. Using a data-driven approach, we segmented the EEG into a time series of transient microstate intervals based on clustering of topographic voltage patterns. Participants also provided daily reports of felt mindful awareness and mental quiescence, and reported daily on four experiential qualities of their meditation practice during training. We found that meditation training led to increases in mindful qualities of awareness, which corroborate contemplative accounts of deepening mental calm and attentional focus. We also observed reductions in the strength and duration of EEG microstates across both interventions. Importantly, changes in the dynamic sequencing of microstates were associated with daily increases in felt attentiveness and serenity during training. Our results connect shifts in subjective qualities of meditative experience with the large-scale dynamics of whole brain functional EEG networks at rest.


Identification of Burkholderia mallei and Burkholderia pseudomallei adhesins for human respiratory epithelial cells.
Burkholderia pseudomallei and Burkholderia mallei cause the diseases melioidosis and glanders, respectively. A well-studied aspect of pathogenesis by these closely-related bacteria is their ability to invade and multiply within eukaryotic cells. In contrast, the means by which B. pseudomallei and B. mallei adhere to cells are poorly defined. The purpose of this study was to identify adherence factors expressed by these organisms. Comparative sequence analyses identified a gene product in the published genome of B. mallei strain ATCC23344 (locus # BMAA0649) that resembles the well-characterized Yersinia enterocolitica autotransporter adhesin YadA. The gene encoding this B. mallei protein, designated boaA, was expressed in Escherichia coli and shown to significantly increase adherence to human epithelial cell lines, specifically HEp2 (laryngeal cells) and A549 (type II pneumocytes), as well as to cultures of normal human bronchial epithelium (NHBE). Consistent with these findings, disruption of the boaA gene in B. mallei ATCC23344 reduced adherence to all three cell types by ~50%. The genomes of the B. pseudomallei strains K96243 and DD503 were also found to contain boaA and inactivation of the gene in DD503 considerably decreased binding to monolayers of HEp2 and A549 cells and to NHBE cultures.A second YadA-like gene product highly similar to BoaA (65% identity) was identified in the published genomic sequence of B. pseudomallei strain K96243 (locus # BPSL1705). The gene specifying this protein, termed boaB, appears to be B. pseudomallei-specific. Quantitative attachment assays demonstrated that recombinant E. coli expressing BoaB displayed greater binding to A549 pneumocytes, HEp2 cells and NHBE cultures. Moreover, a boaB mutant of B. pseudomallei DD503 showed decreased adherence to these respiratory cells. Additionally, a B. pseudomallei strain lacking expression of both boaA and boaB was impaired in its ability to thrive inside J774A.1 murine macrophages, suggesting a possible role for these proteins in survival within professional phagocytic cells. The boaA and boaB genes specify adhesins that mediate adherence to epithelial cells of the human respiratory tract. The boaA gene product is shared by B. pseudomallei and B. mallei whereas BoaB appears to be a B. pseudomallei-specific adherence factor.


Ultra-wideband three-dimensional optoacoustic tomography.
Broadband optoacoustic waves generated by biological tissues excited with nanosecond laser pulses carry information corresponding to a wide range of geometrical scales. Typically, the frequency content present in the signals generated during optoacoustic imaging is much larger compared to the frequency band captured by common ultrasonic detectors, the latter typically acting as bandpass filters. To image optical absorption within structures ranging from entire organs to microvasculature in three dimensions, we implemented optoacoustic tomography with two ultrasound linear arrays featuring a center frequency of 6 and 24 MHz, respectively. In the present work, we show that complementary information on anatomical features could be retrieved and provide a better understanding on the localization of structures in the general anatomy by analyzing multi-bandwidth datasets acquired on a freshly excised kidney.


