Purification and characterization of a novel restricted antigen expressed by normal and transformed human colonic epithelium.
A cell surface antigen that is expressed by normal and 95% of transformed colonic epithelium and is recognized by the monoclonal antibody A33 (Welt, S., Divgi, C. R., Real, F. X., Yeh, S. D., Garin-Chesa, P., Finstad, C. L., Sakamoto, J., Cohen, A., Sigurdson, E. R., Kemeny, N., Carswell, E. A., Oettgen, H. F., and Old, L. J. (1990) J. Clin. Oncol. 8, 1894-1906) has been purified to homogeneity from the human colonic carcinoma cell line LIM1215. The A33 protein was purified from Triton X-114 extracts of LIM1215 cells under nondenaturing conditions. These extracts were applied sequentially to Green-Sepharose HE-4BD, Mono-Q HR 10/10, Superose 12 HR 10/30, and micropreparative Brownlee Aquapore RP 300. The purification was monitored by biosensor analysis using surface plasmon resonance detection with a F(ab')2 fragment of the humanized A33 monoclonal antibody immobilized on the sensor surface and Western blot analysis following SDS-polyacrylamide gel electrophoresis (PAGE) under nonreducing conditions using humanized A33 monoclonal antibody. The purified A33 antigen has a Mr on SDS-PAGE of 43,000 under nonreducing conditions. By contrast, the purified protein displayed a Mr of approximately 180,000 under native conditions on both size exclusion chromatography and native PAGE, possibly due to the formation of a homotetramer. N-terminal amino acid sequence analysis of the purified protein identified 34 amino acid residues of a unique sequence: ISVETPQDVLRASQGKSVTLPXTYHTSXXXREGLIQWD. A polyclonal antibody was raised against a synthetic peptide corresponding to residues 2-20 of this sequence. The antipeptide serum recognized the purified protein using Western blot analysis under both nonreducing (Mr 43,000) and reducing (Mr 49,000) conditions.


Repeated thermal therapy improves impaired vascular endothelial function in patients with coronary risk factors.
We sought to determine whether sauna therapy, a thermal vasodilation therapy, improves endothelial function in patients with coronary risk factors such as hypercholesterolemia, hypertension, diabetes mellitus and smoking. Exposure to heat is widely used as a traditional therapy in many different cultures. We have recently found that repeated sauna therapy improves endothelial and cardiac function in patients with chronic heart failure. Twenty-five men with at least one coronary risk factor (risk group: 38 +/- 7 years) and 10 healthy men without coronary risk factors (control group: 35 +/- 8 years) were enrolled. Patients in the risk group were treated with a 60 degrees C far infrared-ray dry sauna bath for 15 min and then kept in a bed covered with blankets for 30 min once a day for two weeks. To assess endothelial function, brachial artery diameter was measured at rest, during reactive hyperemia (flow-mediated endothelium-dependent dilation [%FMD]), again at rest and after sublingual nitroglycerin administration (endothelium-independent vasodilation [%NTG]) using high-resolution ultrasound. The %FMD was significantly impaired in the risk group compared with the control group (4.0 +/- 1.7% vs. 8.2 +/- 2.7%, p < 0.0001), while %NTG was similar (18.7 +/- 4.2% vs. 20.4 +/- 5.1%). Two weeks of sauna therapy significantly improved %FMD in the risk group (4.0 +/- 1.7% to 5.8 +/- 1.3%, p < 0.001). In contrast, %NTG did not change after two weeks of sauna therapy (18.7 +/- 4.2% to 18.1 +/- 4.1%). Repeated sauna treatment improves impaired vascular endothelial function in the setting of coronary risk factors, suggesting a therapeutic role for sauna treatment in patients with risk factors for atherosclerosis.


The 5' and 3' extremities of the satellite tobacco necrosis virus translational enhancer domain contribute differentially to stimulation of translation.
The translational enhancer domain (TED) of satellite tobacco necrosis virus (STNV) RNA stimulates translation of uncapped RNAs autonomously. Here we set out to identify the 5' and 3' extremities of TED and features of these sequences with respect to translation. We found that both in wheat germ extract and in tobacco protoplasts, the 5' border is confined to 3 nt. Mutational analysis revealed that the autonomous function of TED is sensitive to 5' flanking sequences. At the 3' end of TED, 23 nt have a cumulative, quantitative effect on translation in wheat germ extract, whereas in tobacco protoplasts, the most 3' 14 nt of these 23 nt do not enhance translation. The 5' and 3' sequence requirements triggered the development of a new secondary structure model. In this model, TED folds into a phylogenetically conserved stem-loop structure in which the essential 5' nucleotides base-pair with the 3' nucleotides that stimulate translation both in vitro and in vivo. Importantly, the 14 3' nucleotides in TED that stimulate translation in the wheat germ extract only do not require the predicted base-pairing in order to function. The discrepancy between in vitro and in vivo sequence requirements thus correlates with potential base-pairing requirements, opening the possibility that TED contains two functional domains.


Sex differences in adult rat insulin and glucose responses to arginine: programming effects of neonatal separation, hypoxia, and hypothermia.
Acute neonatal hypoxia, a common stressor, causes a spontaneous decrease in body temperature which may be protective. There is consensus that hypothermia should be prevented during acute hypoxia in the human neonate; however, this may be an additional stress with negative consequences. We hypothesize that maintaining body temperature during hypoxia in the first week of postnatal life alters the subsequent insulin, glucose, and glucagon secretion in adult rats. Rat pups were separated from their dam daily from postnatal days (PD) 2-6 for the following 90 min experimental treatments: (1) normoxic separation (control), (2) hypoxia (8% O2) allowing spontaneous hypothermia, (3) normoxic hypothermia with external cold, and (4) exposure to 8% O2 while maintaining body temperature using external heat. An additional normoxic non-separated control group was performed to determine if separation per se changed the adult phenotype. Plasma insulin, glucose, and glucagon responses to arginine stimulation were evaluated from PD105 to PD133. Maternal separation (compared to non-separated neonates) had more pronounced effects on the adult response to arginine compared to the hypoxic, hypothermic, and hypoxic-isothermic neonatal treatments. Adult males exposed to neonatal maternal separation had augmented insulin and glucose responses to arginine compared to unseparated controls. Additionally, neonatal treatment had a significant effect on body weight gain; adults exposed to neonatal maternal separation were significantly heavier. Female adults had significantly smaller insulin and glucose responses to arginine regardless of neonatal treatment. Neonatal maternal separation during the first week of life significantly altered adult beta-cell function in a sexually dimorphic manner.


Sperm Diffusion Models and In Situ Confirmation of Long-Distance Fertilization in the Free-Spawning Asteroid Acanthaster planci.
This study was undertaken to compare fertilization rates of the sea star Acanthaster planci that were predicted using sperm diffusion models with those that were determined under natural conditions in the field. During experimentally induced spawnings, measured fertilization rates for broadcast eggs were high. More than 70% of the eggs were fertilized at distances as great as 8 m downstream from a single spawning male starfish, and more than 20% were fertilized at separations of more than 60 m. Fertilization was still measurable, at 5.8%, 100 m downstream. Lateral diffusion of sperm away from the axis of flow produced fertilization rates of 13.8% at 8 m normal to the flow and 32 m downstream. The large volumes of sperm released by male A. planci are the primary cause of high rates of fertilization for eggs derived from widely spaced individuals. Models of sperm diffusion using high sperm release rates such as those found in this starfish accurately confirmed the fertilization rates measured in situ for two populations of A. planci with widely differing rates of sperm release. We observed some changes in starfish density and degree of aggregation in the study population for spawning periods during two spawning seasons, though these were not striking. High levels of aggregation may not be necessary for fertilization success in this starfish, due to the potential for long-distance fertilization and the probability that, for any spawning starfish, the total number of zygotes formed will be greater at some distance from the point of spawning. Although fertilization rates in areas distant from the sperm source were relatively low, the total area for potential gamete encounters is much greater and may make a large contribution to net fertilization. We predict that other behaviors, such as migration to shallow water, commonly associated with spawning in A. planci and other marine invertebrates will have measurable impacts on fertilization success. The potential for high levels of fertilization in A. planci was realized during natural spawnings. Fertilization rates as high as 99% were recorded when levels of spawning synchrony were high.


Situation analysis for delivering integrated comprehensive sexual and reproductive health services in humanitarian crisis condition for Rohingya refugees in Cox's Bazar, Bangladesh: protocol for a mixed-method study.
Rohingya diaspora are one of the most vulnerable groups seeking refuge in camps of Cox's Bazar, Bangladesh, arising an acute humanitarian crisis. More than half of the Rohingya refugees are women and adolescent girls requiring quality sexual and reproductive health (SRH) services. Minimum initial service package of SRH are being rendered in the refugee camps; however, WHO is aiming to provide integrated comprehensive SRH services to meet the unmet needs of this most vulnerable group. For sustainable and successful implementation of such comprehensive SRH service packages, a critical first step is to undertake a situation analysis and understand the current dimensions and capture the lessons learnt on their SRH-specific needs and implementation challenges. This situation analysis is pertinent in current humanitarian condition and will provide an overview of the needs, availability and delivery of SRH services for adolescent girls and women, barriers in accessing and providing those services in Rohingya refugee camps in Cox's Bazar, Bangladesh, and similar humanitarian contexts. A concurrent mixed-methods design will be used in this study. A community-based household survey coupled with facility assessments as well as qualitative in-depth interviews, key informant interviews and focus group discussions will be conducted with community people of Rohingya refugee camps and relevant stakeholders providing SRH services to Rohingya population in Cox's Bazar, Bangladesh. Survey data will be analysed using univariate, bivariate and multivariable regression statistics. Descriptive analysis will be done for facility assessment and thematic analysis will be conducted with qualitative data. Ethical approval from Institutional Review Board of BRAC James P Grant School of Public Health (2018-017-IR) has been obtained. Findings from this research will be disseminated through presentations in local, national and international conferences, workshops, peer-reviewed publications, policy briefs and interactive project report.


