3D pose estimation of ground rigid target based on ladar range image.
In the target recognition of laser radar (ladar), the accurate estimation of target pose can effectively simplify the recognition process. To achieve 3D pose estimation of rigid objects on the ground and simplify the complexity of the algorithm, a novel pose estimation method is proposed in this paper. In this approach, based on the feature that most rigid objects on the ground have large planar areas which are horizontal on the top of the targets and vertical sides and combined with the 3D geometric characteristics of ladar range images, the planar normals of rigid targets were adopted as the vectors in the positive direction of the axes in the model coordinate system to estimate the 3D pose angles of targets. The simulation experiments were performed with six military vehicle models and the performance in self-occlusion, occlusion, and noise was investigated. The results show that the estimation errors are less than 2° in self-occlusion. For the tank LECRERC model, as long as the upper and side planes of the target are not completely occluded, even though the occlusion reaches 80%, the pose angles can be estimated with the estimation error less than 2.5°. Moreover, the proposed method is robust to noise and effective.


Mitochondrial genomes reveal an explosive radiation of extinct and extant bears near the Miocene-Pliocene boundary.
Despite being one of the most studied families within the Carnivora, the phylogenetic relationships among the members of the bear family (Ursidae) have long remained unclear. Widely divergent topologies have been suggested based on various data sets and methods. We present a fully resolved phylogeny for ursids based on ten complete mitochondrial genome sequences from all eight living and two recently extinct bear species, the European cave bear (Ursus spelaeus) and the American giant short-faced bear (Arctodus simus). The mitogenomic data yield a well-resolved topology for ursids, with the sloth bear at the basal position within the genus Ursus. The sun bear is the sister taxon to both the American and Asian black bears, and this clade is the sister clade of cave bear, brown bear and polar bear confirming a recent study on bear mitochondrial genomes. Sequences from extinct bears represent the third and fourth Pleistocene species for which complete mitochondrial genomes have been sequenced. Moreover, the cave bear specimen demonstrates that mitogenomic studies can be applied to Pleistocene fossils that have not been preserved in permafrost, and therefore have a broad application within ancient DNA research. Molecular dating of the mtDNA divergence times suggests a rapid radiation of bears in both the Old and New Worlds around 5 million years ago, at the Miocene-Pliocene boundary. This coincides with major global changes, such as the Messinian crisis and the first opening of the Bering Strait, and suggests a global influence of such events on species radiations.


A limit on the variation of the speed of light arising from quantum gravity effects.
A cornerstone of Einstein's special relativity is Lorentz invariance-the postulate that all observers measure exactly the same speed of light in vacuum, independent of photon-energy. While special relativity assumes that there is no fundamental length-scale associated with such invariance, there is a fundamental scale (the Planck scale, l(Planck) approximately 1.62 x 10(-33) cm or E(Planck) = M(Planck)c(2) approximately 1.22 x 10(19) GeV), at which quantum effects are expected to strongly affect the nature of space-time. There is great interest in the (not yet validated) idea that Lorentz invariance might break near the Planck scale. A key test of such violation of Lorentz invariance is a possible variation of photon speed with energy. Even a tiny variation in photon speed, when accumulated over cosmological light-travel times, may be revealed by observing sharp features in gamma-ray burst (GRB) light-curves. Here we report the detection of emission up to approximately 31 GeV from the distant and short GRB 090510. We find no evidence for the violation of Lorentz invariance, and place a lower limit of 1.2E(Planck) on the scale of a linear energy dependence (or an inverse wavelength dependence), subject to reasonable assumptions about the emission (equivalently we have an upper limit of l(Planck)/1.2 on the length scale of the effect). Our results disfavour quantum-gravity theories in which the quantum nature of space-time on a very small scale linearly alters the speed of light.


Use of SGLT-2 Inhibitors in Patients with Type 2 Diabetes Mellitus and Abdominal Obesity: An Asian Perspective and Expert Recommendations.
The prevalence of obesity in Asia is of epidemic proportions, with an estimated 1 billion overweight/obese individuals in the region. The majority of patients with type 2 diabetes mellitus (T2DM) are overweight/obese, which increases the risk of cardiorenal outcomes in these patients; hence, sustained reductions in body weight and visceral adiposity are important management goals. However, most of the glucose-lowering therapies such as insulin, sulfonylureas, glinides, and thiazolidinediones induce weight gain, which makes the management of overweight/obese T2DM patients challenging. Sodium-glucose cotransporter-2 (SGLT-2) inhibitors are the only oral glucose-lowering agents that have been shown to reduce body weight and visceral adiposity. In addition, SGLT-2 inhibitors therapy reduces ectopic fat deposition and improves adipose tissue function and weight-related quality of life. In this article, we aim to consolidate the existing literature on the effects of SGLT-2 inhibitors in Asian patients with T2DM and to produce clinical recommendations on their use in overweight or obese patients with T2DM. Recommendations from international and regional guidelines, as well as published data from clinical trials in Asian populations and cardiovascular outcomes trials are reviewed. Based on the available data, SGLT-2 inhibitors represent an evidence-based therapeutic option for the management of overweight/obese patients with T2DM.


Validation of the force and frequency characteristics of the activator adjusting instrument: effectiveness as a mechanical impedance measurement tool.
To determine the dynamic force-time and force-frequency characteristics of the Activator Adjusting Instrument and to validate its effectiveness as a mechanical impedance measurement device; in addition, to refine or optimize the force-frequency characteristics of the Activator Adjusting Instrument to provide enhanced dynamic structural measurement reliability and accuracy. An idealized test structure consisting of a rectangular steel beam with a static stiffness similar to that of the human thoracolumbar spine was used for validation of a method to determine the dynamic mechanical response of the spine. The Activator Adjusting Instrument equipped with a load cell and accelerometer was used to measure forces and accelerations during mechanical excitation of the steel beam. Driving point and transfer mechanical impedance and resonant frequency of the beam were determined by use of a frequency spectrum analysis for different force settings, stylus masses, and stylus tips. Results were compared with beam theory and transfer impedance measurements obtained by use of a commercial electronic PCB impact hammer. The Activator Adjusting Instrument imparted a very complex dynamic impact comprising an initial high force (116 to 140 N), short duration pulse (<0.1 ms) followed by several lower force (30 to 100 N), longer duration impulses (1 to 5 ms). The force profile was highly reproducible in terms of the peak impulse forces delivered to the beam structure (<8% variance). Spectrum analysis of the Activator Adjusting Instrument impulse indicated that the Activator Adjusting Instrument has a variable force spectrum and delivers its peak energy at a frequency of 20 Hz. Added masses and different durometer stylus tips had very little influence on the Activator Adjusting Instrument force spectrum. The resonant frequency of the beam was accurately predicted by both the Activator Adjusting Instrument and electronic PCB impact hammer, but variations in the magnitude of the driving point impedance at the resonant frequency were high (67%) compared with the transfer impedance measurements obtained with the electronic PCB impact hammer, which had a more uniform force spectrum and was more repeatable (<10% variation). The addition of a preload-control frame to the Activator Adjusting Instrument improved the characteristics of the force frequency spectrum and repeatability of the driving point impedance measurements. These findings indicate that the Activator Adjusting Instrument combined with an integral load cell and accelerometer was able to obtain an accurate description of a steel beam with readily identifiable geometric and dynamic mechanical properties. These findings support the rationale for using the device to assess the dynamic mechanical behavior of the vertebral column. Such information would be useful for SMT and may ultimately be used to evaluate the [corrected] biomechanical effectiveness of various manipulative, surgical, and rehabilitative spinal procedures.


Senataxin associates with replication forks to protect fork integrity across RNA-polymerase-II-transcribed genes.
Transcription hinders replication fork progression and stability. The ATR checkpoint and specialized DNA helicases assist DNA synthesis across transcription units to protect genome integrity. Combining genomic and genetic approaches together with the analysis of replication intermediates, we searched for factors coordinating replication with transcription. We show that the Sen1/Senataxin DNA/RNA helicase associates with forks, promoting their progression across RNA polymerase II (RNAPII)-transcribed genes. sen1 mutants accumulate aberrant DNA structures and DNA-RNA hybrids while forks clash head-on with RNAPII transcription units. These replication defects correlate with hyperrecombination and checkpoint activation in sen1 mutants. The Sen1 function at the forks is separable from its role in RNA processing. Our data, besides unmasking a key role for Senataxin in coordinating replication with transcription, provide a framework for understanding the pathological mechanisms caused by Senataxin deficiencies and leading to the severe neurodegenerative diseases ataxia with oculomotor apraxia type 2 and amyotrophic lateral sclerosis 4.


Randomized, placebo-controlled phase II study of vandetanib plus docetaxel in previously treated non small-cell lung cancer.
Vandetanib is a once-daily oral inhibitor of vascular endothelial growth factor receptor-2 and epidermal growth factor receptor kinase activity. The activity of vandetanib plus docetaxel was assessed in patients with previously treated non-small-cell lung cancer (NSCLC). This two-part study comprised an open-label run-in phase and a double-blind randomized phase. Eligible patients had locally advanced or metastatic (stage IIIB/IV) NSCLC after failure of first-line platinum-based chemotherapy. The primary objective of the randomized phase was to prolong progression-free survival (PFS) in patients receiving vandetanib (100 or 300 mg/d) plus docetaxel (75 mg/m2 intravenous infusion every 21 days) versus placebo plus docetaxel. The study was designed to have more than 75% power to detect 50% prolongation at a one-sided significance level of P < .20. Secondary objectives included objective response rate, overall survival, safety and tolerability. In the randomized phase (n = 127), median PFS was 18.7 weeks for vandetanib 100 mg plus docetaxel (n = 42; hazard ratio v docetaxel = 0.64; one-sided P = .037); 17.0 weeks for vandetanib 300 mg plus docetaxel (n = 44; hazard ratio v docetaxel = 0.83; one-sided P = .231); and 12 weeks for docetaxel (n = 41). There was no statistically significant difference in overall survival among the three treatment arms. Common adverse events included diarrhea, rash, and asymptomatic prolongation of corrected QT (QTC) interval. The primary objective was achieved, with vandetanib 100 mg plus docetaxel demonstrating a significant prolongation of PFS compared with docetaxel in relation to the prespecified significance level. On the basis of these encouraging data, phase III evaluation of vandetanib 100 mg plus docetaxel in second-line NSCLC has been initiated.


