The measurement of immediate and persistent radiation-induced chromosome damage in rodent primary cells using premature chromosome condensation.
The chromosome damage observed in cultured peripheral blood lymphocytes at their metaphase is considered to be the most reliable basis for biological dosimetry. However, physical and biological factors such as irradiation conditions and chromosome repair could make difficult the interpretation of chromosome aberration yield in terms of absorbed dose. To establish correction factors to take into account partial body exposure, dose rate, and post-irradiation time requires in-vivo studies. Yet, such studies may be hampered by the laborious techniques required to culture the blood of the more readily available laboratory animals. In this paper, a cytogenetic approach based on a simple method for polyethylene-glycol-mediated cell fusion and premature chromosome condensation induction in primary cells is shown to be useful for in-vivo studies involving rodents. This method enables the direct assessment of radiation-induced chromosome damage without having the cells proceed to mitosis. The applicability of the method is demonstrated in particular for the measurement of immediate and persistent radiation-induced chromosome damage in rat peripheral blood, spleen, and thymus lymphocytes.


Recurrent spinal meningeal melanocytoma at lumbar spine level: a case report.
Spinal meningeal melanocytoma is an extremely rare tumour with an estimated annual incidence of 1 per 10 million people. It usually arises from the intradural extramedullary compartment at the cervical levels. Although these tumours are histologically benign, they may behave aggressively. Local recurrence could occur even after total tumour excision. We report a case of a 33-year-old Asian male who developed progressive weakness and numbness of the bilateral lower extremities as well as urinary retention five years after complete tumour resection of lumbar spinal meningeal melanocytoma. Magnetic resonance imaging of the lumbar spine revealed a mass with thecal sac compression which was hypointense on T2-weighted images and hyperintense on T1-weighted images. The patient underwent total tumour removal. Histologic examination was compatible with recurrent meningeal melanocytoma. After a 4-week inpatient rehabilitation programme, he was able to ambulate without assistance and to do clean intermittent catheterisation for micturition on a regular basis. This is the first reported case of intradural extramedullary meningeal melanocytoma located at the lumbar region. Clinicians should consider the possibility of these rare tumours at any level of the spine, and be aware of sphincter dysfunction in addition to motor and sensory deficits of extremities.


Stimulation of prostaglandin production in rabbit ileal mucosa by bradykinin.
When rabbit ileal mucosa was incubated with exogenous [3H]arachidonic acid (AA), its major metabolites, identified by comigration with known standards on thin-layer chromatography, were prostaglandin (PG) E2, 6-keto-PGF1 alpha and to a lesser extent PGF2 alpha and PGD2. The rate of prostanoid release from the serosal surface of the mucosa only was increased after incubation th either bradykinin, lys-bradykinin, melittin or the calcium ionophore A 23187, in a rapid and dose-dependent fashion. Peptide concentrations as low as 10(-9) M were effective. Kinin-induced release of AA or its metabolites required the presence of Ca++ in the incubation medium. Stimulation of prostanoid release by lys-bradykinin was completely blocked by indomethacin. The combined lipoxygenase/cyclooxygenase inhibitors BW 755 and eicosa-5,8,11,14-tetraynoic acid and the lipoxygenase inhibitor nordihydroguaiaretic acid also blocked the stimulation of PG synthesis by lys-bradykinin. These inhibitors caused an increase in levels of AA released from the tissue by lys-bradykinin. The phospholipase inhibitors, mepacrine and U- 10029, inhibited the lys-bradykinin-stimulated release of both prostanoids and AA. At higher concentrations, U- 10029 inhibited the stimulation of transepithelial potential difference and short-circuit current across rabbit ileal mucosa produced by lys-bradykinin. These results support the hypothesis that bradykinin-stimulated intestinal secretion may be mediated by PGs.


Characterization of myenteric sensory neurons in the mouse small intestine.
We recorded from myenteric AH/Dogiel type II cells, demonstrated mechanosensitive responses, and characterized their basic properties. Recordings were obtained using the mouse longitudinal muscle myenteric plexus preparation with patch-clamp and sharp intracellular electrodes. The neurons had an action potential hump and a slow afterhyperpolarization (AHP) current. The slow AHP was carried by intermediate conductance Ca2+ -dependent K+ -channel currents sensitive to charybdotoxin and clotrimazole. All possessed a hyperpolarization-activated current that was blocked by extracellular cesium. They also expressed a TTX-resistant Na+ current with an onset near the resting potential. Pressing on the ganglion containing the patched neuron evoked depolarizing potentials in 17/18 cells. The potentials persisted after synaptic transmission was blocked. Volleys of presynaptic electrical stimuli evoked slow excitatory postsynaptic potentials (EPSPs) in 9/11 sensory neurons, but 0/29 cells received fast EPSP input. The slow EPSP was generated by removal of a voltage-insensitive K+ current. Patch-clamp recording with a KMeSO4-containing, but not a conventional KCl-rich, intracellular solution reproduced the single-spike slow AHPs and low input resistances seen with sharp intracellular recording. Cell-attached recording of intermediate conductance potassium channels supported the conclusion that the single-spike slow AHP is an intrinsic property of intestinal AH/sensory neurons. Unitary current recordings also suggested that the slow AHP current probably does not contribute significantly to the high resting background conductance seen in these cells. The characterization of mouse myenteric sensory neurons opens the way for the study of their roles in normal and pathological physiology.


Virtual Reality Exposure Simulation for Student Veteran Social Anxiety and PTSD: A Case Study.
Exposure based exercises are a common element of many gold standard treatments for anxiety disorders and post-traumatic stress disorder and virtual reality simulations have been evaluated as a platform for providing clients with opportunities for repeated exposure during treatment. Although research on virtual reality exposure therapy (VRET) indicates effectiveness and high levels of user satisfaction, VRETs require a participant to complete exposure exercises in-offices with specialized equipment. The current exploratory case method study evaluates the experience and outcomes of one student veteran with social anxiety disorder and PTSD completing twelve sessions of VRET exposure using a mobile phone simulation of a virtual grocery store. The participant reported decreases in psychological symptoms, improvements in neurological connectivity, and better sleep quality upon completing the trial. Results suggest that VRET using a mobile application is feasible and warrants further research to evaluate effectiveness more fully. Implications include the use of a mobile based virtual reality simulation for intervening in social anxiety for student veterans.


Induction of apoptosis in HeLa cells by chloroform fraction of seed extracts of Nigella sativa.
Cancer remains one of the most dreaded diseases causing an astonishingly high death rate, second only to cardiac arrest. The fact that conventional and newly emerging treatment procedures like chemotherapy, catalytic therapy, photodynamic therapy and radiotherapy have not succeeded in reverting the outcome of the disease to any drastic extent, has made researchers investigate alternative treatment options. The extensive repertoire of traditional medicinal knowledge systems from various parts of the world are being re-investigated for their healing properties. This study progresses in the direction of identifying component(s) from Nigella sativa with anti cancer activity. In the present study we investigated the efficacy of Organic extracts of Nigella sativa seed powder for its clonogenic inhibition and induction of apoptosis in HeLa cancer cell. Methanolic, n-Hexane and chloroform extracts of Nigella sativa seedz effectively killed HeLa cells. The IC50 values of methanolic, n-hexane, and chloroform extracts of Nigella sativa were 2.28 microg/ml, 2.20 microg/ml and 0.41 ng/ml, respectively. All three extracts induced apoptosis in HeLa cells. Apoptosis was confirmed by DNA fragmentation, western blot and terminal transferase-mediated dUTP-digoxigenin-end labeling (TUNEL) assay. Western Blot and TUNEL results suggested that Nigella sativa seed extracts regulated the expression of pro- and anti- apoptotic genes, indicating its possible development as a potential therapeutic agent for cervical cancer upon further investigation.


Sickle-cell trait in human biological and cultural evolution. Development of agriculture causing increased malaria is bound to gene-pool changes causing malaria reduction.
The particular agricultural adaptation we have been considering is the ultimate determinant of the presence of malaria parasites in the intracellular environment of the human red blood cell. This change in the cellular environment is deleterious for normal individuals, but individuals with the sickle-cell gene are capable of changing their red-cell environment so that intense parasitism never develops. Normal individuals suffer higher mortality rates and lower fertility rates in a malarious environment than individuals with the sickle-cell trait do, so the latter contribute proportionately more people to succeeding generations.


The FLAME-accelerated signalling tool (FaST) for facile parallelisation of flexible agent-based models of cell signalling.
Agent-based modelling is particularly adept at modelling complex features of cell signalling pathways, where heterogeneity, stochastic and spatial effects are important, thus increasing our understanding of decision processes in biology in such scenarios. However, agent-based modelling often is computationally prohibitive to implement. Parallel computing, either on central processing units (CPUs) or graphical processing units (GPUs), can provide a means to improve computational feasibility of agent-based applications but generally requires specialist coding knowledge and extensive optimisation. In this paper, we address these challenges through the development and implementation of the FLAME-accelerated signalling tool (FaST), a software that permits easy creation and parallelisation of agent-based models of cell signalling, on CPUs or GPUs. FaST incorporates validated new agent-based methods, for accurate modelling of reaction kinetics and, as proof of concept, successfully converted an ordinary differential equation (ODE) model of apoptosis execution into an agent-based model. We finally parallelised this model through FaST on CPUs and GPUs resulting in an increase in performance of 5.8× (16 CPUs) and 53.9×, respectively. The FaST takes advantage of the communicating X-machine approach used by FLAME and FLAME GPU to allow easy alteration or addition of functionality to parallel applications, but still includes inherent parallelisation optimisation. The FaST, therefore, represents a new and innovative tool to easily create and parallelise bespoke, robust, agent-based models of cell signalling.


New Paradigms in the Psychology of Reasoning.
The psychology of verbal reasoning initially compared performance with classical logic. In the last 25 years, a new paradigm has arisen, which focuses on knowledge-rich reasoning for communication and persuasion and is typically modeled using Bayesian probability theory rather than logic. This paradigm provides a new perspective on argumentation, explaining the rational persuasiveness of arguments that are logical fallacies. It also helps explain how and why people stray from logic when given deductive reasoning tasks. What appear to be erroneous responses, when compared against logic, often turn out to be rationally justified when seen in the richer rational framework of the new paradigm. Moreover, the same approach extends naturally to inductive reasoning tasks, in which people extrapolate beyond the data they are given and logic does not readily apply. We outline links between social and individual reasoning and set recent developments in the psychology of reasoning in the wider context of Bayesian cognitive science.


