An image understanding system using attributed symbolic representation and inexact graph-matching.
This paper presents a powerful image understanding system that utilizes a semantic-syntactic (or attributed-synibolic) representation scheme in the form of attributed relational graphs (ARG's) for comprehending the global information contents of images. Nodes in the ARG represent the global image features, while the relations between those features are represented by attributed branches between their corresponding nodes. The extraction of ARG representation from images is achieved by a multilayer graph transducer scheme. This scheme is basically a rule-based system that uses a combination of model-driven and data-driven concepts in performing a hierarchical symbolic mapping of the image information content from the spatial-domain representation into a global representation. Further analysis and inter-pretation of the imagery data is performed on the extracted ARG representation. A distance measure between images is defined in terms of the distance between their respective ARG representations. The distance between two ARG's and the inexact matching of their respective components are calculated by an efficient dynamic programming technique. The system handles noise, distortion, and ambiguity in real-world images by two means, namely, through modeling and embedding them into the transducer's mapping rules, as well as through the appropriate cost of error-transformation for the inexact matching of the ARG image representation. Two illustrative experiments are presented to demonstrate some capabilities of the proposed system. Experiment I deals with locating objects in multiobject scenes, while Experiment II is concerned with target detection in SAR images.


Hyperbaric oxygen in the treatment of acute retinal artery occlusion.
Acute retinal artery occlusion (ARAO) is a major cause of sudden, painless visual loss, often leaving no useful vision in the affected eye. Its incidence is cited at 0.85 per 100,000 persons per year but may be higher because of under-reporting. The natural history is difficult to study, but a spontaneous resolution rate of < 1-8% for acute, non-arteritic ARAO has been cited. Occurrence in an only eye is devastating for the patient. There is currently no consensus regarding management of ARAO and little evidence to support any treatment modality. Despite only limited case series, hyperbaric oxygen treatment (HBOT) is recommended for ARAO by the Undersea and Hyperbaric Medical Society (UHMS) and by the European Committee for Hyperbaric Medicine. Between early 2003 and December 2012, all ARAO patients presenting to Christchurch Hospital were referred for consideration of HBOT. These 31 consecutive patients' medical records were reviewed retrospectively. The time delay from onset of visual loss to commencing HBOT; the presenting visual acuity; various demographic data; the HBOT administered and the outcome visual acuity were documented. All 31 patients underwent at least one HBOT (median 4, range 1-7) at a pressure of 203-284 kPa for 1.5 to 2.0 h. One patient's treatment was terminated after 60 min at their request; another declined further HBOT and one suffered middle ear barotrauma. Thirteen patients also received anticoagulants at the discretion of the referring ophthalmologist. Twenty three patients had temporarily improved vision with the first HBOT. Seven patients had permanent, good visual recovery (6/18 or better; Snellen chart); and two only modest improvement (6/60). All nine patients who improved permanently were treated within 10 hours of symptom onset. Where available, HBOT is indicated for ARAO. Our protocol may not have been aggressive enough and the UHMS protocol is recommended. A multi-centre, randomised controlled trial is feasible, but would be logistically difficult and expensive and may be ethically unsupportable given the lack of alternative, effective treatments.


Colorectal cancer screening among African American church members: a qualitative and quantitative study of patient-provider communication.
A healthcare provider's recommendation to undergo screening has been shown to be one of the strongest predictors of completing a colorectal cancer (CRC) screening test. We sought to determine the relationship between the general quality of self-rated patient-provider communication and the completion of CRC screening. A formative study using qualitative data from focus groups and quantitative data from a cross-sectional survey of church members about the quality of their communication with their healthcare provider, their CRC risk knowledge, and whether they had completed CRC screening tests. Focus group participants were a convenience sample of African American church members. Participants for the survey were recruited by telephone from membership lists of 12 African American churches located in rural counties of North Carolina to participate in the WATCH (Wellness for African Americans Through Churches) Project. Focus Groups. Six focus groups (n = 45) were conducted prior to the baseline survey. Discussions focused on CRC knowledge, and perceived barriers/motivators to CRC screening. A theme that emerged during each groups' discussion about CRC screening was the quality of the participants' communication with their health care provider. Survey. Among the 397 participants over age 50, 31% reported CRC screening within the recommended guidelines. Participants who self-rated their communication as good were more likely to have been screened (36%) within the recommended guidelines than were participants with poor communication (17%) (OR = 2.8, 95% CI 1.2, 6.4; p = 0.013). Participants who had adequate CRC knowledge completed CRC screening at a higher rate than those with inadequate knowledge (p = 0.011). The percentage of participants with CRC screening in the recommended guidelines, stratified by communication and knowledge group were: 42% for good communication/adequate knowledge; 27% for good communication/inadequate knowledge; 29% for poor communication/adequate knowledge; and 5% for poor communication/inadequate knowledge. Participants who rated their patient-provider communication as good were more likely to have completed CRC screening tests than those reporting poor communication. Among participants reporting good communication, knowledge about colorectal cancer was also associated with test completion. Interventions to improve patient-provider communication may be important to increase low rates of CRC screening test completion among African Americans.


The rise of 3-d single-ion magnets in molecular magnetism: towards materials from molecules?
Single-molecule magnets (SMMs) that contain one spin centre (so-called single-ion magnets) theoretically represent the smallest possible unit for spin-based electronic devices. The realisation of this and related technologies, depends on first being able to design systems with sufficiently large energy barriers to magnetisation reversal, Ueff, and secondly, on being able to organise these molecules into addressable arrays. In recent years, significant progress has been made towards the former goal - principally as a result of efforts which have been directed towards studying complexes based on highly anisotropic lanthanide ions, such as Tb(iii) and Dy(iii). Since 2013 however, and the remarkable report by Long and co-workers of a linear Fe(i) system exhibiting Ueff = 325 K, single-ion systems of transition metals have undergone something of a renaissance in the literature. Not only do they have important lessons to teach us about anisotropy and relaxation dynamics in the quest to enhance Ueff, the ability to create strongly coupled spin systems potentially offers access to a whole of host of 1, 2 and 3-dimensional materials with interesting structural and physical properties. This perspective summarises recent progress in this rapidly expanding sub-genre of molecular magnetism from the viewpoint of the synthetic chemist, with a particular focus on the lessons that have so far been learned from single-ion magnets of the d-block, and, the future research directions which we feel are likely to emerge in the coming years.


A Case of Descemet's Membrane Endothelial Keratoplasty in a failed Penetrating Keratoplasty.
Descemet's membrane endothelial keratoplasty (DMEK) is an evolving option for failed penetrating keratoplasty (PK) as it has shown promising results compared to repeat PK. A 22 years young gentleman presented with gradual diminution of vision in the left eye (LE) for six months following therapeutic PK for a perforated fungal corneal ulcer. His best spectacle corrected visual acuity (BSCVA) was 6/60 in LE. Slit-lamp examination of LE revealed circumcorneal congestion with full thickness corneal graft with micro and macrobullae, diffuse stromal edema, DM folds and seven well-buried sutures. Intraocular pressure was 20 mmHg in the right eye whereas it could not be assessed in the left eye. The diagnosis of LE failed PK was made and the patient underwent DMEK. At four months postoperative follow up visit, BSCVA improved to 6/9. This is the first reported case of DMEK after a failed PK in a young patient from Nepal which was successfully treated without any complications. DMEK has a higher potential for better visual rehabilitation compared to DSEK and PK. It has also proved its efficacy in the treatment of endothelial graft failure following PK as it is a less invasive procedure.


Apparent versus true gene expression changes of three hypoxia-related genes in autopsy derived tissue and the importance of normalisation.
The aim of our work was to show how a chosen normal-isation strategy can affect the outcome of quantitative gene expression studies. As an example, we analysed the expression of three genes known to be upregulated under hypoxic conditions: HIF1A, VEGF and SLC2A1 (GLUT1). Raw RT-qPCR data were normalised using two different strategies: a straightforward normalisation against a single reference gene, GAPDH, using the 2(-ΔΔCt) algorithm and a more complex normalisation against a normalisation factor calculated from the quantitative raw data from four previously validated reference genes. We found that the two different normalisation strategies revealed contradicting results: normalising against a validated set of reference genes revealed an upregulation of the three genes of interest in three post-mortem tissue samples (cardiac muscle, skeletal muscle and brain) under hypoxic conditions. Interestingly, we found a statistically significant difference in the relative transcript abundance of VEGF in cardiac muscle between donors who died of asphyxia versus donors who died from cardiac death. Normalisation against GAPDH alone revealed no upregulation but, in some instances, a downregulation of the genes of interest. To further analyse this discrepancy, the stability of all reference genes used were reassessed and the very low expression stability of GAPDH was found to originate from the co-regulation of this gene under hypoxic conditions. We concluded that GAPDH is not a suitable reference gene for the quantitative analysis of gene expression in hypoxia and that validation of reference genes is a crucial step for generating biologically meaningful data.


Response to atrial natriuretic peptide in dogs with acute selective intrahepatic hypertension.
Acute selective intrahepatic hypertension (IHH) is associated with the renal tubular retention of sodium in volume expanded dogs. To determine if acute selective IHH per se, without volume loading or ascites sequestration, would blunt the natriuretic response to i.v. infusions of atrial natriuretic peptide (ANP), 175 ng/kg/min of the 1-28 rat peptide was infused into 6 hydropaenic dogs where intrahepatic hydrostatic pressures were normal, and again, following the portal infusion of histamine, a maneuver known to selectively increase postsinusoidal resistance, within the hepatic microcirculation and so raise intrahepatic sinusoidal pressure. In 6 healthy dogs, while histamine 4.0 micrograms/min free base on average was being infused into a femoral vein, the infusion of ANP increased sodium excretion by 168 microEq/min, compared to 160 microEq/min when the same dose of histamine was being infused into the portal vein (portal pressure increased by 46% or 6 cm H2O). These changes in sodium excretion were not significantly different. Urine flow rate increased by 1.5 ml/min in the control phase and by 1.4 ml/min during intrahepatic hypertension (NS). Although the ANP infusion did not alter GFR or CPAH during the control phase, during IHH, ANP caused GFR to rise significantly by 20%, while there was no change to CPAH. Despite the increment in GFR, the natriuretic response during IHH was not different from that observed during the control phase of the study. We conclude that acute IHH per se does not blunt the renal tubular natriuretic response to ANP.