Humoral immune response to alpha-methylacyl-CoA racemase and prostate cancer.
Although prostate-specific antigen (PSA) is a prototypic biomarker for prostate cancer, it has poor specificity. Expression of alpha-methylacyl-CoA racemase (AMACR), which is involved in the conversion of R-stereoisomers of branched-chain fatty acids to S-stereoisomers, has been shown to be specifically increased in prostate cancer epithelia. However, attempts to detect AMACR in circulation have not been successful. Hence, we determined whether an immune response to AMACR could be used as a serum biomarker for prostate cancer. Sera from patients with biopsy-proven prostate cancer and from control subjects were screened for a humoral immune response to selected tumor antigens, including AMACR, by using protein microarrays (46 patients, 28 control subjects). Humoral immune response to AMACR was then validated using high-throughput immunoblot analysis (151 patients, 259 control subjects) and enzyme-linked immunosorbent assay (ELISA) (54 patients, 55 control subjects). Receiver operating characteristic curves were used to determine the sensitivity and specificity of the immune response to AMACR to detect prostate cancer. Immunoreactivity against AMACR was statistically significantly higher in sera from patients with prostate cancer than in control subjects by all three techniques (P(protein microarray) =.009, P(immunoblot)<.001, P(ELISA) =.011). High-throughput immunoblot analysis revealed that, in subjects with intermediate PSA levels (4-10 ng/mL), the immune response against AMACR was more sensitive and specific than was PSA in distinguishing sera from prostate cancer patients relative to control subjects (sensitivity and specificity of 77.8% and 80.6% versus 45.6% and 50%, respectively; area under the curve of 0.789 versus 0.492; P<.001). Assays to detect a humoral immune response against AMACR may have the potential to supplement PSA screening in identifying patients with clinically significant prostate cancer, especially those with intermediate PSA levels.


Obstructive sleep apnoea in diabetes: Does it matter?
Obstructive sleep apnoea (OSA) is very common in patients with Type 2 diabetes (T2D), which is not surprising considering that obesity is a common risk factor for both conditions. In general population studies, OSA has been shown to be associated with several comorbidities including increased risk of road traffic accidents, T2D, hypertension and lack of nocturnal dipping of blood pressure, hyperlipidaemia, increased inflammation, increased risk of cardiovascular disease and mortality, increased risk of atrial fibrillation, worse quality of life, and erectile dysfunction. However, the impact of OSA on diabetes-related vascular and metabolic outcomes remains unclear. Furthermore, the impact of continuous positive airway pressure (CPAP) treatment in patients with T2D is also unclear. This unclarity regarding the impact of OSA and CPAP in patients with T2D has possibly contributed to the lack of screening for OSA in patients with T2D in the UK despite the high prevalence of OSA in patients with T2D. In this commentary, I provide an overview about OSA with a particular focus on its role and impact in patients with T2D.


A failure of high level verbal response selection in progressive dynamic aphasia.
Different theoretical interpretations have been offered in order to account for a specific language impairment termed dynamic aphasia. We report a patient (CH) who presented with a dynamic aphasia in the context of nonfluent progressive aphasia. CH had the hallmark of reduced spontaneous speech in the context of preserved naming, reading, and single word repetition and comprehension. Articulatory and grammatical difficulties were also present. CH had a very severe verbal generation impairment despite being able to describe pictorial scenes and action sequences well. In the experimental investigations CH was severely impaired in word, phrase, and sentence generation tasks when many competing responses were activated by a stimulus. By contrast, he could generate verbal responses satisfactorily when a dominant response was activated by a stimulus. For the first time, we demonstrated that the verbal generation impairment was specific to the production of language. Strikingly, our patient was unimpaired on a number of nonverbal generation tasks (e.g., design fluency, gesture fluency, and motor movement generation). MRI revealed focal left frontal atrophy that predominantly affected Brodmann's Areas 44 and 45. Our findings are discussed with reference to alternative accounts of dynamic aphasia and models of speech production. We interpret our patient's impairment as being underpinned by an inability to select between competing verbal response options. This interpretation converges with evidence from the neuroimaging literature, which implicates the left inferior frontal gyrus in the selection of a response among competing information. We conclude that the left posterior inferior frontal gyrus is involved in the generation of verbal output, and specifically in the selection between competing verbal responses.


Complexity and Simplification in Understanding Travel Preferences Among Tourists.
Travel preferences are complex phenomena, and thus cumbersome to deal with in full width in diagnostic and strategic planning processes. The aim of the present investigation was to explore to what extent individual preferences can be simplified into structures, and if tourists can be grouped into preference clusters that are viable and practically applicable for tourism planning. Building on prior studies that have validated survey instruments designed to measure different tourist role orientations, we used a factor analytical approach to develop a simplified structure of individual preferences, and a standard clustering technique for grouping tourists into preference clusters. Further analyses indicated that preference clusters based on reduced factor preference-data are to some extent related to context-specific valuations, perceptions, and revisit intentions; however, the magnitude of differences between groups was rather small. Overall findings provide reason to suggest that the identified preference clusters are insufficient when it comes to explaining variability in which aspects tourists emphasize as part of their vacation. Possible managerial implications and methodological limitations of the present investigation are noted.


The functional and morphological consequences of balloon catheter injury in veins.
Endovascular procedures are now used to treat occlusive arterial and venous diseases. However, after arterial catheter injury, intimal hyperplasia develops and there is altered endothelial and smooth muscle cell vasoreactivity of the vessel segment. This study examines the morphology and vasoreactivity of veins which have undergone balloon catheter injury. Under direct vision, 22 jugular veins of 11 New Zealand White rabbits were catheterized with a Fogarty balloon catheter (4Fr, three passes, 0.75 ml H2O inflation). Ten uninstrumented jugular veins from 5 additional animals were used as controls. Immediately after balloon injury, 10 jugular veins were harvested. Scanning electron microscopy of these vessels confirmed complete endothelial denudation and no in vitro endothelium-dependent relaxation was observed. After 28 days, 12 veins were harvested. There were no occlusions. Histologically, the 28-day balloon-injured veins had an almost confluent endothelium. However, there was medial hypertrophy but no development of intimal hyperplasia. There were no significant changes in the contractile responses of the balloon-injured veins after 28 days to norepinephrine. Bradykinin and histamine responses were markedly attenuated. Following bradykinin precontraction, balloon-injured veins showed diminished acetylcholine sensitivity with a reduced maximal response, and a small decrease in serotonin sensitivity with a markedly decreased maximal response. In contrast, there was an increased sensitivity in response to sodium nitroprusside without a change in maximal response in precontracted balloon-injured veins. This study shows that after 28 days venous balloon injury does not induce the development of intimal hyperplasia but does reduce overall endothelial dependent relaxation (acetylcholine- and serotonin-mediated) with increased sensitivity to endothelial independent relaxation (sodium nitroprusside-mediated). These findings would suggest that when compared to arterial balloon injury, venous balloon injury does not carry the same histological and vasoconstrictive sequelae but does demonstrate similarly impaired endothelial cell function.


Thickness measurement of transparent glass plates using a lateral shearing cyclic path optical configuration setup and polarization phase shifting interferometry.
We present a measurement technique to determine the thickness of a transparent glass plate (GP) by using a lateral shearing cyclic path optical configuration (CPOC) setup and polarization phase shifting interferometry (PPSI). In the technique, the GP introduces a longitudinal shift in the focus of the beam and, as a result, a spherical wavefront emerges from the lens, which is otherwise set for producing a collimated beam. Using CPOC, two laterally sheared orthogonally polarized beams are generated from the incident spherical wavefront. By applying PPSI, the slope of the optical path difference variation between the laterally sheared interfering beams is evaluated, and the radius of the spherical wavefront and the longitudinal shift of the beam focus are calculated. The thickness of the GP is determined from the standard relation between the longitudinal shift of the focus introduced by the GP and the thickness of the GP. Results obtained for a GP of 9.810mm thickness are presented.


Complex formation of amphotericin B in sterol-containing membranes as evidenced by surface plasmon resonance.
Amphotericin B (AmB) is a membrane-active antibiotic that increases the permeability of fungal membranes. Thus, the dynamic process of its interaction with membranes poses intriguing questions, which prompted us to elaborate a quick and reliable method for real-time observation of the drug's binding to phospholipid liposomes. We focused on surface plasmon resonance (SPR) and devised a new modification method of sensor chips, which led to a significant reduction in the level of nonspecific binding of the drug in a control lane. With this method in hand, we examined the affinity of AmB for various membrane preparations. As expected, AmB exhibited much higher affinity for sterol-containing palmitoyloleoylphosphatidylcholine membranes than those without sterol. The sensorgrams recorded under various conditions partly fitted theoretical curves, which were based on three interaction models. Among those, a two-state reaction model reproduced well the sensorgram of AmB binding to an ergosterol-containing membrane; in this model, two states of membrane-bound complexes, AB and AB*, are assumed, which correspond to a simple binding to the surface of the membrane (AB) and formation of another assembly in the membrane (AB*) such as an ion channel complex. Kinetic analysis demonstrated that the association constant in ergosterol-containing POPC liposomes is larger by 1 order of magnitude than that in the cholesterol-containing counterpart. These findings support the previous notion that ergosterol stabilizes the membrane-bound assembly of AmB.


Climate, landscape, and the risk of orbivirus exposure in cattle in Illinois and western Indiana.
Climate and environmental data were used to estimate the risk of testing positive for antibodies to bluetongue (BTV) and epizootic hemorrhagic disease viruses (EHDV) in cattle in Illinois and western Indiana over three transmission seasons (2000-2002). The risks of BTV and EHDV seropositivity were positively associated with temperature during every year of the study. The EHDV seropositivity was also positively associated with forest patchiness in two of the years. During 2002, a year with unusually high spring rainfall, forest patchiness was not significantly associated with EHDV but spring rainfall did have a moderating effect on temperature. Maps of predicted probability of exposure to BTV or EHDV were created using these best-fitting models and show distinctly different spatial patterns within the same cattle population.


