Vector analyses of linearly and circularly polarized Bessel beams using Hertz vector potentials.
Using the transverse Hertz vector potentials, vector analyses of linearly and circularly polarized Bessel beams of arbitrary orders are presented in this paper. Expressions for the electric and magnetic fields of vector Bessel beams in free space that are rigorous solutions to the vector Helmholtz equation are derived. Their respective time averaged energy density and Poynting vector are also obtained, in order to exhibit their non-diffracting properties. Polarization patterns and magnitude profiles with different parameters are displayed. Particular emphasis is placed on the cases where the ratio of wave number over its transverse component k/kt approximately equals to one and largely exceeds it, which corresponding to the nonparaxial and paraxial condition, respectively. These results allow us to recognize that the vector Bessel beams exhibit new and important features, compared with the scalar fields.


Prognostic factors in adults with knee pain in general practice.
To predict the 1-year outcome of incident nontraumatic knee symptoms in adults presenting in general practice. Adults age >35 years with nontraumatic knee symptoms (n = 480) were followed for 1 year. At baseline, data on knee symptoms and demographics were collected and a physical examination performed. Knee symptoms were assessed by self-report questionnaires at 3-month intervals. After 1 year the physical examination was repeated. Multivariate prognostic regression models of patient characteristics, symptom characteristics, and physical examination were used to predict persisting knee symptoms after 1 year. Areas under receiving operating characteristic curves (AUCs) were used to determine the predictive value of the model. To assess the added predictive value of symptom characteristics and physical examination, these models were added to the model of patient characteristics. The improvement was expressed as the difference between the 2 AUCs. In the multivariate prognostic model of patient characteristics, age >60 years, educational level, kinesophobia, and comorbidity of the skeletal system were associated with persistent knee symptoms after 1 year (AUC 0.67). Of the symptom characteristics, history of nontraumatic knee symptoms, bilateral symptoms, and duration of symptoms >3 months were associated (AUC 0.73). For determinants of physical examination, crepitus of passive extension was associated (AUC 0.55). The added value of the symptom characteristics model to the patient characteristics model was 0.09 (AUC 0.76). Physical examination added no further value. Symptom characteristics are the strongest predictors of persisting knee symptoms at 1-year followup. Physical examination has no added value in predicting persistent knee symptoms in general practice.


Polypoid nodular histiocytic hyperplasia associated with endometrioid adenocarcinoma of the endometrium: report of a case.
A 45 year old woman underwent Laparoscopy-assisted total hysterectomy with staging procedure following a diagnosis of endometrial endometrioid adenocarcinoma on her endometrial biopsy. The hysterectomy specimen showed a FIGO I stage 1a, endometrioid carcinoma. A separate polypoid lesion in the endometrium, distinct from the carcinoma, was also identified. Microscopically the polypoid lesion was "nodular histiocytic hyperplasia". The H&E, immunohistochemical staining findings and the differential diagnoses are discussed in this report. Although description of similar lesions is available in the literature, the current lesion is unique as it is identified in a hysterectomy specimen in its entirety and its association with an endometrial endometrioid carcinoma. The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1060511915121922.


Constraint-induced movement therapy in the adult rat after unilateral corticospinal tract injury.
Smaller spinal cord injuries often allow some degree of spontaneous behavioral improvements because of structural rearrangements within different descending fiber tracts or intraspinal circuits. In this study, we investigate whether rehabilitative training of the forelimb (forced limb use) influences behavioral recovery and plastic events after injury to a defined spinal tract, the corticospinal tract (CST). Female adult Lewis rats received a unilateral CST injury at the brainstem level. Use of the contralateral impaired forelimb was either restricted, by a cast, or forced, by casting the unimpaired forelimb immediately after injury for either 1 or 3 weeks. Forced use of the impaired forelimb was followed by full behavioral recovery on the irregular horizontal ladder, whereas animals that could not use their affected side remained impaired. BDA (biotinylated dextran amine) labeling of the intact CST showed lesion-induced growth across the midline where CST collaterals increased their innervation density and extended fibers toward the ventral and the dorsal horn in response to forced limb use. Gene chip analysis of the denervated ventral horn revealed changes in particular for growth factors, adhesion and guidance molecules, as well as components of synapse formation suggesting an important role for these factors in activity-dependent intraspinal reorganization after unilateral CST injury.


Genetic organization of the Pantoea stewartii subsp. stewartii hrp gene cluster and sequence analysis of the hrpA, hrpC, hrpN, and wtsE operons.
The hrp/wts gene cluster of Pantoea stewartii subsp. stewartii is required for pathogenicity on sweet corn and the ability to elicit a hypersensitive response (HR) in tobacco. Site-directed transposon mutagenesis and nucleotide sequencing were used to identify hrp/wts genes within the left 20 kb of this cluster. Seventeen open reading frames (ORFs) comprise seven genetic complementation groups. These ORFs share homology with hrp and dsp genes from Erwinia amylovora, Erwinia chrysanthemi, and Pseudomonas syringae pathovars and have been designated, in map order, wtsF, wtsE, hrpN, hrpV, hrpT, hrcC, hrpG, hrpF, hrpE, hrpD, hrcJ, hrpB, hrpA, hrpS, hrpY, hrpX, and hrpL. Putative hrp consensus promoter sequences were identified upstream of hrpA, hrpF, hrpN, and wtsE. Expression of the hrpA, hrpC, and wtsE operons was regulated by HrpS. Transposon mutations in all of the hrp operons abolished pathogenicity and HR elicitation, except for the hrpN and hrpV mutants, which were still pathogenic. hrpS, hrpXY, and hrpL regulatory mutations abolished HrpN synthesis, whereas secretory mutations in the hrpC, hrpA, and hrpJ operons permitted intracellular HrpN synthesis. wtsEF mutants were not pathogenic but still produced HrpN and elicited the HR. wtsE encodes a 201-kDa protein that is similar to DspE in E. amylovora and AvrE in P. syringae pv. tomato, suggesting that this protein is a major virulence factor involved in the elicitation of water-soaked lesions.


Cell-to-cell variability analysis dissects the plasticity of signaling of common γ chain cytokines in T cells.
Natural variability in the abundance of signaling regulators can lead to divergence in cell fate, even within genetically identical cells that share a common differentiation state. We introduce cell-to-cell variability analysis (CCVA), an experimental and computational methodology that quantifies the correlation between variability in signaling regulator abundance and variation in the sensitivity of cells to stimuli. With CCVA, we investigated the unexpected effects of the interleukin 2 (IL-2) receptor α chain (IL-2Rα) on the sensitivity of primary mouse T lymphocytes to cytokines that signal through receptors that have the common γ chain (γ(c)). Our work showed that increased IL-2Rα abundance decreased the concentration of IL-2 required for a half-maximal activation (EC(50)) of the downstream effector signal transducer and activator of transcription 5 (STAT5), but reduced the responsiveness to IL-7 or IL-15, without affecting the EC(50) values of other γ(c) cytokines. To investigate the mechanism of the effect of IL-2Rα on γ(c) cytokine signaling, we introduced a Bayesian-inference computational framework that models the formation of receptor signaling complexes with data from previous biophysical measurements. With this framework, we found that a model in which IL-2Rα drives γ(c) depletion through the assembly of functional IL-2R complexes was consistent with both the CCVA data and experimental measurements. The combination of CCVA and computational modeling produced quantitative understanding of the crosstalk between γ(c) cytokine receptor signaling in T lymphocytes.


A comparison of the occurrence of common dental abnormalities in stabled and free-grazing horses.
The purpose of this study was to gain evidence on the prevalence of dental abnormalities in stable-kept horses in comparison with free-living horses. It is expected that free-living horses that graze for as much as 16 h/day will have fewer dental abnormalities than stable-kept horses. In this study, the latter group was fed a diet that was based on a relatively high-energy, cereal-based feed. This was thought to be a representative of common practice in domesticated, stable-kept horses. Compound diets such as this have previously been shown to increase the frequency of chewing cycles and decrease mediolateral excursion. The occurrence of 10 named dental abnormalities present in the dentition of 60 Thoroughbred-type horses was recorded. Half of the population was at grass all year round in New South Wales, Australia. The remainder were stabled for 24 h/day in Gloucestershire, UK. All horses were between 5 and 15 years of age. The same, experienced, equine dental technician examined all horses using a full-mouth speculum and produced a routine dental chart. Stable-kept horses had a significantly higher total occurrence of abnormalities (P < 0.001) than free-living horses. The stable-kept group had a significantly higher prevalence of exaggerated transverse ridging across the occlusal surface of the cheek teeth, focal or ramped overgrowths of the cheek teeth and periodontal disease (P < 0.01 in all cases). All horses in both groups had some occurrence of sharp edges of the buccal and lingual edges of the cheek teeth. The results are in agreement with anecdotal evidence that a fibre-based, grazed diet results in fewer dental abnormalities. However, sharp edges may occur even with what is perceived as a 'natural' diet.


ABCA7 p.G215S as potential protective factor for Alzheimer's disease.
Genome-wide association studies (GWASs) have been effective approaches to dissect common genetic variability underlying complex diseases in a systematic and unbiased way. Recently, GWASs have led to the discovery of over 20 susceptibility loci for Alzheimer's disease (AD). Despite the evidence showing the contribution of these loci to AD pathogenesis, their genetic architecture has not been extensively investigated, leaving the possibility that low frequency and rare coding variants may also occur and contribute to the risk of disease. We have used exome and genome sequencing data to analyze the single independent and joint effect of rare and low-frequency protein coding variants in 9 AD GWAS loci with the strongest effect sizes after APOE (BIN1, CLU, CR1, PICALM, MS4A6A, ABCA7, EPHA1, CD33, and CD2AP) in a cohort of 332 sporadic AD cases and 676 elderly controls of British and North-American ancestry. We identified coding variability in ABCA7 as contributing to AD risk. This locus harbors a low-frequency coding variant (p.G215S, rs72973581, minor allele frequency = 4.3%) conferring a modest but statistically significant protection against AD (p-value = 0.024, odds ratio = 0.57, 95% confidence interval = 0.41-0.80). Notably, our results are not driven by an enrichment of loss of function variants in ABCA7, recently reported as main pathogenic factor underlying AD risk at this locus. In summary, our study confirms the role of ABCA7 in AD and provides new insights that should address functional studies.