The Level of NMDA Receptor in the Membrane Modulates Amyloid-β Association and Perforation.
Alzheimer's disease is a neurodegenerative disorder that affects mostly the elderly. The main histopathological markers are the senile plaques formed by amyloid-β peptide (Aβ) aggregates that can perforate the plasma membrane of cells, increasing the intracellular calcium levels and releasing synaptic vesicles that finally lead to a delayed synaptic failure. Several membrane proteins and lipids interact with Aβ affecting its toxicity in neurons. Here, we focus on NMDA receptors (NMDARs) as proteins that could be modulating the association and neurotoxic perforation induced by Aβ on the plasma membrane. In fact, our results showed that decreasing NMDARs, using enzymatic or siRNA approaches, increased the association of Aβ to the neurons. Furthermore, overexpression of NMDARs also resulted in an enhanced association between NMDA and Aβ. Functionally, the reduction in membrane NMDARs augmented the process of membrane perforation. On the other hand, overexpressing NMDARs had a protective effect because Aβ was now unable to cause membrane perforation, suggesting a complex relationship between Aβ and NMDARs. Because previous studies have recognized that Aβ oligomers are able to increase membrane permeability and produce amyloid pores, the present study supports the conclusion that NMDARs play a critical protective role on Aβ actions in hippocampal neurons. These results could explain the lack of correlation between brain Aβ burden and clinically observed dementia.


X-Linked Alport Dogs Demonstrate Mesangial Filopodial Invasion of the Capillary Tuft as an Early Event in Glomerular Damage.
X-linked Alport syndrome (XLAS), caused by mutations in the type IV collagen COL4A5 gene, accounts for approximately 80% of human Alport syndrome. Dogs with XLAS have a similar clinical progression. Prior studies in autosomal recessive Alport mice demonstrated early mesangial cell invasion as the source of laminin 211 in the glomerular basement membrane (GBM), leading to proinflammatory signaling. The objective of this study was to verify this process in XLAS dogs. XLAS dogs and WT littermates were monitored with serial clinicopathologic data and kidney biopsies. Biopsies were obtained at set milestones defined by the onset of microalbuminuria (MA), overt proteinuria, onset of azotemia, moderate azotemia, and euthanasia. Kidney biopsies were analyzed by histopathology, immunohistochemistry, and electron microscopy. XLAS dogs showed progressive decrease in renal function and progressive increase in interstitial fibrosis and glomerulosclerosis (based on light microscopy and immunostaining for fibronectin). The only identifiable structural abnormality at the time of microalbuminuria was ultrastructural evidence of mild segmental GBM multilamination, which was more extensive when overt proteinuria developed. Co-localization studies showed that mesangial laminin 211 and integrin α8β1 accumulated in the GBM at the onset of overt proteinuria and coincided with ultrastructural evidence of mild cellular interpositioning, consistent with invasion of the capillary loops by mesangial cell processes. In a large animal model, the induction of mesangial filopodial invasion of the glomerular capillary loop leading to the irregular deposition of laminin 211 is an early initiating event in Alport glomerular pathology.


Effect of sex on symptoms associated with gastroesophageal reflux.
Previous research suggests that females have a poorer outcome than do males after surgery for gastroesophageal reflux. To evaluate reflux and esophageal symptoms in males and females in a community sample and in patients undergoing antireflux surgery. Face-to-face interview. A South Australian community. Random sample of 2973 individuals from the community and 2153 patients presenting for antireflux surgery. In a random sample of 2973 individuals from the community, the prevalence of reflux and other esophageal symptoms was determined and compared with symptoms in 2153 patients presenting for antireflux surgery. Identical questions were used to assess frequency and severity of heartburn and dysphagia and medication use. Analog scales assessed heartburn and dysphagia (0 indicating no symptoms and 10, severe symptoms). Outcomes for males vs females were compared across both groups. In the community, females were more likely to report heartburn, and when reported, symptom severity was higher. The prevalence of dysphagia was similar for males and females, although females reported higher dysphagia scores for solid foods. A similar proportion of males and females took antireflux medications. Females presenting for antireflux surgery were, on average, 7 years older than males, had a higher body mass index, and had higher heartburn and dysphagia symptom scores. At endoscopy, men were more likely to have ulcerative esophagitis and Barrett esophagus, and at surgery they were less likely to have a hiatal hernia. Significant differences were noted between males and females in the frequency and severity of gastroesophageal reflux-associated symptoms in the community and in patients presenting for surgery. These might reflect differences in symptom perception, which explain previously reported better outcomes in men undergoing antireflux surgery.


A Pilot Study of a Grape Seed Procyanidin Extract for Lung Cancer Chemoprevention.
Grape seed procyanidin extract (GSE) had been reported to exert antineoplastic properties in preclinical studies. A modified phase I, open-label, dose-escalation clinical study was conducted to evaluate the safety, tolerability, MTD, and potential chemopreventive effects of leucoselect phytosome (LP), a standardized GSE complexed with soy phospholipids to enhance bioavailability, in heavy active and former smokers. Eight subjects ages 46-68 years were enrolled into the study and treated with escalating oral doses of LP for 3 months. Bronchoscopies with bronchoalveolar lavage and bronchial biopsies were performed before and after 3 months of LP treatment. Hematoxylin and eosin stain for histopathology grading and IHC examination for Ki-67 proliferative labeling index (Ki-67 LI) were carried out on serially matched bronchial biopsy samples from each subject to determine responses to treatment. Two subjects were withdrawn due to issues unrelated to the study medication, and a total of 6 subjects completed the full study course. In general, 3 months of LP, reaching the highest dose per study protocol was well tolerated and no dosing adjustment was necessary. Such a treatment regimen significantly decreased bronchial Ki-67 LI by an average of 55% (P = 0.041), with concomitant decreases in serum miR-19a, -19b, and -106b, which were oncomirs previously reported to be downregulated by GSE, including LP, in preclinical studies. In spite of not reaching the original enrollment goal of 20, our findings nonetheless support the continued clinical translation of GSE as an antineoplastic and chemopreventive agent against lung cancer.


Fixation of mandibular angle fractures: in vitro biomechanical assessments and computer-based studies.
The purpose of this study was to review the literature regarding the evolution of current thoughts on fixation of mandibular angle fractures (MAFs), based on in vitro biomechanical assessments and computer-based studies. An electronic search in PubMed was undertaken in August 2012. The titles and abstracts from these results were read to identify studies within the selection criteria. Eligibility criteria included studies from the last 30 years (from 1983 onwards). The search strategy initially identified 767 studies. Thirty-one studies were identified without repetition within the selection criteria. Two articles showing significance in the development of treatment techniques was included. Additional hand searching yielded five additional papers. Thus, a total of 38 studies were included. The osteosynthesis positions as well as the plating technique play important roles in the stability of MAF repair. The only in vitro study evaluating the use of wire osteosynthesis concluded that wires placed through the lower border approach would provide greater stability than those at the upper border. Many studies indicate that the use of two miniplates avoids (or decreases) lateral displacement of the lower mandibular border and opening of the inferior fracture gap. Some studies even suggest that the use of two miniplates may be considered a more "rigid" fixation technique for MAFs than the use of a reconstruction plate. When using two miniplates, the biplanar plate orientation provides greater biomechanical stability than the monoplanar one. However, despite its greater biomechanical stability, the two-miniplate technique has some disadvantages that should also be taken into account. Studies with biodegradable plates suggest the use of at least two plates for each MAF. There are few studies with compression plates, and they have not yet reached a consensus. The solitary lag screw proved to withstand the functional loading of the mandible; however, only few biomechanical assessments were performed. In vitro studies have shown good biomechanical stability with the use of 3-D grid plates. The use of malleable miniplates alone is not sufficient to withstand the early postoperative bite force. Some studies suggest that the segment of the tension band miniplate located at the distal fragment of the MAF should be fixed with three screws. The studies also showed some limitations. None considered the stabilization of the fracture site afforded by the masseter-pterygoid muscle pouch. Most of the studies did not evaluate plating system strength in the long term and therefore did not observe the effect of resorption on the strength of the different biodegradable plating systems. Another limitation of many studies is the absence of a control group. A confounding factor that could not be tested in in vitro investigations is the additional resistance to displacement of jagged fracture margins present in the human fracture.


Advancing Eucalyptus genomics: identification and sequencing of lignin biosynthesis genes from deep-coverage BAC libraries.
Eucalyptus species are among the most planted hardwoods in the world because of their rapid growth, adaptability and valuable wood properties. The development and integration of genomic resources into breeding practice will be increasingly important in the decades to come. Bacterial artificial chromosome (BAC) libraries are key genomic tools that enable positional cloning of important traits, synteny evaluation, and the development of genome framework physical maps for genetic linkage and genome sequencing. We describe the construction and characterization of two deep-coverage BAC libraries EG_Ba and EG_Bb obtained from nuclear DNA fragments of E. grandis (clone BRASUZ1) digested with HindIII and BstYI, respectively. Genome coverages of 17 and 15 haploid genome equivalents were estimated for EG_Ba and EG_Bb, respectively. Both libraries contained large inserts, with average sizes ranging from 135 Kb (Eg_Bb) to 157 Kb (Eg_Ba), very low extra-nuclear genome contamination providing a probability of finding a single copy gene ≥ 99.99%. Libraries were screened for the presence of several genes of interest via hybridizations to high-density BAC filters followed by PCR validation. Five selected BAC clones were sequenced and assembled using the Roche GS FLX technology providing the whole sequence of the E. grandis chloroplast genome, and complete genomic sequences of important lignin biosynthesis genes. The two E. grandis BAC libraries described in this study represent an important milestone for the advancement of Eucalyptus genomics and forest tree research. These BAC resources have a highly redundant genome coverage (> 15×), contain large average inserts and have a very low percentage of clones with organellar DNA or empty vectors. These publicly available BAC libraries are thus suitable for a broad range of applications in genetic and genomic research in Eucalyptus and possibly in related species of Myrtaceae, including genome sequencing, gene isolation, functional and comparative genomics. Because they have been constructed using the same tree (E. grandis BRASUZ1) whose full genome is being sequenced, they should prove instrumental for assembly and gap filling of the upcoming Eucalyptus reference genome sequence.