Comparison of antiemetic efficacy and safety of palonosetron vs ondansetron in the prevention of chemotherapy-induced nausea and vomiting in children.
Chemotherapy-induced nausea and vomiting (CINV) in children is a major side effect despite the use of combination antiemetic drugs. To compare the efficacy and safety profile of palonosetron, a second-generation 5-hydroxytryptamine-3 (5-HT3) receptor antagonist, with ondansetron in the prevention of CINV in children. A prospective, randomized, crossover study was conducted in patients aged 2-18 years. 160 chemotherapy cycles, consisting of chemotherapy drugs with moderate- and high-emetogenic potential, were studied. The study group received a single dose of intravenous (IV) palonosetron 5 mcg/kg, and the standard group received IV ondansetron 5 mg/m2 every 8 hours while receiving chemotherapy. The patients were observed for vomiting, use of rescue antiemetic medications, and nausea from Day 1 0-72 hours after completion of each chemotherapy cycle. All adverse events during the study period were recorded. The overall percentage of patients with complete response (CR) in the palonosetron and ondansetron groups were 60% and 56.2%, respectively (𝑃 = .631). The CR rates in the palonosetron and ondansetron groups were 75% and 70%, respectively, in the acute phase (𝑃 = .479), and 68.8% and 65%, respectively, in the delayed phase (𝑃 = .614). There was no statistically significant difference in the CR rates cross both groups. A single dose of palonosetron is noninferior to ondansetron in the prevention of CINV in children and can be considered as an alternative antiemetic drug. There was no significant difference in adverse effects between the palonosetron and ondansetron group.


Needle-guided ablation of locally advanced pancreatic cancer: cytoreduction or immunomodulation by in vivo vaccination?
Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive forms of cancer with a dismal prognosis. About a third of all patients with pancreatic cancer present with locally advanced inoperable disease and are currently designated to palliative systemic chemotherapy. Despite improved chemotherapeutic treatment by the introduction of FOLFIRINOX, median overall survival remains poor at approximately 14 months. Several needle-guided ablative therapies are investigated as treatment option for patients with locally advanced pancreatic cancer (LAPC). This review aims to give an overview of literature of the following needle-guided ablative techniques for the treatment of patients with LAPC: radiofrequency ablation (RFA), microwave ablation (MWA), cryoablation, and irreversible electroporation (IRE). The immunomodulatory effects of the ablative techniques in PDAC are also discussed. These effects in combination with immunotherapy may provide an opportunity to enhance clinical efficacy and endow local ablation with systemic tumor control. Especially less immunogenic tumors like PDAC may benefit from this new approach. Preclinical and clinical findings, possible mechanisms of action, and future research directions are addressed.


Transcriptome analysis of Pseudomonas aeruginosa PAO1 grown at both body and elevated temperatures.
Functional genomics research can give us valuable insights into bacterial gene function. RNA Sequencing (RNA-seq) can generate information on transcript abundance in bacteria following abiotic stress treatments. In this study, we used the RNA-seq technique to study the transcriptomes of the opportunistic nosocomial pathogen Pseudomonas aeruginosa PAO1 following heat shock. Samples were grown at both the human body temperature (37 °C) and an arbitrarily-selected temperature of 46 °C. In this work using RNA-seq, we identified 133 genes that are differentially expressed at 46 °C compared to the human body temperature. Our work identifies some key P. aeruginosa PAO1 genes whose products have importance in both environmental adaptation as well as in vivo infection in febrile hosts. More importantly, our transcriptomic results show that many genes are only expressed when subjected to heat shock. Because the RNA-seq can generate high throughput gene expression profiles, our work reveals many unanticipated genes with further work to be done exploring such genes products.


(1)H and (19)F spin-lattice relaxation and CH3 or CF3 reorientation in molecular solids containing both H and F atoms.
The dynamics of methyl (CH3) and fluoromethyl (CF3) groups in organic molecular (van der Waals) solids can be exploited to survey their local environments. We report solid state (1)H and (19)F spin-lattice relaxationexperiments in polycrystalline 3-trifluoromethoxycinnamic acid, along with an X-ray diffraction determination of the molecular and crystal structure, to investigate the intramolecular and intermolecular interactions that determine the properties that characterize the CF3 reorientation. The molecule is of no particular interest; it simply provides a motionless backbone (on the nuclear magnetic resonance(NMR) time scale) to investigate CF3 reorientation occurring on the NMR time scale. The effects of (19)F-(19)F and (19)F-(1)H spin-spin dipolar interactions on the complicated nonexponential NMRrelaxation provide independent inputs into determining a model for CF3 reorientation. As such, these experiments provide much more information than when only one spin species (usually (1)H) is present. In Sec. IV, which can be read immediately after the Introduction without reading the rest of the paper, we compare the barrier to CH3 and CF3 reorientation in seven organic solids and separate this barrier into intramolecular and intermolecular components.


Non-conditioned bone marrow chimeric mouse generation using culture-based enrichment of hematopoietic stem and progenitor cells.
Bone marrow (BM) chimeric mice are a valuable tool in the field of immunology, with the genetic manipulation of donor cells widely used to study gene function under physiological and pathological settings. To date, however, BM chimera protocols require myeloablative conditioning of recipient mice, which dramatically alters steady-state hematopoiesis. Additionally, most protocols use fluorescence-activated cell sorting (FACS) of hematopoietic stem/progenitor cells (HSPCs) for ex vivo genetic manipulation. Here, we describe our development of cell culture techniques for the enrichment of functional HSPCs from mouse BM without the use of FACS purification. Furthermore, the large number of HSPCs derived from these cultures generate BM chimeric mice without irradiation. These HSPC cultures can also be genetically manipulated by viral transduction, to allow for doxycycline-inducible transgene expression in donor-derived immune cells within non-conditioned immunocompetent recipients. This technique is therefore expected to overcome current limitations in mouse transplantation models.


An insertion/deletion polymorphism in the angiotensin I-converting enzyme gene accounting for half the variance of serum enzyme levels.
A polymorphism consisting of the presence or absence of a 250-bp DNA fragment was detected within the angiotensin I-converting enzyme gene (ACE) using the endothelial ACE cDNA probe. This polymorphism was used as a marker genotype in a study involving 80 healthy subjects, whose serum ACE levels were concomitantly measured. Allele frequencies were 0.6 for the shorter allele and 0.4 for the longer allele. A marked difference in serum ACE levels was observed between subjects in each of the three ACE genotype classes. Serum immunoreactive ACE concentrations were, respectively, 299.3 +/- 49, 392.6 +/- 66.8, and 494.1 +/- 88.3 micrograms/liter, for homozygotes with the longer allele (n = 14), and heterozygotes (n = 37) and homozygotes (n = 29) with the shorter allele. The insertion/deletion polymorphism accounted for 47% of the total phenotypic variance of serum ACE, showing that the ACE gene locus is the major locus that determines serum ACE concentration. Concomitant determination of the ACE genotype will improve discrimination between normal and abnormal serum ACE values by allowing comparison with a more appropriate reference interval.


Enhancement of electron accepting ability of para-benzoquinone by a single water molecule.
Electron acceptors built upon the para-benzoquinone (pBQ) electrophore are ubiquitous in nature. Here, we present a frequency-resolved photoelectron spectroscopic study of the cold pBQ radical anion, pBQ-, solvated by a single water molecule, as commonly encountered in nature. Our results show that the electron accepting ability is enhanced by the single water molecule and by elevated temperatures.


Epidermal patterning in Arabidopsis: models make a difference.
The leaf and root epidermis in Arabidopsis provide ideal systems in which to explore the mechanisms that underlie the patterned assignment of cell fates during development. Extensive experimental studies have uncovered a complex interlocked feedback network that operates within the epidermis to coordinate the choice between hair and nonhair fates. A number of recent studies using mathematical models have begun to study this network, highlighting new mechanisms that have subsequently been confirmed in model-directed experiments. These studies illustrate the potential of integrated modeling and experimentation to shed new light on developmental processes. Moreover, these models enable systems-level comparative analyses that may help understand the origin and role of properties, such as robustness and redundancy in developmental systems and, concomitantly, the evolution of development itself.


Cancer patients' use of and attitudes towards medicinal cannabis.
Objectives Access to medicinal cannabis is a timely and important issue in cancer care. Recent legislative changes in Australia have increased access to medicinal cannabis, but the views of people with cancer on this topic are poorly understood. The aim of this study was to explore the prevalence of the use of and attitudes towards medicinal cannabis among people with cancer. Methods A cross-sectional study was performed using an anonymous, 15-item study-specific paper-based survey. The survey was administered over a 2-week period in August 2017 in the waiting rooms of a specialist cancer hospital. Results In all, 339 patients completed the survey (mean (±s.d.) age 59±15 years; 52% male). Fourteen respondents (4%) were currently using cannabis medicinally. Only one of these respondents had a prescription for their cannabis product. Most respondents would consider using a medicinal cannabis product if recommended by their doctor (n=271; 80%). Conclusion This study is the first of its kind to survey the use of and attitudes towards medicinal cannabis in a broad sample of Australian people with cancer. Few respondents were currently using cannabis for medicinal purposes, but an overwhelming majority were in favour of increasing access and would consider using a prescribed product. What is known about the topic? Cannabis may have a wide variety of medicinal uses, particularly in the cancer setting. Currently, people with cancer in Victoria have limited access to medicinal cannabis despite recent legislative changes. What does this paper add? In a general sample of people with cancer, few were using cannabis for medicinal purposes, but most were in favour of widening access and would consider using a product their doctor prescribed. What are the implications for practitioners? Despite supporting access, patients indicated that the recommendations of doctors and increasing the evidence base are necessary requirements to their use of medicinal cannabis.


Role of the reverse-Trendelenberg patient position in maintaining low-CVP anaesthesia during liver resections.
The Trendelenberg position is recommended during liver resection, to decrease the risk of venous air embolism. However, this position raises the central venous pressure and may increase blood loss. We propose that the reverse-Trendelenberg position can be safely and effectively used to maintain a low central venous pressure during liver surgery. Fifty consecutive patients underwent elective liver resection at a single centre during a 17-month period. Patients were positioned with a head-up tilt during division of the liver parenchyma. Patients had a mean central venous pressure of 9.2 mmHg when supine, despite fluid restriction. The central venous pressure fell consistently and rapidly when they were tilted head-up, to a mean of 1.7 mmHg. The resections were completed with a median operative blood loss of 600 mL. No patient developed a clinically apparent venous air embolism. Postoperative renal dysfunction that could be attributed to low central venous pressure anaesthesia occurred in only one case. The reverse-Trendelenberg position effectively lowers the CVP during liver surgery. It is easy to monitor, titrate and reverse, and avoids the need for complex pharmacological interventions. We recommend this position to liver surgeons and anaesthetists who have found it difficult to maintain a low CVP with the supine or Trendelenberg positions.