Mode coupling theory for nonequilibrium glassy dynamics of thermal self-propelled particles.
We present a mode coupling theory study for the relaxation and glassy dynamics of a system of strongly interacting self-propelled particles, wherein the self-propulsion force is described by Ornstein-Uhlenbeck colored noise and thermal noises are included. Our starting point is an effective Smoluchowski equation governing the distribution function of particle positions, from which we derive a memory function equation for the time dependence of density fluctuations in nonequilibrium steady states. With the basic assumption of the absence of macroscopic currents and standard mode coupling approximation, we can obtain expressions for the irreducible memory function and other relevant dynamic terms, wherein the nonequilibrium character of the active system is manifested through an averaged diffusion coefficient D[combining macron] and a nontrivial structural function S2(q) with q being the magnitude of wave vector q. D[combining macron] and S2(q) enter the frequency term and the vertex term for the memory function, and thus influence both the short time and the long time dynamics of the system. With these equations obtained, we study the glassy dynamics of this thermal self-propelled particle system by investigating the Debye-Waller factor fq and relaxation time τα as functions of the persistence time τp of self-propulsion, the single particle effective temperature Teff as well as the number density ρ. Consequently, we find the critical density ρc for given τp shifts to larger values with increasing magnitude of propulsion force or effective temperature, in good accordance with previously reported simulation work. In addition, the theory facilitates us to study the critical effective temperature T for fixed ρ as well as its dependence on τp. We find that T increases with τp and in the limit τp → 0, it approaches the value for a simple passive Brownian system as expected. Our theory also well recovers the results for passive systems and can be easily extended to more complex systems such as active-passive mixtures.


Single-port-access surgery with a novel magnet camera system.
In this paper, we designed, built, and tested a novel single-port access laparoscopic surgery (SPA) specific camera system. This device (magnet camera) integrates a light source and video camera into a small, inexpensive, portable package that does not compete for space with the surgical tools during SPA. The device is inserted through a 26-mm incision in the umbilicus, followed by the SPA port, which is used to maintain an insufflation seal and support the insertion of additional tools. The camera, now in vivo, remains separate from the SPA port, thereby removing the need for a dedicated laparoscope, and, thus, allowing for an overall reduction in SPA port size or the use of a third tool through the insertion port regularly reserved for the traditional laparoscope. The SPA camera is mounted to the abdominal ceiling using one of the two methods: fixation to the SPA port through the use of a rigid ring and cantilever bar, or by an external magnetic handle. The purpose of the magnet camera system is to improve SPA by: 1) eliminating the laparoscope SPA channel; 2) increasing the field of view through enhanced camera system mobility; and 3) reducing interference between the camera system and the surgical tools at the port, both in vivo and ex vivo.


Structure of N-formimino-L-glutamate iminohydrolase from Pseudomonas aeruginosa.
N-Formimino-l-glutamate iminohydrolase (HutF), from Pseudomonas aeruginosa with a locus tag of Pa5106 ( gi|15600299 ), is a member of the amidohydrolase superfamily. This enzyme catalyzes the deamination of N-formimino-l-glutamate to N-formyl-l-glutamate and ammonia in the histidine degradation pathway. The crystal structure of Pa5106 was determined in the presence of the inhibitors N-formimino-l-aspartate and N-guanidino-l-glutaric acid at resolutions of 1.9 and 1.4 Å, respectively. The structure of an individual subunit is composed of two domains with the larger domain folding as a distorted (β/α)8-barrel. The (β/α)8-barrel domain is composed of eight β-strands flanked by 11 α-helices, whereas the smaller domain is made up of eight β-strands. The active site of Pa5106 contains a single zinc atom that is coordinated by His-56, His-58, His-232, and Asp-320. The nucleophilic solvent water molecule coordinates with the zinc atom at a distance of 2.0 Å and is hydrogen bonded to Asp-320 and His-269. The α-carboxylate groups of both inhibitors are hydrogen bonded to the imidazole moiety of His-206, the hydroxyl group of Tyr-121, and the side chain amide group of Gln-61. The side chain carboxylate groups of the two inhibitors are ion-paired with the guanidino groups of Arg-209 and Arg-82. Computational docking of high-energy tetrahedral intermediate forms of the substrate, N-formimino-l-glutamate, to the three-dimensional structure of Pa5106 suggests that this compound likely undergoes a re-faced nucleophilic attack at the formimino group by the metal-bound hydroxide. A catalytic mechanism of the reaction catalyzed by Pa5106 is proposed.


Surface plasmon-enhanced lasing in dye-doped cholesteric liquid crystals.
This study shows the results of a photonic band-edge laser using dye-doped cholesteric liquid crystals (CLCs) combined with silver (Ag) nanoparticles. When the Ag nanoparticle surface plasmon resonance wavelength matched the excitation source wavelength, the large optical fields provided by surface plasmons increased the fluorescence of dye molecules by enhancing the molecular excitation rate, achieving a low lasing threshold and high pumping efficiency.


Associated and disassociated patterns in hormones, song, behavior and brain receptor expression between life-cycle stages in male black redstarts, Phoenicurus ochruros.
Testosterone has been suggested to be involved in the regulation of male territorial behavior. For example, seasonal peaks in testosterone typically coincide with periods of intense competition between males for territories and mating partners. However, some species also express territorial behavior outside a breeding context when testosterone levels are low and, thus, the degree to which testosterone facilitates territorial behavior in these species is not well understood. We studied territorial behavior and its neuroendocrine correlates in male black redstarts. Black redstarts defend territories in spring during the breeding period, but also in the fall outside a reproductive context when testosterone levels are low. In the present study we assessed if song output and structure remain stable across life-cycle stages. Furthermore, we assessed if brain anatomy may give insight into the role of testosterone in the regulation of territorial behavior in black redstarts. We found that males sang spontaneously at a high rate during the nonbreeding period when testosterone levels were low; however the trill-like components of spontaneously produced song contained less repetitive elements during nonbreeding than during breeding. This higher number of repetitive elements in trills did not, however, correlate with a larger song control nucleus HVC during breeding. However, males expressed more aromatase mRNA in the preoptic area - a brain nucleus important for sexual and aggressive behavior - during breeding than during nonbreeding. In combination with our previous studies on black redstarts our results suggest that territorial behavior in this species only partly depends on sex steroids: spontaneous song output, seasonal variation in trills and non-vocal territorial behavior in response to a simulated territorial intruder seem to be independent of sex steroids. However, context-dependent song during breeding may be facilitated by testosterone - potentially by conversion of testosterone to estradiol in the preoptic area.


Chronic acetaminophen exposure in pediatric acute liver failure.
Acetaminophen (N-acetyl-p-aminophenol [APAP]) is a widely used medication that can cause hepatotoxicity. We examined characteristics and outcomes of children with chronic exposure (CE) to APAP in the multinational Pediatric Acute Liver Failure (PALF) Study. A total of 895 children enrolled from 2002 to 2009 were grouped by APAP exposure history as: CE (received multiple doses \x{2265}2 days; n = 83), single dose exposure (SE; n = 85), and no exposure (NE; n = 498). CE was the reference group for pairwise comparisons. Median values are shown. Patients with CE compared with those with SE were younger (3.5 vs 15.2 years, P < .0001), less likely to be female (46% vs 82%, P < .0001), and more likely to be Hispanic (25% vs 7%, P = .001), but they did not differ significantly from the NE group. At enrollment, total bilirubin was lower with CE than with NE (3.2 vs 13.1 mg/dL, P < .001). Alanine aminotransferase levels were higher with CE than with NE (2384 vs 855 IU/L, P < .0001), but lower than with SE (5140 IU/L, P < .0001). Survival without liver transplantation at 21 days was worse for CE than for SE (68% vs 92%, P = .0004) but better than for NE (49%, P = .008). Children in the PALF study with CE had lower bilirubin and higher alanine aminotransferase than those with NE. Outcomes with CE were worse than with SE but better than with NE. Potential reasons for this outcomes advantage over non-APAP-exposed subjects should be explored.


Predicting runoff-induced pesticide input in agricultural sub-catchment surface waters: linking catchment variables and contamination.
An urgent need exists for applicable methods to predict areas at risk of pesticide contamination within agricultural catchments. As such, an attempt was made to predict and validate contamination in nine separate sub-catchments of the Lourens River, South Africa, through use of a geographic information system (GIS)-based runoff model, which incorporates geographical catchment variables and physicochemical characteristics of applied pesticides. We compared the results of the prediction with measured contamination in water and suspended sediment samples collected during runoff conditions in tributaries discharging these sub-catchments. The most common insecticides applied and detected in the catchment over a 3-year sampling period were azinphos-methyl (AZP), chlorpyrifos (CPF) and endosulfan (END). AZP was predominantly found in water samples, while CPF and END were detected at higher levels in the suspended particle samples. We found positive (p < 0.002) correlations between the predicted average loss and the concentrations of the three insecticides both in water and suspended sediments (r between 0.87 and 0.94). Two sites in the sub-catchment were identified as posing the greatest risk to the Lourens River mainstream. It is assumed that lack of buffer strips, presence of erosion rills and high slopes are the main variables responsible for the high contamination at these sites. We conclude that this approach to predict runoff-related surface water contamination may serve as a powerful tool for risk assessment and management in South African orchard areas.