Simultaneous dual frequency 1H and 19F open coil imaging of arthritic rabbit knee at 3T.
The combination of sensitive magnetic resonance techniques with a selective site-targeted nanoparticle contrast agent has a demonstrated utility for molecular imaging studies. By detecting a unique signature of the contrast agent, this approach can be employed to identify specific bio-molecular markers and observe cellular-level processes within a large and complex organism (e.g., in vivo rabbit). The objective of the present investigation was to design, fabricate and characterize a radio-frequency (RF) coil for the dual frequency ((1)H and (19)F) simultaneous collection of both nuclei images in a 3T field, in order to facilitate studies of arthritic knee degradation in rabbits. The coil supports both transmit and receive modes. The supporting activities included: 1) establishing a technical database for calculating the required coil parameters, 2) selection of a favorable coil geometry, and 3) adaption of existing RF measurement techniques to the design, development and electrical evaluation of the coil. The coil is used in conjunction with a Philips Medical Systems clinical MRI scanner, requiring all RF simultaneous dual frequency ((1)H and (19)F) coils to operate in both transmit and receive modes. A commercial version of SPICE (simulation program with integrated circuit emphasis) was used to estimate significant operational parameters prior to fabricating the imaging coil. Excellent images were obtained with the fabricated coil and no operational problems were observed that would limit the use of other coil geometries and field strengths.


Label-free in situ SERS imaging of biofilms.
Surface-enhanced Raman scattering (SERS) is a promising technique for the chemical characterization of biological systems. It yields highly informative spectra, can be applied directly in aqueous environment, and has high sensitivity in comparison with normal Raman spectroscopy. Moreover, SERS imaging can provide chemical information with spatial resolution in the micrometer range (chemical imaging). In this paper, we report for the first time on the application of SERS for in situ, label-free imaging of biofilms and demonstrate the suitability of this technique for the characterization of the complex biomatrix. Biofilms, being communities of microorganisms embedded in a matrix of extracellular polymeric substances (EPS), represent the predominant mode of microbial life. Knowledge of the chemical composition and the structure of the biofilm matrix is important in different fields, e.g., medicine, biology, and industrial processes. We used colloidal silver nanoparticles for the in situ SERS analysis. Good SERS measurement reproducibility, along with a significant enhancement of Raman signals by SERS (>10(4)) and highly informative SERS signature, enables rapid SERS imaging (1 s for a single spectrum) of the biofilm matrix. Altogether, this work illustrates the potential of SERS for biofilm analysis, including the detection of different constituents and the determination of their distribution in a biofilm even at low biomass concentration.


Temporal sequence of pulmonary and systemic inflammatory responses to graded polymicrobial peritonitis in mice.
The lungs are the remote organ most commonly affected in human peritonitis. The major goals of this study were to define the dose- and time-dependent relationship between graded septic peritonitis and systemic and pulmonary inflammatory responses in mice. BALB/c mice were treated with intraperitoneal polymicrobial inoculi and sacrificed at 3, 12, and 24 h. The treatment protocol resulted in distinct groups of animals with respect to mortality rate, kinetics, and concentrations of a broad spectrum of pro- and anti-inflammatory endogenous mediators, intrapulmonary bacterial accumulation, and static lung compliance. In sublethally infected mice, pulmonary bacterial proliferation was controlled. Levels of monocyte chemoattractant protein-1 (MCP-1), interleukin-10, interleukin-6, granulocyte colony-stimulating factor (G-CSF), and tumor necrosis factor (TNF) in plasma were elevated 3 h after infection exclusively. At 3 h, MCP-1, gamma interferon, and TNF were detected in extracts of pulmonary tissue or in bronchoalveolar lavage (BAL) fluid. Static lung compliance (C(st)) was transiently decreased at 12 h. In contrast, in lethally infected mice pulmonary bacterial proliferation was not contained. Concentrations of MCP-1, G-CSF, and TNF in plasma were maximal at 24 h, as were pulmonary MCP-1 levels. Lung myeloperoxidase activity was increased at 3, 12, and 24 h. C(st) was reduced after 3 h and did not reach control values at 24 h. Pulmonary cyclooxygenase-2 mRNA and eicosanoids in BAL fluid and plasma were elevated at 3 and 24 h. This study shows that polymicrobial peritonitis in mice leads to dose-dependent systemic and pulmonary inflammation accompanied by a decrease in lung compliance.


Fabrication of cellulose carbamate hydrogel-dressing with rarasaponin surfactant for enhancing adsorption of silver nanoparticles and antibacterial activity.
Bacterial contamination on external wounds is known to be a factor that prevents wound healing and triggers tissue damage. Hydrogel-dressings with antibacterial activity is a useful medical device to avoid this contamination, wherein the antibacterial activity can be provided via incorporation of silver nanoparticles (AgNPs). Contrary to the conventional two-step preparation of an AgNPs-loaded hydrogel (AgNPs@hydrogel), this work aims to establish a new and facile synthesis method employing the adsorption principle. Once AgNO3 adsorbed into active sites of the hydrogels, in situ reductions using NaBH4 was employed to produce AgNPs@hydrogel. The effect of surfactant addition on the AgNO3 loading and the antibacterial activity of the resulting hydrogel dressing was investigated. The outcome of this work indicates that the addition of rarasaponin not only can increase the loading of AgNPs on cellulose carbamate hydrogel (CCH) but also significantly enhance the antibacterial activity of the resulted hydrogel-dressing. Superior to the other studied surfactant, the loading capacity (LC) of AgNPs is found to be 10.15, 9.94, and 7.53 mg/g for CCH modified with rarasaponin, CTAB, and Tween80, respectively. These findings conclude that the addition of surfactant, especially rarasaponin, can effectively improve the loading of AgNPs onto hydrogel-dressing via adsorption and promote the antibacterial activity. Furthermore, the cytotoxic test shows that the hydrogel-dressings have good biocompatibility toward skin fibroblast cells.


Conservative treatment of scald burns is superior to early excision.
Early excision of deep burns has been advocated; however, it is difficult to clinically determine the depth of scald burns during the early postburn period. This prospective, randomized study was designed to determine whether early excision was superior to conservative treatment of scald injuries. Patients with scald injuries (which were not caused by grease) of clinically indeterminant depth were randomized to early (n = 12) or late (n = 12) excision; all patients with obvious superficial and full-thickness injuries were excluded. In the early excision group, all deep wounds were tangentially excised and grafted within 72 hours of admission, whereas in the late treatment group wounds were excised and grafted after 2 weeks had passed since injury. Area excised, postburn day of excision, percent graft take, operating-room time, blood replacement, incidence of infection, and length of hospital stay were compared. No patient experienced a significant wound infection or systemic sepsis. A significantly smaller area of excision was necessary for those patients who were treated with delayed surgery, and concomitant decreases in operating-room time and blood loss were observed. Notably, only one half of the patients who were randomized to the delayed excision group ultimately required surgical intervention to achieve wound closure. Graft take was comparable for both groups, as was length of hospital stay. Early clinical evaluation of scald injuries appears to be equivocal, and later evaluations reveal a less severe injury. Financial gains can be made when surgical excision of scald injuries is delayed until 2 weeks after injury because of a related reduction in hospital expenditures.


Generalizing the wavelet-based multifractal formalism to random vector fields: application to three-dimensional turbulence velocity and vorticity data.
We use singular value decomposition techniques to generalize the wavelet transform modulus maxima method to the multifractal analysis of vector-valued random fields. The method is calibrated on synthetic multifractal 2D vector measures and monofractal 3D fractional Brownian vector fields. We report the results of some application to the velocity and vorticity fields issued from 3D isotropic turbulence simulations. This study reveals the existence of an intimate relationship between the singularity spectra of these two vector fields which are found significantly more intermittent than previously estimated from longitudinal and transverse velocity increment statistics.


Biomechanical studies of double-bundle posterior cruciate ligament reconstructions.
Double-bundle reconstruction of the posterior cruciate ligament has been advocated to better replicate the anatomy of the native ligament and restore normal knee biomechanics. The goal of this study was to measure knee laxities and graft forces following single and double-bundle reconstructions and to compare these values with those for the intact knee in a cadaver model. Forces in the posterior cruciate ligament were measured as the knee was passively extended from 120 degrees to 0 degrees with applied tibial loading. Anterior-posterior laxities were measured as well. An anterolateral tunnel was located at the anterolateral margin of the native ligament footprint, and a posteromedial tunnel was placed at one of two locations within the footprint; one location resulted in a wide bridge separating the tunnels and the other, a narrow bridge. Testing was repeated with a single anterolateral graft tensioned to match, within +/-1 mm, the laxity in the intact knee at 90 degrees of flexion. Double-bundle reconstructions were tested with the addition of a posteromedial graft tensioned at 30 degrees of flexion. Two levels of posteromedial graft tension (10 and 30 N) were studied in both the narrow and the wide-bridge posteromedial tunnels. Mean laxities with a single anterolateral graft were 1.1 to 2.0 mm greater than normal between 0 degrees and 30 degrees of flexion. With the posteromedial graft tensioned to 10 N in the wide-bridge tunnel, the mean laxity of the double grafts was not significantly different from that in the intact knee at any flexion angle. With the posteromedial graft tensioned to 10 N in the narrow-bridge tunnel, the mean laxity at 0 degrees was 0.9 mm greater than that in the intact knee. With the posteromedial graft tensioned to 30 N, the mean laxity at 10 degrees was 1.7 mm less than the intact-knee value in the wide-bridge tunnel and 1.3 mm less than the intact-knee value in the narrow bridge-tunnel. Increasing posteromedial graft tension from 10 to 30 N decreased the mean laxities by 0.5 to 1.1 mm between 0 degrees and 30 degrees . Mean graft forces following a single anterolateral reconstruction were not significantly different from the native posterior cruciate ligament forces under any mode of loading except valgus moment. With the wide-bridge tunnel, the mean forces with the posteromedial graft tensioned to 10 N were somewhat higher than the native posterior cruciate ligament forces at full extension; when the graft was tensioned to 30 N, the mean forces were substantially higher. A single anterolateral graft best reproduced the normal posterior cruciate ligament force profiles, but laxities were greater than normal between 0 degrees and 30 degrees of knee flexion. The addition of a second, posteromedial graft reduced laxity in this flexion range but did so at the expense of higher-than-normal forces in the posteromedial graft.