Mental competence and end-of-life decision making: death row volunteering and euthanasia.
This article reports on a qualitative study of defense attorneys' perceptions of the mental competence or rationality of death row inmates' decisions to waive habeas appeals and proceed directly to execution. Interviews were conducted with twenty attorneys who have either directly represented or been closely involved with would-be volunteers. Through analytic comparison with another end-of-life decision, euthanasia, this article reports on four themes from the interviews: (a) attorneys' perceptions of the legal standard of competence, (b) their perceptions of the competency evaluation process, (c) implications of competing interpretive frames (i.e., volunteering vs. suicide), and (d) the rationality of decisions to waive appeals. Implications of research findings, particularly in terms of recent restructured models of competence, are also discussed.


Surface-plasmon polariton solutions at a lossy slab in a symmetric surrounding.
A rigorous theoretical formulation based on electromagnetic plane waves is utilized to construct a unified framework and identification of all possible surface-plasmon polariton solutions at an absorptive slab in a symmetric, lossless dielectric surrounding. In addition to the modes reported in literature, sets of entirely new mode solutions are presented. The corresponding fields are classified into different categories and examined in terms of bound and leaky modes, as well as forward-and backward-propagating modes, both outside and inside the slab. The results could benefit plasmon based applications in thin-film nanophotonics.


Mismatched anti-predator behavioral responses in predator-naïve larval anurans.
Organisms are adept at altering behaviors to balance the tradeoff between foraging and predation risk in spatially and temporally shifting predator environments. In order to optimize this tradeoff, prey need to be able to display an appropriate response based on degree of predation risk. To be most beneficial in the earliest life stages in which many prey are vulnerable to predation, innate anti-predator responses should scale to match the risk imposed by predators until learned anti-predator responses can occur. We conducted an experiment that examined whether tadpoles with no previous exposure to predators (i.e., predator-naive) exhibit innate antipredator behavioral responses (e.g., via refuge use and spatial avoidance) that match the actual risk posed by each predator. Using 7 treatments (6 free-roaming, lethal predators plus no-predator control), we determined the predation rates of each predator on Lithobates sphenocephalus tadpoles. We recorded behavioral observations on an additional 7 nonlethal treatments (6 caged predators plus no-predator control). Tadpoles exhibited innate responses to fish predators, but not non-fish predators, even though two non-fish predators (newt and crayfish) consumed the most tadpoles. Due to a mismatch between innate response and predator consumption, tadpoles may be vulnerable to greater rates of predation at the earliest life stages before learning can occur. Thus, naïve tadpoles in nature may be at a high risk to predation in the presence of a novel predator until learned anti-predator responses provide additional defenses to the surviving tadpoles.


MicroRNA-370 carried by M2 macrophage-derived exosomes alleviates asthma progression through inhibiting the FGF1/MAPK/STAT1 axis.
Emerging evidence has suggested the functions of exosomes in allergic diseases including asthma. By using a mouse model with asthma induced by ovalbumin (OVA), we explored the roles of M2 macrophage-derived exosomes (M2Φ-Exos) in asthma progression. M2Φ-Exos significantly alleviated OVA-induced fibrosis and inflammatory responses in mouse lung tissues, as well as inhibited abnormal proliferation, invasion, and fibrosis-related protein production in platelet derived growth factor (PDGF-BB) treated primary mouse airway smooth muscle cells (ASMCs). The OVA administration in mice or the PDGF-BB treatment in ASMCs reduced the expression of miR-370, which was detected in M2Φ-Exos by miRNA sequencing. However, treating the mice or ASMCs with M2Φ-Exos reversed the inhibitory effect of OVA or PDGF-BB on miR-370 expression. We identified that the target of miR-370 was fibroblast growth factor 1 (FGF1). Downregulation of miR-370 by Lv-miR-370 inhibitor or overexpression of FGF1 by Lv-FGF1 blocked the protective roles of M2Φ-Exos in asthma-like mouse and cell models. M2Φ-Exos were found to inactivate the MAPK signaling pathway, which was recovered by miR-370 inhibition or FGF1 overexpression. Collectively, we conclude that M2Φ-Exos carry miR-370 to alleviate asthma progression through downregulating FGF1 expression and the MAPK/STAT1 signaling pathway. Our study may offer a novel insight into asthma treatment.


Anti-cancer effect of bee venom on colon cancer cell growth by activation of death receptors and inhibition of nuclear factor kappa B.
Bee venom (BV) has been used as a traditional medicine to treat arthritis, rheumatism, back pain, cancerous tumors, and skin diseases. However, the effects of BV on the colon cancer and their action mechanisms have not been reported yet. We used cell viability assay and soft agar colony formation assay for testing cell viability, electro mobility shift assay for detecting DNA binding activity of nuclear factor kappa B (NF-κB) and Western blotting assay for detection of apoptosis regulatory proteins. We found that BV inhibited growth of colon cancer cells through induction of apoptosis. We also found that the expression of death receptor (DR) 4, DR5, p53, p21, Bax, cleaved caspase-3, cleaved caspase-8, and cleaved caspase-9 was increased by BV treatment in a dose dependent manner (0-5 μg/ml). Consistent with cancer cell growth inhibition, the DNA binding activity of nuclear factor kappa B (NF-κB) was also inhibited by BV treatment. Besides, we found that BV blocked NF-κB activation by directly binding to NF-κB p50 subunit. Moreover, combination treatment with BV and p50 siRNA or NF-κB inhibitor augmented BV-induced cell growth inhibition. However, p50 mutant plasmid (C62S) transfection partially abolished BV-induced cell growth inhibiton. In addition, BV significantly suppressed tumor growth in vivo. Therefore, these results suggested that BV could inhibit colon cancer cell growth, and these anti-proliferative effects may be related to the induction of apoptosis by activation of DR4 and DR5 and inhibition of NF-κB.


Theranostic potential of oncolytic vaccinia virus.
Biological cancer therapies, such as oncolytic, or replication-selective viruses have advantages over traditional therapeutics as they can employ multiple different mechanisms to target and destroy cancers (including direct cell lysis, immune activation and vascular collapse). This has led to their rapid recent clinical development. However this also makes their pre-clinical and clinical study complex, as many parameters may affect their therapeutic potential and so defining reason for treatment failure or approaches that might enhance their therapeutic activity can be complicated. The ability to non-invasively image viral gene expression in vivo both in pre-clinical models and during clinical testing will considerably enhance the speed of oncolytic virus development as well as increasing the level and type of useful data produced from these studies. Further, subsequent to future clinical approval, imaging of reporter gene expression might be used to evaluate the likelihood of response to oncolytic viral therapy prior to changes in tumor burden. Here different reporter genes used in conjunction with oncolytic viral therapy are described, along with the imaging modalities used to measure their expression, while their applications both in pre-clinical and clinical testing are discussed. Possible future applications for reporter gene expression from oncolytic viruses in the phenotyping of tumors and the personalizing of treatment regimens are also discussed.


Direct current-excited cw CO(2) metal waveguide laser.
A novel design for a dc-excited cw CO(2) metal waveguide laser has been developed in which a slotted hollow cathode also doubles as a metal waveguide for the cavity modes. This design has been implemented in a compact structure that produces over 1 W of cw 10.6-microm radiation. The discharge characteristics, laser gain, and laser output have been studied as functions of various discharge parameters. The advantages of the transverse discharge of the slotted hollow cathode geometry include low voltage, positive impedance, rugged structure, and high optical gain. Overall efficiency is comparable with that of conventional longitudinal CO(2) lasers. The output laser modes are clean low-order Hermite-Gaussian or Airy function modes.


Basal autophagy induction without AMP-activated protein kinase under low glucose conditions.
When ATP levels in a cell decrease, various homeostatic intracellular mechanisms initiate attempts to restore ATP levels. As a prominent energy sensor, AMP-activated protein kinase (AMPK) represents one molecular gauge that links energy levels to regulation of anabolic and catabolic processes to restore energy balance. Although pharmacological studies have suggested that an AMPK activator, AIC AR (5-aminoimidazole-4-carboxamide ribonucleoside) may link AMPK activation to autophagy, a process that can provide short-term energy within the cell, AICAR can have AMPK-independent effects. Therefore, using a genetic-based approach we investigated the role of AMPK in cellular energy balance. We demonstrate that genetically altered cells, mouse embryonic fibroblasts (MEFs), lacking functional AMPK, display altered energy balance under basal conditions and die prematurely under low glucose-serum starvation challenge. These AMPK mutant cells appear to be abnormally reliant on autophagy under low glucose basal conditions, and therefore cannot rely further on autophagy like wild-type cells during further energetic stress and instead undergo apoptosis. This data suggests that AMPK helps regulate basal energy levels under low glucose. Further, AMPK mutant cells show increased basal phosphorylation of p53 at serine 15, a residue phosphorylated under glucose deprivation. We propose that cells lacking AMPK function have altered p53 activity that may help sensitize these cells to apoptosis under energetic stress.


Learning from the genome sequence of Mycobacterium tuberculosis H37Rv.
Mycobacterium tuberculosis, the scourge of humanity, is one of the most successful and scientifically challenging pathogens of all time. To catalyse the conception of new prophylactic and therapeutic interventions against tuberculosis, and to enhance our understanding of the biology of the tubercle bacillus, the complete genome sequence of the most widely used strain, H37Rv, has been determined. Bioinformatic analysis led to the identification of approximately 4000 genes in the 4.41 Mb genome sequence and provided fresh insight into the biochemistry, physiology. genetics and immunology of this much-feared bacterium. Genomic information is centralised in TubercuList (http://www.pasteur.fr/Bio/TubercuList/).