Fluorescent excimers and exciplexes of the purine base derivative 8-phenylethynyl-guanine in DNA hairpins.
The ground- and excited-state electronic interactions between the nucleobase analog 8-(4'-phenylethynyl)deoxyguanosine, EG, with natural nucleobases and 7-deazaguanine, as well as between adjacent EG base analogs, have been characterized using a combination of steady-state spectroscopy and time-resolved fluorescence, absorption, and stimulated Raman spectroscopies. The properties of the nucleoside EG-H2 are only weakly perturbed upon incorporation into synthetic DNA hairpins in which thymine, cytosine or adenine are the bases flanking EG. Incorporation of the nucleoside to be adjacent to guanine or deazaguanine results in the formation of short-lived (40-80 ps) exciplexes, the charge transfer character of which increases as the oxidation potential of the donor decreases. Hairpins possessing two or three adjacent EG base analogs display exciton-coupled circular dichroism in the ground state and form long-lived fluorescent excited states upon electronic excitation. Incorporation of EG into the helical scaffold of the DNA hairpins places it adjacent to its neighboring nucleobases or a second EG, thus providing the close proximity required for the formation of exciplex or excimer intermediates upon geometric relaxation of the short-lived EG excited state. The three time-resolved spectroscopic methods employed permit both the characterization of the several intermediates and the kinetics of their formation and decay.


Changes in white blood cells and platelets in children with falciparum malaria: relationship to disease outcome.
Little is known about the changes in white blood cells and platelets in children with falciparum malaria in endemic areas. We measured the white cell count (WCC) and platelets of 230 healthy children from the community, 1369 children admitted to hospital with symptomatic malaria, and 1461 children with other medical conditions. Children with malaria had a higher WCC compared with community controls, and leucocytosis was strongly associated with younger age, deep breathing, severe anaemia, thrombocytopenia and death. The WCC was not associated with a positive blood culture. In children with malaria, high lymphocyte and low monocyte counts were independently associated with mortality. A platelet count of less than 150 x 109/l was found in 56.7% of children with malaria, and was associated with age, prostration and parasite density, but not with bleeding problems or mortality. The mean platelet volume was also higher in children with malaria compared with other medical conditions. This may reflect early release from the bone marrow in response to peripheral platelet destruction. Thus, leucocytosis was associated with both severity and mortality in children with falciparum malaria, irrespective of bacteraemia, whereas thrombocytopenia, although very common, was not associated with adverse outcome.


Performance comparison of optical interference cancellation system architectures.
The performance of three optics-based interference cancellation systems are compared and contrasted with each other, and with traditional electronic techniques for interference cancellation. The comparison is based on a set of common performance metrics that we have developed for this purpose. It is shown that thorough evaluation of our optical approaches takes into account the traditional notions of depth of cancellation and dynamic range, along with notions of link loss and uniformity of cancellation. Our evaluation shows that our use of optical components affords performance that surpasses traditional electronic approaches, and that the optimal choice for an optical interference canceller requires taking into account the performance metrics discussed in this paper.


Support for hospital-based HIV testing and counseling: a national survey of hospital marketing executives.
Today, hospitals are involved extensively in social marketing and promotional activities. Recently, investigators from the Centers for Disease Control and Prevention (CDC) estimated that routine testing of hospital patients for human immunodeficiency virus (HIV) could identify more than 100,000 patients with previously unrecognized HIV infections. Several issues are assessed in this paper. These include hospital support for voluntary HIV testing and AIDS education and the impact that treating AIDS patients has on the hospital's image. Also tested is the hypothesis that certain hospitals, such as for-profit institutions and those outside the AIDS epicenters, would be less supportive of hospital-based AIDS intervention strategies. To assess these issues, a national random sample of 193 executives in charge of hospital marketing and public relations were surveyed between December 1992 and January 1993. The survey was part of an ongoing annual survey of hospitals and included questions about AIDS, health education, marketing, patient satisfaction, and hospital planning. Altogether, 12.4 percent of executives indicated their hospital had a reputation for treating AIDS patients. Among hospitals without an AIDS reputation, 34.1 percent believed developing one would be harmful to the hospital's image, in contrast to none in hospitals that had such a reputation (chi 2 = 11.676, df = 1, P = .0006). Although 16.6 percent did not know if large-scale HIV testing should be implemented, a near majority (47.7 percent) expressed some support. In addition, 15 percent reported that HIV-positive physicians on the hospital's medical staff should not be allowed to practice medicine, but 32.1 percent indicated that they should. Also, 33.1 percent thought the hospital should be more involved in AIDS education. Finally, certain hospital characteristics,such as location and for-profit status, were not associated with support for hospital-based AIDS interventions. Contrary to what was hypothesized,however, hospitals in AIDS epicenters were less supportive of the CDC recommendations for some reason (X2 = 7.735, df = 1, P = .005).Support for AIDS education and voluntary testing is significant among hospital marketing and public relations executives. Over the past decade, community marketing and public relations have become an integral part of the hospital's business activities.However, financial pressures now are forcing hospitals to restrict these efforts. Findings reported in this paper suggest that future health care reform may assist public health aims by redirecting these endeavors towards the fight against AIDS and other preventable diseases, not eliminating them. Additional research is needed to determine why executives in AIDS epicenters are less supportive of large-scale hospital HIV testing and counseling in comparison with those outside these areas.


High-Throughput Screening of Patient-Derived Cultures Reveals Potential for Precision Medicine in Glioblastoma.
Identifying drugs for the treatment of glioblastoma (GBM), a rapidly fatal disease, has been challenging. Most screening efforts have been conducted with immortalized cell lines grown with fetal bovine serum, which have little relevance to the genomic features found in GBM patients. Patient-derived neurosphere cultures, while being more physiologically relevant, are difficult to screen and therefore are only used to test a few drug candidates after initial screening efforts. Laminin has been used to generate two-dimensional cell lines from patient tumors, preserving the genomic signature and alleviating some screening hurdles. We present here the first side-by-side comparison of inhibitor sensitivity of laminin and neurosphere-grown patient-derived GBM cell lines and show that both of these culture methods result in the same pattern of inhibitor sensitivity. We used these screening methods to evaluate the dependencies of seven patient-derived cell models: three grown on laminin and four grown as neurospheres, against 56 agents in 17-point dose-response curves in 384-well format in triplicate. This allowed us to establish differential sensitivity of chemotherapeutic agents across the seven patient-derived models. We found that MEK inhibition caused patient-sample-specific growth inhibition and that bortezomib, an FDA-approved proteasome inhibitor, was potently lethal in all patient-derived models. Furthermore, the screening results led us to test the combination of the Bcl-2 inhibitor ABT-263, and the mTOR inhibitor AZD-8055, which we found to be synergistic in a subset of patient-derived GBM models. Thus, we have identified new candidate therapeutics and developed a high-throughput screening system using patient-derived GBM samples.


A qualitative review of existing national and international occupational safety and health policies relating to occupational sedentary behaviour.
Prolonged sedentary time is now recognised as an emergent ergonomics issue. We aimed to review current occupational safety and health policies relevant to occupational sedentary behaviour. An electronic search for documents was conducted on websites of ergonomics and occupational safety and health organisations from 10 countries and six international/pan-European agencies. Additionally, 43 informants (nine countries) were contacted and an international conference workshop held. 119 documents (e.g. legislation, guidelines, codes of practice) were identified. Using a qualitative synthesis, it was observed that many jurisdictions had legal frameworks establishing a duty of care for employers, designers/manufacturers/suppliers and employees. While no occupational authority policies focusing specifically on sedentary behaviour were found, relevant aspects of existing policies were identified. We highlight implications for ergonomics research and practice and recommend the development of policy to specifically address occupational sedentary behaviour and support workplace initiatives to assess and control the risks of this emergent hazard.


Compendium of measures to prevent disease associated with animals in public settings, 2009: National Association of State Public Health Veterinarians, Inc. (NASPHV).
Certain venues encourage or permit the public to be in contact with animals, resulting in millions of human-animal interactions each year. These settings include county or state fairs, petting zoos, animal swap meets, pet stores, zoologic institutions, circuses, carnivals, educational farms, livestock-birthing exhibits, educational exhibits at schools and child-care facilities, and wildlife photo opportunities. Although human-animal contact has many benefits, many human health problems are associated with these settings, including infectious diseases, exposure to rabies, and injuries. Infectious disease outbreaks reported during the previous decade have been caused by Escherichia coli O157:H7, Salmonella species, Cryptosporidium species, Coxiella burnetii, Mycobacterium tuberculosis, ringworm, and other pathogens. Such infections have substantial medical, public health, legal, and economic effects. This report provides recommendations for public health officials, veterinarians, animal venue staff members, animal exhibitors, visitors to animal venues, physicians, and others concerned with minimizing risks associated with animals in public settings. The recommendation to wash hands is the most important prevention step for reducing the risk for disease transmission associated with animals in public settings. Other critical recommendations are that venues prohibit food in animal areas, venues include transition areas between animal areas and nonanimal areas, visitors receive information about disease risk and prevention procedures, and animals be properly cared for and managed. These updated 2009 guidelines also emphasize risks associated with baby poultry, reptiles, and rodents in public settings, and information about aquatic animal zoonoses has been incorporated.


p52 expression enhances lung cancer progression.
While many studies have demonstrated that canonical NF-κB signaling is a central pathway in lung tumorigenesis, the role of non-canonical NF-κB signaling in lung cancer remains undefined. We observed frequent nuclear accumulation of the non-canonical NF-κB component p100/p52 in human lung adenocarcinoma. To investigate the impact of non-canonical NF-κB signaling on lung carcinogenesis, we employed transgenic mice with doxycycline-inducible expression of p52 in airway epithelial cells. p52 over-expression led to increased tumor number and progression after injection of the carcinogen urethane. Gene expression analysis of lungs from transgenic mice combined with in vitro studies suggested that p52 promotes proliferation of lung epithelial cells through regulation of cell cycle-associated genes. Using gene expression and patient information from The Cancer Genome Atlas (TCGA) database, we found that expression of p52-associated genes was increased in lung adenocarcinomas and correlated with reduced survival, even in early stage disease. Analysis of p52-associated gene expression in additional human lung adenocarcinoma datasets corroborated these findings. Together, these studies implicate the non-canonical NF-κB component p52 in lung carcinogenesis and suggest modulation of p52 activity and/or downstream mediators as new therapeutic targets.