The contribution of the exosite residues of plasminogen activator inhibitor-1 to proteinase inhibition.
The binding of plasminogen activator inhibitor-1 (PAI-1) to serine proteinases, such as tissue-type plasminogen activator (tPA) and urokinase-type plasminogen activator (uPA), is mediated by the exosite interactions between the surface-exposed variable region-1, or 37-loop, of the proteinase and the distal reactive center loop (RCL) of PAI-1. Although the contribution of such interactions to the inhibitory activity of PAI-1 has been established, the specific mechanistic steps affected by interactions at the distal RCL remain unknown. We have used protein engineering, stopped-flow fluorimetry, and rapid acid quenching techniques to elucidate the role of exosite interactions in the neutralization of tPA, uPA, and beta-trypsin by PAI-1. Alanine substitutions at the distal P4' (Glu-350) and P5' (Glu-351) residues of PAI-1 reduced the rates of Michaelis complex formation (k(a)) and overall inhibition (k(app)) with tPA by 13.4- and 4.7-fold, respectively, whereas the rate of loop insertion or final acyl-enzyme formation (k(lim)) increased by 3.3-fold. The effects of double mutations on k(a), k(lim), and k(app) were small with uPA and nonexistent with beta-trypsin. We provide the first kinetic evidence that the removal of exosite interactions significantly alters the formation of the noncovalent Michaelis complex, facilitating the release of the primed side of the distal loop from the active-site pocket of tPA and the subsequent insertion of the cleaved reactive center loop into beta-sheet A. Moreover, mutational analysis indicates that the P5' residue contributes more to the mechanism of tPA inhibition, notably by promoting the formation of a final Michaelis complex.


Coping with dementia: evaluation of four nonpharmacologic interventions.
To evaluate nonpharmacologic interventions, caregivers (65 women, 38 men) and their dementia-diagnosed spouses (patients) were randomized to one of four treatment programs (cognitive stimulation, dyadic counseling, dual supportive seminar, and early-stage day care) or to a wait-list control group. Assessments occurred initially and at postintervention (3 months). Patients were evaluated on memory, verbal fluency, and problem-solving ability, and caregivers were assessed on marital interaction, emotional status, and physical health, along with stress, coping, and social support. Caregivers also completed a program evaluation. Repeated measures procedures showed that patients in the cognitive stimulation group demonstrated more improvement over time in cognitive outcomes, and caregivers decreased in depressive symptoms. Early-stage day-care and dual supportive seminar group caregivers reported a decrease in hostility and a decrease in use of negative coping strategies, respectively. Although qualitatively derived benefits differed across groups, similarities in program content reduced the potential for quantitative differentiation among the groups.


Juvenile social experience affects pairing success at adulthood: congruence with the loser effect?
Social interactions with adults are often critical for the development of mating behaviours. However, the potential role of other primary social partners such as juvenile counterparts is rarely considered. Most interestingly, it is not known whether interactions with juvenile females improve males' courtship and whether, similar to the winner and loser effects in a fighting context--outcome of these interactions shapes males' behaviour in future encounters. We investigated the combined effects of male quality and juvenile social experience on pairing success at adulthood in zebra finches (Taeniopygia guttata). We manipulated brood size to alter male quality and then placed males in either same- or mixed-sex juvenile dyads until adulthood. We found that males from reduced broods obtained more copulations and males from mixed-sex dyads had more complete courtships. Furthermore, independent of their quality, males that failed to pair with juvenile females, but not juvenile males, had a lower pairing success at adulthood. Our study shows that negative social experience with peers during adolescence may be a potent determinant of pairing success that can override the effects of early environmental conditions on male attractiveness and thereby supports the occurrence of an analogous process to the loser effect in a mating context.


INDUCTION OF A STEREOSCOPIC DEPTH EFFECT.
Two pairs of horizontal lines of equal length, with different configurations of dots above and below, are presented stereoscopically. Upon fusion, the two lines are perceived as tilting in space in directions opposite to the depth effects of the dots.


Familial Pancreatic Cancer: Current Perspectives.
Pancreatic cancer (PC) is a highly lethal disease, mostly incurable when detected. Thus, despite advances in PC treatments, only around 7% of patients survive 5-years after diagnosis. This morbid outcome is secondary to multifactorial reasons, such as late-stage diagnosis, rapid progression and minimal response to chemotherapy. Based on these factors, it is of special relevance to identify PC high-risk individuals in order to establish preventive and early detection measures. Although most PC are sporadic, approximately 10% cases have a familial basis. No main causative gene of PC has been identified but several known germline pathogenic mutations are related with an increased risk of this tumor. These inherited cancer syndromes represent 3% of all PC. On the other hand, in 7% of cases of PC, there is a strong family history without a causative germline mutation, a situation known as familial pancreatic cancer (FPC). In recent years, there is increasing evidence supporting the benefit of genetic germline analysis in PC patients, and periodic pancreatic screening in PC high-risk patients (mainly those with a lifetime risk greater than 5%), although there is no general agreement in the group of patients and individuals to study and screen. In the present review, we expose an update in the field of hereditary and FPC, with the aim of describing the current strategies and implications in genetic counseling, surveillance and therapeutic interventions.


[A survey of the diagnostic policy in nocturnal enuresis].
A written enquiry regarding the current diagnostic policy in nocturnal enuresis was conducted among paediatricians (response 38%, n = 22) and urologists (response 49%, n = 23) while an enquiry by telephone was held among GPs (n = 40) with the same questions. With reference to three case histories, the question was asked what supplementary examinations, if any, would be carried out. All respondents showed a tendency to perform extensive urological examinations. Since bedwetting in most cases is not associated with demonstrable anatomical or functional abnormalities, this is incorrect for medical reasons, even apart from the costs involved.


Shared muscle synergies in human walking and cycling.
The motor system may rely on a modular organization (muscle synergies activated in time) to execute different tasks. We investigated the common control features of walking and cycling in healthy humans from the perspective of muscle synergies. Three hypotheses were tested: 1) muscle synergies extracted from walking trials are similar to those extracted during cycling; 2) muscle synergies extracted from one of these motor tasks can be used to mathematically reconstruct the electromyographic (EMG) patterns of the other task; 3) muscle synergies of cycling can result from merging synergies of walking. A secondary objective was to identify the speed (and cadence) at which higher similarities emerged. EMG activity from eight muscles of the dominant leg was recorded in eight healthy subjects during walking and cycling at four matched cadences. A factorization technique [nonnegative matrix factorization (NNMF)] was applied to extract individual muscle synergy vectors and the respective activation coefficients behind the global muscular activity of each condition. Results corroborated hypotheses 2 and 3, showing that 1) four synergies from walking and cycling can successfully explain most of the EMG variability of cycling and walking, respectively, and 2) two of four synergies from walking appear to merge together to reconstruct one individual synergy of cycling, with best reconstruction values found for higher speeds. Direct comparison of the muscle synergy vectors of walking and the muscle synergy vectors of cycling (hypothesis 1) produced moderated values of similarity. This study provides supporting evidence for the hypothesis that cycling and walking share common neuromuscular mechanisms.


Combinatorial MAPLE gradient thin film assemblies signalling to human osteoblasts.
There is increased interest in smart bioactive materials to control tissue regeneration for the engineering of cell instructive scaffolds. We introduced combinatorial matrix-assisted pulsed laser evaporation (C-MAPLE) as a new method for the fabrication of organic thin films with a compositional gradient. Synchronized C-MAPLE of levan and oxidized levan was employed to assemble a two-compound biopolymer film structure. The gradient of the film composition was validated by fluorescence microscopy. In this study, we investigated the cell response induced by the compositional gradient using imaging of early osteoblast attachment and analysis of signalling phosphoprotein expression. Cells attached along the gradient in direct proportion to oxidized levan concentration. During this process distinct areas of the binary gradient have been shown to modulate the osteoblasts' extracellular signal-regulated kinase signalling with different propensity. The proposed fabrication method results in the preparation of a new bioactive material, which could control the cell signalling response. This approach can be extended to screen new bioactive interfaces for tissue regeneration.


The chemistry of scrapie infection: implications of the 'ice 9' metaphor.
The transmissible spongiform encephalopathies pose an increasing problem for animal, and perhaps human, health. The infectious agent seems to lack a nucleic acid component, posing the question of how it can reproduce. A model of reproduction by nucleated polymerization suggests a number of novel approaches to the problem.


Heat Emitting Damage in Skin: A Thermal Pathway for Mechanical Algesia.
Mechanical pain (or mechanical algesia) can both be a vital mechanism warning us for dangers or an undesired medical symptom important to mitigate. Thus, a comprehensive understanding of the different mechanisms responsible for this type of pain is paramount. In this work, we study the tearing of porcine skin in front of an infrared camera, and show that mechanical injuries in biological tissues can generate enough heat to stimulate the neural network. In particular, we report local temperature elevations of up to 24°C around fast cutaneous ruptures, which shall exceed the threshold of the neural nociceptors usually involved in thermal pain. Slower fractures exhibit lower temperature elevations, and we characterise such dependency to the damaging rate. Overall, we bring experimental evidence of a novel-thermal-pathway for direct mechanical algesia. In addition, the implications of this pathway are discussed for mechanical hyperalgesia, in which a role of the cutaneous thermal sensors has priorly been suspected. We also show that thermal dissipation shall actually account for a significant portion of the total skin's fracture energy, making temperature monitoring an efficient way to detect biological damages.


Presbyopic patient with hyperopia and moderately steep corneas: December consultation #1.
A 47-year-old woman was referred for refractive surgery evaluation. She has no ocular or medical history. Spectacle dependence is her chief concern. She cannot tolerate contact lens. The corrected near and distance visual acuity (CDVA) are 20/20, easily, in both eyes. The manifest refraction is +3.50 in the right eye and +2.75 in the left eye (dominant eye). The intraocular pressure and the results from a slitlamp examination and a fundus retinoscopy are normal. She has no dry eye and no complaints of halos or glare. She works in an office using a computer most of the day.The Scheimpflug device revealed the thinnest point of corneal thickness to be 497 μm in the right eye and 501 μm in the left eye ( and ). Keratometry (K) values were K1 43.20 diopters (D), K2 45.00 D, and Kmax 45.50 D in the right eye and K1 43.40 D, K2 45.00 D, and Kmax 46.00 D in the left eye. Optical coherence tomography revealed thinnest epithelial thickness of 48 μm in the right eye and 49 μm in the left eye. A Zernike analysis showed low values for coma and spherical aberrations.(Figure is included in full-text article.)(Figure is included in full-text article.)The topographic maps, despite the relative scale, revealed a pattern that seems to truncate in the center, in both eyes, but especially in the right eye. Pachymetric progression indices were within normal limits ().(Figure is included in full-text article.)What kind of surgery (if any) would you recommend to this patient? Would you consider laser in situ keratomileusis (LASIK) with a thin, predictable LASIK flap? And in this case, what are your limits for final, postoperative steep K? How much concern would you have if this patient needed an excimer laser enhancement?Would you offer clear lens extraction (CLE)? And in this case any specific intraocular lens (IOL) model?What data helped you most in making your decision? If you recommend proceeding with surgery, would age have played any significant role in the decision process?