Spontaneous assembly of a hybrid crystal-liquid phase in inverse patchy colloid systems.
Materials with well-defined architectures are heavily sought after in view of their diverse technological applications. Among the desired target architectures, lamellar phases stand out for their exceptional mechanical and optical features. Here we show that charged colloids, decorated on their poles with two oppositely charged regions possess the unusual ability to spontaneously assemble in different morphologies of (semi-)ordered, layered particle arrangements which maintain their structural stability over a surprisingly large temperature range. This remarkable capacity is related to a characteristic bonding mechanism: stable intra-layer bonds guarantee the formation of planar aggregates, while strong inter-layer bonds favor the stacking of the emerging planar assemblies. These two types of bonds together are responsible for the self-healing processes occurring during the spontaneous assembly. The resulting phases are characterized by parallel, densely packed, particle layers connected by a relatively small number of intra-layer particles. We investigate the properties of the (semi-)ordered phases in terms of static and dynamic correlation functions, focusing in particular on a novel hybrid crystal-liquid phase that prevails at intermediate temperatures where the inter-layer particles form a mobile, fluid phase.


Investigation of a clinical PET detector module design that employs large-area avalanche photodetectors.
We investigated the feasibility of designing an Anger-logic PET detector module using large-area high-gain avalanche photodiodes (APDs) for a brain-dedicated PET/MRI system. Using Monte Carlo simulations, we systematically optimized the detector design with regard to the scintillation crystal, optical diffuser, surface treatment, layout of large-area APDs, and signal-to-noise ratio (SNR, defined as the 511 keV photopeak position divided by the standard deviation of noise floor in an energy spectrum) of the APD devices. A detector prototype was built comprising an 8 × 8 array of 2.75 × 3.00 × 20.0 mm3 LYSO (lutetium-yttrium-oxyorthosilicate) crystals and a 22.0 × 24.0 × 9.0 mm3 optical diffuser. From the four designs of the optical diffuser tested, two designs employing a slotted diffuser are able to resolve all 64 crystals within the block with good uniformity and peak-to-valley ratio. Good agreement was found between the simulation and experimental results. For the detector employing a slotted optical diffuser, the energy resolution of the global energy spectrum after normalization is 13.4 ± 0.4%. The energy resolution of individual crystals varies between 11.3 ± 0.3% and 17.3 ± 0.4%. The time resolution varies between 4.85 ± 0.04 (center crystal), 5.17 ± 0.06 (edge crystal), and 5.18 ± 0.07 ns (corner crystal). The generalized framework proposed in this work helps to guide the design of detector modules for selected PET system configurations, including scaling the design down to a preclinical PET system, scaling up to a whole-body clinical scanner, as well as replacing APDs with other novel photodetectors that have higher gain or SNR such as silicon photomultipliers.


Effects of free fatty acids (FFA) on glucose metabolism: significance for insulin resistance and type 2 diabetes.
Most obese individuals have elevated plasma levels of free fatty acids (FFA) which are known to cause peripheral (muscle) insulin resistance. They do this by inhibiting insulin-stimulated glucose uptake and glycogen synthesis. The mechanism involves intramyocellular accumulation of diacylglycerol and activation of protein kinase C. FFAs also cause hepatic insulin resistance. They do this by inhibiting insulin-mediated suppression of glycogenolysis. On the other hand, FFAs support between 30 and 50 % of basal insulin secretion and potentiate glucose-stimulated insulin secretion. The insulin stimulatory action of FFAs is responsible for the fact that the vast majority ( approximately 80 %) of obese insulin resistant people do not develop type 2 diabetes. They are able to compensate for their FFA mediated insulin resistance with increased FFA mediated insulin secretion. Individuals who are unable to do this (probably for genetic reasons) eventually develop type 2 diabetes. FFAs have recently been shown to activate the IkappaB/NFkappaB pathway which is involved in many inflammatory processes. Thus, elevated plasma levels of FFAs are not only a major cause of insulin resistance in skeletal muscle and liver but may, in addition, play a role in the pathogenesis of coronary artery disease.


Regulation of G protein-coupled receptors by palmitoylation and cholesterol.
Due to their membrane location, G protein-coupled receptors (GPCRs) are subject to regulation by soluble and integral membrane proteins as well as membrane components, including lipids and sterols. GPCRs also undergo a variety of post-translational modifications, including palmitoylation. A recent article by Zheng et al. in BMC Cell Biology demonstrates cooperative roles for receptor palmitoylation and cholesterol binding in GPCR dimerization and G protein coupling, underlining the complex regulation of these receptors.


The tumor suppressor p53 regulates polarity of self-renewing divisions in mammary stem cells.
Stem-like cells may be integral to the development and maintenance of human cancers. Direct proof is still lacking, mainly because of our poor understanding of the biological differences between normal and cancer stem cells (SCs). Using the ErbB2 transgenic model of breast cancer, we found that self-renewing divisions of cancer SCs are more frequent than their normal counterparts, unlimited and symmetric, thus contributing to increasing numbers of SCs in tumoral tissues. SCs with targeted mutation of the tumor suppressor p53 possess the same self-renewal properties as cancer SCs, and their number increases progressively in the p53 null premalignant mammary gland. Pharmacological reactivation of p53 correlates with restoration of asymmetric divisions in cancer SCs and tumor growth reduction, without significant effects on additional cancer cells. These data demonstrate that p53 regulates polarity of cell division in mammary SCs and suggest that loss of p53 favors symmetric divisions of cancer SCs, contributing to tumor growth.


Half of 30-Day Hospital Readmissions Among HIV-Infected Patients Are Potentially Preventable.
Thirty-day readmission rates, a widely utilized quality metric, are high among HIV-infected individuals. However, it is unknown how many 30-day readmissions are preventable, especially in HIV patients, who have been excluded from prior potentially preventable readmission analyses. We used electronic medical records to identify all readmissions within 30 days of discharge among HIV patients hospitalized at a large urban safety net hospital in 2011. Two independent reviewers assessed whether readmissions were potentially preventable using both published criteria and detailed chart review, how readmissions might have been prevented, and the phase of care deemed suboptimal (inpatient care, discharge planning, post-discharge). Of 1137 index admissions, 213 (19%) resulted in 30-day readmissions. These admissions occurred among 930 unique HIV patients, with 130 individuals (14%) experiencing 30-day readmissions. Of these 130, about half were determined to be potentially preventable using published criteria (53%) or implicit chart review (48%). Not taking antiretroviral therapy (ART) greatly increased the odds of a preventable readmission (OR 5.9, CI:2.4-14.8). Most of the preventable causes of readmission were attributed to suboptimal care during the index hospitalization. Half of 30-day readmission in HIV patients are potentially preventable. Increased focus on early ART initiation, adherence counseling, management of chronic conditions, and appropriate timing of discharge may help reduce readmissions in this vulnerable population.


Early effects of gliadin on enterocyte intracellular signalling involved in intestinal barrier function.
Despite the progress made in understanding the immunological aspects of the pathogenesis of coeliac disease (CD), the early steps that allow gliadin to cross the intestinal barrier are still largely unknown. The aim of this study was to establish whether gliadin activates a zonulin dependent enterocyte intracellular signalling pathway(s) leading to increased intestinal permeability. The effect of gliadin on the enterocyte actin cytoskeleton was studied on rat intestinal epithelial (IEC-6) cell cultures by fluorescence microscopy and spectrofluorimetry. Zonulin concentration was measured on cell culture supernatants by enzyme linked immunosorbent assay. Transepithelial intestinal resistance (Rt) was measured on ex vivo intestinal tissues mounted in Ussing chambers. Incubation of cells with gliadin led to a reversible protein kinase C (PKC) mediated actin polymerisation temporarily coincident with zonulin release. A significant reduction in Rt was observed after gliadin addition on rabbit intestinal mucosa mounted in Ussing chambers. Pretreatment with the zonulin inhibitor FZI/0 abolished the gliadin induced actin polymerisation and Rt reduction but not zonulin release. Gliadin induces zonulin release in intestinal epithelial cells in vitro. Activation of the zonulin pathway by PKC mediated cytoskeleton reorganisation and tight junction opening leads to a rapid increase in intestinal permeability.


Early cortical thickness change after mild traumatic brain injury following motor vehicle collision.
In a motor vehicle collision (MVC), survivors often receive mild traumatic brain injuries (mTBI). Although there have been some reports of early white matter changes after an mTBI, much less is known about early cortical structural changes. To investigate early cortical changes within a few days after an MVC, we compared cortical thickness of mTBI survivors with non-mTBI survivors, then reexamined cortical thickness in the same survivors 3 months later. MVC survivors were categorized as mTBI or non-mTBI based on concussive symptoms documented in emergency departments (EDs). Cortical thickness was measured from MRI images using FreeSurfer within a few days and again at 3 months after MVC. Post-traumatic stress symptoms and physical conditions were also assessed. Compared with the non-mTBI group (n = 23), the mTBI group (n = 21) had thicker cortex in the left rostral middle frontal (rMFG) and right precuneus gyri, but thinner cortex in the left posterior middle temporal gyrus at 7.2 ± 3.1 days after MVC. After 3 months, cortical thickness had decreased in left rMFG in the mTBI group but not in the non-mTBI group. The cortical thickness of the right precuneus region in the initial scans was positively correlated with acute traumatic stress symptoms for all survivors and with the number of reduced activity days for mTBI survivors who completed the follow-up. The preliminary results suggest that alterations in cortical thickness may occur at an early stage of mTBI and that frontal cortex structure may change dynamically over the initial 3 months after mTBI.


Synthetic retinoid Am80 ameliorates chronic graft-versus-host disease by down-regulating Th1 and Th17.
Chronic GVHD (cGVHD) is a main cause of late death and morbidity after allogeneic hematopoietic cell transplantation, but its pathogenesis remains unclear. We investigated the roles of Th subsets in cGVHD with the use of a well-defined mouse model of cGVHD. In this model, development of cGVHD was associated with up-regulated Th1, Th2, and Th17 responses. Th1 and Th2 responses were up-regulated early after BM transplantation, followed by a subsequent up-regulation of Th17 cells. Significantly greater numbers of Th17 cells were infiltrated in the lung and liver from allogeneic recipients than those from syngeneic recipients. We then evaluated the roles of Th1 and Th17 in cGVHD with the use of IFN-γ-deficient and IL-17-deficient mice as donors. Infusion of IFN-γ(-/-) or IL-17(-/-) T cells attenuated cGVHD in the skin and salivary glands. Am80, a potent synthetic retinoid, regulated both Th1 and Th17 responses as well as TGF-β expression in the skin, resulting in an attenuation of cutaneous cGVHD. These results suggest that Th1 and Th17 contribute to the development of cGVHD and that targeting Th1 and Th17 may therefore represent a promising therapeutic strategy for preventing and treating cGVHD.