A novel system for the production of fully deleted adenovirus vectors that does not require helper adenovirus.
Fully deleted adenovirus vectors (FD-AdVs) would appear to be promising tools for gene therapy. Since these vectors are deleted of all adenoviral genes, they require a helper adenovirus for their propagation. The contamination of the vector preparation by the helper limits the utility of currently existing FD-AdVs in gene therapy applications. We have developed an alternative system for the propagation of FD-AdVs, in which the adenoviral genes essential for replication and packaging of the vector are delivered into producer cells by a baculovirus-adenovirus hybrid. A hybrid baculovirus Bac-B4 was constructed to carry a Cre recombinase-excisable copy of the packaging-deficient adenovirus genome. Although the total size of the DNA insert in Bac-B4 was 38 kb, the genetic structure of this recombinant baculovirus was stable. Bac-B4 gave high yields in Sf9 insect cells, with titers of 5 x 10(8)p.f.u./ml before concentration. Transfection of 293-Cre cells with lacZ-expressing FD-AdV plasmid DNA followed by infection by Bac-B4 at a MOI of 2000 p.f.u./ml resulted in rescue of the helper-free vector. Subsequent passaging of the obtained FD-AdV using Bac-B4 as a helper resulted in approximately 100-fold increases of the vector titer at each passage. This resulting vector was completely free of helper virus and was able to transduce cultured 293 cells. However, scaling-up of FD-AdV production was prevented by the eventual emergence of replication-competent adenovirus (RCA). Experiments are underway to optimize this system for the large-scale production of helper virus-free FD-AdVs and to minimize the possibility of generation of replication-competent adenovirus (RCA) during vector production. This baculovirus-based system will be a very useful alternative to current methods for the production of FD-AdVs.


Contribution of NMDA Receptors to Synaptic Function in Rat Hippocampal Interneurons.
The ability of neurons to produce behaviorally relevant activity in the absence of pathology relies on the fine balance of synaptic inhibition to excitation. In the hippocampal CA1 microcircuit, this balance is maintained by a diverse population of inhibitory interneurons that receive largely similar glutamatergic afferents as their target pyramidal cells, with EPSCs generated by both AMPA receptors (AMPARs) and NMDA receptors (NMDARs). In this study, we take advantage of a recently generated GluN2A-null rat model to assess the contribution of GluN2A subunits to glutamatergic synaptic currents in three subclasses of interneuron found in the CA1 region of the hippocampus. For both parvalbumin-positive and somatostatin-positive interneurons, the GluN2A subunit is expressed at glutamatergic synapses and contributes to the EPSC. In contrast, in cholecystokinin (CCK)-positive interneurons, the contribution of GluN2A to the EPSC is negligible. Furthermore, synaptic potentiation at glutamatergic synapses on CCK-positive interneurons does not require the activation of GluN2A-containing NMDARs but does rely on the activation of NMDARs containing GluN2B and GluN2D subunits.


Engineering a leucine zipper-TRAIL homotrimer with improved cytotoxicity in tumor cells.
Successful cancer therapies aim to induce selective apoptosis in neoplastic cells. The current suboptimal efficiency and selectivity drugs have therapeutic limitations and induce concomitant side effects. Recently, novel cancer therapies based on the use of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) have emerged. TRAIL, a key component of the natural antitumor immune response, selectively kills many tumor cell types. Earlier studies with recombinant TRAIL, however, revealed its many shortcomings including a short half-life, off-target toxicity, and existence of TRAIL-resistant tumor cells. We improved the efficacy of recombinant TRAIL by redesigning its structure and the expression and purification procedures. The result is a highly stable leucine zipper (LZ)-TRAIL chimera that is simple to produce and purify. This chimera functions as a trimer in a manner that is similar to natural TRAIL. The formulation of the recombinant LZ-TRAIL we have developed has displayed high specific activity in both cell-based assays in vitro and animal tests in vivo. Our results have shown that the half-life of LZ-TRAIL is improved and now exceeds 1 h in mice compared with a half-life of only minutes reported earlier for recombinant TRAIL. We have concluded that our LZ-TRAIL construct will serve as a foundation for a new generation of fully human LZ-TRAIL proteins suitable for use in preclinical and clinical studies and for effective combination therapies to overcome tumor resistance to TRAIL.


A cyclin without cyclin-dependent kinases: cyclin F controls genome stability through ubiquitin-mediated proteolysis.
Cell cycle transitions are driven by the periodic oscillations of cyclins, which bind and activate cyclin-dependent kinases (CDKs) to phosphorylate target substrates. Cyclin F uses a substrate recruitment strategy similar to that of the other cyclins, but its associated catalytic activity is substantially different. Indeed, cyclin F is the founding member of the F-box family of proteins, which are the substrate recognition subunits of Skp1-Cul1-F-box protein (SCF) ubiquitin ligase complexes. Here, we discuss cyclin F function and recently identified substrates of SCF(cyclin)(F) involved in deoxyribonucleotide triphosphate (dNTP) production, centrosome duplication, and spindle formation. We highlight the relevance of cyclin F in controlling genome stability through ubiquitin-mediated proteolysis and the implications for cancer development.


Surveillance of potential hosts and vectors of scrub typhus in Taiwan.
Scrub typhus is a lethal infectious disease vectored by larval trombiculid mites (i.e. chiggers) infected with Orientia tsutsugamushi (OT) and recent decades have witnessed an emergence of scrub typhus in several countries. Identification of chigger species and their vertebrate hosts is fundamental for the assessment of human risks to scrub typhus under environmental changes, but intensive and extensive survey of chiggers and their hosts is still lacking in Taiwan. Chiggers were collected from shrews and rodents in nine counties of Taiwan and were assayed for OT infections with nested polymerase chain reaction (PCR). PCR products were further sequenced to reveal probable OT strains. Rodents were assessed for OT exposure by immunofluorescent antibody assay. Lastly, incidence rate of scrub typhus in each county was associated with loads and prevalence of chigger infestations, seropositivity rate in rodents, and OT positivity rate in chiggers. Rattus losea was the most abundant (48.7% of 1,285 individuals) and widespread (occurred in nine counties) small mammal species and hosted the majority of chiggers (76.4% of 128,520 chiggers). Leptotrombidium deliense was the most common (64.9% of all identified chiggers) and widespread (occurred in seven counties) chigger species but was replaced by Leptotrombidium pallidum or Leptotrombidium scutellare during the cold seasons in two counties (Matsu and Kinmen) where winter temperatures were lower than other study sites. Seropositivity rate for OT exposure in 876 assayed rodents was 43.0% and OT positivity rate in 347 pools of chiggers was 55.9%, with 15 OT strains identified in the 107 successfully sequenced samples. Incidence rate of scrub typhus was positively correlated with chigger loads, prevalence of chigger infestations, seropositivity rate but not OT positivity rate in chiggers. Our study reveals R. losea as the primary host for chiggers and there exists a geographical and seasonal variation in chigger species in Taiwan. It also emphasizes the importance of recognition of chigger vectors and their vertebrate hosts for a better prediction of human risks to scrub typhus under rapid environmental changes.


An efficient learning procedure for deep Boltzmann machines.
We present a new learning algorithm for Boltzmann machines that contain many layers of hidden variables. Data-dependent statistics are estimated using a variational approximation that tends to focus on a single mode, and data-independent statistics are estimated using persistent Markov chains. The use of two quite different techniques for estimating the two types of statistic that enter into the gradient of the log likelihood makes it practical to learn Boltzmann machines with multiple hidden layers and millions of parameters. The learning can be made more efficient by using a layer-by-layer pretraining phase that initializes the weights sensibly. The pretraining also allows the variational inference to be initialized sensibly with a single bottom-up pass. We present results on the MNIST and NORB data sets showing that deep Boltzmann machines learn very good generative models of handwritten digits and 3D objects. We also show that the features discovered by deep Boltzmann machines are a very effective way to initialize the hidden layers of feedforward neural nets, which are then discriminatively fine-tuned.


The maintenance and regeneration of the planarian excretory system are regulated by EGFR signaling.
The maintenance of organs and their regeneration in case of injury are crucial to the survival of all animals. High rates of tissue turnover and nearly unlimited regenerative capabilities make planarian flatworms an ideal system with which to investigate these important processes, yet little is known about the cell biology and anatomy of their organs. Here we focus on the planarian excretory system, which consists of internal protonephridial tubules. We find that these assemble into complex branching patterns with a stereotyped succession of cell types along their length. Organ regeneration is likely to originate from a precursor structure arising in the blastema, which undergoes extensive branching morphogenesis. In an RNAi screen of signaling molecules, we identified an EGF receptor (Smed-EGFR-5) as a crucial regulator of branching morphogenesis and maintenance. Overall, our characterization of the planarian protonephridial system establishes a new paradigm for regenerative organogenesis and provides a platform for exploring its functional and evolutionary homologies with vertebrate excretory systems.


Enzyme replacement therapy and hematopoietic stem cell transplant: a new paradigm of treatment in Wolman disease.
Wolman disease is a rare, lysosomal storage disorder in which biallelic variants in the LIPA gene result in reduced or complete lack of lysosomal acid lipase. The accumulation of the substrates; cholesterol esters and triglycerides, significantly impacts cellular function. Untreated patients die within the first 12 months of life. Clinically, patients present severely malnourished, with diarrhoea and hepatosplenomegaly, many have an inflammatory phenotype, including with hemophagocytic lymphohistiocytosis (HLH). Hematopoietic stem cell transplant (HCT) had been historically the only treatment available but has a high procedure-related mortality because of disease progression and disease-associated morbidities. More recently, enzyme replacement therapy (ERT) with dietary substrate reduction (DSR) has significantly improved patient survival. However, ERT is life long, expensive and its utility is limited by anti-drug antibodies (ADA) and the need for central venous access. We describe five Wolman disease patients diagnosed in infancy that were treated at Royal Manchester Children's Hospital receiving ERT with DSR then HCT-multimodal therapy. In 3/5 an initial response to ERT was attenuated by ADA with associated clinical and laboratory features of deterioration. 1/5 developed anaphylaxis to ERT and the other patient died post HCT with ongoing HLH. All patients received allogeneic HCT. 4/5 patients are alive, and both disease phenotype and laboratory parameters are improved compared to when they were on ERT alone. The gastrointestinal symptoms are particularly improved after HCT, with reduced diarrhoea and vomiting. This allows gradual structured normalisation of diet with improved tolerance of dietary fat. Histologically there are reduced cholesterol clefts, fewer foamy macrophages and an improved villous structure. Disease biomarkers also show improvement with ERT, immunotherapy and HCT. Three patients have mixed chimerism after HCT, indicating a likely engraftment-defect in this condition. We describe combined ERT, DSR and HCT, multimodal treatment for Wolman disease. ERT and DSR stabilises the sick infant and reduces the formerly described prohibitively high, transplant-associated mortality in this condition. HCT abrogates the problems of ERT, namely attenuating ADA, the need for continuing venous access, and continuing high cost drug treatment. HCT also brings improved efficacy, particularly evident in improved gastrointestinal function and histology. Multimodal therapy should be considered a new paradigm of treatment for Wolman disease patients where there is an attenuated response to ERT, and for all patients where there is a well-matched transplant donor, in order to improve long term gut function, tolerance of a normal diet and quality of life.