Spontaneous ventilation and respiratory motor output during carbachol-induced atonia of REM sleep in the decerebrate cat.
Microinjections of carbachol into the pons induce a state that resembles rapid eye movement (REM) sleep in intact cats and, in decerebrate, artificially ventilated cats, produce postural atonia accompanied by a powerful depression of the respiratory motor output. In this study, pontine carbachol was used in decerebrate, spontaneously breathing cats to assess the effects of mechanical and chemical respiratory reflexes on the magnitude and pattern of the carbachol-induced depression of breathing, and to determine whether the depression is altered in those animals in which rapid eye movements are present. Phrenic nerve activity and tidal volume were only transiently depressed at the onset of the carbachol-induced postural atonia, whereas the decrease in respiratory rate and the depressions of hypoglossal and intercostal activities persisted until the response was reversed by a pontine microinjection of atropine 15-101 minutes after the onset of carbachol response. Ventilation was reduced to 70% of control during the steady-state conditions. The irregularity of breathing, characterized by the inter-quartile ranges of the distributions of the peak phrenic nerve activity and respiratory timing, did not increase following pontine carbachol. Neither vagotomy nor vigorous eye movements were associated with increased breathing irregularity. This contrasts with the irregular breathing (with minor average changes in ventilation) typical of natural REM sleep. We propose that the carbachol-injected decerebrate cat provides a useful model of the depressant effects that neural events associated with REM sleep may have on breathing.


Mapping displacement and deformation of the heart with local sine-wave modeling.
The new SinMod method extracts motion from magnetic resonance imaging (MRI)-tagged (MRIT) image sequences. Image intensity in the environment of each pixel is modeled as a moving sine wavefront. Displacement is estimated at subpixel accuracy. Performance is compared with the harmonic-phase analysis (HARP) method, which is currently the most common method used to detect motion in MRIT images. SinMod can handle line tags, as well as speckle patterns. In artificial images (tag distance six pixels), SinMod detects displacements accurately (error < 0.02 pixels). Effects of noise are suppressed effectively. Sharp transitions in motion at the boundary of an object are smeared out over a width of 0.6 tag distance. For MRIT images of the heart, SinMod appears less sensitive to artifacts, especially later in the cardiac cycle when image quality deteriorates. For each pixel, the quality of the sine-wave model in describing local image intensity is quantified objectively. If local quality is low, artifacts are avoided by averaging motion over a larger environment. Summarizing, SinMod is just as fast as HARP, but it performs better with respect to accuracy of displacement detection, noise reduction, and avoidance of artifacts.


Adding web-based behavioural support to exercise referral schemes for inactive adults with chronic health conditions: the e-coachER RCT.
There is modest evidence that exercise referral schemes increase physical activity in inactive individuals with chronic health conditions. There is a need to identify additional ways to improve the effects of exercise referral schemes on long-term physical activity. To determine if adding the e-coachER intervention to exercise referral schemes is more clinically effective and cost-effective in increasing physical activity after 1 year than usual exercise referral schemes. A pragmatic, multicentre, two-arm randomised controlled trial, with a mixed-methods process evaluation and health economic analysis. Participants were allocated in a 1 : 1 ratio to either exercise referral schemes plus e-coachER (intervention) or exercise referral schemes alone (control). Patients were referred to exercise referral schemes in Plymouth, Birmingham and Glasgow. There were 450 participants aged 16-74 years, with a body mass index of 30-40 kg/m2, with hypertension, prediabetes, type 2 diabetes, lower limb osteoarthritis or a current/recent history of treatment for depression, who were also inactive, contactable via e-mail and internet users. e-coachER was designed to augment exercise referral schemes. Participants received a pedometer and fridge magnet with physical activity recording sheets, and a user guide to access the web-based support in the form of seven 'steps to health'. e-coachER aimed to build the use of behavioural skills (e.g. self-monitoring) while strengthening favourable beliefs in the importance of physical activity, competence, autonomy in physical activity choices and relatedness. All participants were referred to a standard exercise referral scheme. Minutes of moderate and vigorous physical activity in ≥ 10-minute bouts measured by an accelerometer over 1 week at 12 months, worn ≥ 16 hours per day for ≥ 4 days including ≥ 1 weekend day. Other accelerometer-derived physical activity measures, self-reported physical activity, exercise referral scheme attendance and EuroQol-5 Dimensions, five-level version, and Hospital Anxiety and Depression Scale scores were collected at 4 and 12 months post randomisation. Participants had a mean body mass index of 32.6 (standard deviation) 4.4 kg/m2, were referred primarily for weight loss and were mostly confident self-rated information technology users. Primary outcome analysis involving those with usable data showed a weak indicative effect in favour of the intervention group (n = 108) compared with the control group (n = 124); 11.8 weekly minutes of moderate and vigorous physical activity (95% confidence interval -2.1 to 26.0 minutes; p = 0.10). Sixty-four per cent of intervention participants logged on at least once; they gave generally positive feedback on the web-based support. The intervention had no effect on other physical activity outcomes, exercise referral scheme attendance (78% in the control group vs. 75% in the intervention group) or EuroQol-5 Dimensions, five-level version, or Hospital Anxiety and Depression Scale scores, but did enhance a number of process outcomes (i.e. confidence, importance and competence) compared with the control group at 4 months, but not at 12 months. At 12 months, the intervention group incurred an additional mean cost of £439 (95% confidence interval -£182 to £1060) compared with the control group, but generated more quality-adjusted life-years (mean 0.026, 95% confidence interval 0.013 to 0.040), with an incremental cost-effectiveness ratio of an additional £16,885 per quality-adjusted life-year. A significant proportion (46%) of participants were not included in the primary analysis because of study withdrawal and insufficient device wear-time, so the results must be interpreted with caution. The regression model fit for the primary outcome was poor because of the considerable proportion of participants [142/243 (58%)] who recorded no instances of ≥ 10-minute bouts of moderate and vigorous physical activity at 12 months post randomisation. The design and rigorous evaluation of cost-effective and scalable ways to increase exercise referral scheme uptake and maintenance of moderate and vigorous physical activity are needed among patients with chronic conditions. Adding e-coachER to usual exercise referral schemes had only a weak indicative effect on long-term rigorously defined, objectively assessed moderate and vigorous physical activity. The provision of the e-coachER support package led to an additional cost and has a 63% probability of being cost-effective based on the UK threshold of £30,000 per quality-adjusted life-year. The intervention did improve some process outcomes as specified in our logic model. Current Controlled Trials ISRCTN15644451. This project was funded by the National Institute for Health Research (NIHR) Health Technology Assessment programme and will be published in full in Health Technology Assessment; Vol. 24, No. 63. See the NIHR Journals Library website for further project information.