Modulation of biomechanical properties of hyaluronic acid hydrogels by crosslinking agents.
Modulation of both mechanical properties and biocompatibilities of hyaluronic acid (HA) hydrogels is very importance for their applications in biomaterials. Pure HA solution was converted into a hydrogel by using butanediol diglycidyl ether (BDDE) as a crosslinking agent. Mechanical properties of the HA hydrogels have been evaluated by adding up different amount of BDDEs. While the mechanical properties of the obtained HA hydrogels were evaluated by measuring their crosslinking degrees, elastic modulus and viscosity, their in vitro biocompatibilities were done by measuring the degrees of anti-inflammatory reactions, cell viabilities and cytotoxicity. The degrees of anti-inflammatory reactions were determined by measuring the amount of nitric oxides (NOs) released from lipopolysaccharide(LPS)(+)-induced macrophages; cell viability was evaluated by observing differences in the behaviors of fibroblasts covered with the HA hydrogels, compared with those covered with the films of Teflon and Latex. Cytotoxicity of the HA hydrogels was also evaluated by measuring the degrees of viability of the cells exposed on the extracts of the HA hydrogels over those of Teflon, Latex and pure HA solutions by the assays of thiazoly blue tetrazolium bromide (MTT), neutral reds, and bromodeoxyuridine (BrdU). The results showed that employment of BDDEs beyond critical amounts showed lower biocompatibility of the crosslinked HA hydrogels but higher crosslinking degrees and mechanical properties, indicating the importance of controlling the HA concentrations, BDDE amounts and their reaction times for the synthesis of the crosslinked HA hydrogels for their clinical applications as biomaterials.


Evolution by flight and fight: diverse mechanisms of adaptation by actively motile microbes.
Evolutionary adaptation can be achieved by mechanisms accessible to all organisms, including faster growth and interference competition, but self-generated motility offers additional possibilities. We tested whether 55 populations of the bacterium Myxococcus xanthus that underwent selection for increased fitness at the leading edge of swarming colonies adapted by swarming faster toward unused resources or by other means. Populations adapted greatly but diversified markedly in both swarming phenotypes and apparent mechanisms of adaptation. Intriguingly, although many adapted populations swarm intrinsically faster than their ancestors, numerous others do not. Some populations evolved interference competition toward their ancestors, whereas others gained the ability to facultatively increase swarming rate specifically upon direct interaction with ancestral competitors. Our results both highlight the diverse range of mechanisms by which actively motile organisms can adapt evolutionarily and help to explain the high levels of swarming-phenotype diversity found in local soil populations of M. xanthus.


Intra-connected three-dimensionally isotropic bulk negative index photonic metamaterial.
Isotropic negative index metamaterials (NIMs) are highly desired, particularly for the realization of ultra-high resolution lenses. However, existing isotropic NIMs function only two-dimensionally and cannot be miniaturized beyond microwaves. Direct laser writing processes can be a paradigm shift toward the fabrication of three-dimensionally (3D) isotropic bulk optical metamaterials, but only at the expense of an additional design constraint, namely connectivity. Here, we demonstrate with a proof-of-principle design that the requirement connectivity does not preclude fully isotropic left-handed behavior. This is an important step towards the realization of bulk 3D isotropic NIMs at optical wavelengths.


Potential role of blood biomarkers in the management of nontraumatic intracerebral hemorrhage.
Intracerebral hemorrhage (ICH), a subtype of stroke associated with high mortality and disability, accounts for 13% of all strokes. Basic and clinical research has contributed to our understanding of the complex pathophysiology of neuronal injury in ICH. Outcome rates, however, remain stable, and questions regarding acute management of ICH remain unanswered. Newer research is aiming at matching measured levels of serum proteins, enzymes, or cells to different stages of brain damage, suggesting that blood biomarkers may assist in acute diagnosis, therapeutic decisions, and prognostication. This paper provides an overview on the most promising blood biomarkers and their potential role in the diagnosis and management of spontaneous ICH. Information was collected from studies, reviews, and guidelines listed in PubMed up to November 2013 on blood biomarkers of nontraumatic ICH in humans. We describe the potential role and limitations of GFAP, S100B/RAGE, and ApoC-III as diagnostic biomarkers, β-Amyloid as a biomarker for etiological classification, and 27 biomarkers for prognosis of mortality and functional outcome. Within the group of prognostic markers we discuss markers involved in coagulation processes (e.g., D-Dimers), neuroendocrine markers (e.g., copeptin), systemic metabolic markers (e.g., blood glucose levels), markers of inflammation (e.g., IL-6), as well as growth factors (e.g., VEGF), and others (e.g., glutamate). Some of those blood biomarkers are agents of pathologic processes associated with hemorrhagic stroke but also other diseases, whereas others play more distinct pathophysiological roles and help in understanding the basic mechanisms of brain damage and/or recovery in ICH. Numerous blood biomarkers are associated with different pathophysiological pathways in ICH, and some of them promise to be useful in the management of ICH, eventually contributing additional information to current tools for diagnosis, therapy monitoring, risk stratification, or intervention. Up to date, however, no blood biomarker of ICH has been studied sufficiently to find its way into clinical routine yet; well-designed, large-scale, clinical studies addressing relevant clinical questions are needed. We suggest that the effectiveness of biomarker research in ICH might be improved by international cooperation and shared resources for large validation studies, such as provided by the consortium on stroke biomarker research (http://stroke-biomarkers.com/page.php?title=Resources).


Birth weight and grip strength in young Swedish males: a longitudinal matched sibling analysis and across all body mass index ranges.
Low birth weight is associated with a lower grip strength later in life. However, associations between birth weight among infants born at-term and factors driving associations between birth weight and grip strength are largely unknown. A cohort of 144,369 young men born at-term, including 10,791 individuals who had at least one male sibling/s, were followed until conscription where they performed a grip strength test. We used linear and non-linear regression analyses in the full cohort, and fixed-effects regression analyses in the sibling cohort, to address confounding by factors that are shared between siblings. After adjustment, each unit increase in birth weight z-score was associated with increases of 17.7 (95% CI, 17.2-18.2) and 13.4 (10.1-16.6) newton grip strength, which converts to approximately 1.8 and 1.4 kilogram-force in the full and within-families cohorts, respectively. The associations did not vary with young adulthood BMI. Birth weight, within the at-term range, is robustly positively associated with grip strength in young adulthood among men across all BMI categories and associations appears to be mainly driven by factors that are not shared between siblings. These findings underline the importance of recognizing the influence of low birth weight, also within the at-term-range, on young adulthood muscle strength.


Effect of three NiTi files on transportation of the apical foramen.
To compare landed and nonlanded rotary file overinstrumentation on transportation of the apical foramen in the curved canals of extracted teeth. Severely curved molar root canals (n = 45) were distributed into three equal groups (n = 15) according to angle (mean 54°) and radius of curvature (mean 5 mm). Canals were overinstrumented 0.5 mm beyond the foramen to a size 35 master apical file using landed (ProFile ISO), nonlanded (ProFile Vortex) or nonlanded, reduced shape memory (Vortex Blue) files. Post-instrumentation images of the apical foramen were compared with pre-instrumentation control images for differences in area, circularity and ratio of Feret's diameters. Groups were compared using anova or Kruskal-Wallis tests with significance of P < 0.05. There were no differences between pre-treatment groups in the parameters tested. All groups demonstrated alterations in the geometry of the apical foramen. There were no significant differences between ProFile ISO, ProFile Vortex or Vortex Blue in area, circularity and ratio of Feret's diameters. Landed, nonlanded and nonlanded reduced shape memory files all produced transportation of the apical foramen when overinstrumented by 0.5 mm in severely curved canals. There was no difference between these file systems with regard to the degree of this effect.


Selected amplification of the cell division genes ftsQ-ftsA-ftsZ in Escherichia coli.
Rapidly growing Escherichia coli is unable to divide in the presence of the antibiotic mecillinam, whose direct target is penicillin-binding protein 2 (PBP2), responsible for the elongation of the cylindrical portion of the cell wall. Division can be restored in the absence of PBP2 activity by increasing the concentration of the cell division proteins FtsQ, FtsA, and FtsZ. We tried to identify regulators of the ftsQ-ftsA-ftsZ operon among mecillinam-resistant mutants, which include strains overexpressing these genes. By insertional mutagenesis with mini-Tn10 elements, we selected for insertions that conferred mecillinam resistance. Among 15 such mutants, 7 suppressed the thermosensitivity of the ftsZ84(Ts) mutant, strongly suggesting that they had increased FtsZ activity. In all 7 cases, however, the mutants resulted from a duplication of the ftsQAZ region. These duplications seemed to result from multiple events, suggesting that no simple insertional inactivation can result in a mutant with sufficiently amplified ftsQAZ expression to confer mecillinam resistance. The structure of the duplications suggests a general method for constructing directed duplications of precise sequences.


Epileptogenic high-frequency oscillations skip the motor area in children with multilobar drug-resistant epilepsy.
Subtotal hemispherectomy involves the resection of multiple lobes in children with drug-resistant epilepsy, skipping the motor area (MA). We determined epileptogenicity using the occurrence rate (OR) of high-frequency oscillations (HFOs) and the modulation index (MI), demonstrating strength of coupling between HFO and slow wave. We hypothesized that epileptogenicity increased over the multiple lobes but skipped the MA. We analyzed 23 children (14 subtotal hemispherectomy; 9 multilobar resections). Scalp video-EEG and magnetoencephalography were performed before surgery. We analyzed the OR(HFO) and MI(5 phases=0.5-8 Hz) on electrodes of total area, resection areas, and MA. We compared the data between good [International League Against Epilepsy (ILAE) class I-II] and poor (III-VI) seizure outcome groups. ILAE class Ia outcome was achieved in 18 children. Among the MI(5 phases) in the resection areas, MI(3-4 Hz) was the highest. The OR(HFO) and MI(3-4 Hz) in both total area and resection areas were significantly higher in the good seizure outcome group than in the poor outcome group. The OR(HFO) and MI(3-4 Hz) in resection areas were significantly higher than in the MA. Our patients with multilobar drug-resistant epilepsy showed evidence of multifocal epileptogenicity that specifically skipped the MA. This is the first study demonstrating that the electrophysiological phenotype of multifocal epilepsy specifically skips the MA using OR(HFO) and MI(3-4 Hz).