Prediction of prostate cancer extracapsular extension with high spatial resolution dynamic contrast-enhanced 3-T MRI.
To assess the value of dynamic contrast-enhanced (DCE) combined with T2-weighted (T2W) endorectal coil (ERC) magnetic resonance imaging (MRI) at 3 T for determining extracapsular extension (ECE) of prostate cancer. In this IRB-approved study, ERC 3-T MRI of the prostate was performed in 108 patients before radical prostatectomy. T2W fast spin-echo and DCE 3D gradient echo images were acquired. The interpretations of readers with varied experience were analysed. MRI-based staging results were compared with radical prostatectomy histology. Descriptive statistics were generated for prediction of ECE and staging accuracies were determined by the area under the receiver-operating characteristic curve. The overall sensitivity, specificity, positive predictive value and negative predictive value for ECE were 75 %, 92 %, 79 % and 91 %, respectively. Diagnostic accuracy for staging was 86 %, 80 % and 91 % for all readers, experienced and less experienced readers, respectively. ERC 3-T MRI of the prostate combining DCE and T2W imaging is an accurate pretherapeutic staging tool for assessment of ECE in clinical practice across varying levels of reader experience. • Endorectal coil (ERC) magnetic resonance imaging is widely used for imaging prostatic disease. • ERC 3-T MRI is reasonably accurate for local prostate cancer staging. • High diagnostic accuracy is achievable across different levels of reader experience. • MRI facilitates therapeutic decisions in patients with prostate cancer.


Comparison of CellSearch and Circulating Tumor Cells (CTC)-Biopsy Systems in Detecting Peripheral Blood Circulating Tumor Cells in Patients with Gastric Cancer.
BACKGROUND The purpose of this study was to compare circulating tumor cells (CTCs)/circulating tumor microemboli (CTM) detection rates of the CellSearch and CTC-Biopsy systems in patients with gastric cancer (GC). We also investigated potential correlations between clinicopathological characteristics and prognosis in patients with GC. MATERIAL AND METHODS This prospective study was conducted at the Shandong Institute of Cancer Prevention and Control in China. Fifty-nine patients with GC and 22 healthy volunteers were recruited and their peripheral blood samples were examined by the CTC-Biopsy system and CellSearch system for CTC. RESULTS The rate of detection of CTCs/CTM was significantly higher with the CTC-Biopsy system than with the CellSearch system (59.32% vs. 27.12%, P<0.001). The Kappa value was 0.179, indicating poor consistency. CTCs detected with the CellSearch system in patients with stage III/IV GC was significantly correlated with neutrophil count (P=0.020), neutrophil/lymphocyte ratio (N/L ratio) (P=0.009), CA19-9 (P=0.049), tumor size (P=0.026), and the extent of vascular invasion (P=0.007). CTCs detected with the CTC-Biopsy system correlated with tumor differentiation (P=0.010). CTM in patients with stage I/II GC and stage II/IV GC correlated with CEA (P=0.004) and tumor differentiation (P=0.030), respectively. A CTC count >3 detected with the CellSearch system, and not the CTC-Biopsy system, correlated with reduced progression-free survival and overall survival. CONCLUSIONS The CTC-Biopsy system was superior to the CellSearch system for detecting CTCs in GC patients. CTM were detected with the CTC-Biopsy system but not with the CellSearch system. CTCs detected with the CellSearch system correlated with various clinicopathological factors and long-term survival outcomes.


Revisiting the T2 spectrum imaging inverse problem: Bayesian regularized non-negative least squares.
Multi-echo T2 magnetic resonance images contain information about the distribution of T2 relaxation times of compartmentalized water, from which we can estimate relevant brain tissue properties such as the myelin water fraction (MWF). Regularized non-negative least squares (NNLS) is the tool of choice for estimating non-parametric T2 spectra. However, the estimation is ill-conditioned, sensitive to noise, and highly affected by the employed regularization weight. The purpose of this study is threefold: first, we want to underline that the apparently innocuous use of two alternative parameterizations for solving the inverse problem, which we called the standard and alternative regularization forms, leads to different solutions; second, to assess the performance of both parameterizations; and third, to propose a new Bayesian regularized NNLS method (BayesReg). The performance of BayesReg was compared with that of two conventional approaches (L-curve and Chi-square (X2) fitting) using both regularization forms. We generated a large dataset of synthetic data, acquired in vivo human brain data in healthy participants for conducting a scan-rescan analysis, and correlated the myelin content derived from histology with the MWF estimated from ex vivo data. Results from synthetic data indicate that BayesReg provides accurate MWF estimates, comparable to those from L-curve and X2, and with better overall stability across a wider signal-to-noise range. Notably, we obtained superior results by using the alternative regularization form. The correlations reported in this study are higher than those reported in previous studies employing the same ex vivo and histological data. In human brain data, the estimated maps from L-curve and BayesReg were more reproducible. However, the T2 spectra produced by BayesReg were less affected by over-smoothing than those from L-curve. These findings suggest that BayesReg is a good alternative for estimating T2 distributions and MWF maps.


Natural radiation exposure in the vicinity of Spanish nuclear power stations.
Nationwide and regional surveys have been conducted to evaluate natural radiation exposure of people in the vicinity of the Spanish nuclear power stations. In these surveys, indoor radon, external gamma dose rates outdoors and indoors and radioactivity in soils were measured. Measurements of indoor radon concentrations were performed in 214 dwellings by using nuclear track-etched detectors. External exposure from terrestrial gamma rays outdoors and indoors were measured in a total of 596 points. Radioactivity in soil samples collected at 293 sampling sites are also reported in this paper. By taking into account all sources of natural radiation exposure, mean annual effective doses to the population in the surroundings of the six Spanish nuclear power stations have been estimated and compared with those coming from their normal operation.


Magnetic Resonance Imaging of Gases: A Single-Point Ramped Imaging with T1 Enhancement (SPRITE) Study.
A pure phase-encoding MRI technique, single-point ramped imaging with T1 enhancement, SPRITE, is introduced for the purpose of gas phase imaging. The technique utilizes broadband RF pulses and stepped phase encode gradients to produce images, substantially free of artifacts, which are sensitive to the gas T1 and T&ast:2 relaxation times. Images may be acquired from gas phase species with transverse relaxation times substantially less than 1 ms. Methane gas images, 1H, were acquired in a phantom study. Sulfur hexafluoride, 19F, images were acquired from a gas-filled porous coral sample. High porosity regions of the coral are observed in both the MRI image and an X-ray image. Sensitivity and resolution effects due to signal modulation during the time-efficient acquisition are discussed. A method to increase the image sensitivity is discussed, and the predicted improvement is shown through 1D images of the methane gas phantom. Copyright 1999 Academic Press.


A general approach to multicomponent metal-decorated crumpled reduced graphene oxide nanocomposites using a flame-based process.
We introduce a general approach for synthesizing multicomponent metal-decorated crumpled reduced graphene oxide nanocomposites using a one-step, continuous flame-based process. Crumpled reduced graphene oxide balls (CGB) were produced from graphene oxide (GO) in a High Temperature Reducing Jet (HTRJ) reactor. Moreover, CGBs were simultaneously decorated with different transition metal nanoparticles, including cobalt (Co), nickel (Ni), iron (Fe), and palladium (Pd). Various metal alloy-decorated crumpled reduced graphene oxide balls (M-CGBs) including CoPd-, CoNi-, CoPdNi-, and CoNiFe-CGBs were successfully synthesized using a general recipe. The key advantage of the HTRJ system over common flame-based aerosol synthesis methods is the separation of flame and product formation zones, which allows production and/or reduction of nanomaterials that can be reduced by H2 in the presence of H2O. Nanomaterials are produced from aqueous precursors containing low-cost metal salts and dispersed GO. Electron microscopy and other characterization methods show the decoration of the CGBs with sub-4 nm diameter binary and ternary alloy, non-oxide transition metal nanoparticles of controlled compositions. The nanostructures made by this process can potentially be used as electrocatalysts for fuel cells, electrodes in batteries and supercapacitors, conductive inks for printed electronics, catalysts in wastewater treatment, and many other applications where a graphitized carbon-metal nanomaterial is needed.


Effect of cholecalciferol replacement on vascular calcification and left ventricular mass index in dialysis patients.
The aim of this study was to determine the effect of oral cholecalciferol treatment on vascular calcification, left ventricular mass index (LVMI) and other cardiac functions in dialysis patients. A six-month course of oral cholecalciferol treatment was recommended to dialysis patients with vitamin D insufficiency. While 26 patients were given cholecalciferol treatment, 17 patients who could not tolerate to therapy received standard therapy. Initial biochemical parameters were measured, and they were measured again after 6 months of treatment. Echocardiographic measurements were also performed, and the vascular calcification score (VCS) was calculated at baseline and at the 6th month. The cholecalciferol replacement group showed no significant change in LVMI and VCS values (p > 0.05). However, while LVMI was similar between groups at initial evaluation, it was lower in the cholecalciferol group at the 6th month when compared to the standard treatment group (141.8 ± 40.2 g/m(2) vs. 166.3 ± 31.4 g/m(2); p = 0.04). Likewise, left ventricular diastolic diameters (48.8 ± 5.1 mm vs. 47.5 ± 4.6 mm; p = 0.023) and left atrial diameters (41.2 ± 8.9 mm vs. 38.9 ± 8.1 mm; p = 0.006) decreased in the cholecalciferol group. Additionally, significant increases were observed in serum 25-hydroxyvitamin D (25(OH)D) and albumin levels, with a significant decrease in serum C-reactive protein levels. A lesser increase in left ventricular mass and better diastolic functions was observed in dialysis patients after 6 months of cholecalciferol treatment.