Treatment efficacy of sacral nerve stimulation in slow transit constipation: a two-phase, double-blind randomized controlled crossover study.
Sacral nerve stimulation (SNS) is a potential treatment for constipation refractory to standard therapies. However, there have been no randomized controlled studies examining its efficacy. In patients with slow transit constipation, we evaluated the efficacy of suprasensory and subsensory SNS compared with sham, in a prospective, 18-week randomized, double-blind, placebo-controlled, two-phase crossover study. The primary outcome measure was the proportion of patients who, on more than 2 days/week for at least 2 of 3 weeks, reported a bowel movement associated with a feeling of complete evacuation. After 3 weeks of temporary peripheral nerve evaluation (PNE), all patients had permanent implantation and were randomized to subsensory/sham (3 weeks each) and then re-randomized to suprasensory/sham (3 weeks each) with a 2-week washout period between each arm. Daily stool dairies were kept, and quality of life (QoL; SF36) was measured at the end of each arm. Between November 2006 and March 2012, 234 constipated patients were assessed, of whom 59 were willing and deemed eligible to participate (4 male; median age 42 years). Of the 59 patients, 16 (28%) responded to PNE. Fifty-five patients went on to permanent SNS implantation. The proportion of patients satisfying the primary outcome measure did not differ between suprasensory (30%) and sham (21%) stimulations, nor between subsensory (25%) and sham (25%) stimulations. There were no significant changes in QoL scores. In patients with refractory slow transit constipation, SNS did not improve the frequency of complete bowel movements over the 3-week active period.


Antigenic variation in clones of Trypanosoma brucei grown in immune-deficient mice.
We have produced monoclonal antibodies against six variant surface glycoproteins from early variant antigen types (VATs) of the IsTaR 1 serodeme of Trypanosoma brucei brucei. We have used these in fixed cell immunofluorescence assays to follow the VAT composition of populations of each early VAT when passaged through irradiated mice. The IsTat 1.A and 1.7a populations were stable for more than 30 days (approximately 150 generations), but 1.1a, 1.3a, 1.5a, and 1.11a all changed to 1.A within this time. The time and rate of this antigenic switch were characteristic for each VAT. Growth rates of the VATs were determined when they were both grown separately and grown with 1.A. It appeared that the order of growth rates was 1.7a greater than 1.A = 1.1a greater than 1.11a greater than 1.5a greater than 1.3a. We have generated theoretical curves for the replacement of one VAT by another based on differences in their growth rates and the rate at which one VAT switches to another (switch frequency). These curves closely match those derived experimentally. We postulate that the differences in growth rates between VATs and the different switch frequencies for VATs may be sufficient to generate the loosely defined sequence of VATs seen in chronic infections.


Phosphoregulation promotes release of kinetochores from dynamic microtubules via multiple mechanisms.
During mitosis, multiprotein complexes called kinetochores orchestrate chromosome segregation by forming load-bearing attachments to dynamic microtubule tips, and by participating in phosphoregulatory error correction. The conserved kinase Aurora B phosphorylates the major microtubule-binding kinetochore subcomplexes, Ndc80 and (in yeast) Dam1, to promote release of erroneous attachments, giving another chance for proper attachments to form. It is unknown whether Aurora B phosphorylation promotes release directly, by increasing the rate of kinetochore detachment, or indirectly, by destabilizing the microtubule tip. Moreover, the relative importance of phosphorylation of Ndc80 vs. Dam1 in the context of whole kinetochores is unclear. To address these uncertainties, we isolated native yeast kinetochore particles carrying phosphomimetic mutations on Ndc80 and Dam1, and applied advanced laser-trapping techniques to measure the strength and stability of their attachments to individual dynamic microtubule tips. Rupture forces were reduced by phosphomimetic mutations on both subcomplexes, in an additive manner, indicating that both subcomplexes make independent contributions to attachment strength. Phosphomimetics on either subcomplex reduced attachment lifetimes under constant force, primarily by accelerating detachment during microtubule growth. Phosphomimetics on Dam1 also increased the likelihood of switches from microtubule growth into shortening, further promoting release in an indirect manner. Taken together, our results suggest that, in vivo, Aurora B releases kinetochores via at least two mechanisms: by weakening the kinetochore-microtubule interface and also by destabilizing the kinetochore-attached microtubule tip.


Water pulses and biogeochemical cycles in arid and semiarid ecosystems.
The episodic nature of water availability in arid and semiarid ecosystems has significant consequences on belowground carbon and nutrient cycling. Pulsed water events directly control belowground processes through soil wet-dry cycles. Rapid soil microbial response to incident moisture availability often results in almost instantaneous C and N mineralization, followed by shifts in C/N of microbially available substrate, and an offset in the balance between nutrient immobilization and mineralization. Nitrogen inputs from biological soil crusts are also highly sensitive to pulsed rain events, and nitrogen losses, particularly gaseous losses due to denitrification and nitrate leaching, are tightly linked to pulses of water availability. The magnitude of the effect of water pulses on carbon and nutrient pools, however, depends on the distribution of resource availability and soil organisms, both of which are strongly affected by the spatial and temporal heterogeneity of vegetation cover, topographic position and soil texture. The 'inverse texture hypothesis' for net primary production in water-limited ecosystems suggests that coarse-textured soils have higher NPP than fine-textured soils in very arid zones due to reduced evaporative losses, while NPP is greater in fine-textured soils in higher rainfall ecosystems due to increased water-holding capacity. With respect to belowground processes, fine-textured soils tend to have higher water-holding capacity and labile C and N pools than coarse-textured soils, and often show a much greater flush of N mineralization. The result of the interaction of texture and pulsed rainfall events suggests a corollary hypothesis for nutrient turnover in arid and semiarid ecosystems with a linear increase of N mineralization in coarse-textured soils, but a saturating response for fine-textured soils due to the importance of soil C and N pools. Seasonal distribution of water pulses can lead to the accumulation of mineral N in the dry season, decoupling resource supply and microbial and plant demand, and resulting in increased losses via other pathways and reduction in overall soil nutrient pools. The asynchrony of resource availability, particularly nitrogen versus water due to pulsed water events, may be central to understanding the consequences for ecosystem nutrient retention and long-term effects on carbon and nutrient pools. Finally, global change effects due to changes in the nature and size of pulsed water events and increased asynchrony of water availability and growing season will likely have impacts on biogeochemical cycling in water-limited ecosystems.


SLC22A3 polymorphisms do not modify pancreatic cancer risk, but may influence overall patient survival.
Expression of the solute carrier (SLC) transporter SLC22A3 gene is associated with overall survival of pancreatic cancer patients. This study tested whether genetic variability in SLC22A3 associates with pancreatic cancer risk and prognosis. Twenty four single nucleotide polymorphisms (SNPs) tagging the SLC22A3 gene sequence and regulatory elements were selected for analysis. Of these, 22 were successfully evaluated in the discovery phase while six significant or suggestive variants entered the validation phase, comprising a total study number of 1,518 cases and 3,908 controls. In the discovery phase, rs2504938, rs9364554, and rs2457571 SNPs were significantly associated with pancreatic cancer risk. Moreover, rs7758229 associated with the presence of distant metastases, while rs512077 and rs2504956 correlated with overall survival of patients. Although replicated, the association for rs9364554 did not pass multiple testing corrections in the validation phase. Contrary to the discovery stage, rs2504938 associated with survival in the validation cohort, which was more pronounced in stage IV patients. In conclusion, common variation in the SLC22A3 gene is unlikely to significantly contribute to pancreatic cancer risk. The rs2504938 SNP in SLC22A3 significantly associates with an unfavorable prognosis of pancreatic cancer patients. Further investigation of this SNP effect on the molecular and clinical phenotype is warranted.


The variation in the eating quality of beef from different sexes and breed classes cannot be completely explained by carcass measurements.
Delivering beef of consistent quality to the consumer is vital for consumer satisfaction and will help to ensure demand and therefore profitability within the beef industry. In Australia, this is being tackled with Meat Standards Australia (MSA), which uses carcass traits and processing factors to deliver an individual eating quality guarantee to the consumer for 135 different 'cut by cooking methods' from each carcass. The carcass traits used in the MSA model, such as ossification score, carcass weight and marbling explain the majority of the differences between breeds and sexes. Therefore, it was expected that the model would predict with eating quality of bulls and dairy breeds with good accuracy. In total, 8128 muscle samples from 482 carcasses from France, Poland, Ireland and Northern Ireland were MSA graded at slaughter then evaluated for tenderness, juiciness, flavour liking and overall liking by untrained consumers, according to MSA protocols. The scores were weighted (0.3, 0.1, 0.3, 0.3) and combined to form a global eating quality (meat quality (MQ4)) score. The carcasses were grouped into one of the three breed categories: beef breeds, dairy breeds and crosses. The difference between the actual and the MSA-predicted MQ4 scores were analysed using a linear mixed effects model including fixed effects for carcass hang method, cook type, muscle type, sex, country, breed category and postmortem ageing period, and random terms for animal identification, consumer country and kill group. Bulls had lower MQ4 scores than steers and females and were predicted less accurately by the MSA model. Beef breeds had lower eating quality scores than dairy breeds and crosses for five out of the 16 muscles tested. Beef breeds were also over predicted in comparison with the cross and dairy breeds for six out of the 16 muscles tested. Therefore, even after accounting for differences in carcass traits, bulls still differ in eating quality when compared with females and steers. Breed also influenced eating quality beyond differences in carcass traits. However, in this case, it was only for certain muscles. This should be taken into account when estimating the eating quality of meat. In addition, the coefficients used by the Australian MSA model for some muscles, marbling score and ultimate pH do not exactly reflect the influence of these factors on eating quality in this data set, and if this system was to be applied to Europe then the coefficients for these muscles and covariates would need further investigation.