Gene expression disruptions of organism versus organ in Drosophila species hybrids.
Hybrid dysfunctions, such as sterility, may result in part from disruptions in the regulation of gene expression. Studies of hybrids within the Drosophila simulans clade have reported genes expressed above or below the expression observed in their parent species, and such misexpression is associated with male sterility in multigenerational backcross hybrids. However, these studies often examined whole bodies rather than testes or had limited replication using less-sensitive but global techniques. Here, we use a new RNA isolation technique to re-examine hybrid gene expression disruptions in both testes and whole bodies from single Drosophila males by real-time quantitative RT-PCR. We find two early-spermatogenesis transcripts are underexpressed in hybrid whole-bodies but not in assays of testes alone, while two late-spermatogenesis transcripts seem to be underexpressed in both whole-bodies and testes alone. Although the number of transcripts surveyed is limited, these results provide some support for a previous hypothesis that the spermatogenesis pathway in these sterile hybrids may be disrupted sometime after the expression of the early meiotic arrest genes.


Epidemiology of sudden cardiac death in rural South India - insights from the andhra pradesh rural health initiative.
Sudden cardiac death (SCD) is a common initial presentation of coronary artery disease (CAD). Despite the growing epidemic of CAD in India, the epidemiology of SCD is largely unknown. The objective of the study was to define the prevalence and determinants of sudden cardiac deaths in rural South India. Prospective mortality surveillance was conducted in 45 villages (180,162 subjects) in rural South India between January 2006 and October 2007. Trained multipurpose health workers sought to do verbal autopsies within 4 weeks of any death. Detailed questionnaires including comorbidities and circumstances surrounding death were recorded. SCD was adjudicated using the modified Hinkle-Thaler classification. A total of 1916 deaths occurred in the study population over the 22 month time period and verbal autopsy was obtained in 1827 (95%) subjects. Overall mean age of the deceased was 62 ± 20 years and 1007 (55%) were men. Cardiovascular and cerebrovascular diseases together accounted for 559 deaths (31%), followed by infectious disease (163 deaths, 9%), cancer (126 deaths, 7%) and suicide (93 deaths, 5%). Of the 1827 deaths, after excluding accidental deaths (89 deaths), 309 deaths (17%) met criteria for SCD. Cardiovascular disease was the underlying causes in the majority of the SCD events (231/309 (75%)). On multivariate analyses, previous MI/CAD (p < 0.001, OR 14.25), hypertension (p < 0.001, OR 1.84), and age groups between 40-60 yrs (p=0.029) were significantly associated with SCD. Sudden cardiac death accounted for up to half of the cardiovascular deaths in rural Southern India. Traditional cardiovascular risk factors were strongly associated with SCD.


Social determinants in an Australian urban region: a 'complexity' lens.
Area-based strategies have been widely employed in efforts to improve population health and take action on social determinants of health (SDH) and health inequities, including in urban areas where many of the social, economic and environmental factors converge to influence health. Increasingly, these factors are recognized as being part of a complex system, where population health outcomes are shaped by multiple, interacting factors operating at different levels of social organization. This article reports on research to assess the extent to which an alliance of health and human service networks is able to promote action on SDH within an Australian urban region, using a complex systems frame. We found that such an alliance was able to promote some effective action which takes into account complex interactions between social factors affecting health, but also identified significant potential barriers to other forms of desired action identified by alliance members. We found that a complex systems lens was useful in assessing a collaborative intervention to address SDH within an urban region.


A peptide antagonist of the ErbB1 receptor inhibits receptor activation, tumor cell growth and migration in vitro and xenograft tumor growth in vivo.
The epidermal growth factor family of receptor tyrosine kinases (ErbBs) plays essential roles in tumorigenesis and cancer disease progression, and therefore has become an attractive target for structure-based drug design. ErbB receptors are activated by ligand-induced homo- and heterodimerization. Structural studies have revealed that ErbB receptor dimers are stabilized by receptor-receptor interactions, primarily mediated by a region in the second extracellular domain, termed the "dimerization arm". The present study is the first biological characterization of a peptide, termed Inherbin3, which constitutes part of the dimerization arm of ErbB3. Inherbin3 binds to the extracellular domains of all four ErbB receptors, with the lowest peptide binding affinity for ErbB4. Inherbin3 functions as an antagonist of epidermal growth factor (EGF)-ErbB1 signaling. We show that Inherbin3 inhibits EGF-induced ErbB1 phosphorylation, cell growth, and migration in two human tumor cell lines, A549 and HN5, expressing moderate and high ErbB1 levels, respectively. Furthermore, we show that Inherbin3 inhibits tumor growth in vivo and induces apoptosis in a tumor xenograft model employing the human non-small cell lung cancer cell line A549. The Inherbin3 peptide may be a useful tool for investigating the mechanisms of ErbB receptor homo- and heterodimerization. Moreover, the here described biological effects of Inherbin3 suggest that peptide-based targeting of ErbB receptor dimerization is a promising anti-cancer therapeutic strategy.


Monitoring and modeling of emissions from concentrated animal feeding operations: overview of methods.
Accurate monitors are required to determine ambient concentration levels of contaminants emanating from concentrated animal feeding operations (CAFOs), and accurate models are required to indicate the spatial variability of concentrations over regions affected by CAFOs. A thorough understanding of the spatial and temporal variability of concentration levels could then be associated with locations of healthy individuals or subjects with respiratory ailments to statistically link the presence of CAFOs to the prevalence of ill health effects in local populations. This workgroup report, which was part of the Conference on Environmental Health Impacts of Concentrated Animal Feeding Operations: Anticipating Hazards-Searching for Solutions, describes instrumentation currently available for assessing contaminant concentration levels in the vicinity of CAFOs and reviews plume dispersion models that may be used to estimate concentration levels spatially. Recommendations for further research with respect to ambient air monitoring include accurately determining long-term average concentrations for a region under the influence of CAFO emissions using a combination of instruments based on accuracy, cost, and sampling duration. In addition, development of instruments capable of accurately quantifying adsorbed gases and volatile organic compounds is needed. Further research with respect to plume dispersion models includes identifying and validating the most applicable model for use in predicting downwind concentrations from CAFOs. Additional data are needed to obtain reliable emission rates from CAFOs.


Inverted direction of wave propagation (IDWP) in the cochlea.
The "classical" view on wave propagation is that propagating waves are possible in both directions along the length of the basilar membrane and that they have identical properties. Results of several recently executed experiments [T. Ren, Nat. Neurosci. 2, 333-334 (2004) and W. X. He, A. L. Nuttall, and T. Ren, Hear. Res., 228, 112-122 (2007)] appear to contradict this view. In the current work measurements were made of the velocity of the guinea-pig basilar membrane (BM). Distortion products (DPs) were produced by presenting two primary tones, with frequencies below the characteristic frequency f(0) of the BM location at which the BM measurements were made, with a constant frequency ratio. In each experiment the phase of the principal DP, with frequency 2f(1)-f(2), was recorded as a function of the DP frequency. The results indicate that the DP wave going from the two-tone interaction region toward the stapes is not everywhere traveling in the reverse direction, but also in the forward direction. The extent of the region in which the forward wave occurs appears larger than is accounted for by classical theory. This property has been termed "inverted direction of wave propagation." The results of this study confirm the wave propagation findings of other authors. The experimental data are compared to theoretical predictions for a classical three-dimensional model of the cochlea that is based on noise-response data of the same animal. Possible physical mechanisms underlying the findings are discussed.


Bottom-up, not top-down, modulation of imitation by human and robotic models.
Visual observation of human actions provokes more motor activation than observation of robotic actions. We investigated the extent to which this visuomotor priming effect is mediated by bottom-up or top-down processing. The bottom-up hypothesis suggests that robotic movements are less effective in activating the 'mirror system' via pathways from visual areas via the superior temporal sulcus to parietal and premotor cortices. The top-down hypothesis postulates that beliefs about the animacy of a movement stimulus modulate mirror system activity via descending pathways from areas such as the temporal pole and prefrontal cortex. In an automatic imitation task, subjects performed a prespecified movement (e.g. hand opening) on presentation of a human or robotic hand making a compatible (opening) or incompatible (closing) movement. The speed of responding on compatible trials, compared with incompatible trials, indexed visuomotor priming. In the first experiment, robotic stimuli were constructed by adding a metal and wire 'wrist' to a human hand. Questionnaire data indicated that subjects believed these movements to be less animate than those of the human stimuli but the visuomotor priming effects of the human and robotic stimuli did not differ. In the second experiment, when the robotic stimuli were more angular and symmetrical than the human stimuli, human movements elicited more visuomotor priming than the robotic movements. However, the subjects' beliefs about the animacy of the stimuli did not affect their performance. These results suggest that bottom-up processing is primarily responsible for the visuomotor priming advantage of human stimuli.


Total hip arthroplasty versus hemiarthroplasty for independently mobile older adults with intracapsular hip fractures.
Displaced intracapsular hip fractures are typically treated with hemiarthroplasty (HA) or total hip arthroplasty (THA). A number of professional bodies recommend considering THA for patients that were independently mobile and cognitively intact before injury. The aim of this study was to compare the outcomes between HA and THA for independently mobile older adults with hip fractures. A systematic review and meta-analysis of RCTs was undertaken alongside analysis of a propensity score matched national cohort of older adults (aged > 60) with hip fractures. Participants were identified for the propensity score matched cohort from the National Hip Fracture Database (NHFD), which was linked to Hospital Episode Statistics (HES) and civil death registration data. The primary outcomes were 12-month dislocation, revision, and mortality. The secondary outcomes were length of stay, discharge home, unplanned re-admission, functional outcomes, and health-related quality of life. Five RCTs reported higher THA dislocation but this was not statistically significant (THA risk ratio [RR] 2.77, 95% CI 0.81 to 9.48). However, THA dislocation was significantly higher in the national observational dataset (sub-distribution hazard ratio [SHR] 1.73, 95% CI 1.24 to 2.41). Meta-analysis of data from four RCTs did not identify a significant difference in terms of revision (RR 1.52, 95% CI 0.56 to 4.14). However, THA revision was significantly lower in the national dataset (SHR 0.66, 95% CI 0.48 to 0.90). Meta-analysis of data from 5 RCTs suggested higher mortality amongst patients undergoing HA (RR 0.63, 95% CI 0.38 to 1.04), which was also observed within the national registry dataset (hazard ratio 0.45, 95% CI 0.37 to 0.54). National clinical registries can provide important context when interpreting RCT data, which may alone be inadequate for comparing the safety profile of surgical interventions. These data suggest that THA is at significantly higher risk of dislocation but lower risk of revision within 12 months. The finding from both RCT and clinical registry data that THA is associated with lower 12-month mortality amongst the fittest patients with hip fractures requires urgent further study to determine whether or not this can be replicated in other balanced populations.