Detecting in vivo urokinase plasminogen activator activity with a catalyCEST MRI contrast agent.
Urokinase plasminogen activator (uPA) promotes tumor invasion and metastasis. The monitoring of uPA activity using molecular imaging may have prognostic value and be predictive for response to anti-cancer therapies. However, the detection of in vivo enzyme activity with molecular imaging remains a challenge. To address this problem, we designed a nonmetallic contrast agent, GR-4Am-SA, that can be detected with chemical exchange saturation transfer (CEST) MRI. This agent has a peptide that is cleaved by uPA, which causes a CEST signal at 5.0 ppm to decrease, and also has a salicylic acid moiety that can produce a CEST signal at 9.5 ppm, which is largely unresponsive to enzyme activity. The two CEST signals were used to determine a reaction coordinate, representing the extent of enzyme-catalyzed cleavage of the GR-4Am-SA agent during an experimental study. Initial biochemical studies showed that GR-4Am-SA could detect uPA activity in reducing conditions. Subsequently, we used our catalyCEST MRI protocol with the agent to detect the uPA catalysis of GR-4Am-SA in a flank xenograft model of Capan-2 pancreatic cancer. The results showed an average reaction coordinate of 80% ± 8%, which was strongly dependent on the CEST signal at 5.0 ppm. The relative independence of the reaction coordinate on the CEST signal at 9.5 ppm showed that the detection of enzyme activity was largely independent of the concentration of GR-4Am-SA within the tumor tissue. These results demonstrated the advantages of a single CEST agent with biomarker-responsive and unresponsive signals for reliably assessing enzyme activity during in vivo cancer studies.


Endovascular treatment of pericallosal aneurysms.
Pericallosal artery aneurysms are uncommon. Their treatment strategy, surgical or endovascular, will present specific challenges. The objective of the study was to compare risks of coil therapy and the recurrence rate of pericallosal artery aneurysms with aneurysms in other intradural locations. The authors examined data that were stored in a prospectively collected database for pericallosal artery aneurysms in patients who underwent coil placement between 1992 and 2005. Hemorrhagic and thromboembolic complications as well as clinical and angiographic outcomes were reviewed. Angiographically documented recurrences were classified as minor or major. These lesions were compared with a historical cohort of non-pericallosal artery aneurysms in patients who underwent coil therapy between 1992 and 2002. The known risk factors for recurrence and procedure-related hemorrhagic complications were evaluated in both groups to assess baseline imbalances. During a 13-year period, 25 pericallosal artery aneurysms were treated with coils in 25 patients. The non-pericallosal artery lesion group included 488 aneurysms of which 344 underwent follow-up imaging. Procedure-related perforations were more frequent for pericallosal artery aneurysms than those in other intradural locations (three of 25 compared with eight of 476, respectively; risk ratio 7.1, 95% confidence interval [CI] 2.1-22.5, p = 0.03). Follow-up imaging studies (obtained at a mean 28 months) were available for 19 patients with pericallosal artery aneurysms. The recurrence rate was not significantly higher in these patients (22.9/100 person-years of observation) than in those with non-pericallosal artery aneurysms (17.9/100 person-years of observation) (incidence rate ratio 1.3, 95% CI 0.6-2.4, p = 0.46). Pericallosal artery aneurysms were associated with significantly higher periprocedural rupture than non-pericallosal artery lesions. No significant intergroup difference was found for aneurysm recurrence.


Folate status during pregnancy in women is improved by long-term high vegetable intake compared with the average western diet.
The effect of increasing dietary folate on folate status during pregnancy is controversial. The aim of this study was to compare folate intake and folate status during pregnancy of women with high long-term vegetable intake and those eating an average Western diet. In a prospective study that included 109 participants, pregnant women adhering to a predominant vegetarian diet with high vegetable intake for 8 +/- 0.5 y with subgroups of ovo-lacto vegetarians (n = 27) and low meat eaters (n = 43) and women eating an average Western diet (control group, n = 39) were compared with regard to dietary intake and plasma and red blood cell (RBC) folate concentrations during wk 9-12, 20-22 and 36-38 of gestation. Plasma and RBC folate concentrations were highest in ovo-lacto vegetarians, followed by low meat eaters and lowest in the controls. Ovo-lacto vegetarians and low meat eaters showed a lower risk for folate deficiency, with RBC folate concentrations of <320 nmol/L resulting in odds ratios of 0.10 (95% confidence interval, 0.01-0.56) and 0.52 (95% confidence interval, 0.20-1.34), respectively. In ovo-lacto vegetarians, the RBC folate concentration was positively related to the intake of vitamin B-12 (r = 0.51, P: < 0.0001). The results of the study suggest that long-term high vegetable intake favorably affects plasma folate as well as RBC folate concentrations throughout pregnancy and reduces the risk of folate deficiency if an adequate vitamin B-12 supply is ensured.


[Early relaparotomies in relation to mechanical intestinal obstruction].
The article is concerned with the clinical picture, diagnosis and nearest results of the treatment of 51 patients with acute mechanical ileus resulting from different surgical interventions on organs of the abdominal cavity and small pelvis within the first 3 weeks after operations. Fourty four patients recovered, seven patients died (13,7%) due to intoxication and resulting cardio-pulmonary complications. Relaparatomy for acute mechanical ileus is believed to be an important link in the complex of postoperative measures.


Investigation into the influence of a high fat diet on antipsychotic-induced weight gain in female rats.
Atypical antipsychotic drug therapy may result in substantial weight gain, increased adiposity and the promotion of metabolic abnormalities. The mechanism(s) which underlie such effects remain unclear. Previous studies in our laboratory have demonstrated significant weight gain in female rats maintained on a standard laboratory diet after sub-chronic administration of olanzapine and risperidone, but not ziprasidone. The aim of this paper is to investigate the effect of antipsychotic drugs on body weight, ingestive behaviour and adiposity in female rats with access to a high fat diet. Adult female rats given free access to a high fat diet received either olanzapine (2 mg/kg), risperidone (0.5 mg/kg), ziprasidone (2.5 mg/kg) or vehicle for 28 days. Body weight, food and water intake in addition to intra-abdominal fat deposition were assessed. Olanzapine initially increased body weight but by the end of the study olanzapine animals appeared to have lost weight compared to the vehicle-treated group. Olanzapine-induced reductions in body weight were accompanied by a significant hypophagia during weeks 3 and 4. Risperidone increased body weight during week 1 only and reduced intake of a high fat diet during weeks 3 and 4. Ziprasidone was without effect on indices of body weight and ingestive behaviour. There were no effects of antipsychotic drugs on intra-abdominal fat deposition. Access to a diet high in fat attenuated weight gain induced by olanzapine and risperidone in female rats.


Effect of different levels of dietary trans fat or corn oil on azoxymethane-induced colon carcinogenesis in F344 rats.
The effect of various levels of dietary corn oil or trans fat on azoxymethane (AOM; CAS: 25843-45-2)-induced carcinogenesis was investigated in female F344 rats fed the AIN-76 semipurified diets. Starting at 5 weeks of age, groups of rats were fed the low-fat diet containing 5% corn oil (designated as low-fat control diet). At 7 weeks of age, all animals except the vehicle-treated controls, were given sc injections of AOM (15 mg/kg body wt, once weekly) for 3 weeks. After 1 week, groups of animals were transferred to semipurified diets containing 13.6% corn oil and 23.5% corn oil or high-fat diets containing 5.9% corn oil plus 5.9% trans fat plus 11.8% Oleinate (low trans fat), 5.9% corn oil plus 11.8% trans fat plus 5.9% Oleinate (intermediate trans fat), and 5.9% corn oil plus 17.6% trans fat (high trans fat). Fecal bile acids were measured in vehicle-treated rats. All animals were necropsied 34 weeks after the last AOM injection. The animals fed the 23.5% corn oil diet had a higher incidence of colon tumors than did those in the groups fed the 5 and 13.6% corn oil diets. There was no difference in colon tumor incidence between the 5 and 13.6% corn oil diet groups. The animals fed the high-fat diets containing low trans fat, intermediate trans fat, and high trans fat developed significantly fewer liver and colon tumors and more small intestinal tumors than did the rats fed 23.5% corn oil diet. The excretion of fecal deoxycholic acid, lithocholic acid, and 12-ketolithocholic acid was higher in animals fed the 23.5% corn oil diet compared to the excretion in animals fed the other diets.


Mitochondrial 16S rRNA Is Methylated by tRNA Methyltransferase TRMT61B in All Vertebrates.
The mitochondrial ribosome, which translates all mitochondrial DNA (mtDNA)-encoded proteins, should be tightly regulated pre- and post-transcriptionally. Recently, we found RNA-DNA differences (RDDs) at human mitochondrial 16S (large) rRNA position 947 that were indicative of post-transcriptional modification. Here, we show that these 16S rRNA RDDs result from a 1-methyladenosine (m1A) modification introduced by TRMT61B, thus being the first vertebrate methyltransferase that modifies both tRNA and rRNAs. m1A947 is conserved in humans and all vertebrates having adenine at the corresponding mtDNA position (90% of vertebrates). However, this mtDNA base is a thymine in 10% of the vertebrates and a guanine in the 23S rRNA of 95% of bacteria, suggesting alternative evolutionary solutions. m1A, uridine, or guanine may stabilize the local structure of mitochondrial and bacterial ribosomes. Experimental assessment of genome-edited Escherichia coli showed that unmodified adenine caused impaired protein synthesis and growth. Our findings revealed a conserved mechanism of rRNA modification that has been selected instead of DNA mutations to enable proper mitochondrial ribosome function.


Metabolic Switching of Tumor Cells under Hypoxic Conditions in a Tumor-on-a-chip Model.
Hypoxia switches the metabolism of tumor cells and induces drug resistance. Currently, no therapeutic exists that effectively and specifically targets hypoxic cells in tumors. Development of such therapeutics critically depends on the availability of in vitro models that accurately recapitulate hypoxia as found in the tumor microenvironment. Here, we report on the design and validation of an easy-to-fabricate tumor-on-a-chip microfluidic platform that robustly emulates the hypoxic tumor microenvironment. The tumor-on-a-chip model consists of a central chamber for 3D tumor cell culture and two side channels for medium perfusion. The microfluidic device is fabricated from polydimethylsiloxane (PDMS), and oxygen diffusion in the device is blocked by an embedded sheet of polymethyl methacrylate (PMMA). Hypoxia was confirmed using oxygen-sensitive probes and the effect on the 3D tumor cell culture investigated by a pH-sensitive dual-labeled fluorescent dextran and a fluorescently labeled glucose analogue. In contrast to control devices without PMMA, PMMA-containing devices gave rise to decreases in oxygen and pH levels as well as an increased consumption of glucose after two days of culture, indicating a rapid metabolic switch of the tumor cells under hypoxic conditions towards increased glycolysis. This platform will open new avenues for testing anti-cancer therapies targeting hypoxic areas.