OST4 is a subunit of the mammalian oligosaccharyltransferase required for efficient N-glycosylation.
The eukaryotic oligosaccharyltransferase (OST) is a membrane-embedded protein complex that catalyses the N-glycosylation of nascent polypeptides in the lumen of the endoplasmic reticulum (ER), a highly conserved biosynthetic process that enriches protein structure and function. All OSTs contain a homologue of the catalytic STT3 subunit, although in many cases this is assembled with several additional components that influence function. In S. cerevisiae, one such component is Ost4p, an extremely small membrane protein that appears to stabilise interactions between subunits of assembled OST complexes. OST4 has been identified as a putative human homologue, but to date neither its relationship to the OST complex, nor its role in protein N-glycosylation, have been directly addressed. Here, we establish that OST4 is assembled into native OST complexes containing either the catalytic STT3A or STT3B isoforms. Co-immunoprecipitation studies suggest that OST4 associates with both STT3 isoforms and with ribophorin I, an accessory subunit of mammalian OSTs. These presumptive interactions are perturbed by a single amino acid change in the transmembrane region of OST4. Using siRNA knockdowns and native gel analysis, we show that OST4 plays an important role in maintaining the stability of native OST complexes. Hence, upon OST4 depletion well-defined OST complexes are partially destabilised and a novel ribophorin I-containing subcomplex can be detected. Strikingly, cells depleted of either OST4 or STT3A show a remarkably similar defect in the N-glycosylation of endogenous prosaposin. We conclude that OST4 most likely promotes co-translational N-glycosylation by stabilising STT3A-containing OST isoforms.


Views of health professionals on climate change and health: a multinational survey study.
Climate change arguably represents one of the greatest global health threats of our time. Health professionals can advocate for global efforts to reduce emissions and protect people from climate change; however, evidence of their willingness to do so remains scarce. In this Viewpoint, we report findings from a large, multinational survey of health professionals (n=4654) that examined their views of climate change as a human health issue. Consistent with previous research, participants in this survey largely understood that climate change is happening and is caused by humans, viewed climate change as an important and growing cause of health harm in their country, and felt a responsibility to educate the public and policymakers about the problem. Despite their high levels of commitment to engaging in education and advocacy on the issue, many survey participants indicated that a range of personal, professional, and societal barriers impede them from doing so, with time constraints being the most widely reported barrier. However, participants say various resources-continuing professional education, communication training, patient education materials, policy statements, action alerts, and guidance on how to make health-care workplaces sustainable-can help to address those barriers. We offer recommendations on how to strengthen and support health professional education and advocacy activities to address the human health challenges of climate change.


Fibrinogen as a therapeutic target for bleeding: a review of critical levels and replacement therapy.
Fibrinogen plays a critical role in achieving and maintaining hemostasis and is fundamental to effective clot formation. There is increasing awareness of the important role of fibrinogen as a key target for the treatment and prevention of acquired bleeding. Fibrinogen is the first coagulation factor to fall to critically low levels (<1.0 g/L) during major hemorrhage (normal plasma fibrinogen levels range from 2.0 to 4.5 g/L), and current guidelines recommend maintaining the plasma fibrinogen level above 1.5 g/L. Fibrinogen supplementation can be achieved using plasma or cryoprecipitate; however, there are a number of safety concerns associated with these allogeneic blood products and there is a lack of high-quality evidence to support their use. Additionally, there is sometimes a long delay associated with the preparation of frozen products for infusion. Fibrinogen concentrate provides a promising alternative to allogeneic blood products and has a number of advantages: it allows a standardized dose of fibrinogen to be rapidly administered in a small volume, has a very good safety profile, and is virally inactivated as standard. Administration of fibrinogen concentrate, often guided by point-of-care viscoelastic testing to allow individualized dosing, has been successfully used as hemostatic therapy in a range of clinical settings, including cardiovascular surgery, postpartum hemorrhage, and trauma. Results show that fibrinogen concentrate is associated with a reduction or even total avoidance of allogeneic blood product transfusion. Fibrinogen concentrate represents an important option for the treatment of coagulopathic bleeding; further studies are needed to determine precise dosing strategies and thresholds for fibrinogen supplementation.


Human hopping on very soft elastic surfaces: implications for muscle pre-stretch and elastic energy storage in locomotion.
During hopping in place and running, humans maintain similar center of mass dynamics by precisely adjusting leg mechanics to compensate for moderate changes in surface stiffness. We investigated the limits of this precise control by asking humans to hop in place on extremely soft elastic surfaces. We found that hoppers drastically altered leg mechanics and maintained similar center of mass dynamics despite a sevenfold change in surface stiffness (11-81 kN m(-1)). On the stiffest surfaces, the legs compressed in early stance and then extended in late stance in the pattern that is typical for normal bouncing gaits. On the softest surfaces, however, subjects reversed this pattern so that the legs extended up to 8 cm in early stance and then compressed by a similar distance in late stance. Consequently, the center of mass moved downward during stance by 5-7 cm less than the surface compressed and by a similar distance as on the stiffest surfaces. This unique leg action probably reduced extensor muscle pre-stretch because the joints first extended and then flexed during stance. This interpretation is supported by the observation that hoppers increased muscle activation by 50% on the softest surface despite similar joint moments and mechanical leg work as on the stiffest surface. Thus, the extreme adjustment to leg mechanics for very soft surfaces helps maintain normal center of mass dynamics but requires high muscle activation levels due to the loss of the normal extensor muscle stretch-shorten cycle.


Dynamics of the reaction of O(3P) atoms with alkylthiol self-assembled monolayers.
We have studied the dynamics of the reactions of O((3)P) atoms with alkylthiol self-assembled monolayers (SAMs). Superthermal O((3)P) atoms, with a fairly broad distribution of laboratory-frame kinetic energies (mean = 16 kJ mol(-1), fwhm = 26 kJ mol(-1)), were generated by 355 nm photolysis of NO(2) introduced at a low pressure above the SAM surface. Nascent OH v' = 0 products were detected by laser-induced fluorescence. SAMs of two different alkyl chain lengths, C(6) and C(18), were studied. The existence of SAM layers, and their robustness under our experimental conditions during the relevant measurement period, were confirmed by scanning-tunneling microscopy (STM). Reaction at the SAM surface was verified as the authentic source of the hydroxyl radicals using a perdeuterated C(6)D(13)-SAM sample. The OH appearance profiles as a function of photolysis-probe delay, and the rotational-state distributions at their peaks, were compared with those of liquid squalane (C(30)H(62), 2,6,10,15,19,23-hexamethyltetracosane). The reactivity of the SAMs and of squalane was found to be comparable. We conclude that the O((3)P) atoms must be able to access the more reactive secondary hydrogen atoms along the alkyl chains of the SAMs. We find no perceptible differences in reactivity or product energy disposal between the two SAM chain lengths. Both produce a substantial fraction of the OH with relatively high velocities, which must result from direct, impulsive reaction. There is also a slower component, with velocities consistent with a thermal, trapping-desorption mechanism. The proportion of this component appears to be lower for SAMs than for squalane. This would be compatible with the expected greater smoothness of the SAM surface at the molecular scale. We find little evidence for significant rotational excitation of the OH products, although the details of any correlation between translational and rotational energy release require further investigation. We compare our results with the limited available prior theoretical modeling of O((3)P) + SAM systems.


Female and male transgender quality of life: socioeconomic and medical differences.
Studies show a positive impact of gender reassignment treatment on the quality of life (QOL) of transgender persons, but little is known about the influence of their socioeconomic status. First, to assess health-related QOL of transgender men and women and compare it with a general population sample, second, to investigate the differences between transgender men and transgender women, and third, to analyze how their levels of QOL differ according to socioeconomic and transition data. One hundred forty-eight current and former transgender patients of a gender identity clinic participated in a large QOL study. Health-related QOL was measured using the Short Form 36-Item Questionnaire. The QOL of transgender women did not differ significantly from the general Dutch female population, although transgender men showed reduced mental health-related QOL compared with the general Dutch male sample. Transgender women had a lower QOL than transgender men for the subscales physical functioning and general health, but better QOL for bodily pain. Time since start of hormone use was positively associated for transgender women with subscales bodily pain and general health, and negatively associated for transgender men with the subscale role limitations due to physical health problems. There was no significant difference in QOL between the group who had undergone genital surgery or surgical breast augmentation and the group who did not have these surgeries. Transgender men with an erection prosthesis scored significantly better on the subscales vitality and (at trend level) on role limitations due to emotional problems. A series of univariate analyses revealed significantly lower QOL scores for transgender persons that were older, low educated, unemployed, had a low household income, and were single. Specific social indicators are important in relation to health-related QOL of transgenders in a context of qualitative and adequate medical care.


Literature syntheses for the Council on Chiropractic Guidelines and Practice Parameters: methodology.
The purpose of this project was to initiate an iterative process for systematic review of the literature involving a broad spectrum of individuals with experience across multiple domains (clinicians, educators, clinical scientists, and politically active) within the chiropractic profession. The Scientific Commission of the Council on Chiropractic Guidelines and Practice Parameters (CCGPP) was charged with developing literature syntheses, organized by anatomical region, to evaluate and report on the evidence-based values for chiropractic care. Content and process-experienced team leaders were selected to manage 8 domains based on regional disorders: low back and related lower extremity conditions; neck pain, headache, and related upper extremity conditions; costovertebral and thoracic conditions; upper extremity disorders; lower extremity disorders; nonmusculoskeletal disorders; and subluxation. Team efforts in review, rating, and reporting of literature synthesis were guided, as best possible, by the widely accepted Appraisal of Guidelines for Research and Evaluation process. The main features included (1) review by a panel of experts; (2) detailed topic selection based on literature of most common conditions and procedures; (3) structured instruments for rating the quality of and results from the literature; (4) formal consensus process to adjudicate differences in professional opinion; and (5) wide stakeholder review by patients, professionals, policymakers, and third-party payers. As part of the CCGPP process, preliminary drafts of these articles were posted on the CCGPP Web site www.ccgpp.org (2006-2008) to allow for an open process and the broadest possible mechanism for stakeholder input. Reports on findings from this process are being published. The reports from each domain summarize methodological challenges and their unique content. Although all literature in health care is challenged by complex methodological issues that limit how the information may be generalized, the preponderance of evidence in any of the domains can be informative to the clinician as well as give guidance to new scientific efforts to improve the quality of care.