Coverage of atypical antipsychotics among medicare drug plans in the state of washington for fiscal year 2007.
To examine drug coverage and patient costs for 6 atypical antipsychotics (olanzapine, quetiapine, ziprasidone, aripiprazole, clozapine, and risperidone) in Medicare Part D formularies using health plan data from the state of Washington. Fiscal year 2007 coverage and cost-sharing characteristics for 57 prescription drug plans (PDPs) and 43 Medicare advantage prescription drug plans (MAPDs) were collected from the Centers for Medicare and Medicaid Services. Plans were compared in terms of formulary restrictions, out-of-pocket costs, and premium charges. Medicare released plan information for fiscal year 2007 in October 2006. Data were collected for this study in February 2007. Almost all plans covered the 6 atypical antipsychotics. The PDPs were more likely to restrict coverage than the MAPDs. Prior authorization requirements were enforced in 5% to 21% of plans, depending on plan type and medication. Monthly drug plan premiums were higher for PDPs than MAPDs, but the MAPDs had concurrent monthly health premiums. About 80% of MAPDs and 60% of PDPs also had no annual deductible for medications. The patient out-of-pocket cost for atypical antipsychotics varied depending on the stage of coverage-median monthly drug costs ranged from $5 to $50 during the initial period, but if costs exceeded the annual cap, patients were responsible for the full cost of the drug, which ranged from $292 to $665. Patients with low incomes and those who exceeded the annual spending limit ($3850 in fiscal year 2007) had a median monthly cost of $17 to $33. There is considerable variation across health plans in terms of patients' out-of-pocket drug costs. Given the significant needs and vulnerabilities of Medicare beneficiaries with mental illness, changes for atypical antipsychotic coverage should be monitored carefully, and the complexity of Medicare drug plans should be minimized.


Effects of Response to 2014-2015 Ebola Outbreak on Deaths from Malaria, HIV/AIDS, and Tuberculosis, West Africa.
Response to the 2014-2015 Ebola outbreak in West Africa overwhelmed the healthcare systems of Guinea, Liberia, and Sierra Leone, reducing access to health services for diagnosis and treatment for the major diseases that are endemic to the region: malaria, HIV/AIDS, and tuberculosis. To estimate the repercussions of the Ebola outbreak on the populations at risk for these diseases, we developed computational models for disease transmission and infection progression. We estimated that a 50% reduction in access to healthcare services during the Ebola outbreak exacerbated malaria, HIV/AIDS, and tuberculosis mortality rates by additional death counts of 6,269 (2,564-12,407) in Guinea; 1,535 (522-2,8780) in Liberia; and 2,819 (844-4,844) in Sierra Leone. The 2014-2015 Ebola outbreak was catastrophic in these countries, and its indirect impact of increasing the mortality rates of other diseases was also substantial.


Vitamin D receptor pathway is required for probiotic protection in colitis.
Low expression of vitamin D receptor (VDR) and dysfunction of vitamin D/VDR signaling are reported in patients with inflammatory bowel disease (IBD); therefore, restoration of VDR function to control inflammation in IBD is desirable. Probiotics have been used in the treatment of IBD. However, the role of probiotics in the modulation of VDR signaling to effectively reduce inflammation is unknown. We identified a novel role of probiotics in activating VDR activity, thus inhibiting inflammation, using cell models and VDR knockout mice. We found that the probiotics Lactobacillus rhamnosus strain GG (LGG) and Lactobacillus plantarum (LP) increased VDR protein expression in both mouse and human intestinal epithelial cells. Using the VDR luciferase reporter vector, we detected increased transcriptional activity of VDR after probiotic treatment. Probiotics increased the expression of the VDR target genes, such as antimicrobial peptide cathelicidin, at the transcriptional level. Furthermore, the role of probiotics in regulating VDR signaling was tested in vivo using a Salmonella-colitis model in VDR knockout mice. Probiotic treatment conferred physiological and histologic protection from Salmonella-induced colitis in VDR(+/+) mice, whereas probiotics had no effects in the VDR(-/-) mice. Probiotic treatment also enhanced numbers of Paneth cells, which secrete AMPs for host defense. These data indicate that the VDR pathway is required for probiotic protection in colitis. Understanding how probiotics enhance VDR signaling and inhibit inflammation will allow probiotics to be used effectively, resulting in innovative approaches to the prevention and treatment of chronic inflammation.


Working in the Field of Complex Psychological Trauma: A Framework for Personal and Professional Growth, Training, and Supervision.
The aim of this article is to explore the positive and negative impacts of working therapeutically in complex psychological trauma (CPT), particularly the field of gender-based violence (GBV) and childhood sexual abuse (CSA), from the clinicians' perspective. The focus was on the prospect of positive gains and growth for therapists. Twenty-one clinicians (n = 21; counselors/psychotherapists and psychologists) from National Health Service (NHS) specialist trauma services, a community mental health team, and specialist sexual assault counseling organization participated. Interpretative phenomenological analysis (IPA) was utilized to conduct single one-off interviews and analysis. Six themes were identified: Called to the work; Connection, Separation, and Oneness; Into and out of the darkness; Chaos into meaning; Reparation not repetition; and Expansion and growth. The first "Therapist Led Framework of Growth in Trauma Work" is presented. Vicarious posttraumatic growth (VPTG) was a key finding, with CPT therapists experiencing a "challenge/benefit/change" growth process. Adoption of actively relational strategies to enhance clinicians' growth process through trauma work is being proposed. The benefits of conceptualizing both the positive and negative impacts of such work for supervision, training, shaping the formal curricula, service management, and continuing professional development (CPD) are being discussed. The need for good practice guidelines on self-care internationally is highlighted.


Activity of NaV1.2 promotes neurodegeneration in an animal model of multiple sclerosis.
Counteracting the progressive neurological disability caused by neuronal and axonal loss is the major unmet clinical need in multiple sclerosis therapy. However, the mechanisms underlying irreversible neuroaxonal degeneration in multiple sclerosis and its animal model experimental autoimmune encephalomyelitis (EAE) are not well understood. A long-standing hypothesis holds that the distribution of voltage-gated sodium channels along demyelinated axons contributes to neurodegeneration by increasing neuroaxonal sodium influx and energy demand during CNS inflammation. Here, we tested this hypothesis in vivo by inserting a human gain-of-function mutation in the mouse NaV1.2-encoding gene Scn2a that is known to increase NaV1.2-mediated persistent sodium currents. In mutant mice, CNS inflammation during EAE leads to elevated neuroaxonal degeneration and increased disability and lethality compared with wild-type littermate controls. Importantly, immune cell infiltrates were not different between mutant EAE mice and wild-type EAE mice. Thus, this study shows that increased neuronal NaV1.2 activity exacerbates inflammation-induced neurodegeneration irrespective of immune cell alterations and identifies NaV1.2 as a promising neuroprotective drug target in multiple sclerosis.


Regulation of expression of zebrafish (Danio rerio) insulin-like growth factor 2 receptor: implications for evolution at the IGF2R locus.
The insulin-like growth factor 2 receptor (IGF2R) is an unusual multifunctional receptor that interacts with a diverse variety of ligands. While the receptor has been well-characterized in mammals, little is known of its biology in other vertebrates. In this report, we characterize the expression of the zebrafish (Danio rerio) ortholog of the IGF2R gene. We show that two distinct, cell-type-specific promoters drive transcription of zebrafish igf2r and that these encode receptor isoforms that differ in their amino termini. Both promoters are active in adult fish and during embryonic development, but the proximal promoter generates more abundant transcripts. The 5'-UTR of the more abundantly expressed transcript contains several AUGs upstream of the main start codon, and these negatively regulate translation of a downstream reporter gene. Comparative sequence analysis shows that upstream AUGs (uAUGs) are a feature of IGF2R genes in several other vertebrates, including Xiphophorus, Xenopus, chicken, platypus, and opossum, but not in eutherian mammals. The IGF2R is imprinted in marsupial and placental mammals, and this transcriptional control of receptor abundance was proposed to have evolved following acquisition of an insulin-like growth factor 2 (IGF2) binding site by the ancestral receptor. Our observations suggest that receptor abundance was regulated at translation in ancestral vertebrates, before acquisition of an IGF2 binding site. We propose that evolution of imprinting at the mammalian IGF2R may have facilitated the loss of negative regulation of translation conferred by uAUGs.


Patients infected by tuberculosis and human immunodeficiency virus facing their disease, their reactions to disease diagnosis and its implication about their families and communities, in Burkina Faso: a mixed focus group and cross sectional study.
Patients facing tuberculosis (TB) and human immunodeficiency virus (HIV) infection receive particular care. Despite efforts in the care, misconceptions about TB and HIV still heavily impact patients, their families and communities. This situation severely limits achievement of TB and HIV programs goals. This study reports current situation of TB patients and patients living with HIV/AIDS (PLWHA) facing their disease and its implications, by comparing results from both qualitative and quantitative study design. Cross sectional study using mixed methods was used and excluded patients co-infected by TB and HIV. Focus group included 96 patients (6 patients per group) stratified by setting, disease profile and gender; from rural (Orodara Health District) and urban (Bobo Dioulasso) areas, all from Hauts-Bassins region in Burkina Faso. Quantitative study included 862 patients (309 TB patients and 553 PLWHA) attending TB and HIV care facilities in two main regions (Hauts-Bassins and Centre) of Burkina Faso. A content analysis of reports found TB patients and PLWHA felt discriminated and stigmatized because of misconceptions with its aftermaths (rejection, emotional and financial problems), mainly among PLWHA and women patients. PLWHA go to healers when facing limited solutions in health system. There are fewer associations for TB patients, and less education and sensitization sessions to give them opportunity for sharing disease status and learning from other TB patients. TB patients and PLWHA still need to better understand their disease and its implication. Access to care (diagnosis and treatment) remains one of the key issues in health system, especially for PLWHA. Individual counseling is centered among PLWHA but not for TB patients. With research progress and experiences sharing, TB patients and PLWHA have some hope to implement their life project, and to receive psychosocial and nutritional support. Despite international aid, TB patients and PLWHA are facing misconceptions effects. There is a need to reinforce health education towards patients and healers, inside community, health centers and associations, and for specific settings. International aid must be adapted to specific targets and strategies implementing programs. Maintaining psychosocial and nutritional support is crucial for better outcomes of medication adherence. Individual counseling has to be centered among TB patients and PLWHA.