LAMINA: a tool for rapid quantification of leaf size and shape parameters.
An increased understanding of leaf area development is important in a number of fields: in food and non-food crops, for example short rotation forestry as a biofuels feedstock, leaf area is intricately linked to biomass productivity; in paleontology leaf shape characteristics are used to reconstruct paleoclimate history. Such fields require measurement of large collections of leaves, with resulting conclusions being highly influenced by the accuracy of the phenotypic measurement process. We have developed LAMINA (Leaf shApe deterMINAtion), a new tool for the automated analysis of images of leaves. LAMINA has been designed to provide classical indicators of leaf shape (blade dimensions) and size (area), which are typically required for correlation analysis to biomass productivity, as well as measures that indicate asymmetry in leaf shape, leaf serration traits, and measures of herbivory damage (missing leaf area). In order to allow Principal Component Analysis (PCA) to be performed, the location of a chosen number of equally spaced boundary coordinates can optionally be returned. We demonstrate the use of the software on a set of 500 scanned images, each containing multiple leaves, collected from a common garden experiment containing 116 clones of Populus tremula (European trembling aspen) that are being used for association mapping, as well as examples of leaves from other species. We show that the software provides an efficient and accurate means of analysing leaf area in large datasets in an automated or semi-automated work flow.


pH prediction and control in bioprocesses using mid-infrared spectroscopy.
An on-line pH monitoring method based on mid-infrared spectroscopy relevant to bioprocesses is presented. This approach is non-invasive and does not require the addition of indicators or dyes, since it relies on the analysis of species of common buffers used in culture media, such as phosphate buffer. Starting with titrations of phosphoric and acetic acid solutions over almost the entire pH range (2-12), it was shown that the infrared spectra of all samples can be expressed as a linear combination of the molar absorbance of the acids and their deprotonated forms. In other words, pH had no direct influence on the molar infrared spectra themselves, but only on deprotonation equilibria. Accurate prediction (standard error of prediction for pH < 0.15 pH units) was achieved by taking into account the non-ideal behavior of the solutions, using the Debye-Hückel theory to estimate the activity coefficients. Batch cultures of E. coli were chosen as a case study to show how this approach can be applied to bioprocess monitoring. The discrepancy between the spectroscopic prediction and the conventional electrochemical probe never exceeded 0.12 pH units, and the technique was fast enough to implement a feedback controller to maintain the pH constant during cultivation.


Longitudinal mouse-PET imaging: a reliable method for estimating binding parameters without a reference region or blood sampling.
Longitudinal mouse PET imaging is becoming increasingly popular due to the large number of transgenic and disease models available but faces challenges. These challenges are related to the small size of the mouse brain and the limited spatial resolution of microPET scanners, along with the small blood volume making arterial blood sampling challenging and impossible for longitudinal studies. The ability to extract an input function directly from the image would be useful for quantification in longitudinal small animal studies where there is no true reference region available such as TSPO imaging. Using dynamic, whole-body 18F-DPA-714 PET scans (60 min) in a mouse model of hippocampal sclerosis, we applied a factor analysis (FA) approach to extract an image-derived input function (IDIF). This mouse-specific IDIF was then used for 4D-resolution recovery and denoising (4D-RRD) that outputs a dynamic image with better spatial resolution and noise properties, and a map of the total volume of distribution (VT) was obtained using a basis function approach in a total of 9 mice with 4 longitudinal PET scans each. We also calculated percent injected dose (%ID) with and without 4D-RRD. The VT and %ID parameters were compared to quantified ex vivo autoradiography using regional correlations of the specific binding from autoradiography against VT and %ID parameters. The peaks of the IDIFs were strongly correlated with the injected dose (Pearson R = 0.79). The regional correlations between the %ID estimates and autoradiography were R = 0.53 without 4D-RRD and 0.72 with 4D-RRD over all mice and scans. The regional correlations between the VT estimates and autoradiography were R = 0.66 without 4D-RRD and 0.79 with application of 4D-RRD over all mice and scans. We present a FA approach for IDIF extraction which is robust, reproducible and can be used in quantification methods for resolution recovery, denoising and parameter estimation. We demonstrated that the proposed quantification method yields parameter estimates closer to ex vivo measurements than semi-quantitative methods such as %ID and is immune to tracer binding in tissue unlike reference tissue methods. This approach allows for accurate quantification in longitudinal PET studies in mice while avoiding repeated blood sampling.


Gene landscape and correlation between B-cell infiltration and programmed death ligand 1 expression in lung adenocarcinoma patients from The Cancer Genome Atlas data set.
Tumor-infiltrating lymphocytes are related to positive clinical prognoses in numerous cancer types. Programmed death ligand 1 (PD-L1), a mediator of the PD-1 receptor, plays an inhibitory role in cancer immune responses. PD-L1 upregulation can impede infiltrating T-cell functions in lung adenocarcinoma (LUAD), a lung cancer subtype. However, associations between the expression of PD-L1 and infiltration of B cells (a major immunoregulatory cell) remain unknown. Therefore, we investigated the role of infiltrating B cells in LUAD progression and its correlation with PD-L1 expression. The Cancer Genome Atlas (TCGA) LUAD data set was used to explore associations among B-cell infiltration, PD-L1 expression, clinical outcome, and gene landscape. Gene set enrichment analysis was used to explore putative signaling pathways and candidate genes. The drug enrichment analysis was used to identify candidate genes and the related drugs. We found that high B-cell infiltration was correlated with better prognoses; however, PD-L1 may interfere with the survival advantage in patients with high B-cell infiltration. The gene landscape was characterized comprehensively, with distinct PD-L1 levels in cell populations with high B-cell infiltration. We obtained five upregulated signaling pathways from the gene landscape: apoptosis, tumor necrosis factor (TNF)-α signaling via nuclear factor (NF)-κB, apical surface, interferon-α response, and KRAS signaling. Moreover, four candidate genes and their related target drugs were also identified, namely interleukin-2β receptor (IL2RB), IL-2γ receptor (IL2RG), Toll-like receptor 8 (TLR8), and TNF. These findings suggest that tumor-infiltrating B cells could act as a clinical factor in anti-PD-L1 immunotherapy for LUAD.


Presynaptic alpha-synuclein aggregation in a mouse model of Parkinson's disease.
Parkinson's disease and dementia with Lewy bodies are associated with abnormal neuronal aggregation of α-synuclein. However, the mechanisms of aggregation and their relationship to disease are poorly understood. We developed an in vivo multiphoton imaging paradigm to study α-synuclein aggregation in mouse cortex with subcellular resolution. We used a green fluorescent protein-tagged human α-synuclein mouse line that has moderate overexpression levels mimicking human disease. Fluorescence recovery after photobleaching (FRAP) of labeled protein demonstrated that somatic α-synuclein existed primarily in an unbound, soluble pool. In contrast, α-synuclein in presynaptic terminals was in at least three different pools: (1) as unbound, soluble protein; (2) bound to presynaptic vesicles; and (3) as microaggregates. Serial imaging of microaggregates over 1 week demonstrated a heterogeneous population with differing α-synuclein exchange rates. The microaggregate species were resistant to proteinase K, phosphorylated at serine-129, oxidized, and associated with a decrease in the presynaptic vesicle protein synapsin and glutamate immunogold labeling. Multiphoton FRAP provided the specific binding constants for α-synuclein's binding to synaptic vesicles and its effective diffusion coefficient in the soma and axon, setting the stage for future studies targeting synuclein modifications and their effects. Our in vivo results suggest that, under moderate overexpression conditions, α-synuclein aggregates are selectively found in presynaptic terminals.


The detection of communicative signals directed at the self in infant prefrontal cortex.
A precondition for successful communication between people is the detection of signals indicating the intention to communicate, such as eye contact or calling a person's name. In adults, establishing communication by eye contact or calling a person's name results in overlapping activity in right prefrontal cortex, suggesting that, regardless of modality, the intention to communicate is detected by the same brain region. We measured prefrontal cortex responses in 5-month-olds using near-infrared spectroscopy (NIRS) to examine the neural basis of detecting communicative signals across modalities in early development. Infants watched human faces that either signaled eye contact or directed their gaze away from the infant, and they also listened to voices that addressed them with their own name or another name. The results revealed that infants recruit adjacent but non-overlapping regions in the left dorsal prefrontal cortex when they process eye contact and own name. Moreover, infants that responded sensitively to eye contact in the one prefrontal region were also more likely to respond sensitively to their own name in the adjacent prefrontal region as revealed in a correlation analysis, suggesting that responding to communicative signals in these two regions might be functionally related. These NIRS results suggest that infants selectively process and attend to communicative signals directed at them. However, unlike adults, infants do not seem to recruit a common prefrontal region when processing communicative signals of different modalities. The implications of these findings for our understanding of infants' developing communicative abilities are discussed.


Minimal impact of a de novo-expressed beta-cell autoantigen on spontaneous diabetes development in NOD mice.
During an autoimmune process, the autoaggressive response spreads from the initiating autoantigen to other antigens expressed in the target organ. Based on evidence from experimental models for multiple sclerosis, such "antigenic spreading" can play an important role in the exacerbation of clinical disease. We evaluated whether pathogenesis of spontaneous diabetes in NOD mice could be accelerated in a similar way when a novel autoantigen was expressed in pancreatic beta-cells. Unexpectedly, we found that the expression of the lymphocytic choriomeningitis virus nucleoprotein only led to marginal enhancement of diabetes, although such NOD-nucleoprotein mice were not tolerant to nucleoprotein. Although the frequency of nucleoprotein-specific CD8 T-cells in the pancreatic draining lymph node was comparable with the frequency of islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP)-specific T-cells, more IGRP-specific CD8 T-cells were found both systemically and in the islets where there was a fourfold increase. Interestingly, and in contrast to nucleoprotein-specific CD8 T-cells, IGRP-specific T-cells showed increased CXCR3 expression. Thus, autoreactivity toward de novo-expressed beta-cell autoantigens will not accelerate autoimmunity unless large numbers of antigen-experienced autoreactive T-cells expressing the appropriate chemokine receptors are present.