Skill mastery inhibits adoption of observed alternative solutions among chimpanzees (Pan troglodytes).
Geographic variation in socially transmitted skills and signals, similar to human culture, has been well documented for great apes. The rules governing the adoption of novel behaviours, however, are still largely unknown. We conducted an innovation-and-transmission experiment with two groups of chimpanzees living at hopE Primate Sanctuary Gänserndorf, Austria, presenting a board on which food had to be manoeuvred around obstacles to be acquired. Most chimpanzees used sticks to acquire the food, but five adults independently invented a novel technique, rattling, which was subsequently tested by almost all group members. However, individuals who had become proficient with sticks were reluctant to switch to rattling, despite it being more efficient. Similarly, after rattling was prevented, rattle specialists kept trying to rattle and made no attempt to use the stick technique, despite their knowledge about its existence. We conclude that innovators stimulate others to experiment with the solutions they display, but that chimpanzees are nevertheless conservative; mastery of a skill inhibits further exploration, and hence adoption of alternative techniques even if these are more efficient. Consequently, conformity among group members should not be expected in great apes when individuals develop proficiency at different techniques. Conservatism thus joins conformity as a mechanism to bring about cultural uniformity and stability.


The effect of Gu-Sui-Bu (Drynaria fortunei J. Sm) on bone cell activities.
In the traditional Chinese medicine, Gu-Sui-Bu [Drynaria fortunei (kunze) J. Sm] has been reported as a good enhancer for bone healing. In this experiment, we investigate the biochemical effects of this traditional Chinese medicine on the bone cells culture. Different concentrations of crude extract of Gu-Sui-Bu were added to rat bone cells culture. The mitochondria activity of the bone cells after exposure was determined by colorimetric assay. Biochemical markers such as alkaline phosphatase (ALP), acid phosphatase (ACP) titer, prostaglandin E2 (PGE2) titer and the expression of both osteopontin and osteonectin mRNA were evaluated. The effect on the osteoclasts differentiation was evaluated by tartrate-resistant acid phosphatase (TRAP) stain. The most effective concentration of Gu-Sui-Bu on bone cells was 1 mg/ml. The addition of 1 mg/ml Gu-Sui-Bu to bone cells culture for 7 days can statistically increase the intracellular ALP amount; while the ACP and PGE2 amount in culture medium were significantly increased. In Northern blot analysis, the expression of both osteopontin and osteonectin mRNA were down-regulated after adding Gu-Sui-Bu into bone cells culture. The formation of multi-nucleated osteoclasts was more active than that of the control group, but no giant osteoclasts formation was observed. In this study, we demonstrated that Gu-Sui-Bu has potential effects on the bone cells culture. One of the major effects of Gu-Sui-Bu on the bone cells is probably mediated by its effect on the osteclasts activities. Continued and advanced study on the alterations in gene expression of bone cells by Chinese medicines will provide a basis for understanding the observed bone cell responses to various pharmacological interventions.


Long noncoding RNA Hotair facilitates retinal endothelial cell dysfunction in diabetic retinopathy.
Retinal endothelial cell (REC) dysfunction induced by diabetes mellitus (DM) is an important pathological step of diabetic retinopathy (DR). Long noncoding RNAs (lncRNAs) have emerged as novel modulators in DR. The present study aimed to investigate the role and mechanism of lncRNA Hotair in regulating DM-induced REC dysfunction. The retinal vascular preparations and immunohistochemical staining assays were conducted to assess the role of Hotair in retinal vessel impairment in vivo. The EdU, transwell, cell permeability, CHIP, luciferase activity, RIP, RNA pull-down, and Co-IP assays were employed to investigate the underlying mechanism of Hotair-mediated REC dysfunction in vitro. Hotair expression was significantly increased in diabetic retinas and high glucose (HG)-stimulated REC. Hotair knockdown inhibited the proliferation, invasion, migration, and permeability of HG-stimulated REC in vitro and reduced the retinal acellular capillaries and vascular leakage in vivo. Mechanistically, Hotair bound to LSD1 to inhibit VE-cadherin transcription by reducing the H3K4me3 level on its promoter and to facilitate transcription factor HIF1α-mediated transcriptional activation of VEGFA. Furthermore, LSD1 mediated the effects of Hotair on REC function under HG condition. The Hotair exerts its role in DR by binding to LSD1, decreasing VE-cadherin transcription, and increasing VEGFA transcription, leading to REC dysfunction. These findings revealed that Hotair is a potential therapeutic target of DR.


Molecular assessment of drug-phospholipid interactions consequent to cancer treatment: a study of anthracycline-induced cardiotoxicity.
Cardiotoxicity limits the use of anthracyclines as potent chemotherapeutics. We employ classical molecular dynamics to explore anthracycline interactions with a realistic myocardial membrane and compare to an ideal membrane widely used in literature. The interaction of these two membranes with four anthracyclines; doxorubicin, epirubicin, daunorubicin, and idarubicin are studied. Careful analysis was conducted on three forms of each drug; pristine, primary metabolite, and cationic salt. By examining the molecular residence time near the membrane's surface, the average number of molecule/membrane hydrogen bonds, the immobilization of the molecules near the membrane, and the location of those molecules relative to the mid-plane of the membrane we found out that salt forms exhibit the highest cardiotoxic probability, followed by the metabolites and pristine forms. Additionally, all forms have more affinity to the upper layer of the realistic myocardial membrane. Meanwhile, an ideal membrane consisting of a single type of phospholipids is not capable of capturing the specific interactions of each drug form. These findings confirm that cardiotoxic mechanisms are membrane-layer and drug-form dependent.


Biogenic stabilization of intertidal sediments: the importance of extracellular polymeric substances produced by benthic diatoms.
The sediment-stabilizing effect of benthic diatoms was investigated in a laboratory setting. Axenic cultures of the benthic diatoms Nitzschia cf. brevissima and Cylindrotheca closterium were inoculated in Petri dishes containing sand and incubated under axenic conditions. By ensuring aseptic routines throughout the experiments, interference from other organisms occurring with diatoms in natural photothrophic biofilms was avoided. This allowed the examination of the role of benthic diatoms in sediment stabilization. Increases in the critical erosion shear stress of the sediment were observed in the presence of both diatom taxa relative to sterile sediment. However, N. cf. brevissima was more effective than C. closterium. Values of critical shear stress in the experimental system were in the same range as those observed in natural biofilms, which indicates that diatoms are important agents for biogenic stabilization. Extracellular carbohydrate contents in the microcosms were similar for both diatom species. However, in the presence of N cf. brevissima, extracellular carbohydrate correlated significantly to critical shear stress, explaining up to 80% of the variation, whereas this was not the case for C. closterium. Therefore, it was concluded that the quantity of extracellular polymeric substances (EPS) alone did not explain the biogenic stabilization. Observed adsorption of EPS to sediment particles depended on the relative amount of uronic acids in the exopolymers. Using fluorescently labeled lectins, confocal laser scanning microscopy showed that EPS secretion by N. cf. brevissima resulted in ordered three-dimensional matrix structures. It is suggested that the structuring of EPS plays an prominent role in the process of biostabilization, and that diatoms such as N. cf. brevissima are actively involved in producing the structure of EPS, whereas others such as C. closterium do not do so to the same extent.


Potential pathological and functional links between long noncoding RNAs and hematopoiesis.
Differential abundance and activity of long noncoding RNAs (lncRNAs) are recognized as the hallmark features in various diseases. We highlight the lncRNAs that play a functional role in the development of blood cells. Many lncRNAs and the protein complexes within which they interact have been implicated in various types of cancers. Multiple lncRNAs participate in normal and may be implicated in malignant hematopoiesis associated with blood cell cancers, such as leukemia, by regulating gene expression through such mechanisms as redirecting chromatin remodeling complexes and activating epigenetic silencing, either of which can inactivate tumor suppressor genes or activate oncogenes. Because of their potential importance in cancers of the blood, lncRNAs may be useful as diagnostic and prognostic markers, and it may be possible to develop lncRNA-mediated therapy.


General involvement of hypoxia-inducible factor 1 in transcriptional response to hypoxia.
Transcription of the human erythropoietin (EPO) gene is activated in Hep3B cells exposed to hypoxia. Hypoxia-inducible factor 1 (HIF-1) is a nuclear factor whose DNA binding activity is induced by hypoxia in Hep3B cells, and HIF-1 binds at a site in the EPO gene enhancer that is required for hypoxic activation of transcription. In this paper, we demonstrate that HIF-1 DNA binding activity is also induced by hypoxia in a variety of mammalian cell lines in which the EPO gene is not transcribed. The composition of the HIF-1 DNA binding complex and its isolated DNA binding subunit and the mechanism of HIF-1 activation appear to be similar or identical in EPO-producing and non-EPO-producing cells. Transcription of reporter genes containing the EPO gene enhancer is induced by hypoxia in non-EPO-producing cells and mutations that eliminate HIF-1 binding eliminate inducibility. These results provide evidence that HIF-1 and its recognition sequence are common components of a general mammalian cellular response to hypoxia.


Non-Coding RNAs as Potential Neuroprotectants against Ischemic Brain Injury.
Over the past decade, scientific discoveries have highlighted new roles for a unique class of non-coding RNAs. Transcribed from the genome, these non-coding RNAs have been implicated in determining the biological complexity seen in mammals by acting as transcriptional and translational regulators. Non-coding RNAs, which can be sub-classified into long non-coding RNAs, microRNAs, PIWI-interacting RNAs and several others, are widely expressed in the nervous system with roles in neurogenesis, development and maintenance of the neuronal phenotype. Perturbations of these non-coding transcripts have been observed in ischemic preconditioning as well as ischemic brain injury with characterization of the mechanisms by which they confer toxicity. Their dysregulation may also confer pathogenic conditions in neurovascular diseases. A better understanding of their expression patterns and functions has uncovered the potential use of these riboregulators as neuroprotectants to antagonize the detrimental molecular events taking place upon ischemic-reperfusion injury. In this review, we discuss the various roles of non-coding RNAs in brain development and their mechanisms of gene regulation in relation to ischemic brain injury. We will also address the future directions and open questions for identifying promising non-coding RNAs that could eventually serve as potential neuroprotectants against ischemic brain injury.