Constitutive expression of the Saccharomyces cerevisiae CUP1 gene in Kluyveromyces lactis.
Shuttle plasmids, pE1.CUP1B and pE1.CUP1E of 10.6 kb, have been constructed between the metallothionein-encoding CUP1 gene of Saccharomyces cerevisiae and a vector capable of replication in Kluyveromyces lactis. Introduction of these plasmids into K. lactis confers resistance to copper as well as to cadmium and silver. Resistance to these latter metal ions, in the absence of induction by copper, suggested that the CUP1 gene is constitutively expressed in the foreign background. Introduction of the lacZ reporter gene from Escherichia coli into a cloning site downstream from the CUP1 promoter showed that expression of this gene is constitutive in K. lactis but in S. cerevisiae induction by copper is necessary. Sequences upstream from the CUP1 promoter are involved in the constitutive expression since deletion of 91 nucleotides from this region abolishes metal resistance. It is suggested that a K. lactis protein, normally involved in activating transcription of the resident CUP1 gene in the presence of copper, can promote transcription in the absence of metal ion by binding to the upstream activation sequence of the introduced CUP1 gene.


The impact of bladder neck sparing on urinary continence during laparoscopic radical prostatectomy; Results from a high volume centre.
To evaluate the effects of bladder neck reconstruction techniques on early continence after laparoscopic radical prostatectomy (LRP). This non-randomized retrospective study analyzed prospectively collected data concerning LRP. In total, 3107 patients underwent LRP between March 1999 and December 2016. Exclusion criteria were preoperative urinary incontinence, previous history of external beam radiotherapy, co-morbities which may affect urinary continence such as diabetes mellitus and/or neurogenic disorders, irregular followup, and follow-up shorter than 24 months. All patients were divided into one of three groups, posterior reconstruction being performed in Group 1 (n = 112), anterior reconstruction in Group 2 (n = 762), and bladder neck sparing (BNS) in Group 3 (n = 987). Demographic and pre-, peri-, and postoperative data were collected. Multivariate analyses were performed to determine factors affecting early continence after LRP. 1861 patients were enrolled in the study. The mean follow-up period was 48.12 ± 29.8 months, and subjects' mean age was 63.6 ± 6.2 years. There was no significant difference among the groups in terms of demographic or preoperative data. Postoperative data, including oncological outcomes, were similar among the groups. The level of early continence was higher in Group 3 than in the other groups (p < 0.001). Multivariate analyses identified BNS and age as parameters significantly affecting early continence levels after LRP (p < 0.001 and p < 0.001, respectively). Bladder neck reconstruction provided less earlier continence than BNS.


Origin and consequences of the relationship between protein mean and variance.
Cell-to-cell variance in protein levels (noise) is a ubiquitous phenomenon that can increase fitness by generating phenotypic differences within clonal populations of cells. An important challenge is to identify the specific molecular events that control noise. This task is complicated by the strong dependence of a protein's cell-to-cell variance on its mean expression level through a power-law like relationship (σ2∝μ1.69). Here, we dissect the nature of this relationship using a stochastic model parameterized with experimentally measured values. This framework naturally recapitulates the power-law like relationship (σ2∝μ1.6) and accurately predicts protein variance across the yeast proteome (r2 = 0.935). Using this model we identified two distinct mechanisms by which protein variance can be increased. Variables that affect promoter activation, such as nucleosome positioning, increase protein variance by changing the exponent of the power-law relationship. In contrast, variables that affect processes downstream of promoter activation, such as mRNA and protein synthesis, increase protein variance in a mean-dependent manner following the power-law. We verified our findings experimentally using an inducible gene expression system in yeast. We conclude that the power-law-like relationship between noise and protein mean is due to the kinetics of promoter activation. Our results provide a framework for understanding how molecular processes shape stochastic variation across the genome.


Nurses who work in rural and remote communities in Canada: a national survey.
In Canada, as in other parts of the world, there is geographic maldistribution of the nursing workforce, and insufficient attention is paid to the strengths and needs of those providing care in rural and remote settings. In order to inform workforce planning, a national study, Nursing Practice in Rural and Remote Canada II, was conducted with the rural and remote regulated nursing workforce (registered nurses, nurse practitioners, licensed or registered practical nurses, and registered psychiatric nurses) with the intent of informing policy and planning about improving nursing services and access to care. In this article, the study methods are described along with an examination of the characteristics of the rural and remote nursing workforce with a focus on important variations among nurse types and regions. A cross-sectional survey used a mailed questionnaire with persistent follow-up to achieve a stratified systematic sample of 3822 regulated nurses from all provinces and territories, living outside of the commuting zones of large urban centers and in the north of Canada. Rural workforce characteristics reported here suggest the persistence of key characteristics noted in a previous Canada-wide survey of rural registered nurses (2001-2002), namely the aging of the rural nursing workforce, the growth in baccalaureate education for registered nurses, and increasing casualization. Two thirds of the nurses grew up in a community of under 10 000 people. While nurses' levels of satisfaction with their nursing practice and community are generally high, significant variations were noted by nurse type. Nurses reported coming to rural communities to work for reasons of location, interest in the practice setting, and income, and staying for similar reasons. Important variations were noted by nurse type and region. The proportion of the rural nursing workforce in Canada is continuing to decline in relation to the proportion of the Canadian population in rural and remote settings. Survey results about the characteristics and practice of the various types of nurses can support workforce planning to improve nursing services and access to care.


Effects of Sustained Treatment With Lixisenatide on Gastric Emptying and Postprandial Glucose Metabolism in Type 2 Diabetes: A Randomized Controlled Trial.
Tachyphylaxis for slowing of gastric emptying is seen with continuous exposure to glucagon-like peptide 1 (GLP-1). We therefore aimed to establish whether prolonged use of a "short-acting" GLP-1 receptor agonist, lixisenatide, achieves sustained slowing of gastric emptying and reduction in postprandial glycemia. A total of 30 patients with metformin-treated type 2 diabetes underwent assessment of gastric emptying (scintigraphy) and glucose metabolism (dual tracer technique) after a 75-g glucose drink, before and after 8 weeks' treatment with lixisenatide (20 μg subcutaneously daily) or placebo, in a double-blind randomized parallel design. Gastric retention of the glucose drink was markedly increased after lixisenatide versus placebo (ratio of adjusted geometric means for area under the curve [AUC] over 240 min of 2.19 [95% CI 1.82, 2.64], P < 0.001), associated with substantial reductions in the rate of systemic appearance of oral glucose (P < 0.001) and incremental AUC for blood glucose (P < 0.001). Lixisenatide suppressed both glucagon (P = 0.003) and insulin (P = 0.032), but not endogenous glucose production, over 120 min after oral glucose intake. Postprandial glucose lowering over 240 min was strongly related to the magnitude of slowing of gastric emptying by lixisenatide (r = -0.74, P = 0.002) and to the baseline rate of emptying (r = 0.52, P = 0.048) but unrelated to β-cell function (assessed by β-cell glucose sensitivity). Eight weeks' treatment with lixisenatide is associated with sustained slowing of gastric emptying and marked reductions in postprandial glycemia and appearance of ingested glucose. Short-acting GLP-1 receptor agonists therefore potentially represent an effective long-term therapy for specifically targeting postprandial glucose excursions.


Loss of heterozygosity on chromosome 22q in gastrointestinal stromal tumors (GISTs): a study on 50 cases.
Mutational activation of KIT or PDGFRA is considered an early step in pathogenesis of gastrointestinal stromal tumors (GISTs); however, other nonrandom genetic changes have also been identified. At least three common regions of deletions on chromosome 22q, which may harbor putative tumor suppressor genes, have been defined. However, mapping of these regions has been inconsistent. It has also been speculated that GI autonomous nerve tumors (GANTs), GISTs with ultrastructural features suggestive of autonomic nerve differentiation, are characterized by a specific deletion involving 22q13 cytogenetic region. This study was undertaken to evaluate loss of heterozygosity (LOH) on chromosome 22q in 50 GISTs, including 10 GANTs. Four tumors were incidental minimal lesions <or=10 mm in diameter. LOH was evaluated using 20 PCR-based microsatellite markers and capillary gel electrophoresis. In all, 15 (30%) cases showed LOH of more than 75% of informative markers, suggesting loss of chromosome 22q. A total of 24 GISTs (50%) revealed LOH of one to seven informative markers clustered in different loci suggesting simultaneous involvement of different regions. The highest frequency of LOH was seen at D22S922 and D22S425, mapped to 22q13.33 and 22q11.22, respectively. However, LOH at other regions including IL2RB and NF2 locus was also found. No NF2 mutations were identified in four analyzed tumors. LOH on chromosome 22q was more frequent among intestinal than among gastric GISTs; however, there was no difference between LOH pattern seen in tumors defined by different histologic, ultrastructural (GANT) and molecular features (KIT and PDGFRA mutations). Although minimal GISTs revealed LOH on chromosome 22q, there was a higher LOH frequency in malignant than in benign tumors. An isolated LOH at D22S425 was equally found in both benign and malignant tumors. These observations may suggest that LOHs on chromosome 22q in GISTs play a role in early stages of tumor formation as well as in late tumor progression.


Analysis of Caesarean Section Rates Using the Robson Classification System at a University Hospital in Spain.
The WHO recommends the use of the Robson ten-group classification system (RTGCS) as an effective monitoring and analysis tool to assess the use of caesarean sections (CS). The present study aimed to conduct an analysis of births using the RTGCS in La Ribera University Hospital over nine years and to assess the levels and trends of CS births. Retrospective study between January 1, 2010, and December 31, 2018. All eligible women were allocated in RTGCS to determine the absolute and relative contribution made by each group to the overall CS rate; linear regression and weighted least squares regression analysis were used to analyze trends over time. The risk of CS of women with induced versus spontaneous onset of labor was calculated with an odds ratio (OR) with a 95% CI. 16,506 women gave birth during the study period, 19% of them by CS. Overall, 20.4% of women were in group 1 (nulliparous, singleton cephalic, term, spontaneous labor), 29.4% in group 2 (nulliparous, singleton cephalic, term, induced labor or caesarean before labor), and 12.8% in group 4 (multiparous, singleton cephalic, term, induced or caesarean delivery before labor) made the most significant contributions to the overall rate of CS; Conclusions: In our study, Robson Groups 1, 2, and 4, were identified as the main contributors to the hospital's overall CS rate. The RTGCS provides an easy way of collecting information about the CS rate, is a valuable clinical method that allows standardized comparison of data, and time point, and identifies the groups driving changes in CS rates.