Codesigning a Measure of Person-Centred Coordinated Care to Capture the Experience of the Patient: The Development of the P3CEQ.
Person-centred coordinated care (P3C) is a priority for stakeholders (ie, patients, carers, professionals, policy makers). As a part of the development of an evaluation framework for P3C, we set out to identify patient-reported experience measures (PREMs) suitable for routine measurement and feedback during the development of services. A rapid review of the literature was undertaken to identity existing PREMs suitable for the probing person-centred and/or coordinated care. Of 74 measures identified, 7 met our inclusion criteria. We critically examined these against core domains and subdomains of P3C. Measures were then presented to stakeholders in codesign workshops to explore acceptability, utility, and their strengths/weaknesses. The Long-Term Condition 6 questionnaire was preferred for its short length, utility, and tone. However, it lacked key questions in each core domain, and in response to requests from our codesign group, new questions were added to cover consideration as a whole person, coordination, care plans, carer involvement, and a single coordinator. Cognitive interviews, on-going codesign, and mapping to core P3C domains resulted in the refinement of the questionnaire to 11 items with 1 trigger question. The 11-item modified version was renamed the P3C Experiences Questionnaire. Due to a dearth of brief measures available to capture people's experience of P3C for routine practice, an existing measure was modified using an iterative process of adaption and validation through codesign workshops. Next steps include psychometric validation and modification for people with dementia and learning difficulties.


Forkhead transcription factor HFH-4 expression is temporally related to ciliogenesis.
Members of the forkhead/winged-helix family of transcription factors are expressed in tissue-specific patterns and play critical roles in development and cell differentiation. The expression of forkhead family member hepatocyte nuclear factor-3/forkhead homologue 4 (HFH-4) has been localized by RNA-blot analysis and in situ hybridization to the proximal airway of the lung (trachea, bronchi, and bronchioles) with onset at mouse embryonic day (E) 14.5 and is present in the choroid plexus, ependymal cells, oviduct, and testis. We hypothesized that the restricted expression of HFH-4 messenger RNA suggests a function common to these tissues and therefore a cell-specific role for HFH-4. Accordingly, an anti-HFH-4 antibody was generated and used for cell-specific localization of protein expression to begin to identify the functions of HFH-4. We found HFH-4 expression in proximal airway ciliated epithelial cells, but not Clara cells or alveolar epithelial cells. HFH-4 was also expressed in ciliated epithelial cells of the nose and paranasal sinuses, choroid plexus, ependyma, and oviduct. In developing mouse lung, HFH-4 expression was initially detected in airway epithelial cells at E15.5, before the appearance of cilia, and at later stages was localized to epithelial cells with cilia. In the testis, HFH-4 expression in spermatids was coincident with stage-specific generation of flagella. The temporal relationship of HFH-4 expression to the development of cilia and flagella, and the restricted expression in ciliated epithelial cells, suggest that this transcription factor has a role in regulation and maintenance of the ciliated cell phenotype in epithelial cells.


The critical care nurse work environment, physician staffing, and risk for ventilator-associated pneumonia.
We examined the relationship between intensivist physician staffing, nurse work environment, and ventilator-associated pneumonia (VAP) in 25 intensive care units. We found a significant interaction between the nurse work environment, intensivist physician staffing, and VAP. Future work may need to focus on fostering organizational collaboration between nursing and medicine to leverage skills of both clinician groups to reduce risk for VAP in critically ill patients.


Early diagnosis of intra-abdominal inflammation and sepsis by neutrophil CD64 expression in newborns.
Newborn infants with intra-abdominal inflammation/sepsis often present with nonspecific signs in the early stages of the disease, but can rapidly develop life-threatening complications. A reliable 'early' biomarker would be invaluable. To evaluate the effectiveness of neutrophil CD64 as an 'early' biomarker of intra-abdominal inflammation/sepsis. Blood was collected from newborns with suspected intra-abdominal pathology for neutrophil CD64 and C-reactive protein (CRP) determination at the onset of clinical presentation and 24 h later. They were classified into three groups: intra-abdominal inflammation/sepsis (group 1), extra-abdominal sepsis (group 2) and nonsepsis (group 3). Between-group comparisons were made by Kruskal-Wallis and χ(2) tests. Receiver-operating characteristic curves and diagnostic utilities for single and combination of tests were determined. 310 infants were recruited (102, 34 and 174 in groups 1, 2 and 3, respectively). CD64 (conventional cutoff = 6,010 antibody-PE molecules bound/cell) had substantially better sensitivity (0.81 vs. 0.56) and negative predictive value (0.90 vs. 0.79) for diagnosing intra-abdominal sepsis than CRP, at presentation. Pairing CD64 with routine abdominal radiograph (AXR) substantially increased the sensitivity and negative predictive value for group 1 to 0.99 and 0.99, respectively. By adjusting the CD64 cutoff to 12,500 units, a substantial improvement in specificity could be achieved (0.62 to 0.80) without significantly compromising sensitivity (0.99 to 0.97). CD64 is a sensitive and 'early' biomarker for diagnosing intra-abdominal inflammation/sepsis. Intra-abdominal catastrophes, including necrotizing enterocolitis, intestinal necrosis, perforation and peritonitis can confidently be excluded using CD64 and AXR early in the course of the disease.


Homing to central nervous system vasculature by antigen-specific lymphocytes. I. Localization of 14C-labeled cells during acute, chronic, and relapsing experimental allergic encephalomyelitis.
Chronic relapsing experimental allergic encephalomyelitis was induced by the passive transfer of [14C]thymidine-labeled myelin basic protein (MBP)-sensitized lymphocytes from MBP-immunized mice to naive syngeneic recipients. Labeled lymphocytes were localized and quantitated in the central nervous system during acute and chronic disease and clinical relapses. The results have shown that MBP-immune T cells home to the central nervous system endothelium 24 hours prior to and during initial clinical disease (5 to 7 days posttransfer). Unexpectedly, labeled MBP-immune cells never migrated far from blood vessels and, despite the presence of massive parenchymal inflammatory cell infiltration, almost invariably remained within the perivascular area. Quantitation revealed that labeled cells represented a minority (usually 1% to 4%) of the inflammatory cells during acute and early chronic disease. Furthermore, labeled cells could not be demonstrated in the central nervous system at the time of clinical relapse. We conclude that in this model, MBP-immune lymphocytes act exclusively from a perivascular location to orchestrate the influx of inflammatory cells that are predominantly of recipient derivation.


Polymorphism of the Pv200L fragment of merozoite surface protein-1 of Plasmodium vivax in clinical isolates from the Pacific coast of Colombia.
Merozoite surface protein 1 (MSP-1) is a polymorphic malaria protein with functional domains involved in parasite erythrocyte interaction. Plasmodium vivax MSP-1 has a fragment (Pv200L) that has been identified as a potential subunit vaccine because it is highly immunogenic and induces partial protection against infectious parasite challenge in vaccinated monkeys. To determine the extent of genetic polymorphism and its effect on the translated protein, we sequenced the Pv200L coding region from isolates of 26 P. vivax-infected patients in a malaria-endemic area of Colombia. The extent of nucleotide diversity (π) in these isolates (0.061 ± 0.004) was significantly lower (P ≤ 0.001) than that observed in Thai and Brazilian isolates; 0.083 ± 0.006 and 0.090 ± 0.006, respectively. We found two new alleles and several previously unidentified dimorphic substitutions and significant size polymorphism. The presence of highly conserved blocks in this fragment has important implications for the development of Pv200L as a subunit vaccine candidate.


Serum levels of an isoform of apolipoprotein A-II as a potential marker for prostate cancer.
We recently showed that protein expression profiling of serum using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) has potential as a diagnostic approach for detection of prostate cancer. As a parallel effort, we have been pursuing the identification of the protein(s) comprising the individual discriminatory "peaks" and evaluating their utility as potential biomarkers for prostate disease. We employed liquid chromatography, gel electrophoresis and tandem mass spectroscopy to isolate and identify a protein that correlates with observed SELDI-TOF MS mass/charge (m/z) values. Immunodepletion, immunoassay, and Western analysis were used to verify that the identified protein generated the observed SELDI peak. Subsequent immunohistochemistry was used to examine the expression of the proteins in prostate tumors. An 8,946 m/z SELDI-TOF MS peak was found to retain discriminatory value in each of two separate data sets with an increased expression in the diseased state. Sequence identification by liquid chromatography-MS/MS and subsequent immunoassays verified that an isoform of apolipoprotein A-II (ApoA-II) is the observed 8,946 m/z SELDI peak. Immunohistochemistry revealed that ApoA-II is overexpressed in prostate tumors. SELDI-based immunoassay revealed that an 8.9-kDa isoform of ApoA-II is specifically overexpressed in serum from individuals with prostate cancer. ApoA-II was also overexpressed in the serum of individuals with prostate cancer who have normal prostate-specific antigen (0-4.0 ng/mL). We have identified an isoform of ApoA-II giving rise to an 8.9K m/z SELDI "peak" that is specifically overexpressed in prostate disease. The ability of ApoA-II to detect disease in patients with normal prostate-specific antigen suggests potential utility of the marker in identifying indolent disease.


Bugs in the program: can pregnancy drugs and smoking disturb molecular reprogramming of the fetal germline, increasing heritable risk for autism and neurodevelopmental disorders?
In a seeming paradox, the prevalence of autism spectrum disorder (ASD) has surged, while at the same time research has pointed to the strong heritability of this neurodevelopmental pathology. Here an autism research philanthropist suggests a biological phenomenon of exogenously induced 'gamete disruption' that could reconcile these seemingly contradictory observations. Mining information from her own family history and that of her fellow autism parents, while also engaging with the scientific community, she proposes that a subset of the autisms may be rooted in a variety of molecular glitches in parental gametes induced by certain acute exposures during the parents' own fetal or neonatal development. These exposures include but are not limited to synthetic hormone drugs, tobacco, and general anesthesia. Consistent with this hypothesis, animal models have demonstrated adverse neurobehavioral outcomes in grandoffspring of gestating dams exposed to hormone-disrupting compounds, tobacco components, and general anesthesia. A recent epidemiological study showed a link between grandmaternal smoking and risk for ASD in grandoffspring through the maternal line. Given the urgency of the autism crisis, combined with the biological plausibility of this mostly unexplored paradigm, the writer contends that questions of nongenetic inheritance should be a priority in autism research.