A low-cost, sustainable, second generation system for surveillance of people living with HIV in Spain: 10-year trends in behavioural and clinical indicators, 2002 to 2011.
A second-generation surveillance system of people infected with human immunodeficiency virus (HIV) has been implemented in Spain. Behavioural and clinical data were collected between 2002 and 2011 through an annual one-day, cross-sectional survey in public hospitals, including all in- and outpatients receiving HIVrelated care on the survey day. Mean age increased over time (from 38.7 years in 2002 to 43.8 years in 2011) and 68.4% of the 7,205 subjects were male. The proportion of migrants increased from 6.1% to 15.9%, while people who inject or used to inject drugs (PWID and Ex-PWID) decreased and men who have sex with men (MSM) and heterosexuals increased. Unprotected intercourse at last sex increased among MSM and PWID/Ex-PWID. Patients receiving antiretroviral treatment increased significantly from 76.0% to 88.2% as did those with CD4 T-cell counts ≥350 (from 48.2% to 66.9%) and viral copies <200 (from 47.0% to 85.2%). HIV-infected people with hepatitis C virus RNA decreased from 36.0% in 2004 to 29.9% in 2011, while those with HBsAg remained stable at around 4.4%. Implementation of a low-cost, sustainable system for second-generation surveillance in people living with HIV is feasible. In Spain, the information obtained has helped to define and refine public health policy and document treatment effectiveness.


High efficiency transformation of cultured tobacco cells.
Tobacco calli were transformed at levels up to 50% by cocultivation of tobacco cultured cells with Agrobacterium tumefaciens harboring the binary transfer-DNA vector, pGA472, containing a kanamycin resistance marker. Transformation frequency was dependent on the physiological state of the tobacco cells, the nature of Agrobacterium strain and, less so, on the expression of the vir genes of the tumor-inducing plasmid. Maximum transformation frequency was obtained with exponentially growing plant cells, suggesting that rapid growth of plant cells is an essental factor for efficient transformation of higher plants.


Observation of dynamical crater-shaped charge distribution in the space-time imaging of monolayer graphene.
A better understanding of charge carrier dynamics in graphene is key to improvement of its electronic performance. Here, we present direct space-time visualization of carrier relaxation and diffusion in monolayer graphene using time-resolved scanning electron microscopy techniques. We observed striking fluence-dependent dynamic images, changing from a Gaussian shape to a novel crater-shaped pattern with increasing laser fluence. Such direct observation of dynamic changes in spatial charge distribution is not readily available from the conventional spectroscopic approaches, which reflect essentially overall effective carrier temperature and density. According to our analysis, for this crater-shaped carrier density to occur in aggregated electron-hole pairs in the high fluence regime there exists an unconventional Auger-assisted carrier recombination process to provide effective relaxation channels, most likely involving emission of optical phonons and plasmons, which is dynamically accessible due to a strong temporal overlap among them. The presented model allows us to successfully account for these unexpected phenomena and to quantitatively analyze the observed spatiotemporal behavior.


All in the Family: How the APPs Regulate Neurogenesis.
Recent intriguing evidence suggests that metabolites of amyloid precursor protein (APP), mutated in familial forms of Alzheimer's disease (AD), play critical roles in developmental and postnatal neurogenesis. Of note is soluble APPα (sAPPα) that regulates neural progenitor cell proliferation. The APP family encompasses a group of ubiquitously expressed and evolutionarily conserved, type I transmembrane glycoproteins, whose functions have yet to be fully elucidated. APP can undergo proteolytic cleavage by mutually exclusive pathways. The subtle structural differences between metabolites generated in the different pathways, as well as their equilibrium, may be crucial for neuronal function. The implications of this new body of evidence are significant. Miscleavage of APP would readily impact developmental and postnatal neurogenesis, which might contribute to cognitive deficits characterizing Alzheimer's disease. This review will discuss the implications of the role of the APP family in neurogenesis for neuronal development, cognitive function, and brain disorders that compromise learning and memory, such as AD.


FTIR imaging of the molecular burden around Aβ deposits in an early-stage 3-Tg-APP-PSP1-TAU mouse model of Alzheimer's disease.
Alzheimer's disease is one of the major causes of dementia in the elderly. The disease is caused by the misfolding of water soluble alpha-helical proteins, which leads to the accumulation of β-sheets in the form of amyloid plaques, which can subsequently affect surrounding tissue areas by oxidative stress neurotoxicity. The aim of the present study was to design a novel methodology to analyze the extent to the neuronal burden around protein-rich Aβ plaques suspected to affect molecular components by oxidative stress induced by inflammatory states. To do so, sagittal brain tissue sections from triple transgenic APPxPSP1xTAU mice were used to carry high magnification FTIR-FPA bench-top chemical imaging. The study used the combination of chemometric procedures involving spectral curve fitting and image processing to study the molecular changes occurring around the plaques. The study shows the performance of the approach by demonstrating its usefulness to co-localize molecular changes to different areas around the plaques. The results, although very preliminary, point to the strong interplay between the distance from the plaque and co-accumulation of molecular components indicative of inflammatory states.


The concentration of hydrogen peroxide in exhaled air depends on expiratory flow rate.
Hydrogen peroxide (H2O2) is known to be detectable in exhaled air. The present study aimed to determine whether the concentration of exhaled H2O2 depends on expiratory flow rate in order to make inferences on the site of its production within the lung. Breath condensate was collected in cooled Teflon tubes, at three different expiratorv flow rates, in 15 healthy or mild asthmatic subjects. Tests were repeated 2-5 times to assess reproducibility. Mean+/-SEM concentrations of H2O2 at flow rates of 140, 69 and 48 mL.s(-1) were 0.12+/-0.02, 0.19+/-0.02 and 0.32+/-0.03 microM, respectively. These values differed significantly from each other (p<0.001). For comparison, average coefficients of variability within repeated measurements at each of the three flow rates were 68, 62 and 82%, respectively. These data demonstrate that the concentration of exhaled hydrogen peroxide depends on expiratory flow rate. Since flow dependence is an indicator of production within the airways, this result suggests that, to a large extent, the exhaled hydrogen peroxide originates within the airways. However, even under strictly controlled conditions, a high degree of variability persists, which may limit the usefulness of exhaled hydrogen peroxide as a marker of airway inflammation.


Plasmin is the major protease responsible for processing PDGF-C in the vitreous of patients with proliferative vitreoretinopathy.
Proliferative vitreoretinopathy (PVR) is the primary cause of failure of retinal reattachment surgery. Growth factors such as platelet-derived growth factor (PDGF) are strongly associated with PVR. Of the five PDGF family members, PDGF-C predominates in the vitreous of experimental and clinical PVR. PDGF-C is secreted as a latent protein that requires proteolytic processing for activation. Although tissue plasminogen activator (tPA) is primarily responsible for processing PDGF-C in cultured cells, it constitutes a minority of the processing activity in the vitreous of experimental animals and in patients with PVR. Identifying the major PDGF-C processing protease was the purpose of this study. The presence of serum proteins in the vitreous was detected by Coomassie blue staining and Western blotting. PDGF-C processing activity was detected in an in vitro processing assay using either native or recombinant PDGF-C as the substrate. Plasmin activity was blocked using alpha(2)-plasmin inhibitor. Phosphorylation of the PDGF receptor (PDGFR) was monitored by antiphosphotyrosine Western blotting. Vitreous specimens were collected from experimental rabbits or from patients undergoing vitrectomy to repair retinal detachment or for other reasons. A number of prominent serum proteins (albumin and IgG) were detected in the vitreous of all patients undergoing retinal surgery. The level of these proteins markedly increased in the vitreous of rabbits as they developed PVR. These observations suggested that serum-borne proteases are also likely to be present in the vitreous. Indeed, plasmin (a protease capable of processing PDGF-C) was present in the vitreous from PVR rabbits and retinal surgery patients. Plasmin was dramatically more effective than tPA in processing PDGF-C in an in vitro assay. Blocking plasmin activity eliminated most of the processing activity in the vitreous of patients and rabbits with PVR. Plasmin was the major PDGF-C processing protease in the vitreous of PVR rabbits and patients undergoing retinal surgery. Blocking plasmin prevented the generation of active PDGF-C, which is the major PDGF isoform relevant for PVR. These observations are the first report of an in vivo protease responsible for processing PDGF-C. In addition, plasmin was identified as a novel therapeutic target for patients with PVR.


How telomeres solve the end-protection problem.
The ends of eukaryotic chromosomes have the potential to be mistaken for damaged or broken DNA and must therefore be protected from cellular DNA damage response pathways. Otherwise, cells might permanently arrest in the cell cycle, and attempts to "repair" the chromosome ends would have devastating consequences for genome integrity. This end-protection problem is solved by protein-DNA complexes called telomeres. Studies of mammalian cells have recently uncovered the mechanism by which telomeres disguise the chromosome ends. Comparison to unicellular eukaryotes reveals key differences in the DNA damage response systems that inadvertently threaten chromosome ends. Telomeres appear to be tailored to these variations, explaining their variable structure and composition.