The role of material, psychosocial and behavioral factors in mediating the association between socioeconomic position and allostatic load (measured by cardiovascular, metabolic and inflammatory markers).
Lower socioeconomic position (SEP), both accumulated across the life course and at different life-stages, has been found to be associated with higher cumulative physiological burden, as measured by allostatic load. This study aimed to identify what factors mediate the association between SEP and allostatic load, as measured through combining cardiovascular, metabolic and inflammatory markers. We explored the role of material, psychological and behavioral factors, accumulated across two periods in time, in mediating the association between SEP and allostatic load. Data are from the West of Scotland Twenty-07 Study, with respondents followed over five waves of data collection from ages 35 to 55 (n=999). Allostatic load was measured by summing nine binary biomarker scores ('1'=in the highest-risk quartile) measured when respondents were 55years old (wave 5). SEP was measured by a person's accumulated social class over two periods All mediators and SEP were measured at baseline in 1987 and 20years later and combined to form accumulated measures of risk. Material mediators included car and home ownership, and having low income. The General Health Questionnaire (GHQ-12) was used as the psychosocial mediator. Behavioral mediators included smoking, alcohol consumption, physical activity and diet. Path analysis using linear regressions adjusting for sex were performed for each of the potential mediators to assess evidence of attenuation in the association between lower SEP and higher allostatic load. Analyses by mediator type revealed that renting one's home (approximately 78% attenuation) and having low income (approx. 62% attenuation) largely attenuated the SEP-allostatic load association. GHQ did not attenuate the association. Smoking had the strongest attenuating effect of all health behaviors (by 33%) with no other health behaviors attenuating the association substantially. Material factors, namely home tenure and income status, and smoking have important roles in explaining socioeconomic disparities in allostatic load, particularly when accumulated over time.


Effects of chemical smokes on flora and fauna under field and laboratory exposures.
Various types of obscurant smokes are used routinely in training by the U.S. Army. Because continued routine use of the smokes could be detrimental to the native flora and fauna at training sites, a preliminary biological and chemical field study of fogoil, hexachloroethane, and tank diesel smokes was conducted. Smoke plumes were sampled and chemically analyzed at distances of 15-150 m from the smoke source where Tradescantia clones 4430 and 03 and the native plant Ambrosia dumosa and the native rodent Dipodomys merriami were exposed for 30 min. In addition, Tradescantia clone 4430 was exposed to tank diesel in the laboratory at concentration levels equivalent to exposure at 15 and 50 m. Tradescantia clones were examined for mutagenic effects indicated by micronuclei induction in developing pollen and pink somatic mutations in stamen hairs. Photosynthetic perturbations were measured in Tradescantia and A. dumosa using variable fluorescence induction. Animals were examined for sister chromatid exchanges and chromosome aberrations. It was found that all of the smokes tested exerted varying degrees of physiological and mutagenic effects in one or more assay system at one or more exposure distance. The studies reported here indicate that exposed ecological systems, or at least components of these systems, are at a higher risk than are unexposed components (e.g., organisms) for several types of damage attributed to obscurant smoke exposure.


Mismatches between youth aspirations and participatory HIV/AIDSprogrammes in South Africa.
Although youth participation is a pillar of international HIV/AIDS policy, it is notoriously difficult to facilitate. We explore this challenge through a case study of a community-led HIV/AIDS management project in a South African rural area, in which anticipated youth participation failed to materialise. We take a social psychological view, examining ways in which opportunities offered by the project failed to resonate with the social identities and aspirations of local young people. Interviews and focus group discussions were conducted with 37 young people prior to the programme's establishment and with 21 young people four years later. In response to questions about what they wanted to achieve in life, the young people emphasized: career success through migrating to urban areas to seek education and paid work, non-tokenistic involvement in community affairs, and 'having fun.' We look at how the project unintentionally evolved in ways that undermined these goals. Its strong local focus was inappropriately tailored to young people whose views of the future focused on getting away to urban areas as quickly as possible. The volunteer nature of the work held little appeal for ambitious young people who instead saw paid work as their way out of poverty and were reluctant to take unpaid time out from schoolwork. The project failed to develop new and democratic ways of operating-quickly becoming mired in traditional, adult-dominated social relations, in which young people with initiative and independent views were sometimes belittled by adults as being 'smart' or 'clever.' Finally, the project's focus on sexual abstinence held little interest for young people who took an enthusiastic interest in sex. The article concludes with a discussion of the complexities of implementing youth-friendly projects in communities steeped in top-down adult-dominated social interactions, and recommends ways in which similar projects might seek to involve youths more effectively.


Stride length regulation in Parkinson's disease. Normalization strategies and underlying mechanisms.
Results of our previous studies have shown that the slow, shuffling gait of Parkinson's disease patients is due to an inability to generate appropriate stride length and that cadence control is intact and is used as a compensatory mechanism. The reason for the reduced stride length is unclear, although deficient internal cue production or inadequate contribution to cortical motor set by the basal ganglia are two possible explanations. In this study we have examined the latter possibility by comparing the long-lasting effects of visual cues in improving stride length with that of attentional strategies. Computerized stride analysis was used to measure the spatial (distance) and temporal (timing) parameters of the walking pattern in a total of 54 subjects in three separate studies. In each study Parkinson's disease subjects were trained for 20 min by repeated 10 m walks set at control stride length (determined from control subjects matched for age, sex and height), using either visual floor markers or a mental picture of the appropriate stride size. Following training, the gait patterns were monitored (i) every 15 min for 2 h; (ii) whilst interspersing secondary tasks of increasing levels of complexity; (iii) covertly, when subjects were unaware that measurement was taking place. The results demonstrated that training with both visual cues and attentional strategies could maintain normal gait for the maximum recording time of 2 h. Secondary tasks reduced stride length towards baseline values as did covert monitoring. The findings confirm that the ability to generate a normal stepping pattern is not lost in Parkinson's disease and that gait hypokinesia reflects a difficulty in activating the motor control system. Normal stride length can be elicited in Parkinson's disease using attentional strategies and visual cues. Both strategies appear to share the same mechanism of focusing attention on the stride length. The effect of attention appears to require constant vigilance to prevent reverting to more automatic control mechanisms.


Breast cancer screening: are we seeing the benefit?
A decline in breast cancer mortality has been observed in western European Countries since the middle of the 1990s. Different methodological approaches, including case-control studies, incidence-based mortality studies, and trend studies, have been used to assess the effectiveness of mammography screening programmes in reducing breast cancer mortality. However, not all methods succeed in distinguishing the relative contributions of service screening and taking correctly into consideration the potential source of bias that might affect the estimate. Recently, a review of six case-control studies confirmed a breast cancer mortality reduction ranging from 38% to 70% among screened women. This figure is in accordance with the estimate obtained from incidence-based mortality studies if screening compliance is taken into account. We will describe the methodological constraints of mortality trend studies in predicting the impact of screening on mortality and the necessary caution that must be applied when interpreting the results of such studies. In conclusion, when appropriate methodological approaches are used, it is evident that mammographic screening programmes have contributed substantially to the observed decline in breast cancer mortality.


Ultra-small Ge1-xSnx quantum dots with visible photoluminescence.
Ge1-xSnx alloy quantum dots (QDs) were synthesized with sizes ranging from 1-3 nm exhibiting visible orange-red photoluminescence. Composition dependent optical properties were characterized and supported by theoretical calculations. Structural analysis suggests the QDs are diamond cubic phase, characteristic of Ge1-xSnx thin films and nanocrystals (NCs) reported to date.


A gene linked to the Igh-C locus controls the production of rheumatoid factor in the mouse.
In certain specific pathogen-free colonies, mice, upon aging, produce autoantibodies (RF) specific for the Fc portion of their IgG. In our colony, 129/Sv mice (H-2bvl; Igh-1a) have 10-20 times higher RF levels than C5BL/6 (h-2b; Igh-1b). In addition, the 129 have mainly IgA anti-IgG2a, and the B6 have mainly IgM anti-IgGl. We analyzed the genetic factors that control these differences. The high RF-producer phenotype of strain 129 was inherited as a recessive trait as indicated by the low RF levels of (129 X B6) F1 mice. About 1 of 4 129 X F1 (129 X B6) backcrosses and 1 of 10 (129 X B6) F2 mice had high RF levels, suggesting the involvement of two recessive genes in the control of this RF production. All F2 mice and all but one backcross with high IgA anti-IgG2a levels were homozygous for the Ihg-1a allele of the 129 mouse. In contrast, the B6-type RF was eight times more frequent in Igh-1bb than in Igh-1ab or Igh-1aa mice. High RF titers of either type were suppressed in Igh-1ab mice.


The selective conservative management of penetrating thoracic trauma is still appropriate in the current era.
Traumatic pleural collections secondary to penetrating chest trauma are generally managed by intercostal chest drainage (ICD), but these protocols were developed a few decades ago when stabs (SWs) predominated over gunshot wounds (GSWs). This study reviews the outcome of a selective conservative approach to penetrating thoracic trauma to establish if it is still appropriate in the current era. We reviewed 827 patients over a four-year period with penetrating unilateral non-cardiac wounds of the chest in order to review the efficacy of our policy and to define the differences in the spectrum of injury between SWs and GSWs. Ninety-two per cent (764/827) were males, and the median age was 24 years. Seventy-six per cent (625/827) sustained SWs and twenty-four per cent (202/827) GSWs. Chest pathologies were: pneumothorax (PTX): 362 (44%), haemothorax (HTX): 150 (18%) and haemopneumothorax (HPTX): 315 (38%). Ninety-six per cent of patients were managed non-operatively. Four per cent (36/827) were subjected to a thoracotomy [31 SWs and 5 GSWs]. No difference was observed in terms of the need for operative intervention: 5% vs. 3% [p=0.202]. PTX was seen exclusively in SWs: 58% vs. 0% and there were significantly more HPTXs seen in the GSWs: HPTX: 24% vs. 81% [p<0.001]. The median days of ICD in situ were significantly longer in GSWs compared to SWs for all pathologies. For HTX: 4.5 (interquartile range [IQR]: 3-6) vs. 3.5 (IQR: 0-5) days, p=0.001 and HPTX: 4 (IQR: 3-5) vs. 3.0 (IQR: 3-4) days, p<0.001. There were seven (15%) complications. A total of five (13%) patients died and all deaths were confined to the operative group. SWs continue to predominate over GSWs. PTXs were more commonly associated with SWs, whilst HPTX are more commonly associated with GSWs. A policy of selective conservatism is still applicable to the management of traumatic pleural collections.