A robust and rapid xenograft model to assess efficacy of chemotherapeutic agents for human acute myeloid leukemia.
Relevant preclinical mouse models are crucial to screen new therapeutic agents for acute myeloid leukemia (AML). Current in vivo models based on the use of patient samples are not easy to establish and manipulate in the laboratory. Our objective was to develop robust xenograft models of human AML using well-characterized cell lines as a more accessible and faster alternative to those incorporating the use of patient-derived AML cells. Five widely used AML cell lines representing various AML subtypes were transplanted and expanded into highly immunodeficient non-obese diabetic/LtSz-severe combined immunodeficiency IL2Rγc(null) mice (for example, cell line-derived xenografts). We show here that bone marrow sublethal conditioning with busulfan or irradiation has equal efficiency for the xenotransplantation of AML cell lines. Although higher number of injected AML cells did not change tumor engraftment in bone marrow and spleen, it significantly reduced the overall survival in mice for all tested AML cell lines. On the basis of AML cell characteristics, these models also exhibited a broad range of overall mouse survival, engraftment, tissue infiltration and aggressiveness. Thus, we have established a robust, rapid and straightforward in vivo model based on engraftment behavior of AML cell lines, all vital prerequisites for testing new therapeutic agents in preclinical studies.


Timing of antenatal corticosteroids and neonatal pulmonary mechanics.
We sought to compare lung mechanics in infants treated with multiple courses of antenatal corticosteroids with those in matched control infants delivered >7 days from dosing and those of matched untreated infants. Eighteen infants who received multiple courses of corticosteroids and were delivered within 7 days of dosing were matched with 18 infants who received 1 course of corticosteroids >7 days before delivery (remote) and 18 untreated infants. Respiratory compliance and functional residual capacity were measured within 36 hours. Differences were compared by analysis of variance. Infant demographics were similar. Respiratory compliance was higher in the multiple-course group than in the remote or untreated group (P <.02). Functional residual capacity was higher in the multiple-course group than in the untreated group (P <.05) but similar to that found in the remote group. Babies delivered after multiple courses of corticosteroids and within 7 days of dosing demonstrated improved respiratory compliance compared with untreated and remotely treated infants. This suggests that the enzyme system responsible for surfactant production can be repetitively induced despite prior treatment with corticosteroids. The increased functional residual capacity in remotely treated infants may reflect a maturation of lung architecture independent of surfactant production.


Use of A-type CpG oligodeoxynucleotides as an adjuvant in allergen-specific immunotherapy in humans: a phase I/IIa clinical trial.
B-type CpG oligodeoxynucleotides (ODN) is currently used in clinical trials because of its prolonged half-life, which is due to its phosphorothioate backbone. A-type CpG ODN is a stronger inducer of IFN but has an unstable phosphodiester backbone that has so far prohibited its clinical use. However, upon association with virus-like particles (VLP) consisting of the bacteriophage Qbeta coat protein, A-type CpG ODN can be stabilized and can become an efficient adjuvant in mice. Therefore, the phase I/IIa study presented represents the first test of A-type CpGs in humans. To test the safety, tolerability and clinical efficacy of QbG10 as an adjuvant for subcutaneous immunotherapy with a house dust mite (HDM) allergen extract in allergic patients. A single centre, open-label phase I/IIa study evaluated the safety, tolerability and clinical efficacy of QbG10 as an adjuvant to immunotherapy with a subcutaneous HMD allergen extract in 20 patients suffering from HDM allergy. Twenty-one patients were enrolled between March and July 2005. Individual immunotherapy lasted 10 weeks. Clinical end-points included questionnaires, conjunctival provocation, skin prick tests and the measurement of allergen-specific IgG and IgE. QbG10 was well tolerated. Almost complete tolerance to the allergen was observed in conjunctival provocation testing after treatment with QbG10, and symptoms of rhinitis and allergic asthma were significantly reduced. Within 10 weeks of therapy, patients were nearly symptom-free and this amelioration lasted for at least 38 weeks post-treatment. Following injections of QbG10 and HDM allergen extract, allergen-specific IgG increased, while there was a transient increase in allergen-specific IgE titres. Skin reactivity to HDM was reduced. The subcutaneous application of HDM allergen, together with A-type CpG ODN packaged into VLP, was safe. All patients achieved practically complete alleviation of allergy symptoms after 10 weeks of immunotherapy. This promising clinical outcome calls for larger placebo-controlled phase II studies.


Acute physiopathological effects of honeybee (Apis mellifera) envenoming by subcutaneous route in a mouse model.
Bee stings are a health concern in the Americas, where fatal envenomings due to massive attacks by Africanized honeybees have been documented in the last decades. Most studies on the toxic effects of honeybee venom in experimental animals have been performed using the intravenous or intraperitoneal injection routes. The aim of this study was to develop a mouse model that would better resemble a massive honeybee attack by using the subcutaneous (s.c.) route to induce a severe, sublethal systemic envenoming. An array of acute venom effects were characterized, including biochemical, hematological, histological, and inflammatory alterations, after the s.c. injection of 0.5 median lethal dose of venom. Rapid increases in serum alanine (ALT) and aspartate (AST) transaminases, creatinine, urea nitrogen, uric acid, sodium and chloride electrolytes, and creatine kinase (CK) were recorded, indicating damage to liver, kidneys, and skeletal muscle. Also, coagulation disturbances (fibrinogen decrease, and moderate delay in prothrombin and partial thromboplastin times) were demonstrated. Circulating platelet and leukocyte numbers remained unaltered, but a hemoconcentration effect (hematocrit and hemoglobin increase) was observed. This effect might be related to the marked edema induced by the venom. In addition, this inflammatory response included a systemic increase in cytokines (IL-1 beta, IL-6, TNF-alpha), together with an elevation of serum malondialdehyde and nitric oxide. The myotoxic effects of venom, melittin, and phospholipase A(2) were demonstrated after injection by s.c. route. No synergistic myotoxicity between melittin and PLA(2) was observed. Moreover, these two components, when injected at equivalent concentrations to those present in venom, induced a lower increase in serum CK than venom, suggesting that other components also contribute to its strong systemic toxicity towards skeletal muscle. The model here presented may be useful in preclinical studies to assess therapeutic antivenoms developed to cope with the problem of massive bee attacks.


Curcumin stimulates reactive oxygen species production and potentiates apoptosis induction by the antitumor drugs arsenic trioxide and lonidamine in human myeloid leukemia cell lines.
Arsenic trioxide (ATO, Trisenox) is an important antileukemic drug, but its efficacy is frequently low when used as a single agent. Here, we demonstrate that the apoptotic action of ATO is greatly increased when combined with subcytotoxic curcumin concentrations in U937 and HL60 human acute myeloid leukemia cells, and with lower efficacy in K562 chronic myelogenous leukemia cells. Curcumin exerts similar cooperative effect with the mitochondria-targeting drug lonidamine, whereas the response is negligible in combination with the DNA-targeting drug cisplatin. Curcumin plus ATO or lonidamine stimulates typical events of the mitochondrial executioner pathway (Bax and Bid activation, cytochrome c release, X-linked inhibitor of apoptosis down-regulation, and caspase-9/-3 activation) and causes mitochondrial transmembrane potential dissipation, which nevertheless represents a late event in the apoptotic response. Curcumin increases anion superoxide production, and its proapoptotic action in combination with ATO and lonidamine is mimicked by pro-oxidant agents (2-methoxyestradiol and H(2)O(2)) and prevented by antioxidant agents [Mn(III)tetrakis(4-benzoic acid)porphyrin chloride and N-acetyl-l-cysteine]. Within the assayed time period (16-24 h), curcumin does not significantly modify p38-mitogen-activated protein kinase and c-Jun NH(2)-terminal kinase phosphorylation/activation or nuclear factor-κB activity, but it greatly stimulates extracellular signal-regulated kinase (ERK) phosphorylation, and decreases Akt phosphorylation. Experiments using mitogen-activated protein kinase kinase/ERK inhibitors [2'-amino-3'-methoxyflavone (PD98059) and 1,4-diamino-2,3-dicyano-1,4-bis(2-aminophenylthio)butadiene (U0126)] and phosphatidylinositol 3-kinase inhibitor 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002) indicate that ERK activation does not mediate and even restrains apoptosis potentiation, whereas Akt down-regulation facilitates apoptosis generation. In summary, cotreatment with curcumin may represent a useful manner of increasing the efficacy of ATO and lonidamine as antitumor drugs in myeloid leukemia cells.


Plasma microRNA levels in male and female children with cystic fibrosis.
A gender gap exists in cystic fibrosis (CF). Here we investigate whether plasma microRNA expression profiles differ between the sexes in CF children. MicroRNA expression was quantified in paediatric CF plasma (n = 12; six females; Age range:1-6; Median Age: 3; 9 p.Phe508del homo- or heterozygotes) using TaqMan OpenArray Human miRNA Panels. Principal component analysis indicated differences in male versus female miRNA profiles. The miRNA array analysis revealed two miRNAs which were significantly increased in the female samples (miR-885-5p; fold change (FC):5.07, adjusted p value: 0.026 and miR-193a-5p; FC:2.6, adjusted p value: 0.031), although only miR-885-5p was validated as increased in females using specific qPCR assay (p < 0.0001). Gene ontology analysis of miR-885-5p validated targets identified cell migration, motility and fibrosis as processes potentially affected, with RAC1-mediated signalling featuring significantly. There is a significant increase in miR-885-5p in plasma of females versus males with CF under six years of age.