Sesamol, a lipid lowering agent, ameliorates aluminium chloride induced behavioral and biochemical alterations in rats.
Sesame oil from the seeds of Sesamum indicum Linn. (Pedaliaceae) has been used traditionally in Indian medical practice of Ayurveda in the treatment of central nervous system disorders and insomnia. A few published reports favor the anti-dementia effect of sesamol (SML), an active constituent of sesame oil. Thus, the present study was aimed to explore the anti-dementia effect and possible mechanism (s) of SML in aluminium chloride (AlCl3)-induced cognitive dysfunction model in rodents with special emphasis on memory centers viz., hippocampus and frontal cortex. Male Wistar rats were exposed to AlCl3 (175 mg/kg p.o.) for 60 days. SML (10 and 20 mg/kg) and rivastigmine (1 mg/kg) were administered orally 45 min before administration of AlCl3 for 60 days. Spatial memory was assessed using Morris water maze test. After 60 days of treatment animals were sacrificed, hippocampus and frontal cortex were collected and analyzed for acetylcholinesterase (AChE) activity, tumor necrosis factor (TNF-α) level, antioxidant enzymes (Glutathione, catalase), lipid peroxidation, and nitrite level. The circulating triglycerides, total cholesterol, low-density lipoprotein (LDL) and high-density lipoprotein (HDL) levels were also analyzed. SML significantly prevented behavioral impairments in aluminium-exposed rats. Treatment with SML reversed the increased cholesterol, triglycerides and LDL while raised the HDL levels. SML significantly corrected the effect of AlCl3 on AChE activity. Further, SML reversed the elevated nitric oxide, TNF-α and reduced antioxidant enzymes in hippocampus and frontal cortex. The present study suggests the neuro-protection by SML against cognitive dysfunction induced by environmental toxin (AlCl3) in hippocampus and frontal cortex.