Biomarkers of secondhand smoke exposure in automobiles.
The objectives of this study were: (1) to characterise the exposure of non-smokers exposed to secondhand smoke (SHS) in a vehicle using biomarkers, (2) to describe the time course of the biomarkers over 24 h, and (3) to examine the relationship between tobacco biomarkers and airborne concentrations of SHS markers. Eight non-smokers were individually exposed to SHS in cars with fully open front windows and closed back windows over an hour from a smoker who smoked three cigarettes at 20 min intervals. The non-smokers sat in the back seat on the passenger side, while the smoker sat in the driver's seat. Plasma cotinine and urine cotinine, 3-hydroxycotinine (3HC) and 4-(methylnitrosoamino)-(3-pyridyl)-1-butanol (NNAL) were compared in samples taken at baseline (BL) and several time-points after exposure. Nicotine, particulate matter (PM2.5) and carbon monoxide (CO) were measured inside and outside the vehicle and ventilation rates in the cars were measured. Average plasma cotinine and the molar sum of urine cotinine and 3HC (COT+3HC) increased four-fold, urine cotinine increased six-fold and urine NNAL increased ∼27 times compared to BL biomarker levels. Plasma cotinine, urine COT+3HC and NNAL peaked at 4-8 h post-exposure while urine cotinine peaked within 4 h. Plasma cotinine was significantly correlated to PM2.5 (Spearman correlation rs=0.94) and CO (rs=0.76) but not to air nicotine. The correlations between urine biomarkers, cotinine, COT+3HC and NNAL, and air nicotine, PM2.5 and CO were moderate but non-significant (rs range = 0.31-0.60). Brief SHS exposure in cars resulted in substantial increases in levels of tobacco biomarkers in non-smokers. For optimal characterisation of SHS exposure, tobacco biomarkers should be measured within 4-8 h post-exposure. Additional studies are needed to better describe the relationship between tobacco biomarkers and environmental markers of SHS.


Intimate Partner Violence Survivors' Reports of Their Children's Exposure to Companion Animal Maltreatment: A Qualitative Study.
Children living in households where intimate partner violence (IPV) is present are at increased risk of being exposed to concomitant maltreatment of companion animals. Recent research suggests that childhood exposure to maltreatment of companion animals is associated with compromised socioemotional well-being in childhood and adulthood. To date, there is a dearth of qualitative research examining how children experience animal maltreatment in the context of IPV. The current qualitative study explored the following research question in an ethnically diverse sample of IPV survivors: How do maternal caregivers convey the ways in which their children experience animal maltreatment in IPV-affected households? Sixty-five women with at least one child (age 7-12 years) were recruited from domestic violence agencies and described their child(ren)'s experiences of animal maltreatment in the home. Template analysis was used to analyze interview data (KALPHA = .90). Three themes emerged related to children's experiences of animal maltreatment: (a) direct exposure to animal maltreatment and related threats, (b) emotional and behavioral responses to animal maltreatment exposure, and (c) animal maltreatment as coercive control of the child. Results suggest that children's exposure to animal maltreatment is multifaceted and may exacerbate children's risk of negative psychosocial outcomes in the context of co-occurring IPV. Intervention programs designed to assist children exposed to IPV should consider the extent of children's awareness of the abuse of their pets and their strong and deleterious reactions to it.


Comparative community burden and severity of seasonal and pandemic influenza: results of the Flu Watch cohort study.
Assessment of the effect of influenza on populations, including risk of infection, illness if infected, illness severity, and consultation rates, is essential to inform future control and prevention. We aimed to compare the community burden and severity of seasonal and pandemic influenza across different age groups and study years and gain insight into the extent to which traditional surveillance underestimates this burden. Using preseason and postseason serology, weekly illness reporting, and RT-PCR identification of influenza from nasal swabs, we tracked the course of seasonal and pandemic influenza over five successive cohorts (England 2006-11; 5448 person-seasons' follow-up). We compared burden and severity of seasonal and pandemic strains. We weighted analyses to the age and regional structure of England to give nationally representative estimates. We compared symptom profiles over the first week of illness for different strains of PCR-confirmed influenza and non-influenza viruses using ordinal logistic regression with symptom severity grade as the outcome variable. Based on four-fold titre rises in strain-specific serology, on average influenza infected 18% (95% CI 16-22) of unvaccinated people each winter. Of those infected there were 69 respiratory illnesses per 100 person-influenza-seasons compared with 44 per 100 in those not infected with influenza. The age-adjusted attributable rate of illness if infected was 23 illnesses per 100 person-seasons (13-34), suggesting most influenza infections are asymptomatic. 25% (18-35) of all people with serologically confirmed infections had PCR-confirmed disease. 17% (10-26) of people with PCR-confirmed influenza had medically attended illness. These figures did not differ significantly when comparing pandemic with seasonal influenza. Of PCR-confirmed cases, people infected with the 2009 pandemic strain had markedly less severe symptoms than those infected with seasonal H3N2. Seasonal influenza and the 2009 pandemic strain were characterised by similar high rates of mainly asymptomatic infection with most symptomatic cases self-managing without medical consultation. In the community the 2009 pandemic strain caused milder symptoms than seasonal H3N2. Medical Research Council and the Wellcome Trust.


Financial ties of principal investigators and randomized controlled trial outcomes: cross sectional study.
To examine the association between the presence of individual principal investigators' financial ties to the manufacturer of the study drug and the trial's outcomes after accounting for source of research funding. Cross sectional study of randomized controlled trials (RCTs). Studies published in "core clinical" journals, as identified by Medline, between 1 January 2013 and 31 December 2013. Random sample of RCTs focused on drug efficacy. Association between financial ties of principal investigators and study outcome. A total of 190 papers describing 195 studies met inclusion criteria. Financial ties between principal investigators and the pharmaceutical industry were present in 132 (67.7%) studies. Of 397 principal investigators, 231 (58%) had financial ties and 166 (42%) did not. Of all principal investigators, 156 (39%) reported advisor/consultancy payments, 81 (20%) reported speakers' fees, 81 (20%) reported unspecified financial ties, 52 (13%) reported honorariums, 52 (13%) reported employee relationships, 52 (13%) reported travel fees, 41 (10%) reported stock ownership, and 20 (5%) reported having a patent related to the study drug. The prevalence of financial ties of principal investigators was 76% (103/136) among positive studies and 49% (29/59) among negative studies. In unadjusted analyses, the presence of a financial tie was associated with a positive study outcome (odds ratio 3.23, 95% confidence interval 1.7 to 6.1). In the primary multivariate analysis, a financial tie was significantly associated with positive RCT outcome after adjustment for the study funding source (odds ratio 3.57 (1.7 to 7.7). The secondary analysis controlled for additional RCT characteristics such as study phase, sample size, country of first authors, specialty, trial registration, study design, type of analysis, comparator, and outcome measure. These characteristics did not appreciably affect the relation between financial ties and study outcomes (odds ratio 3.37, 1.4 to 7.9). Financial ties of principal investigators were independently associated with positive clinical trial results. These findings may be suggestive of bias in the evidence base.


Loss of function of the nuclear envelope protein LEMD2 causes DNA damage-dependent cardiomyopathy.
Mutations in nuclear envelope proteins (NEPs) cause devastating genetic diseases, known as envelopathies, that primarily affect the heart and skeletal muscle. A mutation in the NEP LEM domain-containing protein 2 (LEMD2) causes severe cardiomyopathy in humans. However, the roles of LEMD2 in the heart and the pathological mechanisms responsible for its association with cardiac disease are unknown. We generated knockin (KI) mice carrying the human c.T38>G Lemd2 mutation, which causes a missense amino acid exchange (p.L13>R) in the LEM domain of the protein. These mice represent a preclinical model that phenocopies the human disease, as they developed severe dilated cardiomyopathy and cardiac fibrosis leading to premature death. At the cellular level, KI/KI cardiomyocytes exhibited disorganization of the transcriptionally silent heterochromatin associated with the nuclear envelope. Moreover, mice with cardiac-specific deletion of Lemd2 also died shortly after birth due to heart abnormalities. Cardiomyocytes lacking Lemd2 displayed nuclear envelope deformations and extensive DNA damage and apoptosis linked to p53 activation. Importantly, cardiomyocyte-specific Lemd2 gene therapy via adeno-associated virus rescued cardiac function in KI/KI mice. Together, our results reveal the essentiality of LEMD2 for genome stability and cardiac function and unveil its mechanistic association with human disease.


Human thrombomodulin regulates complement activation as well as the coagulation cascade in xeno-immune response.
With the introduction of the α1, 3-galactosyltransferase gene-knockout (GT-KO) pig and its pivotal role in preventing hyperacute rejection (HAR), coagulation remains a considerable obstacle yet to be overcome in order to provide long-term xenograft survival. Thrombomodulin (TBM) plays a critical anticoagulant and anti-inflammatory role in its part of the protein C pathway. Many studies have demonstrated the strong anticoagulant effects of TBM in xenotransplantation, but its complement regulatory effects have not been appropriately examined. Here, we investigate whether TBM can regulate complement activation as well as coagulation in response to xenogeneic stimuli. We transfected porcine endothelial cells (MPN-3) with adenovirus vectors containing the human TBM gene (ad-hTBM), or a control gene containing GFP (ad-GFP). The expression level of ad-hTBM was measured by flow cytometry. To confirm the anticoagulant effect of TBM, coagulation time was measured after treatment with recalcified human plasma in ad-hTBM-transfected MPN-3, and a thrombin activity assay was performed after treatment with 50% human serum in ad-hTBM-infected MPN-3. Thrombin generation was significantly decreased in a dose-dependent manner in ad-TBM group, and coagulation time was increased in the ad-hTBM group when compared to the ad-GFP group. Complement-dependent serum toxicity assays were performed after treatment with 20% human serum or heat-inactivated human serum by LDH assay. Complement-dependent toxicity was significantly attenuated in the ad-hTBM group, but complement-independent toxicity was not attenuated in the ad-hTBM group. These results suggest that human thrombomodulin (hTBM) has complement regulatory effects as well as anticoagulant effects. To further investigate the mechanisms of complement regulation by hTBM, we deleted the EGF5, 6 domains that are involved in thrombin generation or the lectin-like domain involved in inflammation of TBM and functional tests were performed using these modified forms. We showed that the EGF5, 6 domain of TBM principally inhibits complement activation rather than the lectin domain. The EGF5, 6 domains of TBM appear to be the major domains for down-regulating the complement system rather than the lectin-like domain during xenogenic stimuli. The role of EGF5, 6 domains of hTBM may be due to inhibition of thrombin as thrombin can cleave C3a and C5a directly and hTBM may also be involved in complement regulation. Clearly then human TBM has complement regulatory effects as well as anticoagulant effects in xeno-immune response, and it is a promising target for attenuating xenograft rejection.