Outcomes of transcatheter and surgical aortic valve replacement in high-risk patients with aortic stenosis and left ventricular dysfunction: results from the Placement of Aortic Transcatheter Valves (PARTNER) trial (cohort A).
The Placement of Aortic Transcatheter Valves (PARTNER) trial demonstrated similar survival after transcatheter and surgical aortic valve replacement (TAVR and SAVR, respectively) in high-risk patients with symptomatic, severe aortic stenosis. The aim of this study was to evaluate the effect of left ventricular (LV) dysfunction on clinical outcomes after TAVR and SAVR and the impact of aortic valve replacement technique on LV function. The PARTNER trial randomized high-risk patients with severe aortic stenosis to TAVR or SAVR. Patients were stratified by the presence of LV ejection fraction (LVEF) <50%. All-cause mortality was similar for TAVR and SAVR at 30-days and 1 year regardless of baseline LV function and valve replacement technique. In patients with LV dysfunction, mean LVEF increased from 35.7±8.5% to 48.6±11.3% (P<0.0001) 1 year after TAVR and from 38.0±8.0% to 50.1±10.8% after SAVR (P<0.0001). Higher baseline LVEF (odds ratio, 0.90 [95% confidence interval, 0.86, 0.95]; P<0.0001) and previous permanent pacemaker (odds ratio, 0.34 [95% confidence interval, 0.15, 0.81]) were independently associated with reduced likelihood of ≥10% absolute LVEF improvement by 30 days; higher mean aortic valve gradient was associated with increased odds of LVEF improvement (odds ratio, 1.04 per 1 mm Hg [95% confidence interval, 1.01, 1.08]). Failure to improve LVEF by 30 days was associated with adverse 1-year outcomes after TAVR but not SAVR. In high-risk patients with severe aortic stenosis and LV dysfunction, mortality rates and LV functional recovery were comparable between valve replacement techniques. TAVR is a feasible alternative for patients with symptomatic severe aortic stenosis and LV dysfunction who are at high risk for SAVR. URL: http://www.clinicaltrials.gov. Unique identifier: NCT00530894.


Convergent synthesis and cellular uptake of multivalent cell penetrating peptides derived from Tat, Antp, pVEC, TP10 and SAP.
Cell penetrating peptides (CPP) are peptides of 10 to 30 residues derived from natural translocating proteins. Multivalency is known to enhance cellular uptake for the Tat peptide and closely related polycationic sequences. To test whether multivalency effects on cellular uptake might also occur with other CPP types, we prepared multivalent versions of the strongly cationic Tat, the amphipathic sequences Antp, pVEC and TP10, and the polyproline helix SAP by convergent thioether ligation of the linear CPP onto multivalent scaffolds, and evaluated their uptake in HeLa and CHO cells, intracellular localization, cytotoxicity and hemolysis. While multivalency did not increase the cellular uptake of pVEC or SAP, multivalency effects on uptake comparable to Tat were observed with TP10 and Antp, which are attributable to their polycationic nature. The efficient synthetic protocol for these divalent CPP and their localization in the cytoplasm suggest that CPP might be useful for application in cargo delivery into cells.


Overview of robotic thyroidectomy.
With the advancement and adaptation of technology, there has been a tremendous evolution in the surgical approaches for thyroidectomy. Robotic thyroidectomy has become increasingly popular worldwide attracting both surgeons and patients searching for new and innovative techniques for thyroidectomy with a superior cosmetic result when compared to the conventional open procedures. In this review, we describe the following surgical approaches for robotic thyroidectomy: transaxillary, retroauricular (facelift) and transoral. The advantages and disadvantages as well as limitations of each approach are examined, and future directions of robotic thyroidectomy are discussed.


Geriatric Interdisciplinary Team Training 2.0: A collaborative team-based approach to delivering care.
Interprofessional collaborative education and practice has become a cornerstone of optimal person-centered management in the current complex health care climate. This is especially important when working with older adults, many with multiple chronic conditions and challenging health care needs. This paper describes a feasibility study of the Geriatric Interdisciplinary Team Training 2.0 (GITT 2.0) program focused on providing interprofessional care to complex and frail older adults with multiple chronic conditions. A concurrent triangulation mixed-methods design facilitated program implementation and evaluation. Over three years (2013-2016), 65 graduate students from nursing, midwifery, social work, and pharmacy participated along with 25 preceptors. Participants were surveyed on their attitudes toward interprofessional collaboration pre and post-intervention and participated in focus groups. While attitudes toward interprofessional collaboration did not change quantitatively, focus groups revealed changes in language and enhanced perspectives of participants. Based on the evaluation data, the GITT 2.0 Toolkit was refined for use in interprofessional education and practice activities related to quality initiatives.


Effect of routine assessment of specific psychosocial problems on personalized communication, counselors' awareness, and distress levels in cancer genetic counseling practice: a randomized controlled trial.
This study evaluated the efficacy of a cancer genetics–specific questionnaire in facilitating communication about, awareness of, and management of psychosocial problems, as well as in lowering distress levels. Individuals referred to genetic counseling for cancer at two family cancer clinics in The Netherlands were randomly assigned to an intervention or a control group. All participants completed the psychosocial questionnaire before counseling. In the intervention group, the counselors received the results of this questionnaire before the counseling session. All sessions were audiotaped for content analysis. Primary outcomes were the frequency with which psychosocial problems were discussed, the genetic counselors' awareness of these problems, and their management. Secondary outcomes included cancer worries and psychological distress, duration and dynamics of the counseling, and satisfaction. The frequency with which psychosocial problems were discussed with 246 participating counselees was significantly higher in the intervention group (n = 127) than in the control group (n =119; P = .004), as was the counselors' awareness of psychosocial problems regarding hereditary predisposition (P < .001), living with cancer (P = .01), and general emotions (P < .001). Counselors initiated more discussion of psychosocial problems in the intervention group (P < .001), without affecting the length of the counseling session. No significant differences were found on management (P = .19). The intervention group reported significantly lower levels of cancer worries (p = .005) and distress (p = .02) after counseling. The routine assessment of psychosocial problems by questionnaire facilitates genetic counselors' recognition and discussion of their clients' psychosocial problems and reduces clients' distress levels.


[Biosafety education: educational contributions to the health professional education].
The research, accomplished in the period 2004-2005, had as general objective to analyse teachers and students' perceptions of the biosafety teaching-learning process in secondary courses of the health area. The data were collected at Oswaldo Cruz Foundation courses in Rio de Janeiro. A total of 82 students and 12 teachers have participated on the study. It was a teorical-empirical research with qualitative basis and quantitative data that emerged throughout the working process. The data were analyzed in the multireferenciality context. This investigation is justified by discrepancy of the biosafety in relation to the work world and the school world and the current demands of the technical-scientific progress and of the own social evolution, in relation to the biosafety at the spaces of the health. The obtained results, pointed at a necessity of improvement on the teaching processes in biosafety in secondary courses of the health area. Then, the results may contribute to the inclusion of the biosafety in the health professional education.


Activation of mitochondrial apoptotic pathways in human renal allografts after ischemiareperfusion injury.
Ischemia-reperfusion injury in cadaveric (CAD) kidney allografts is associated with tubular cell injury, delayed graft function, and an increased incidence of acute and chronic rejection. We tested the hypothesis that activation of specific apoptotic pathways represents a mechanism for tubular cell death after CAD kidney transplantation. Serial tissue sections from paraffin-embedded needle biopsy specimens obtained at approximately 1 hr of reperfusion after transplantation of 13 CAD and 12 living-related donor (LRD) renal allografts were examined by using the terminal deoxynucleotide transferase-mediated dUTP nick-end labeling assay to detect apoptosis and by immunohistochemistry for expression of key pro-apoptotic molecules (Bax, Bak, tumor necrosis factor receptor [TNFR]-1, Fas, and cytochrome c). Apoptosis was detected primarily in tubular cells, with a mean+/-standard deviation of 6.8+/-2.2 apoptotic cells per 100 cells examined in CAD renal allografts compared with 1.8+/-2.7 cells per 100 in LRD (P<0.001) renal allografts. There was a significant correlation between apoptosis rate and cold ischemia time in CAD (r=0.86, P<0.001) renal allografts. Bax was expressed in 100% of CAD versus 17% of LRD renal allografts (P<0.001), Bak in 92% of CAD versus 17% of LRD renal allografts (P<0.001), and TNFR-1 in 100% of CAD versus 58% of LRD renal allografts (P<0.05). Fas was expressed in only a small number of samples (23% of CAD and 17% of LRD renal allografts, P=not significant). Bax and Bak were expressed predominantly in apoptotic cells. Cytochrome c was detected as a mitochondrial pattern in LRD renal allografts, but in a diffuse cytosolic distribution in CAD renal allografts. Ischemia-reperfusion injury in CAD kidney transplants is associated with a duration-dependent increase in tubular cell apoptosis, mediated at least in part by activation of mitochondrial pathways.


An association of cervical inflammation with high-grade cervical neoplasia in women infected with oncogenic human papillomavirus (HPV).
Previous reports of genital conditions, such as nonspecific genital infection/sore or vaginal discharge associated with cervical cancer (L. A. Brinton et al., J. Natl. Cancer Inst. (Bethesda), 79: 23-30, 1987; C. J. Jones et al., Cancer Res., 50: 3657-3662, 1990), suggest a possible link between either genital tract inflammation or changes in bacteria flora consistent with bacterial vaginosis (BV) and cervical cancer. To test whether changes in vaginal bacterial flora or the degree of cervical inflammation are associated with women having a human papillomavirus (HPV) infection or with women infected with oncogenic HPV having high-grade cervical lesions (high-grade squamous intraepithelial lesions or cancer), we conducted a case-control study of women <50 years old enrolled in the Costa Rican natural history study of HPV and cervical neoplasia. To test whether BV and inflammation were associated with HPV DNA positivity, Analysis 1 was restricted to women with no or mild (low-grade or equivocal) cytological abnormalities, and the degree of inflammation and Nugent score (a measure of BV) were compared between women infected (n = 220) and not infected (n = 130) with HPV. To test whether BV and inflammation were associated with high-grade lesions, Analysis 2 was restricted to women infected with oncogenic HPV, and the degree of inflammation and Nugent score were compared between women with (n = 95) and without (n = 158) high-grade cervical lesions. In Analysis 1, BV and cervical inflammation were not associated with HPV infection. In Analysis 2, BV was not associated with high-grade lesions. However, we found a marginally significant positive trend of increasing cervical inflammation associated with high-grade lesions in oncogenic HPV-infected women, (P(trend) = 0.05). Overt cervicitis was associated with a 1.9-fold increase in risk of high-grade lesions (95% confidence interval, 0.90-4.1). The results of this study suggest that cervical inflammation may be associated with high-grade lesions and may be a cofactor for high-grade cervical lesions in women infected with oncogenic HPV.