Are Helicobacter pylori and other Helicobacter species infection associated with human biliary lithiasis? A meta-analysis.
Since the isolation of Helicobacter species in biliary system, a hypothetical question was raised about the role of these agents in the development of cholelithiasis. This meta-analysis is to explore the association between the Helicobacter infection and biliary lithiasis. A systematic literature search was performed to identify all eligible articles. Meta-analysis which was carried out using odds ratio and random effect model, 95% confidence intervals for odds ratio was calculated. Quantitative assessment of heterogeneity was explored by chi-square test with significance set at P value 0.10 and was measured using I(2) statistic. Eighteen studies published between 1998 and 2011 were finally eligible for meta-analysis. H. pylori, H. bilis, H. hepaticus, H. pullorum and H. ganmani were studied. With heterogeneity (I(2) = 69.5%, P<0.0001), significantly higher pooled infection rates of H. pylori (OR: 2.59, 35.82% versus 26.75%, P = 0.01) and H. hepaticus (OR: 3.13, 31.30% versus 12.12%, P = 0.02) were observed in lithiasis group. Higher prevalence of H. pylori in cholelithiasis patients were reported by studies from East Asia, South Asia and South America. Evidences supporting the higher presence of H. pylori in cholelithiasis patients could be found by PCR for detecting 16s rRNA in bile, 26 kDa protein gene in biliary tissue and immunohistochemistry. Using multiple detection tests could increase the detection rate of H. pylori. Our meta-analysis suggests a trend of higher presence of H. pylori in cholelithiasis patients than control group and this trend was significant in the regions with higher prevalence of this agent. Evidences supporting the association between Helicobacter and cholelithiasis could be found by using different tests but the gold standard for the identification of these bacteria in biliary system has yet to be established. Considering obvious heterogeneity, a large multi-center study will facilitate us to further clarify the association between the Helicobacter infection and cholelithiasis.


Production and Structural Diversification of Withanolides by Aeroponic Cultivation of Plants of Solanaceae: Cytotoxic and Other Withanolides from Aeroponically Grown Physalis coztomatl.
Withanolides constitute one of the most interesting classes of natural products due to their diversity of structures and biological activities. Our recent studies on withanolides obtained from plants of Solanaceae including Withania somnifera and a number of Physalis species grown under environmentally controlled aeroponic conditions suggested that this technique is a convenient, reproducible, and superior method for their production and structural diversification. Investigation of aeroponically grown Physalis coztomatl afforded 29 withanolides compared to a total of 13 obtained previously from the wild-crafted plant and included 12 new withanolides, physacoztolides I-M (9-13), 15α-acetoxy-28-hydroxyphysachenolide C (14), 28-oxophysachenolide C (15), and 28-hydroxyphysachenolide C (16), 5α-chloro-6β-hydroxy-5,6-dihydrophysachenolide D (17), 15α-acetoxy-5α-chloro-6β-hydroxy-5,6-dihydrophysachenolide D (18), 28-hydroxy-5α-chloro-6β-hydroxy-5,6-dihydrophysachenolide D (19), physachenolide A-5-methyl ether (20), and 17 known withanolides 3-5, 8, and 21-33. The structures of 9-20 were elucidated by the analysis of their spectroscopic data and the known withanolides 3-5, 8, and 21-33 were identified by comparison of their spectroscopic data with those reported. Evaluation against a panel of prostate cancer (LNCaP, VCaP, DU-145, and PC-3) and renal carcinoma (ACHN) cell lines, and normal human foreskin fibroblast (WI-38) cells revealed that 8, 13, 15, and 17-19 had potent and selective activity for prostate cancer cell lines. Facile conversion of the 5,6-chlorohydrin 17 to its 5,6-epoxide 8 in cell culture medium used for the bioassay suggested that the cytotoxic activities observed for 17-19 may be due to in situ formation of their corresponding 5β,6β-epoxides, 8, 27, and 28.


Polycomb YY1 is a critical interface between epigenetic code and miRNA machinery after exposure to hypoxia in malignancy.
Yin Yang 1 (YY1) is a member of polycomb protein family involved in epigenetic modifications and transcriptional controls. We have shown that YY1 acts as positive regulator of tumor growth and angiogenesis by interfering with the VEGFA network. Yet, the link between polycomb chromatin complex and hypoxia regulation of VEGFA is still poorly understood. Here, we establish that hypoxia impairs YY1 binding to VEGFA mRNA 3'UTR (p<0.001) in bone malignancy. Moreover, RNA immunoprecipitation reveals the formation of triplex nuclear complexes among YY1, VEGFA DNA, mRNA, and unreached about 200 fold primiRNA 200b and 200c via Dicer protein. In this complex, YY1 is necessary to maintain the steady-state level of VEGFA expression while its silencing increases VEGFA mRNA half-life at 4 h and impairs the maturation of miRNA 200b/c. Hypoxia promotes histone modification through ubiquitination both of YY1 and Dicer proteins. Hypoxia-mediated down-regulation of YY1 and Dicer changes post-transcriptional VEGFA regulation by resulting in the accumulation of primiRNA200b/c in comparison to mature miRNAs (p<0.001). Given the regulatory functions of VEGFA on cellular activities to promote neoangiogenesis, we conclude that YY1 acts as novel critical interface between epigenetic code and miRNAs machinery under chronic hypoxia in malignancy.


Huntingtin cleavage product A forms in neurons and is reduced by gamma-secretase inhibitors.
The mutation in Huntington's disease is a polyglutamine expansion near the N-terminus of huntingtin. Huntingtin expressed in immortalized neurons is cleaved near the N-terminus to form N-terminal polypeptides known as cleavage products A and B (cpA and cpB). CpA and cpB with polyglutamine expansion form inclusions in the nucleus and cytoplasm, respectively. The formation of cpA and cpB in primary neurons has not been established and the proteases involved in the formation of these fragments are unknown. Delivery of htt cDNA into the mouse striatum using adeno-associated virus or into primary cortical neurons using lentivirus generated cpA and cpB, indicating that neurons in brain and in vitro can form these fragments. A screen of small molecule protease inhibitors introduced to clonal striatal X57 cells and HeLa cells identified compounds that reduced levels of cpA and are inhibitors of the aspartyl proteases cathepsin D and cathepsin E. The most effective compound, P1-N031, is a transition state mimetic for aspartyl proteases. By western blot analysis, cathepsin D was easily detected in clonal striatal X57 cells, mouse brain and primary neurons, whereas cathepsin E was only detectible in clonal striatal X57 cells. In primary neurons, levels of cleavage product A were not changed by the same compounds that were effective in clonal striatal cells or by mRNA silencing to partially reduce levels of cathepsin D. Instead, treating primary neurons with compounds that are known to inhibit gamma secretase activity either indirectly (Imatinib mesylate, Gleevec) or selectively (LY-411,575 or DAPT) reduced levels of cpA. LY-411,575 or DAPT also increased survival of primary neurons expressing endogenous full-length mutant huntingtin. We show that cpA and cpB are produced from a larger huntingtin fragment in vivo in mouse brain and in primary neuron cultures. The aspartyl protease involved in forming cpA has cathepsin-D like properties in immortalized neurons and gamma secretase-like properties in primary neurons, suggesting that cell type may be a critical factor that specifies the aspartyl protease responsible for cpA. Since gamma secretase inhibitors were also protective in primary neurons, further study of the role of gamma-secretase activity in HD neurons is justified.


A clinical evaluation of treatments that target cell cycle machinery in breast cancer.
Introduction: The dysregulation of cell cycle control can lead to cancer development. In breast cancer, cyclin D, CDK 4,6 and the retinoblastoma protein play a central role in the control of cell proliferation, in crosstalk with the estrogen receptor and Her2 pathways. Although the mechanisms by which the CDK4/6 complex is involved in the control of cell growth in triple negative breast cancer (TNBC) are still unclear, some TNBCs might be sensitive to CDK4/6 inhibitors.Areas covered: The authors provide an overview of the treatments that target cell cycle machinery in breast cancer and provide their perspectives for the future.Expert opinion: CDK 4/6 inhibitors are active drugs in HR+ MBC, but some unresolved issues remain. We need to identify biomarkers of response. Moreover, we need to determine the optimal timing for the incorporation of CDK 4/6 inhibitors in the current treatment algorithm. In the Her2 positive subtype, the triple combination of anti Her2 therapies with CDK4/6 inhibitors and endocrine therapy seems to be a promising chemotherapy free approach. Efforts must still be made for the treatment of the TNBC subtype, even though new CDK 4/6 combinations are emerging as promising approaches to selected patients.


Vaginal aerobic bacteria of healthy bitches and those with fertility problems.
The most common problems in veterinary practice in bitches are bacterial infections of the reproductive tract associated with fertility problems. Research to determine the correlation between the health status of female dogs and bacterial flora of the genital tract has been ongoing for years, but the results obtained by different authors are often contradictory, and do not always concern breeding bitches. Our study identified the most common aerobic bacteria in the genital tract of numerous breeding bitches population. A total of 275 breeding dogs in anestrous phase of the estrous cycle were included in this study. 198 were qualified to the first group with no genital tract infections and no reproductive disorders. 68 bitches were qualified to the second group with complications such as: infertility, abortion, foetus resorptions and newborn mortality. The type of bacterial isolates was almost the same in the healthy bitches and the group with fertility problems. The most common bacteria obtained from the vaginal tract of the tested dogs were Streptococcus spp., Staphylococcus spp., Mycoplasma canis and Escherichia coli. There were no significant differences in bacterial prevalence in the group with reproductive problems versus healthy dogs; however, we found a statistically significant difference between both groups when the numbers of bacterial strains were compared. The number of one-strain bitches was statistically higher in the problematic group than in the non-problematic one. Bacterial culturing of vaginal swab specimens from breeding bitches without clinical signs of genital disease is of little value. Furthermore, it should always be preceded by an examination (clinical, cytological or vaginoscopy etc.). The request or requirement to perform vaginal cultures that is made by some breeders, while common, is not diagnostic for any pathologic condition and the results of these cultures should never be used to determine if antibiotic therapy is indicated.


Increased phototoxicity of hydrochlorothiazide by photodegradation.
The photodegradation products of hydrochlorothiazide produced by ultraviolet (UV) radiation were investigated for their phototoxicity utilizing the photohemolysis and Candida albicans tests. Hydrochlorothiazide was irradiated for 30, 60, 90 and 120 min with a 250 W xenon arc lamp using a WG295 cut-off filter. Irradiation of hydrochlorothiazide resulted in the gradual decrease of all three absorption bands (225, 270 and 320 nm), the blue shift of the 225 nm band, and the appearance of a new band around 290 nm. Since previous results demonstrated that photosubstitution of chloride could occur, the main product of this photolysis most likely is ethoxyhydrochlorothiazide. The photohemolysis test revealed a significant increase in photohemolysis observed in the photodegradation products produced after 60, 90 and 120 min of UV irradiation. This increase in hemolysis value directly correlated with the UV-irradiation time. However, there was no significant phototoxic killing of yeast in the Candida albicans test. This suggests photodegradation products of hydrochlorothiazide may play an important role in phototoxicity by acting on the cell membrane, but not on DNA. Considering the high in vitro phototoxicity observed in bendroflumethiazide and the data presented here, substitution of chloride seems to be responsible for the increased phototoxicity of hydrochlorothiazide.