Hyperactivation of JAK1 tyrosine kinase induces stepwise, progressive pruritic dermatitis.
Skin homeostasis is maintained by the continuous proliferation and differentiation of epidermal cells. The skin forms a strong but flexible barrier against microorganisms as well as physical and chemical insults; however, the physiological mechanisms that maintain this barrier are not fully understood. Here, we have described a mutant mouse that spontaneously develops pruritic dermatitis as the result of an initial defect in skin homeostasis that is followed by induction of a Th2-biased immune response. These mice harbor a mutation that results in a single aa substitution in the JAK1 tyrosine kinase that results in hyperactivation, thereby leading to skin serine protease overexpression and disruption of skin barrier function. Accordingly, treatment with an ointment to maintain normal skin barrier function protected mutant mice from dermatitis onset. Pharmacological inhibition of JAK1 also delayed disease onset. Together, these findings indicate that JAK1-mediated signaling cascades in skin regulate the expression of proteases associated with the maintenance of skin barrier function and demonstrate that perturbation of these pathways can lead to the development of spontaneous pruritic dermatitis.


Gemcitabine plus celecoxib in patients with advanced or metastatic pancreatic adenocarcinoma: results of a phase II trial.
Cycloxygenase-2 (COX-2) is overexpressed in pancreatic tumors where it may be involved in inflammation, carcinogenesis, and the regulation of neoangiogenesis. The purpose of this trial was to evaluate the combination of intravenous gemcitabine with selective COX-2 inhibitor, celecoxib for effect on survival, disease progression, and tolerability in patients with advanced pancreatic cancer. In addition, limited pharmacokinetic and pharmacodynamic analyses were preformed. Eligible patients included those with locally advanced or metastatic pancreatic cancer with no prior chemotherapy and ECOG performance status 0-2. The treatment consisted of intravenous gemcitabine 1000 mg/m weekly x 7 weeks and concurrent daily oral celecoxib 400 mg orally twice a day. Daily oral low-dose aspirin 81 mg was administered throughout the study as a precaution for increased risk of thrombotic events. Those with stable or responsive disease were continued on intravenous gemcitabine 1000 mg/m weekly x 3 weeks and concurrent oral celecoxib. Twenty five patients have been enrolled at 3 centers. Five patients had locally advanced cancer; 20 had metastatic disease. The most common grade 3/4 hematological toxicities were neutropenia (32%) and anemia (20%). Four patients (17%) had partial response and 7 (35%) demonstrated stable disease. The estimated 12-month survival rate was 15%, which did not reach the predetermined efficacy end point. There was a trend suggestive of correlation between a decrease in serum vascular endothelial growth factor and patient survival. The addition of celecoxib to gemcitabine therapy did not demonstrate significant improvement in measured clinical outcomes, in patients with advanced pancreatic cancer. Higher doses of celecoxib may be needed to observe significant antitumor activity.


Efficient homologous recombination-mediated genome engineering in zebrafish using TALE nucleases.
Custom-designed nucleases afford a powerful reverse genetic tool for direct gene disruption and genome modification in vivo. Among various applications of the nucleases, homologous recombination (HR)-mediated genome editing is particularly useful for inserting heterologous DNA fragments, such as GFP, into a specific genomic locus in a sequence-specific fashion. However, precise HR-mediated genome editing is still technically challenging in zebrafish. Here, we establish a GFP reporter system for measuring the frequency of HR events in live zebrafish embryos. By co-injecting a TALE nuclease and GFP reporter targeting constructs with homology arms of different size, we defined the length of homology arms that increases the recombination efficiency. In addition, we found that the configuration of the targeting construct can be a crucial parameter in determining the efficiency of HR-mediated genome engineering. Implementing these modifications improved the efficiency of zebrafish knock-in generation, with over 10% of the injected F0 animals transmitting gene-targeting events through their germline. We generated two HR-mediated insertion alleles of sox2 and gfap loci that express either superfolder GFP (sfGFP) or tandem dimeric Tomato (tdTomato) in a spatiotemporal pattern that mirrors the endogenous loci. This efficient strategy provides new opportunities not only to monitor expression of endogenous genes and proteins and follow specific cell types in vivo, but it also paves the way for other sophisticated genetic manipulations of the zebrafish genome.


Complication rate of diagnostic carotid angiography performed by interventional cardiologists.
The aim of this study was to evaluate the complication rate of diagnostic carotid angiography performed by interventional cardiologists and compare it to previously published data. Percutaneous treatment for carotid artery stenosis is increasingly being performed. Previously published data describes the complication rate of diagnostic carotid angiography performed by radiologists and vascular surgeons, yet the information regarding interventional cardiologists is sparse. Currently in the United States, interventional cardiologists perform a great deal of diagnostic carotid angiograms. A retrospective analysis was done on 333 patients who underwent diagnostic carotid angiography at a single medical center from January 2000 to February 2007. Medical records were reviewed for cardiovascular risk factors, indications for the procedure, angiography technique and in-hospital complications. Complications were categorized as neurological and non-neurological. Neurological complications were further grouped into transient (<7 days) or permanent. Non-neurological complications were grouped into major (requiring additional treatment) or minor. Three hundred and thirty-three patients underwent 347 diagnostic carotid angiograms. Twelve (3.5%) complications occurred in 12 patients. No cerebral vascular accidents occurred and only one (0.3%) transient ischemic attack occurred. Two patients required blood transfusions following the index procedure yielding a major non-neurological complication rate of 0.6%. Review of the literature revealed a transient neurological complication rate from 0 to 2.4% and a major non-neurological complication rate of 0.26-4.3%. Neurological and non-neurological complication rates for carotid angiograms performed by interventional cardiologists are low and compare well with the literature. Interventional cardiologists can safely perform diagnostic carotid angiography with low complication rates.


The protective role of melatonin against the effects of different doses of caffeine on the fetus.
Skeletal system and some organs development changes in rat fetuses with 30 and 60 mg/kg caffeine and melatonin's (10 mg/kg) protective role against rat fetuses were investigated. Groups (n = 4) were formed as Control, LDC, HDC, LDC+melatonin, HDC+melatonin and melatonin. Fetuses were taken by cesarean section and stained using dual skeletal staining method and FESEM. TRAP and AP immune-reactivity concentrations were calculated. Oxidative stress and inflammatory markers were also measured by liver, bone and placenta samples. TNF-α, IL-1ß, IL-6, VEGF-A, SOST and Fetuin-A levels were measured in tissue by using ELISA. TBARS, SOD, GSH, GSSG, TOS, TAS, measured by spectrophotometric assay method. The mRNA levels of Agtr2 gene expressed in placental tissues of control rats and in placental tissues of rats exposed to HDC, LDC, MEL, HDC+MEL, LDC+MEL were analyzed by Real-time PCR. The gene expressions of Agtr2 were significantly upregulated in the placentas exposed to HDC, MEL, HDC+MEL and LDC+MEL (P<0,001). No significant difference in samples of the LDC group (P>0,05). According to these data, caffeine used during pregnancy delayed ossification; melatonin, a powerful antioxidant, was found to eliminate this effect. Besides, changes in angiotensin receptor expression observed in response to a caffeine and melatonin exposure result from high dose and join effect.


Oxygen distributions partly explain the radiation response of human squamous cell carcinomas.
The oxygen tension of human tumours has often been thought to alter tumour response to radiation therapy. The purpose of this analysis is to determine to what extent the observed results of radiotherapy fit predictions based on in situ human tumour pO2 distributions. The radiation dose-response curve for patients treated with radiation alone for squamous cell cancers of the cervix and oropharynx were calculated based on published data. pO2 histograms were obtained from 30 women with cervical cancer and 11 patients with neck nodes from head and neck cancers. An average of 76 +/- 35 (range 28-174) measurements were made from each patient. Hypoxia was assumed to be a purely dose-modifying factor with a maximum OER of 2.5. Assuming patients are treated with daily radiation doses of 2 Gy, the squamous cell carcinoma alpha/beta ratio is 10 Gy, and that tumours have a mean of 10(8) clonogens, it was possible to estimate tumour control probability. Tumour oxygenation was an extremely important modifier of the slope of the dose-response curve and alone was sufficient to account for the slope of the clinically observed dose-response curve for neck nodes. The response curve for uterine cervical cancers is very shallow, and the oxygen distribution did not completely account for heterogeneity of response of these tumours. The results support the conclusion that oxygen tension distribution is an important modifier of human radiation treatment response.