Proof of concept non-invasive estimation of peripheral venous oxygen saturation.
Pulse oximeters continuously monitor arterial oxygen saturation. Continuous monitoring of venous oxygen saturation (SvO2) would enable real-time assessment of tissue oxygen extraction (O2E) and perfusion changes leading to improved diagnosis of clinical conditions, such as sepsis. This study presents the proof of concept of a novel pulse oximeter method that utilises the compliance difference between arteries and veins to induce artificial respiration-like modulations to the peripheral vasculature. These modulations make the venous blood pulsatile, which are then detected by a pulse oximeter sensor. The resulting photoplethysmograph (PPG) signals from the pulse oximeter are processed and analysed to develop a calibration model to estimate regional venous oxygen saturation (SpvO2), in parallel to arterial oxygen saturation estimation (SpaO2). A clinical study with healthy adult volunteers (n = 8) was conducted to assess peripheral SvO2 using this pulse oximeter method. A range of physiologically realistic SvO2 values were induced using arm lift and vascular occlusion tests. Gold standard, arterial and venous blood gas measurements were used as reference measurements. Modulation ratios related to arterial and venous systems were determined using a frequency domain analysis of the PPG signals. A strong, linear correlation (r 2 = 0.95) was found between estimated venous modulation ratio (RVen) and measured SvO2, providing a calibration curve relating measured RVen to venous oxygen saturation. There is a significant difference in gradient between the SpvO2 estimation model (SpvO2 = 111 - 40.6*R) and the empirical SpaO2 estimation model (SpaO2 = 110 - 25*R), which yields the expected arterial-venous differences. Median venous and arterial oxygen saturation accuracies of paired measurements between pulse oximeter estimated and gold standard measurements were 0.29 and 0.65%, respectively, showing good accuracy of the pulse oximeter system. The main outcome of this study is the proof of concept validation of a novel pulse oximeter sensor and calibration model to assess peripheral SvO2, and thus O2E, using the method used in this study. Further validation, improvement, and application of this model can aid in clinical diagnosis of microcirculation failures due to alterations in oxygen extraction.


Silica-surface reorganization during organotin grafting evidenced by ¹¹⁹Sn DNP SENS: a tandem reaction of gem-silanols and strained siloxane bridges.
Grafting reactive molecular complexes on dehydroxylated amorphous silica is a strategy to develop "single-site" heterogeneous catalysts. In general, only the reactivity of isolated silanols is invoked for silica dehydroxylated at 700 °C ([SiO(2-700)]), though ca. 10% of the surface silanols are in fact geminal Q2-silanols. Here we report the reaction of allyltributylstannane with [SiO(2-700)] and find that the geminal Q2-silanols react to form products that would formally arise from vicinal Q3-silanols that are not present on [SiO(2-700)], indicating that a surface rearrangement occurs. The reorganization of the silica surface is unique to silica dehydroxylated at 700 °C or above. The findings were identified using Dynamic Nuclear Polarization Surface Enhanced NMR Spectroscopy (DNP SENS) combined with DFT calculations.


Brain abscess developing in a non-operated spontaneous intracerebral haemorrhage: a case report and literature review.
Brain abscesses are a rare but potentially lethal neurological lesions, generally occurring after septic episodes in immunodeficient patients or complicating neurosurgical procedures. Even though they are known complications of surgically treated intracerebral haemorrhages (ICH), the presence of a brain abscess at the site of an untreated ICH is a rare event. Such cases may result from haematogenous spread from distant foci or contiguous sites and are often preceded by episodes of sepsis and local infection. Immunodeficiency, AIDS, age, diabetes mellitus and vitamin-K deficiency are predisposing factors. Abscess formation should be considered in case of clinical deterioration, headache, and any neurological deficit after febrile episodes. Early diagnosis with neuroradiological imaging, infection blood markers and microbiological identification of the causative pathogen is crucial for treatment with surgical drainage or excision and specific antibiotic therapy, which guarantee good outcome and long-term survival. In fact, while prompt diagnosis and treatment guarantee good outcome and long-term survival, morbidity and mortality are very high in case of misdiagnosis. We report a case of a 49-year old man presenting with a brain abscess 13 weeks after a spontaneous ICH, without previous episodes of sepsis and with a suspected septic arthritis 2 weeks after abscess drainage.


Nonparaxial analysis of the far-field radiation patterns of double-heterostructure lasers.
Previous analytical approximations for the far-field radiation patterns of a double-heterostructure laser have been restricted by TE-mode propagation in slab waveguides. A model that is applicable to both TE- and TM-mode propagation either in a slab waveguide or in a multiple-quantum-well structure is developed. Results of computations agree with the measured irradiance profiles of visible diode lasers of wavelengths from 635 to 670 nm. A process for deriving far-field expressions in terms of the diode-laser parameters listed in data books is suggested. Astigmatic aberration present in the wave front is determined, and the final result is expressed in a form convenient for diffraction analysis of truncated focused (or collimated) diode-laser beams.


Experimental infection of human volunteers.
Controlled human infection (CHI) trials, in which healthy volunteers are experimentally infected, can accelerate the development of novel drugs and vaccines for infectious diseases of global importance. The use of CHI models is expanding from around 60 studies in the 1970s to more than 120 publications in this decade, primarily for influenza, rhinovirus, and malaria. CHI trials have provided landmark data for several registered drugs and vaccines, and have generated unprecedented scientific insights. Because of their invasive nature, CHI studies demand critical ethical review according to established frameworks. CHI-associated serious adverse events are rarely reported. Novel CHI models need standardised safety data from comparable CHI models to facilitate evidence-based risk assessments, as well as funds to produce challenge inoculum according to regulatory requirements. Advances such as the principle of controlled colonisation, the expansion of models to endemic areas, and the use of genetically attenuated strains will further broaden the scope of CHI trials.


Genome-wide association identifies OBFC1 as a locus involved in human leukocyte telomere biology.
Telomeres are engaged in a host of cellular functions, and their length is regulated by multiple genes. Telomere shortening, in the course of somatic cell replication, ultimately leads to replicative senescence. In humans, rare mutations in genes that regulate telomere length have been identified in monogenic diseases such as dyskeratosis congenita and idiopathic pulmonary fibrosis, which are associated with shortened leukocyte telomere length (LTL) and increased risk for aplastic anemia. Shortened LTL is observed in a host of aging-related complex genetic diseases and is associated with diminished survival in the elderly. We report results of a genome-wide association study of LTL in a consortium of four observational studies (n = 3,417 participants with LTL and genome-wide genotyping). SNPs in the regions of the oligonucleotide/oligosaccharide-binding folds containing one gene (OBFC1; rs4387287; P = 3.9 x 10(-9)) and chemokine (C-X-C motif) receptor 4 gene (CXCR4; rs4452212; P = 2.9 x 10(-8)) were associated with LTL at a genome-wide significance level (P < 5 x 10(-8)). We attempted replication of the top SNPs at these loci through de novo genotyping of 1,893 additional individuals and in silico lookup in another observational study (n = 2,876), and we confirmed the association findings for OBFC1 but not CXCR4. In addition, we confirmed the telomerase RNA component (TERC) as a gene associated with LTL (P = 1.1 x 10(-5)). The identification of OBFC1 through genome-wide association as a locus for interindividual variation in LTL in the general population advances the understanding of telomere biology in humans and may provide insights into aging-related disorders linked to altered LTL dynamics.


A comprehensive examination of the model underlying acceptance and commitment therapy for chronic pain.
The therapeutic model underlying Acceptance and Commitment Therapy (ACT) is reasonably well-established as it applies to chronic pain. Several studies have examined measures of single ACT processes, or subsets of processes, and have almost uniformly indicated reliable relations with patient functioning. To date, however, no study has performed a comprehensive examination of the entire ACT model, including all of its component processes, as it relates to functioning. The present study performed this examination in 274 individuals with chronic pain presenting for an assessment appointment. Participants completed a battery of self-report questionnaires, assessing multiple aspects of the ACT model, as well as pain intensity, disability, and emotional distress. Initial exploratory factor analyses examined measures of the ACT model and measures of patient functioning separately with each analysis identifying three factors. Next, the fit of a model including ACT processes on the one hand and patient functioning on the other was examined using Structural Equation Modeling. Overall model fit was acceptable and indicated moderate correlations among the ACT processes themselves, as well as significant relations with pain intensity, emotional distress, and disability. These analyses build on the existing literature by providing, to our knowledge, the most comprehensive evaluation of the ACT theoretical model in chronic pain to date.


Thinking inside the bubble: evidence for a new contextual unit in urban mental health.
Previous quantitative research has suggested that there is a link between housing, the urban environment, and mental health. However, methodological and design issues make it difficult to disentangle the relative influence of dwelling specific and wider urban environmental influences on individual mental health. The aim of this study was to explore the link between the dwelling, the immediate urban environment, and mental health to generate a new conceptual framework by which understanding of dwelling and urban environmental influences on mental health can be advanced. Qualitative interviews and focus groups were conducted with 32 inner city residents. Participants, stratified by sex and mental health status, were randomly recruited from a wider quantitative survey. An almost equal number of men and women as well as people with or without mental health problems participated, allowing for comparison of experience. Data were analysed inductively to generate an appropriate theoretical framework regarding dwelling and urban environmental influences on mental health. An inner city neighbourhood of about 6200 people in north west London. Most of that population live in public housing. The principal study finding is that between the dwelling unit and the neighbourhood unit, evidence was found for another meaningful contextual unit of analysis, the "residential bubble" through which effects on mental health can be mediated. The residential bubble describes a limited area of three dimensional space that surrounds a dwelling, encompassing immediate neighbours (above, below, and adjacent) and shared public space bordering the dwelling. Positive events and processes within the bubble had a beneficial influence on mental health whereas negative ones tended to have a damaging influence. These seemed to disproportionately have an impact on people with pre-existing mental health problems. The concept of the "residential bubble" may be a meaningful new contextual unit of analysis in urban mental health. This may have important implications with regards to interventions and measurement development.