Cord blood angiogenic progenitor cells are decreased in bronchopulmonary dysplasia.
Bronchopulmonary dysplasia (BPD), the chronic lung disease of prematurity, is associated with impaired vascular and alveolar growth. Antenatal factors contribute to the risk for developing BPD by unclear mechanisms. Endothelial progenitor cells, such as angiogenic circulating progenitor cells (CPCs) and late-outgrowth endothelial colony-forming cells (ECFCs), may contribute to angiogenesis in the developing lung. We hypothesise that cord blood angiogenic CPCs and ECFCs are decreased in preterm infants with moderate and severe BPD. We quantified ECFCs and the CPC/nonangiogenic-CPC ratio (CPC/non-CPC) in cord blood samples from 62 preterm infants and assessed their relationships to maternal and perinatal risk factors as well as BPD severity. The CPC/non-CPC ratio and ECFC number were compared between preterm infants with mild or no BPD and those with moderate or severe BPD. ECFC number (p<0.001) and CPC/non-CPC ratio (p<0.05) were significantly decreased in cord blood samples of preterm infants who subsequently developed moderate or severe BPD. Gestational age and birth weight were not associated with either angiogenic marker. Circulating vascular progenitor cells are decreased in the cord blood of preterm infants who develop moderate and severe BPD. These findings suggest that prenatal factors contribute to late respiratory outcomes in preterm infants.


Transplantation of a lobe of lung from mother to child following previous transplantation with maternal bone marrow.
A left lower lobe of the lung was transplanted from a mother to her child, who had previously received a maternal bone marrow transplant for an immune defect. Following the bone marrow transplantation, the child had developed severe pulmonary fibrosis. Surgery and the early postoperative course have been uncomplicated. Immunosuppression with corticosteroids was administered for a short period, after which all immunosuppressive treatment was discontinued. The operation and the outcome are described both in the donor and recipient. Rehabilitation was slow, but one year later the patient is doing well.


Local infiltration analgesia for total knee arthroplasty: Does a mixture of ropivacaine and epinephrine have an impact on hemodynamics? An observational cohort study.
High doses of local anesthetic administered intra-articularly and peri-articularly during local infiltration analgesia (LIA) for total knee arthroplasty (TKA) may have potential effects on patient hemodynamics. The aim of this study was to know if hemodynamic changes are associated with LIA in patients undergoing TKA. In a prospective observational design, elective patients undergoing orthopedic surgery for TKA and treated with LIA consisting of a mixture of ropivacaine (300 mg) and epinephrine (1 mg) were investigated for changes in selected hemodynamic parameters: heart rate (HR), non-invasively registered mean arterial blood pressure (MAP), and incidence of arrhythmias during the perioperative course, consisting of the following periods: period 1. from establishment of spinal anesthesia to prior to LIA administration, period 2. from administration of LIA to before release of ischemia tourniquet, period 3. from release ischemia tourniquet to end of surgery, and period 4. from transfer to the post anesthesia care unit to the ward. Statistical analysis was done with ANOVA-RM for the difference in means in repeated measurements, and with the Tukey Test between pairs. Data are presented as mean ± standard deviation. A P value <0.05 was considered significant. Ninety-nine patients (mean age 77 ± 8 years) were included. HR increased from period 2 to period 3 up to 16% (67 ± 14 to 77 ± 13 bpm, P < 0.001), and from period 2 to period 4 up to 21% (67 ± 14 to 81 ± 12 bpm, P < 0.001). MAP showed no significant changes from period 2 to period 3 (89 ± 13 to 87 ± 13 mmHg, P > 0.50), and from period 2 to period 4 (89 ± 13 to 91 ± 11 mmHg, P > 0.50). No arrhythmias were detected during follow-up. A mixture of ropivacaine and epinephrine for LIA, despite the high doses administered, does not have a negative impact on hemodynamics.


Superimposing incident sexually transmitted infections on HIV phylogram to investigate possible misclassification of men who have sex with men as heterosexuals in a cohort in Antwerp, Belgium.
In this study, we assessed if the superimposition of incident sexually transmitted infections (STIs) on HIV phylogenetic analyses could reveal possible sexual behaviour misclassifications in our HIV-infected population. HIV-1 sequences collected between 1997 and 2014 from 1169 individuals attending a HIV clinic in Antwerp, Belgium were analysed to infer a partial HIV transmission network. Individual demographic, clinical and laboratory data collected during routine HIV follow-up were used to compare clustered and non-clustered individuals using logistic regression analyses. In total, 438 (37.5%) individuals were identified in 136 clusters, including 76 transmission pairs and 60 clusters consisting of three or more individuals. Individuals in a cluster were more likely to have a history of syphilis, Chlamydia and/or gonorrhoea (P < 0.05); however, when analyses were stratified by HIV transmission risk groups (heterosexual and men who have sex with men [MSM]), this association only remained significant for heterosexuals with syphilis (P = 0.001). Under closer scrutiny, this association was driven by six heterosexual men who were located in six almost exclusively MSM clusters. A parsimonious conclusion is that these six individuals were potentially misclassified as heterosexual. Improving the accuracy of sexual behaviour reporting could improve care.