Ability of expert physicians to structure clinical guidelines: reality versus perception.
Structuring Textual Clinical Guidelines (GLs) into a formal representation is a necessary prerequisite for supporting their automated application. We had developed a collaborative guideline-structuring methodology that involves expert physicians, clinical editors and knowledge engineers, to produce a machine-comprehensible representation for automated support of evidence-based, guideline-based care. Our goals in the current study were: (1) to investigate the perceptions of the expert physicians and clinical editors as to the relative importance, for the structuring process, of different aspects of the methodology; (2) to assess, for the clinical editors, the inter-correlations among (i) the reported level of understanding of the guideline structuring ontology's (knowledge scheme's) features, (ii) the reported ease of structuring each feature and (iii) the actual objective quality of structuring. A clinical consensus regarding the contents of three guidelines was prepared by an expert in the domain of each guideline. For each guideline, two clinical editors independently structured the guideline into a semi-formal representation, using the Asbru guideline ontology's features. The quality of the resulting structuring was assessed quantitatively. Each expert physician was asked which aspects were most useful for formation of the consensus. Each clinical editor filled questionnaires relating to: (1) the level of understanding of the ontology's features (before the structuring process); (2) the usefulness of various aspects in the structuring process (after the structuring process); (3) the ease of structuring each ontological feature (after the structuring process). Subjective reports were compared with objective quantitative measures of structuring correctness. Expert physicians considered having medical expertise and understanding the ontological features as the aspects most useful for creation of a consensus. Clinical editors considered understanding the ontological features and the use of the structuring tools as the aspects most useful for structuring guidelines. There was a positive correlation (R = 0.87, P < 0.001) between the reported ease of understanding ontological features and the reported ease of structuring those features. However, there was no significant correlation between the reported level of understanding the features - or the reported ease of structuring by using those features - and the objective quality of the structuring of these features in actual guidelines. Aspects considered important for formation of a clinical consensus differ from those for structuring of guidelines. Understanding the features of a structuring ontology is positively correlated with the reported ease of using these features, but neither of these subjective reports correlated with the actual objective quality of the structuring using these features.


Tumor suppressors microRNA-302d and microRNA-16 inhibit human glioblastoma multiforme by targeting NF-κB and FGF2.
Glioblastoma multiforme (GBM) is a highly malignant cancer in the brain with a median survival time of approximately one year. However, the mechanisms underlying GBM development and occurrence are poorly understood. Recently, miRNAs were reported to play important roles in GBM. We performed microRNA profiling by comparing the human GBM cell line T98G and control cell line HCN1A. MicroRNA assays, PCR and Western blot analysis were performed to detect the expressions of microRNAs, mRNAs and proteins of target genes, respectively. Cell migration and invasion assays were conducted. A murine in situ xenograft tumor model was used to evaluate tumor growth in vivo. Glioblastoma tissues were examined to investigate the clinical relevance of our findings. MiR-302d and miR-16 levels were found to be decreased in T98G cells. MiR-302d and miR-16 inhibited the expressions of p65 and FGF2, respectively, by binding to the 3'-UTR of their mRNAs. Over-expression of miR-302d and miR-16 inhibited T98G cell migration and invasion in vitro, and tumorigenesis in the xenograft tumor mouse model in vivo, by suppressing p65 and FGF2. Negative correlations between miR-302d and p65 and between miR-16 and FGF2 were observed in patient glioblastoma tissues. MiR-302d and miR-16 inhibit tumorigenesis by down-regulating p65 and FGF2, which potentially contributes to the treatment of glioblastoma with clinical relevance.


Investigating the coverage dependent behaviour of CO on Gd/Pt(111).
Gd modified Pt(111) single crystals have been prepared in an ultra high vacuum (UHV). By vacuum deposition of ∼200 Å Gd on a sample heated to 800 °C, a Pt5Gd alloy terminated by a single atomic layer of Pt was formed. Subsequently the surfaces were characterized using low energy electron diffraction (LEED), showing that a highly ordered crystal structure had appeared. To study the molecular dynamics on this surface a detailed study of the CO adsorption on the surface was conducted using temperature programmed desorption (TPD) of CO. The TPD spectra show a desorption peak shifted down in temperature compared to those of pure Pt(111). The shape of the desorption peak and the desorption temperature were shown to be strongly dependent on the CO coverage of the surface. A systematic investigation of CO desorption temperature as a function of coverage was consequently performed. A simple simulation of the TPD spectra was carried out, based on adsorption energies from density functional theory (DFT). This simulation reproduces the shift and the narrowing of the desorption spectrum from the experiments and the DFT calculations suggest that the sharp TPD feature arises from cooperative adsorbate interactions, caused by subtle reconstructions occurring at coverages above 1/3 ML CO, whereas the overall temperature shift relative to pure Pt(111) comes from weaker CO binding due to the contraction of the Gd/Pt(111) surface.


Risk factors for hospital readmission of elderly patients.
The aim of this study was to identify which factors were associated with a risk of hospital readmission within 3 months after discharge of a sample of elderly patients admitted to internal medicine and geriatric wards. Of the 1178 patients aged 65 years or more and discharged from one of the 66 wards of the 'Registry Politerapie SIMI (REPOSI)' during 2010, 766 were followed up by phone interview 3 months after discharge and were included in this analysis. Univariate and multivariate logistic regression models were used to evaluate the association of several variables with rehospitalization within 3 months from discharge. Nineteen percent of patients were readmitted at least once within 3 months after discharge. By univariate analysis in-hospital clinical adverse events (AEs), a previous hospital admission, number of diagnoses and drugs, comorbidity and severity index (according to Cumulative Illness Rating Scale-CIRS), vascular and liver diseases with a level of impairment at discharge of 3 or more at CIRS were significantly associated with risk of readmission. Multivariate logistic regression analysis showed that only AEs during hospitalization, previous hospital admission, and vascular and liver diseases were significantly associated with the likelihood of readmission. The results demonstrate the need for increased medical attention towards elderly patients discharged from hospital with characteristics such as AEs during the hospitalization, previous admission, vascular and liver diseases.


Digital Radiography in Total Hip Arthroplasty: Technique and Radiographic Results.
Obtaining the ideal acetabular cup position in total hip arthroplasty remains a challenge. Advancements in digital radiography and image analysis software allow the assessment of the cup position during the surgical procedure. This study describes a validated technique for evaluating cup position during total hip arthroplasty using digital radiography. Three hundred and sixty-nine consecutive patients undergoing total hip arthroplasty were prospectively enrolled. Preoperative supine anteroposterior pelvic radiographs were made. Intraoperative anteroposterior pelvic radiographs were made with the patient in the lateral decubitus position. Radiographic beam angle adjustments and operative table adjustments were made to approximate rotation and tilt of the preoperative radiograph. The target for cup position was 30° to 50° abduction and 15° to 35° anteversion. Intraoperative radiographic measurements were calculated and final cup position was determined after strict impingement and range-of-motion testing. Postoperative anteroposterior pelvic radiographs were made. Two independent observers remeasured all abduction and anteversion angles. Of the cups, 97.8% were placed within 30° to 50° of abduction, with a mean angle (and standard deviation) of 39.5° ± 4.6°. The 2.2% of cups placed outside the target zone were placed so purposefully on the basis of intraoperative range-of-motion testing and patient factors, and 97.6% of cups were placed between 15° and 35° of anteversion, with a mean angle of 26.6° ± 4.7°. Twenty-eight percent of cups were repositioned on the basis of intraoperative measurements. Subluxation during range-of-motion testing occurred in 3% of hips despite acceptable measurements, necessitating cup repositioning. There was 1 early anterior dislocation. Placing the acetabular component within a target range is a critical component to minimizing dislocation and polyethylene wear in total hip arthroplasty. Using digital radiography, we positioned the acetabular component in our desired target zone in 97.8% of cases and outside the target zone, purposefully, in 2.2% of cases. When used in conjunction with strict impingement testing, digital radiography allows for predictable cup placement in total hip arthroplasty.


An urban runoff model designed to inform stormwater management decisions.
We present an urban runoff model designed for stormwater managers to quantify runoff reduction benefits of mitigation actions that has lower input data and user expertise requirements than most commonly used models. The stormwater tool to estimate load reductions (TELR) employs a semi-distributed approach, where landscape characteristics and process representation are spatially-lumped within urban catchments on the order of 100 acres (40 ha). Hydrologic computations use a set of metrics that describe a 30-year rainfall distribution, combined with well-tested algorithms for rainfall-runoff transformation and routing to generate average annual runoff estimates for each catchment. User inputs include the locations and specifications for a range of structural best management practice (BMP) types. The model was tested in a set of urban catchments within the Lake Tahoe Basin of California, USA, where modeled annual flows matched that of the observed flows within 18% relative error for 5 of the 6 catchments and had good regional performance for a suite of performance metrics. Comparisons with continuous simulation models showed an average of 3% difference from TELR predicted runoff for a range of hypothetical urban catchments. The model usually identified the dominant BMP outflow components within 5% relative error of event-based measured flow data and simulated the correct proportionality between outflow components. TELR has been implemented as a web-based platform for use by municipal stormwater managers to inform prioritization, report program benefits and meet regulatory reporting requirements (www.swtelr.com).


AICAR attenuates organ injury and inflammatory response after intestinal ischemia and reperfusion.
Intestinal ischemia and reperfusion (I/R) is encountered in various clinical conditions and contributes to multiorgan failure and mortality as high as 60% to 80%. Intestinal I/R not only injures the intestine, but affects remote organs such as the lung leading to acute lung injury. The development of novel and effective therapies for intestinal I/R are critical for the improvement of patient outcome. AICAR (5-aminoimidazole-4-carboxyamide ribonucleoside) is a cell-permeable compound that has been shown to possess antiinflammatory effects. The objective is to determine that treatment with AICAR attenuates intestinal I/R injury and subsequent acute lung injury (ALI). Male Sprague Dawley rats (275 to 325 g) underwent intestinal I/R injury with blockage of the superior mesenteric artery for 90 min and subsequent reperfusion. At the initiation of reperfusion, vehicle or AICAR (30 mg/kg BW) was given intravenously (IV) for 30 min. At 4 h after reperfusion, blood and tissues were collected for further analyses. Treatment with AICAR significantly decreased the gut damage score and the water content, indicating improvement in histological integrity. The treatment also attenuated tissue injury and proinflammatory cytokines, and reduced bacterial translocation to the gut. AICAR administration after intestinal I/R maintained lung integrity, attenuated neutrophil chemotaxis and infiltration to the lungs and decreased lung levels of tumor necrosis factor (TNF)-α and interleukin (IL)-6. Inflammatory mediators, lung-inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) proteins, were decreased in the lungs and lung apoptosis was significantly reduced after AICAR treatment. These data indicate that AICAR could be developed as an effective and novel therapeutic for intestinal I/R and subsequent ALI.