Control of photoreceptor autophagy after retinal detachment: the switch from survival to death.
To examine whether calpain inhibition following retinal detachment would prolong autophagy and result in reduced photoreceptor apoptosis. Retinal detachments were created in Brown-Norway rats by subretinal injection of 1% hyaluronic acid and simulated in vitro by Fas-receptor activation of 661W cells, a cone cell line. Protein levels of LC3 and autophagy-related gene 5 (Atg5), both of which are involved in the creation of the autophagosome, were assayed by Western blot. Calpain 1, the protease responsible for Atg5 cleavage and transitioning photoreceptors from autophagy to apoptosis, activity was monitored by α-spectrin cleavage. Various calpain inhibitors were added either to the subretinal space or cell culture media. Apoptosis was assessed in vitro by caspase-8 activity assays and in vivo via TUNEL assays. Cell counts were assessed in vivo at 2 months following detachment. Following retinal detachment or Fas-receptor activation of 661W cells, there was an increase in Atg5 and LC3-II that peaked at 3 days and decreased by 7-days postdetachment. Calpain 1 activity level peaked at 7 days and was associated with decreased autophagy. Calpain inhibition led to increased autophagy, a decrease in caspase-8 activation, reduced TUNEL-positive photoreceptors, and increased photoreceptor cell survival. Our data suggest that calpain activation, which peaks at 7-days postdetachment, is a key step in triggering photoreceptors to shift from cell survival to death. Prolonging autophagy through calpain inhibition leads to significantly reduced photoreceptor apoptosis and increased cell survival.


Self-Calibration Phenomenon for Near-Infrared Clinical Measurements: Theory, Simulation, and Experiments.
An irradiated turbid medium scatters the light in accordance to its optical properties. Near-infrared (NIR) clinical methods, which are based on spectral-dependent absorption, suffer from an inherent error due to spectral-dependent scattering. We present here a unique spatial point, that is, iso-pathlength (IPL) point, on the surface of a tissue at which the intensity of re-emitted light remains constant. This scattering-indifferent point depends solely on the medium geometry. On the basis of this natural phenomenon, we suggest a novel optical method for self-calibrated clinical measurements. We found that the IPL point exists in both cylindrical and semi-infinite tissue geometries (Supporting Information, Video file). Finally, in vivo human finger and mice measurements are used to validate the crossing point between the intensity profiles of two wavelengths. Hence, measurements at the IPL point yield an accurate absorption assessment while eliminating the scattering dependence. This finding can be useful for oxygen saturation determination, NIR spectroscopy, photoplethysmography measurements, and a wide range of optical sensing methods for physiological aims.


Natural selection and age-structured populations.
This paper studies the properties of a new class of demographic parameters for age-structured populations and analyzes the effect of natural selection on these parameters. Two new demographic variables are introduced: the entropy of a population and the reproductive potential. The entropy of a population measures the variability of the contribution of the different age classes to the stationary population. The reproductive potential measures the mean of the contribution of the different age classes to the Malthusian parameter. The Malthusian parameter is precisely the difference between the entropy and the reproductive potential. The effect of these demographic variables on changes in gene frequency is discussed. The concept of entropy of a genotype is introduced and it is shown that in a random mating population in Hardy-Weinberg equilibrium and under slow selection, the rate of change of entropy is equal to the genetic variance in entropy minus the covariance in entropy and reproductive potential. This result is an information theoretic analog of Fisher's fundamental theorem of natural selection.


TIPS stent migration into the heart with 6-year follow-up.
The transjugular intrahepatic portosystemic shunt (TIPS) is widely used for the treatment of portal hypertension in adults, but no studies have defined the best approach to treat portal hypertension in pediatric patients. Pediatric use of TIPS is rare even in large centers of adult practice. The migration of stents has also been reported as a complication in adults. There is no standard way to treat this type of complication, and it is not always clear whether immediate removal or watchful waiting is safer for the patient. We report the case of an 11-year-old patient who underwent urgent TIPS implantation due to variceal bleeding, after unsuccessful sclerotherapy. During the procedure, due to the deep impact of the stent, a second, telescopic, stent was inserted. The portal pressure decreased, no further bleeding occurred, and the patient was listed for transplantation. Three weeks later a routine chest X-ray discovered the migration of the second stent into the right ventricle. No interventional radiological removal or open heart surgery was available for the transplant waiting list patient. The patient underwent uneventful combined liver-kidney transplantation. During the 6-year follow-up period the child had no signs of hemodynamic instability, and his somatic and mental development were appropriate. To our knowledge this case is the first publication on a heart-impacted TIPS stent in a child. The watchful waiting was justified by uneventful combined liver-kidney transplantation and long-term follow-up. This case also underlines the need for best practice guidelines in pediatric portal hypertension.


Immunohistochemistry of the canine vomeronasal organ.
The canine's olfactory acuity is legendary, but neither its main olfactory system nor its vomeronasal system has been described in much detail. We used immunohistochemistry on paraffin-embedded sections of male and female adult dog vomeronasal organ (VNO) to characterize the expression of proteins known to be expressed in the VNO of several other mammals. Basal cell bodies were more apparent in each section than in rodent VNO and expressed immunoreactivity to anticytokeratin and antiepidermal growth factor receptor antibodies. The thin layer of neurone cell bodies in the sensory epithelium and axon fascicles in the lamina propria expressed immunoreactivity to neurone cell adhesion molecule, neurone-specific beta tubulin and protein gene product 9.5. Some neurones expressed growth-associated protein 43 (GAP43): and a number of those also expressed neurone-specific beta tubulin-immunoreactivity. Some axon fascicles were double labelled for those two proteins. The G-protein alpha subunits Gi and Go, involved in the signal transduction pathway, showed immunoreactivity in the sensory cell layer. Our results demonstrate that the canine vomeronasal organ contains a population of cells that expresses several neuronal markers. Furthermore, GAP43 immunoreactivity suggests that the sensory epithelium is neurogenic in adult dogs.


8 weeks of 2S-Hesperidin supplementation improves muscle mass and reduces fat in amateur competitive cyclists: randomized controlled trial.
2S-Hesperidin is the main flavonoid of orange (Citrus sinensis). Previous researches have pointed its effects in muscle development and fat accumulation reduction, although most of these results have not been assessed in humans. The objective of this study is to evaluate the effect of chronic (8-weeks) intake of 2S-hesperidin on amateur cyclists' body composition. A double-blind, parallel and randomized trial, was carried out with 40 amateur cyclists that were divided in two groups, one taking 2S-hesperidin (500 mg d-1, n = 20) and another taking placebo (500 mg d-1 microcellulose, n = 20) for 8 weeks. Dual-energy X-ray absorptiometry (DXA) and anthropometric measurements were used to assess the effect of both treatments on body composition. In addition, the resting metabolic rate was measured. In comparison to placebo, DXA analysis showed a decrease in percentage body fat (%BF) (-10.4%; p = 0.035) and lower limb fat mass (-10.5%; p = 0.029) in favour of 2S-hesperidin. After evaluation of anthropometric data, a decrease in %BF (-3.7%; p = 0.006), total body fat (-3.0%; p = 0.047), ∑ of 8 skinfolds (-6.1%; p = 0.008) was observed in 2S-hesperidin group, but not in placebo. Additionally, there was an increase in muscle mass percentage (1.0%; p = <0.001) and total muscle mass (1.7%; p = 0.011) after ingestion of 2S-hesperidin, with no changes in placebo. Chronic intake of 2S-hesperidin decreased fat mass in amateur cyclists, evaluated through different body composition measurement methodologies (DXA and anthropometry). In addition, 2S-hesperidin supplementation showed a promoting effect on muscle development.


The updating of the representation of visual space in parietal cortex by intended eye movements.
Every eye movement produces a shift in the visual image on the retina. The receptive field, or retinal response area, of an individual visual neuron moves with the eyes so that after an eye movement it covers a new portion of visual space. For some parietal neurons, the location of the receptive field is shown to shift transiently before an eye movement. In addition, nearly all parietal neurons respond when an eye movement brings the site of a previously flashed stimulus into the receptive field. Parietal cortex both anticipates the retinal consequences of eye movements and updates the retinal coordinates of remembered stimuli to generate a continuously accurate representation of visual space.


A Nationally consistent NHDPlus framework for identifying interstate waters: implications for integrated assessments and interjurisdictional TMDLs.
The US Environmental Protection Agency's (EPA's) Total Maximum Daily Loads (TMDL) program promotes nationally consistent approaches for documenting the progress in restoring impaired waters. EPA's TMDL program provides tracking systems comprising both database and geographic information systems (GIS) mapping components. The GIS mapping is implemented using the National Hydrography Dataset (NHD). The EPA and the US Geological Survey have developed an enhanced NHD product (NHDPlus) that is applied in this study to define an interstate waters framework for the conterminous United States. This NHDPlus-based framework provides an efficient watershed-oriented approach for selecting interstate waters. Greater consistency in approaches for interstate waters is essential for providing improved techniques for integrated assessment and management programs. Improved analysis tools for interstate waters are clearly important from a federal perspective. Insights based on tools for federal interstate waters are also of interest for state water quality agencies when they deal with complicated interjurisdictional challenges that can require leveraging support from a wide range of stakeholders. Summaries are provided on the degree of consistency documented for inland waters where states have provided TMDL listing GIS information for shared interstate NHD reaches, and summaries are provided on the patterns for interstate assessments organized according to the ecoregions developed for EPA's Wadeable Streams Assessment. The relevance of this interstate waters framework in leveraging the TMDL program to provide enhanced support for watershed oriented management approaches is also explored.