Development of OASYS-2: a system for the analysis of serial measurement of peak expiratory flow in workers with suspected occupational asthma.
Serial peak expiratory flow (PEF) measurement is usually the most appropriate first step in the confirmation of occupational asthma. Visual assessment of the plotted record is more sensitive and specific than statistical methods so far reported. The use of visual analysis is limited by lack of widespread expertise in the methods. A computer assisted diagnostic aid (OASYS-2) has been developed which is based on a scoring system developed from visual analysis. This removes the requirement for an experienced interpreter and should lead to the more widespread use of the technique. PEF records were collected from workers attending an occupational lung disease clinic for investigation of suspected occupational asthma and from workers participating in a study of respiratory symptoms in a postal sorting office. PEF records were divided into two development sets and two gold standard sets. The latter consisted of records from workers in which a final diagnosis had been reached by a method other than PEF recording. An experienced observer scored individual work and rest periods for the two development set PEF records; linear discriminant analysis was used to compare measurements taken from development set 1 records with visual scores. Two equations were produced which allowed prediction of scores for individual work or rest periods. The development set 2 was used to determine how these scores should be used to produce a whole record score. The first gold standard set was used to determine the whole record score which best separated those with and without occupational asthma. The second set determined the sensitivity and specificity of the chosen score. Two hundred and sixty eight PEF records were collected from 169 workers and divided into two development sets (81 and 60 records) and two gold standard sets (60 and 67 records). Linear discriminant analysis produced equations predicting the score for work periods incorporating five indices of PEF change and one for rest periods using seven indices. These equations correctly predicted the score for development set 1 work and rest periods on 61% of occasions (kappa = 0.47). The whole record score for development set 2 records, after weighting for definite or definitely no occupational effect, correlated with the visual score (correlation coefficient 0.86). Comparison with gold standard set 1 identified a cut off which proved to have a sensitivity of 75% and a specificity of 94% for an independent diagnosis of occupational asthma when applied to gold standard set 2. These results suggest that the sensitivity and specificity of analysing PEF records for occupational asthma using OASYS-2 approaches that of visual analysis, but it should be absolutely reproducible. The performance of OASYS-2 is more specific and approaches the sensitivity of other statistical methods of analysis. The evaluation of a large number of PEF records from workers exposed to different sensitising agents suggests that these results should be robust and should be repeatable in clinical practice.


Pilot Study for OCT Guided Design and Fit of a Prosthetic Device for Treatment of Corneal Disease.
Purpose. To assess optical coherence tomography (OCT) for guiding design and fit of a prosthetic device for corneal disease. Methods. A prototype time domain OCT scanner was used to image the anterior segment of patients fitted with large diameter (18.5-20 mm) prosthetic devices for corneal disease. OCT images were processed and analyzed to characterize corneal diameter, corneal sagittal height, scleral sagittal height, scleral toricity, and alignment of device. Within-subject variance of OCT-measured parameters was evaluated. OCT-measured parameters were compared with device parameters for each eye fitted. OCT image correspondence with ocular alignment and clinical fit was assessed. Results. Six eyes in 5 patients were studied. OCT measurement of corneal diameter (coefficient of variation, CV = 0.76%), cornea sagittal height (CV = 2.06%), and scleral sagittal height (CV = 3.39%) is highly repeatable within each subject. OCT image-derived measurements reveal strong correlation between corneal sagittal height and device corneal height (r = 0.975) and modest correlation between scleral and on-eye device toricity (r = 0.581). Qualitative assessment of a fitted device on OCT montages reveals correspondence with slit lamp images and clinical assessment of fit. Conclusions. OCT imaging of the anterior segment is suitable for custom design and fit of large diameter (18.5-20 mm) prosthetic devices used in the treatment of corneal disease.


Distinct magnetic phase transition at the surface of an antiferromagnet.
In the majority of magnetic systems the surface is required to order at the same temperature as the bulk. In the present Letter, we report a distinct and unexpected surface magnetic phase transition at a lower temperature than the Néel temperature. Employing grazing incidence x-ray resonant magnetic scattering, we have observed the near-surface behavior of uranium dioxide. UO2 is a noncollinear, triple-q, antiferromagnet with the U ions on a face-centered cubic lattice. Theoretical investigations establish that at the surface the energy increase-due to the lost bonds-is reduced when the spins near the surface rotate, gradually losing their component normal to the surface. At the surface the lowest-energy spin configuration has a double-q (planar) structure. With increasing temperature, thermal fluctuations saturate the in-plane crystal field anisotropy at the surface, leading to soft excitations that have ferromagnetic XY character and are decoupled from the bulk. The structure factor of a finite two-dimensional XY model fits the experimental data well for several orders of magnitude of the scattered intensity. Our results support a distinct magnetic transition at the surface in the Kosterlitz-Thouless universality class.


ERdj3, a stress-inducible endoplasmic reticulum DnaJ homologue, serves as a cofactor for BiP's interactions with unfolded substrates.
We recently identified ERdj3 as a component of unassembled immunoglobulin (Ig) heavy chain:BiP complexes. ERdj3 also associates with a number of other protein substrates, including unfolded light chains, a nonsecreted Ig light chain mutant, and the VSV-G ts045 mutant at the nonpermissive temperature. We produced an ERdj3 mutant that was unable to stimulate BiP's ATPase activity in vitro or to bind BiP in vivo. This mutant retained the ability to interact with unfolded protein substrates, suggesting that ERdj3 binds directly to proteins instead of via interactions with BiP. BiP remained bound to unfolded light chains longer than ERdj3, which interacted with unfolded light chains initially, but quickly disassociated before protein folding was completed. This suggests that ERdj3 may bind first to substrates and serve to inhibit protein aggregation until BiP joins the complex, whereas BiP remains bound until folding is complete. Moreover, our findings support a model where interactions with BiP help trigger the release of ERdj3 from the substrate:BiP complex.


Reactive nitrogen species augment fibroblast-mediated collagen gel contraction, mediator production, and chemotaxis.
Reactive nitrogen species (RNS) such as peroxynitrite cause cellular injury and tissue inflammation. Excessive production of nitrotyrosine, which is a footprint of RNS, has been observed in the airways of patients with asthma and chronic obstructive pulmonary disease, disorders characterized by tissue remodeling. The aim of this study was to evaluate whether RNS can affect tissue remodeling through direct effects on fibroblasts, and to determine if these effects depend on production of transforming growth factor-beta (TGF-beta). To accomplish this, human fetal lung fibroblasts (HFL-1) were used to assess fibroblast-mediated contraction of floating gels and chemotaxis toward fibronectin. In addition, the ability of fibroblasts to release TGF-beta1, fibronectin, and vascular endothelial growth factor (VEGF) was assessed by enzyme-linked immunosorbent assay. Authentic peroxynitrite significantly augmented gel contraction (P < 0.01) and chemotaxis (P < 0.01) compared with control in a concentration-dependent manner. Similarly, the peroxynitrite donor 3-morpholynosidenonimine hydrochloride (SIN-1) also augmented gel contraction (P < 0.01). RNS also significantly increased TGF-beta1 (P < 0.01), fibronectin (P < 0.01), and VEGF (P < 0.01) release into the media in both 3D gel and monolayer culture. Anti-TGF-beta antibody reversed RNS-augmented gel contraction (P < 0.01) and mediator production (P < 0.01). Anti-TGF-beta antibody also partially, but significantly, reversed RNS-augmented chemotaxis toward fibronectin (P < 0.01). Finally, peroxynitrite enhanced expression of alpha5beta1 integrin, which is a receptor for fibronectin (P < 0.01), and neutralizing anti-TGF-beta antibody suppressed peroxynitrite-augmented alpha5beta1 expression (P < 0.01). These results suggest that RNS can affect the tissue repair process by modulating TGF-beta1.


Travel and the emergence of high-level drug resistance in Plasmodium falciparum in southwest Uganda: results from a population-based study.
The I164L mutation on the dhfr gene confers high level resistance to sulfadoxine-pyrimethamine (SP) but it is rare in Africa except in a cluster of reports where prevalence >10% in highland areas of southwest Uganda and eastern Rwanda. The occurrence of the dhfr I164L mutation was investigated in community surveys in this area and examined the relationship to migration. A cross-sectional prevalence survey was undertaken in among villages within the catchment areas of two health facilities in a highland site (Kabale) and a highland fringe site (Rukungiri) in 2007. Sociodemographic details, including recent migration, were collected for each person included in the study. A total of 206 Plasmodium falciparum positive subjects were detected by rapid diagnostic test; 203 in Rukungiri and 3 in Kabale. Bloodspot samples were taken and were screened for dhfr I164L. Sequence analysis confirmed the presence of the I164L mutations in twelve P. falciparum positive samples giving an estimated prevalence of 8.6% in Rukungiri. Of the three parasite positive samples in Kabale, none had I164L mutations. Among the twelve I164L positives three were male, ages ranged from 5 to 90 years of age. None of those with the I164L mutation had travelled in the 8 weeks prior to the survey, although three were from households from which at least one household member had travelled during that period. Haplotypes were determined in non-mixed infections and showed the dhfr I164L mutation occurs in both as a N51I + S108N + I164L haplotype (n = 2) and N51I + C59R + S108N + I164L haplotype (n = 5). Genotyping of flanking microsatellite markers showed that the I164L occurred independently on the triple mutant (N51I, C59R + S108N) and double mutant (N51I + S108N) background. There is sustained local transmission of parasites with the dhfr I164L mutation in Rukungiri and no evidence to indicate its occurrence is associated with recent travel to highly resistant neighbouring areas. The emergence of a regional cluster of I164L in SW Uganda and Rwanda indicates that transmission of I164L is facilitated by strong drug pressure in low transmission areas potentially catalysed in those areas by travel and the importation of parasites from relatively higher transmission settings.


Evaluation of the Immediate Effect of Auricular Acupuncture on Pain and Electromyographic Activity of the Upper Trapezius Muscle in Patients with Nonspecific Neck Pain: A Randomized, Single-Blinded, Sham-Controlled, Crossover Study.
Background. The aim of the present study was to assess the immediate effects of auricular acupuncture (AA) on the electromyographic (EMG) activity of the upper trapezius muscle and pain in nonspecific neck pain (NS-NP) patients. Twelve patients with NS-NP (NS-NP group) and 12 healthy subjects (HS Group) were enrolled in a randomized, single-blinded, crossover study. Each subject received a single session of AA and sham AA (SAA). Surface EMG activity was measured in the upper trapezius muscle at different "step contractions" of isometric shoulder elevation (15%, 20%, 25%, and 30% MVC). The outcome measure in patients with NS-NP was based on the numerical pain rating scale (NRS). AA treatment led to a significant decrease in EMG activity in both groups (NS-NP group: p = 0.0001; HS group: p < 0.0001-ANOVA test). This was not the case for the SAA treatment (NS-NP group: p = 0.71; HS group: p < 0.54). Significant decreases (p < 0.001) in the NRS were found for both treatments (AA and SAA). This study demonstrated the immediate effect of auricular acupuncture on the electromyographic activity of the upper trapezius muscle but the effect of this intervention on pain symptoms in patients with nonspecific neck pain was inconclusive.