Randomised controlled trial of postnatal sodium supplementation on body composition in 25 to 30 week gestational age infants.
To compare the effects of early and delayed sodium supplementation on body composition and body water compartments during the first two weeks of postnatal life. Preterm infants of 25-30 weeks' gestation were stratified and randomly assigned according to gender and gestational age, to receive a sodium intake of 4 mmol/kg/day beginning either on the second day after birth or when weight loss of 6% of birthweight had been achieved. Daily sodium intake, total fluid intake, energy intake, urine volume, and urinary sodium excretion were recorded. Total body water was measured by H(2)(18)O dilution on days 1, 7, and 14, and extracellular fluid volume by sodium bromide dilution on days 1 and 14. Twenty four infants received early, and 22 delayed, sodium supplementation. There were no significant differences between the groups in body water compartments on day 1. In the delayed group, but not the early group, there was a significant loss of total body water during the first week (delayed -44 ml/kg, p=0. 048; early 6 ml/kg, p=0.970). By day 14 the delayed, but not the early group, also had a significant reduction in extracellular fluid volume (delayed -53 ml/kg, p=0.01; early -37 ml/kg, p=0.2). These changes resulted in a significant alteration in body composition at the end of the first week (total body weight: delayed 791 ml/kg; early 849 ml/kg, p=0.013). By day 14 there were once again no significant differences in body composition between the two groups. Body composition after preterm birth is influenced by the timing of introduction of routine sodium supplements. Early sodium supplementation can delay the physiological loss of body water that is part of normal postnatal adaptation. This is likely to be of particular relevance to babies with respiratory distress syndrome. A tailored approach to clinical management, delaying the introduction of routine sodium supplements until there has been postnatal loss of body water, is recommended.


Advancing Genome-Resolved Metagenomics beyond the Shotgun.
Exploration of environmental microbiomes has shed light on the ecological and evolutionary principles at play in natural ecosystems and has been further accelerated through the reconstruction of population genomes to provide genome-centric context. Yet technical challenges with traditional shotgun metagenomics remain for computationally intense short-read assembly, strain heterogeneity within communities, and depth of coverage required for low-abundance microbes. In this Perspective, we highlight three main avenues for promising future developments, including coupling stable isotope probing and genome-resolved metagenomics, applying fluorescence-activated cell sorting approaches to target mini-metagenomes within a larger community, and utilizing single-molecule long-read and synthetic long-read technology to link mobile elements to host microbial cells. These developments on the horizon will undoubtedly advance genome-resolved metagenomic approaches and enable a better understanding of uncultivated microbes in their natural environments.


Preparation and in vitro/in vivo evaluation of cyclosporin A-loaded nanodecorated ocular implants for subconjunctival application.
In terms of ocular drug delivery, biodegradable implant systems have several advantages including the ability to provide constant drug concentration at the target site, no necessity for surgical removal, and minimum systemic side effects. Cyclosporin A (CsA) is a neutral, hydrophobic, cyclic peptide of amino acids that frequently used for dry eye disease treatment. The aim of this study was to develop a nanoparticle-loaded implant system for sustained-release CsA delivery following subconjunctival implantation. Poly(lactide-co-glycolide) (85:15) or poly-ε-caprolactone (PCL) were used to prepare two different nanoparticle formulations. These nanoparticles loaded into PCL or poly(lactide-co-caprolactone) implant formulations were prepared by two different methods, which were molding and electrospinning. Size and zeta potential of nanoparticles were determined and the morphology of the formulations were investigated by scanning electron microscopy. CsA-loading efficiencies were calculated and the in vitro degradation and in vitro release studies were performed. MTT test was also performed using L929 fibroblast cells to evaluate the cytotoxicity of the formulations. PCL-PCL-NP-I formulation was implanted to Swiss Albino mice with induced dry eye syndrome to evaluate the efficacy. In vitro release studies showed that the release from the formulations continues between 30 and 60 days, and the cell viability was found to be 77.4%-99.0%. In vivo studies showed that healing is significantly faster in the presence of the selected implant formulation. Results indicated that nanodecorated implants are promising ocular carriers for controlled-release CsA application.


Yellow-legged gull eggs (Larus michahellis) as persistent organic pollutants and trace metal bioindicator for two nearby areas with different human impact.
The concentration of different persistent organic pollutants (POPs including chlorinated and brominated compounds) and trace metals and metalloids (As, Cd, Cu, Cr, Pb, Hg, Ni, and Zn) was examined in eggs from two colonies of yellow-legged gulls. The two colonies are established in Ría de Vigo, Northwest Spain, with a distance between them of only 10 km, one in Vigo town (industrial and harbour activities) and the other in the Cíes Islands in a Natural Park and Marine Protected Area -MPA- (with no known anthropogenic inputs). Statistically significant differences for the two colonies were observed for Hg, the sum of 7 CBs, the sum of DDTs y and the sum of 9 PBDEs, with values that could be causing some toxic effects in the area of the most anthropogenically influenced colony. The estimated isotopic niche was also calculated, based on δ15N and δ13C, for the two colonies, pointing to a wider diet in the Cíes colony when compared to the diet in the Vigo colony. The study supports the use of the yellow-legged seagull eggs as a bioindicator of pollution capable of differentiating pollution level even in geographically close areas.


Induced optical activity of DNA-templated cyanine dye aggregates: exciton coupling theory and TD-DFT studies.
Certain cyanine dye molecules have been observed to self-assemble in DNA templates to form large chiral aggregates, which exhibit induced circular dichroism. The structure and circular dichroism (CD) of one such system, aggregates of a cationic DiSC2(5) cyanine dye, are investigated using the time-dependent Kohn-Sham density functional theory (TD-DFT) and exciton coupling model. A series of TD-DFT calculations on the aggregates with one, two, and four dye molecules clearly shows the onset of CD induced by the helically twisted structure compatible with the minor groove of DNA templates. More simplified exciton coupling model analysis successfully reproduces the major positive Cotton effect observed in the experiment as well as the TD-DFT calculations, but it is unable to capture minor features of the CD spectrum that are closely related to absolute configurations of constituent dyes in the complex. We assess the performance of various methods used for evaluation of the electronic coupling energies between interacting chromophores. Our results confirm that the interchromophore interactions in cyanine dye aggregates are primarily electrostatic in nature and indicate that the exciton coupling model is adequate for studying induced CD of self-assembled aggregates of cyanine dye molecules.


Audiometric findings in Waardenburg's syndrome amongst the institutionalised deaf/blind in Kaduna-Nigeria.
Waardenburg's syndrome (WS) is a rare inherited disorder of congenital hearing loss and Pigmentary disturbances of the eyes, hair, skin and neural crest derivatives. 620 students in a deaf/blind school were examined and four had WS with a frequency of 0.65%. 2 males and 2 females with WS and age ranges between 10-19yrs {mean 15.75yrs.} All 4 subjects had complete blue irides, white forelock and sensorineural hearing loss, and thus met the diagnostic criteria. They were then subjected to Audiometric assessment. Otoscopy was essentially normal but Audiometry revealed sensorineural hearing loss [SNHL] in all the subjects ranging from severe to profound with one subject being stone deaf. WS is a rare disorder in our environment although it may be under reported. Two ofthe subjects benefited from amplification and were given hearing Aids. Provision of early amplification cannot be over emphasized.


Inhibition of Stat3 signaling pathway by nifuroxazide improves antitumor immunity and impairs colorectal carcinoma metastasis.
Colorectal carcinoma (CRC) is the one of the most common cancers with considerable metastatic potential, explaining the need for new drug candidates that inhibit tumor metastasis. The signal transducers and activators of the transcription 3 (Stat3) signaling pathway has an important role in CRC and has been validated as a promising anticancer target for CRC therapy. In the present study, we report our findings on nifuroxazide, an antidiarrheal agent identified as an inhibitor of Stat3. Our studies showed that nifuroxazide decreased the viability of three CRC cell lines and induced apoptosis of cancer cells in a concentration-dependent manner. Moreover, western blot analysis demonstrated that the occurrence of its apoptosis was correlated with the activation of Bax and cleaved caspase-3, and decreased the expression of Bcl-2. In addition, nifuroxazide markedly impaired CRC cell migration and invasion by downregulating phosphorylated-Stat3Tyr705, and also impaired the expression of matrix metalloproteinases (MMP-2 and MMP-9). Furthermore, our studies showed that nifuroxazide also significantly inhibited the tumor metastasis in lung and abdomen metastasis models of colon cancer. Meanwhile, nifuroxazide functionally reduced the proliferation index, induced tumor apoptosis and impaired metastasis. Notably, nifuroxazide reduced the number of myeloid-derived suppressor cells in the blood, spleens and tumors, accompanied by the increased infiltration of CD8+ T cells in the tumors. Importantly, a marked decrease in the number of M2-type macrophages in tumor in the abdomen metastasis model was also observed. Taken together, our results indicated that nifuroxazide could effectively inhibit tumor metastasis by mediating Stat3 pathway and it might have a therapeutic potential for the treatment of CRC.


Mechanism of acute silver toxicity in marine invertebrates.
In freshwater crustaceans and in both freshwater and marine fish, the key mechanism of acute silver toxicity involves ionoregulatory impairment. An inhibition of the Na+ ,K+-ATPase located at the basolateral membrane of the gill epithelium seems to be the key site for silver toxicity. However, studies to determine if the same mechanism of toxicity is occurring in marine invertebrates, which also are ionoregulators, had not been done. Thus, the present study was carried out to determine acute silver effects on hemolymph osmo- and ionoregulation in three marine invertebrates: the shrimp Penaeus duorarum, the sea hare Aplysia californica, and the sea urchin Diadema antillarum. Animals were exposed to silver (1 or 10 microg/L), as silver nitrate, in seawater for 48 h. Results show that acute silver exposure did not affect hemolymph osmolality or ion concentration (Na+, Cl-, K+, Ca2+ and Mg2+) in the three species studied. However, silver induced significant changes in the water content in shrimp gill and sea hare gill and hepatopancreas. Silver also caused significant changes in Na+ ,K+-ATPase activity and in both total and intracellular ion (Cl-, Na+, K+, Mg2+, and Ca2+) concentrations in different tissues of the three species studied. Overall, these results show that the key mechanism of acute silver toxicity in marine invertebrates is not associated with an osmotic or ionoregulatory impairment at the hemolymph level, as observed in freshwater fish and crustaceans and in seawater fish. However, they indicate that acute waterborne silver induces significant changes in Na+ ,K(+)-ATPase activity and probably affects other mechanisms involved in water and ion transport at the cell membrane level, inducing impairments in water and ion regulation at the cellular level in different tissues of marine invertebrates. These results indicate the need to consider other "toxic sites" than gills in any future extension of the biotic ligand model (BLM) for seawater.