Al-Anon Intensive Referral (AIR): A qualitative formative evaluation for implementation.
The Al-Anon mutual-help program helps concerned others (COs; e.g., families, friends) of persons with excessive alcohol use. Despite widespread availability of Al-Anon meetings, participation is limited and little is known about how to best facilitate engagement. Al-Anon Intensive Referral (AIR) was developed to facilitate COs' engagement in Al-Anon and is being tested in a randomized controlled trial (RCT). Toward the end of the recruitment for the RCT, a qualitative formative evaluation was conducted to learn about facilitators, barriers, and recommendations for AIR implementation in substance use disorder (SUD) treatment clinics. Thirty-one directors and staff at ten VA and community SUD clinics were interviewed. Semi-structured interviews were based on the Consolidated Framework for Implementation Research and were thematically analyzed to identify facilitators, barriers, and recommendations for AIR implementation. Perceived facilitators of AIR implementation included AIR's face validity, adaptability, and alignment with staff values and skills, requiring only minimal training. Several interviewees in community settings thought AIR would fit with their current practices (e.g., family groups), and some clinics reported having sufficient staff available for delivering AIR. Perceived barriers included limited staff time, and VA clinics having limited resources for providing services to COs. Furthermore, many clients have no COs, or COs who are unwilling or unable to engage with them. Recommendations included fitting AIR within existing workflows and focusing on COs with highest readiness to receive support. Interviewees also thought AIR could be adapted to a website format or smartphone app, which may expand its reach while decreasing staff burden and cost; however, it may not be as effective and appealing to some demographic groups (e.g., older COs). AIR has strong potential for implementation in SUD treatment settings, but clinics vary on implementation capacity. Most clinics could implement AIR partially (e.g., case-by-case basis) while clinics with sufficient capacity (e.g., staff time) could implement it more fully. These findings can also inform implementation of other interventions for concerned others.


Plasma observations near uranus: initial results from voyager 2.
Extensive measurements of low-energy positive ions and electrons in the vicinity of Uranus have revealed a fully developed magnetosphere. The magnetospheric plasma has a warm component with a temperature of 4 to 50 electron volts and a peak density of roughly 2 protons per cubic centimeter, and a hot component, with a temperature of a few kiloelectron volts and a peak density of roughly 0.1 proton per cubic centimeter. The warm component is observed both inside and outside of L = 5, whereas the hot component is excluded from the region inside of that L shell. Possible sources of the plasma in the magnetosphere are the extended hydrogen corona, the solar wind, and the ionosphere. The Uranian moons do not appear to be a significant plasma source. The boundary of the hot plasma component at L = 5 may be associated either with Miranda or with the inner limit of a deeply penetrating, solar wind-driven magnetospheric convection system. The Voyager 2 spacecraft repeatedly encountered the plasma sheet in the magnetotail at locations that are consistent with a geometric model for the plasma sheet similar to that at Earth.


Pediatric blunt abdominal injury: age is irrelevant and delayed operation is not detrimental.
During the past 40 years, management of solid organ injury in pediatric trauma patients has shifted to highly successful nonoperative management. Our purpose was to characterize children requiring operative intervention. We hypothesized that older children would be more likely to require operative intervention. In particular, we wanted to examine potential outcome disparities between children who were operated upon immediately and those in whom attempted nonoperative management failed. Additionally, we asked whether attempted nonoperative management, when failed, put children at higher risk for mortality or morbidities such as increased blood product transfusions or lengths of stays. Retrospective cohorts from seven Level I pediatric trauma centers were identified. Blunt splenic, hepatic, renal, or pancreatic injuries were documented in 2,944 children <1 to 19 years of age from January 1993 to December 2002. Data collected included demographics, hemodynamics, blood transfusions, Glasgow Coma Scale score, Injury Severity Score, hospital length of stay (LOS), intensive care unit (ICU) LOS, and mortality. Analysis involved 140 (4.8%) of 2,944 patients requiring operation. Two cohorts were characterized: (1) immediate operation (IO), defined as laparotomy </=3 hours after arrival (n = 81; 58%) and (2) failed nonoperative management (F-NOM), defined as laparotomy >3 hours after arrival (n = 59; 42%). Comparing the two cohorts, no age differences were found. Compared with F-NOM, IO had significantly worse hemodynamics, Injury Severity Score, and Glasgow Coma Scale score and was associated with liver injuries. Pancreatic injuries were significantly associated with F-NOM. While controlling for injury severity to compare IO versus F-NOM, linear regression revealed equivalent blood transfusions, ICU LOS, hospital LOS, and mortality rates. IO and F-NOM are rare events and independent of age. When operated upon for appropriate physiology, the timing of operation in pediatric solid organ injury is irrelevant and not detrimental with respect to blood transfusion, mortality, ICU and hospital LOS, and resource utilization.


Ecological mechanisms underpinning climate adaptation services.
Ecosystem services are typically valued for their immediate material or cultural benefits to human wellbeing, supported by regulating and supporting services. Under climate change, with more frequent stresses and novel shocks, 'climate adaptation services', are defined as the benefits to people from increased social ability to respond to change, provided by the capability of ecosystems to moderate and adapt to climate change and variability. They broaden the ecosystem services framework to assist decision makers in planning for an uncertain future with new choices and options. We present a generic framework for operationalising the adaptation services concept. Four steps guide the identification of intrinsic ecological mechanisms that facilitate the maintenance and emergence of ecosystem services during periods of change, and so materialise as adaptation services. We applied this framework for four contrasted Australian ecosystems. Comparative analyses enabled by the operational framework suggest that adaptation services that emerge during trajectories of ecological change are supported by common mechanisms: vegetation structural diversity, the role of keystone species or functional groups, response diversity and landscape connectivity, which underpin the persistence of function and the reassembly of ecological communities under severe climate change and variability. Such understanding should guide ecosystem management towards adaptation planning.


Alcoholism research: delivering on the promise.
Prospects for research advances in alcoholism are very promising, because of the explosion in the neurosciences and advances in epidemiology and typology of the disorder. For example, the field is now ready for molecular genetics studies of the early onset form of alcoholism that is transmitted from father to son with high penetrance. Leading neuroscientists are being recruited into alcoholism research. Paradoxically, this time of new hope coincides with challenges to the scientific enterprise, such as the animal rights movement and impatience with the scientific process in the face of the public health emergencies represented by acquired immunodeficiency syndrome (AIDS) and drug abuse. The emergence of genetically based subtypes of alcoholism suggests that at least two discrete illness processes are involved. Mounting evidence from spinal fluid studies has rekindled interest in a key role for serotonin in the early onset form of alcoholism. One hypothesis now being explored is that genetically low brain serotonin function may be part of the predisposition to this form of alcoholism. It is known that acute alcohol intake transiently increases brain serotonin turnover. Thus, drinking might be viewed as an attempt to correct a deficit, only to produce further serotonin depletion as the drug's effect wears off, setting up a vicious cycle of repeated attempts to self-medicate. Impulsive, violent, and suicidal behavior as well as alcohol abuse are associated with the low brain serotonin activity. Persons with these problems suffer from circadian rhythm and glucose metabolism disturbances that may also be mediated by serotonin. New pharmacological probes are now available to tease out the mechanisms of altered serotonin function. The progressively deteriorating course of severe episodic alcoholism in many ways parallels the process of electrically kindled seizures in experimental animals. There is evidence that repeated withdrawal episodes may kindle a worsening course, including phobic disorders, perhaps by triggering a hyper-reactive noradrenalin system.


The effect of cyanobacterial biomass enrichment by centrifugation and GF/C filtration on subsequent microcystin measurement.
Microcystins are cyclic peptides produced by multiple cyanobacterial genera. After accumulation in the liver of animals they inhibit eukaryotic serine/threonine protein phosphatases, causing liver disease or death. Accurate detection/quantification of microcystins is essential to ensure safe water resources and to enable research on this toxin. Previous methodological comparisons have focused on detection and extraction techniques, but have not investigated the commonly used biomass enrichment steps. These enrichment steps could modulate toxin production as recent studies have demonstrated that high cyanobacterial cell densities cause increased microcystin levels. In this study, three microcystin-producing strains were processed using no cell enrichment steps (by direct freezing at three temperatures) and with biomass enrichment (by centrifugation or GF/C filtration). After extraction, microcystins were analyzed using liquid chromatography-tandem mass spectrometry. All processing methods tested, except GF/C filtration, resulted in comparable microcystin quotas for all strains. The low yields observed for the filtration samples were caused by adsorption of arginine-containing microcystins to the GF/C filters. Whilst biomass enrichment did not affect microcystin metabolism over the time-frame of normal sample processing, problems associated with GF/C filtration were identified. The most widely applicable processing method was direct freezing of samples as it could be utilized in both field and laboratory environments.


Universal parameters for carbon nanotube network-based sensors: can nanotube sensors be reproducible?
Carbon nanotube (CNT) network-based sensors have been often considered unsuitable for practical applications due to their unpredictable characteristics. Herein, we report the study of universal parameters which can be used to characterize CNT network-based sensors and make their response predictable. A theoretical model is proposed to explain these parameters, and sensing experiments for mercury (Hg(2+)) and ammonium (NH(4)(+)) ions using CNT network-based sensors were performed to confirm the validity of our model.


Sox2 and Lef-1 interact with Pitx2 to regulate incisor development and stem cell renewal.
Sox2 marks dental epithelial stem cells (DESCs) in both mammals and reptiles, and in this article we demonstrate several Sox2 transcriptional mechanisms that regulate dental stem cell fate and incisor growth. Conditional Sox2 deletion in the oral and dental epithelium results in severe craniofacial defects, including impaired dental stem cell proliferation, arrested incisor development and abnormal molar development. The murine incisor develops initially but is absorbed independently of apoptosis owing to a lack of progenitor cell proliferation and differentiation. Tamoxifen-induced inactivation of Sox2 demonstrates the requirement of Sox2 for maintenance of the DESCs in adult mice. Conditional overexpression of Lef-1 in mice increases DESC proliferation and creates a new labial cervical loop stem cell compartment, which produces rapidly growing long tusk-like incisors, and Lef-1 epithelial overexpression partially rescues the tooth arrest in Sox2 conditional knockout mice. Mechanistically, Pitx2 and Sox2 interact physically and regulate Lef-1, Pitx2 and Sox2 expression during development. Thus, we have uncovered a Pitx2-Sox2-Lef-1 transcriptional mechanism that regulates DESC homeostasis and dental development.