Autoantibodies to Dense Fine Speckles in Pediatric Diseases and Controls.
Autoantibodies to the dense fine speckled 70 kDa antigen (DFS70) are reported to be more common in individuals who do not have an antinuclear antibody (ANA)-associated rheumatic disease (AARD) than in patients with AARD. The frequency of anti-DFS70 antibodies has been thoroughly studied in adult but not in pediatric populations. The primary objective of this observational study was to determine the frequency of anti-DFS70 in pediatric AARD and reference cohorts. Sera from 743 children with AARD and related conditions, and 345 samples from reference cohorts (healthy children and those being investigated for AARD) were studied for anti-DFS70 autoantibodies as measured by a chemiluminescence immunoassay. A de-identified administrative database was used to retrieve demographic, serologic, and clinical data. Anti-DFS70 antibodies were seen in 2.1% of healthy children and in 4.5% of sera from pediatric individuals referred for ANA testing. The frequency of anti-DFS70 was highest in juvenile localized scleroderma (LS; 4/29, 13.8%), juvenile dermatomyositis (JDM; 2/11, 18.2%), childhood systemic lupus erythematosus (cSLE; 19/331, 5.7%), diffuse cutaneous systemic sclerosis (1/22, 4.5%), celiac disease (2/49, 4.1%), and juvenile idiopathic arthritis (JIA; 5/202, 2.5%). Of note, anti-DFS70 antibodies were observed in 3/26 children (11.5%) with uveitis and JIA-associated uveitis. The frequency of anti-DFS70 autoantibodies in healthy pediatric subjects is within the lower range of that reported in adults. Anti-DFS70 antibodies can be found in childhood SSc and cSLE, but has a remarkably high frequency in children with LS, JDM, and uveitis.


Seven million years of glaciation in greenland.
Glacial till, glaciomarine diamictites, and ice-rafted detritus found in marine cores collected off the shore of southeast Greenland record multiple Late Cenozoic glaciations beginning in the Late Miocene. Distinct rock assemblages and seismic stratigraphic control correlate the diamictites with glaciation of the southeast Greenland margin. Glaciers advanced to the sea during several intervals in the Pliocene and Pleistocene. North Atlantic glaciation may have nucleated in southern Greenland rather than further north because of the high mountains and the high levels of precipitation in this region.


Mobility and efficacy of abamectin and imidacloprid against Rhynchophorus ferrugineus in Phoenix canariensis by different application methods.
Rhynchophorus ferrugineus is the most destructive pest of palms. As detection of early infestation stages is difficult, preventive measures, mostly chemical control, are crucial. Stipe injection of insecticides has developed rapidly as a suitable technique. However, pesticide movement within palms and palm reaction to wounding remain controversial. We used abamectin and imidacloprid applied by crown spray, stipe and frond injections to disentangle how these pesticides move within P. canariensis and how tissues wounded by injection heal. Furthermore, we established their lethal doses to larvae of R. ferrugineus. Maximum residues of imidacloprid (0.1 mg kg(-1) ) were detected in crown and frond samples for up to 2 months after stipe injection, whereas maximum residues of abamectin were found in frond tip samples (0.5 mg active substance kg(-1) ) 5 months after stipe injection. Based on the lethal concentrations calculated, these doses could satisfactorily protect palms for up to 3 months after treatment. No significant wound damage was observed 2 years after injection. Stipe injection, irrespective of the active substance considered, resulted in better distribution and higher persistence compared with frond injection and, especially, crown spray. As a consequence, our results point to stipe injection as a good alternative to control R. ferrugineus.