Hepatitis C virus infection in patients with hemophilia in Isfahan, Iran.
Patients with hereditary bleeding disorders are at risk of viral infection such as hepatitis C due to frequent transfusion of blood and blood products. This study aimed to determine the prevalence of hepatitis C and associated risk factors in hemophilic patients in Isfahan, the second big province in Iran. In a descriptive study, patients with hemophilia in Isfahan province were enrolled. A questionnaire, including demographic and risk factors of hepatitis C was completed through a structured interview with closed questions by a trained interviewer for each patient and HCV-Ab test results were extracted from patient records. In this study, 232 of 350 patients with hemophilia A and B (66%) were positive for hepatitis C. Based on Multivariate Logistic Regression model, no independent risk factor was found. Prevalence of hepatitis C in patients with haemophilia A and B in Isfahan is high. Since no independent risk factor for hepatitis C disease was found in this high risk group, it can be concluded that multitransfusion is the only predictor for hepatitis C.


The challenges in developing a finite element injury model of the neck to predict the penetration of explosively propelled projectiles.
Neck injuries sustained by UK service personnel serving on current operations from explosively propelled fragments result in significant mortality and long-term morbidity. Many of these injuries could potentially have been prevented had the soldiers been wearing their issued neck collars at the time of injury. The aim of this research is to develop an accurate method of predicting the resultant damage to cervical neurovascular structures from explosively propelled fragments. A finite element numerical model has been developed based on an anatomically accurate, anthropometrically representative 3D mathematical mesh of cervical neurovascular structures. Currently, the model simulates the passage of a fragment simulating projectile through all anatomical components of the neck using material models based upon 20% ballistic gelatin on the simplification that all tissue types act like homogenous muscle. The material models used to define the properties of each element within the model will be sequentially replaced by ones specific to each individual tissue within an anatomical structure. However, the cumulative effect of so many additional variables will necessitate experimental validation against both animal models and post-mortem human subjects to improve the credibility of any predictions made by the model. We believe this approach will in the future have the potential to enable objective comparisons between the mitigative effects of different body armour systems to be made with resultant time and financial savings.


Combined zinc supplementation with proinsulin C-peptide treatment decreases the inflammatory response and mortality in murine polymicrobial sepsis.
Zinc is a trace element vital for immune function during host response to infection. The proinsulin C-peptide has been shown to exert beneficial effects through activation of the anti-inflammatory peroxisome proliferator-activated receptor γ (PPARγ) in experimental endotoxemia. Some in vitro activities of C-peptide appear dependent on the presence of zinc. We investigated the effect of zinc supplementation before onset of sepsis on the anti-inflammatory properties of C-peptide. Male C57BL/6 mice were subjected to polymicrobial sepsis by cecal ligation and puncture (CLP). Mice received zinc gluconate (1.3 mg/kg) intraperitoneally (i.p.) for 3 days before CLP. One hour after CLP, animals received C-peptide (280 nmol/kg i.p.) or the antimicrobial agent imipenem (25 mg/kg i.p.). Cecal ligation and puncture was associated with an 11% survival rate, pulmonary leukosequestration, and liver injury. Molecular analysis in lungs of septic mice showed increased nuclear activation of the proinflammatory extracellular signal-regulated kinases 1 and 2 and nuclear factor κB, but decreased PPARγ expression, when compared with sham animals. Combination of zinc supplementation with C-peptide posttreatment significantly improved survival rate (61%) similarly to antibiotic treatment (60%), ameliorated lung architecture and liver function, reduced tissue neutrophil infiltration, and increased bacterial clearance when compared with vehicle, C-peptide, or zinc treatment alone. These beneficial effects were associated with restored lung nuclear expression of PPARγ and reduction of phosphorylated extracellular signal-regulated kinases 1 and 2 and nuclear factor κB activities in comparison to vehicle or single treatment protocols. Our data demonstrate that short-term zinc prophylaxis before the infectious insult is a requisite for the anti-inflammatory properties of C-peptide by facilitating modulation of inflammatory pathways.


Good genes, oxidative stress and condition-dependent sexual signals.
The immune and the detoxication systems of animals are characterized by allelic polymorphisms, which underlie individual differences in ability to combat assaults from pathogens and toxic compounds. Previous studies have shown that females may improve offspring survival by selecting mates on the basis of sexual ornaments and signals that honestly reveal health. In many cases the expression of these ornaments appears to be particularly sensitive to oxidative stress. Activated immune and detoxication systems often generate oxidative stress by an extensive production of reactive metabolites and free radicals. Given that tolerance or resistance to toxic compounds and pathogens can be inherited, female choice should promote the evolution of male ornaments that reliably reveal the status of the bearers' level of oxidative stress. Hence, oxidative stress may be one important agent linking the expression of sexual ornaments to genetic variation in fitness-related traits, thus promoting the evolution of female mate choice and male sexual ornamentation, a controversial issue in evolutionary biology ever since Darwin.


Delta Inulin Adjuvant Enhances Plasmablast Generation, Expression of Activation-Induced Cytidine Deaminase and B-Cell Affinity Maturation in Human Subjects Receiving Seasonal Influenza Vaccine.
There is a major need for new adjuvants to improve the efficacy of seasonal and pandemic influenza vaccines. Advax is a novel polysaccharide adjuvant based on delta inulin that has been shown to enhance the immunogenicity of influenza vaccine in animal models and human clinical trials. To better understand the mechanism for this enhancement, we sought to assess its effect on the plasmablast response in human subjects. This pilot study utilised cryopreserved 7 day post-vaccination (7dpv) peripheral blood mononuclear cell samples obtained from a subset of 25 adult subjects from the FLU006-12 trial who had been immunized intramuscularly with a standard dose of 2012 trivalent inactivated influenza vaccine (TIV) alone (n=9 subjects) or combined with 5mg (n=8) or 10mg (n=8) of Advax adjuvant. Subjects receiving Advax adjuvant had increased 7dpv plasmablasts, which in turn exhibited a 2-3 fold higher rate of non-silent mutations in the B-cell receptor CDR3 region associated with higher expression of activation-induced cytidine deaminase (AID), the major enzyme controlling BCR affinity maturation. Together, these data suggest that Advax adjuvant enhances influenza immunity in immunized subjects via multiple mechanisms including increased plasmablast generation, AID expression and CDR3 mutagenesis resulting in enhanced BCR affinity maturation and increased production of high avidity antibody. How Advax adjuvant achieves these beneficial effects on plasmablasts remains the subject of ongoing investigation. Australia New Zealand Clinical Trials Register ACTRN12612000709842 https://www.anzctr.org.au/Trial/Registration/TrialReview.aspx?id=362709.


Parental Perceptions of Weight During the First Year of Life.
More than half of parents underestimate their overweight child's weight; however, previous research focuses on children older than 2 years of age. The objective of this study was to assess whether parents of 2- to 12-month-old infants are able to accurately perceive their children's weight status. We performed a cross-sectional analysis of data collected from the Greenlight study, a cluster randomized obesity prevention trial, at 4 pediatric clinics serving diverse and low-income populations. Infants' length and weight were measured at well-child checks, and parents completed questionnaires including demographic characteristics and perception of their children's weight. Weight-for-length (WFL) percentile at the fifth to ≤95 was considered healthy weight and WFL percentile >95th was considered overweight. We used chi-squared tests to compare accuracy according to weight category and performed logistic regression analysis to assess accuracy at each time point. Approximately 85% to 90% of infants (n = 853 at 2 months, n = 563 at 12 months) were at a healthy WFL at all measurement times, and parents of these infants were more likely to have an accurate perception of their child's weight (accuracy 89%-95%) than overweight children (accuracy 7%-26%; P < .001 across time points). Approximately 10% of healthy weight infants were perceived as underweight by their parents at all time points. At 12 months, mothers who were overweight were significantly more likely to underestimate their child's weight status (P = .008). In our diverse and low-income sample, parents of overweight infants infrequently know that their infants are overweight. Future studies should examine how perception is related to feeding habits and weight status over time.


Quantitative Susceptibility Mapping for Carotid Atherosclerotic Plaques: A Pilot Study.
Identifying plaque components such as intraplaque hemorrhage, lipid rich necrosis, and calcification is important to evaluate vulnerability of carotid atherosclerotic plaque; however, conventional vessel wall MR imaging may fail to discriminate plaque components. We aimed to evaluate the components of plaques using quantitative susceptibility mapping (QSM), a newly developed post-processing technique to provide voxel-based quantitative susceptibilities. Seven patients scheduled for carotid endarterectomy were enrolled. Magnitude and phase images of five-echo 3D fast low angle shot (FLASH) were obtained using a 3T MRI, and QSM was calculated from the phase images. Conventional carotid vessel wall images (black-blood T1-weighted images [T1WI], T2-weighted images [T2WI], proton-density weighted images [PDWI], and time-of-flight images [TOF]) were also obtained. Pathological findings including intraplaque hemorrhage, calcification, and lipid rich necrosis at the thickest plaque section were correlated with relative susceptibility values with respect to the sternocleidomastoid muscle on QSM. On conventional vessel wall images, the contrast-noise ratio (CNR) between the three components and sternocleidomastoid muscle was measured respectively. Wilcoxon signed-rank test analyses were performed to assess the relative susceptibility values and CNR. Pathologically, lipid rich necrosis was proved in all of seven cases, and intraplaque hemorrhage in five of seven cases. Mean relative susceptibility value of hemorrhage was higher than lipid rich necrosis unexceptionally (P = 0.0313). There were no significant differences between CNR of hemorrhage and lipid rich necrosis on all sequences. In all six cases with plaque calcification, susceptibility value of calcification was significantly lower than lipid rich necrosis unexceptionally (P = 0.0156). There were significant differences between CNRs of lipid rich necrosis and calcification on T1WI, PDWI, TOF (P < 0.05). QSM of carotid plaque would provide a novel quantitative MRI contrast that enables reliable differentiation among intraplaque hemorrhage, lipid rich necrosis, and calcification, and be useful to identify vulnerable plaques.