Insights into Brain Architectures from the Homological Scaffolds of Functional Connectivity Networks.
In recent years, the application of network analysis to neuroimaging data has provided useful insights about the brain's functional and structural organization in both health and disease. This has proven a significant paradigm shift from the study of individual brain regions in isolation. Graph-based models of the brain consist of vertices, which represent distinct brain areas, and edges which encode the presence (or absence) of a structural or functional relationship between each pair of vertices. By definition, any graph metric will be defined upon this dyadic representation of the brain activity. It is however unclear to what extent these dyadic relationships can capture the brain's complex functional architecture and the encoding of information in distributed networks. Moreover, because network representations of global brain activity are derived from measures that have a continuous response (i.e., interregional BOLD signals), it is methodologically complex to characterize the architecture of functional networks using traditional graph-based approaches. In the present study, we investigate the relationship between standard network metrics computed from dyadic interactions in a functional network, and a metric defined on the persistence homological scaffold of the network, which is a summary of the persistent homology structure of resting-state fMRI data. The persistence homological scaffold is a summary network that differs in important ways from the standard network representations of functional neuroimaging data: (i) it is constructed using the information from all edge weights comprised in the original network without applying an ad hoc threshold and (ii) as a summary of persistent homology, it considers the contributions of simplicial structures to the network organization rather than dyadic edge-vertices interactions. We investigated the information domain captured by the persistence homological scaffold by computing the strength of each node in the scaffold and comparing it to local graph metrics traditionally employed in neuroimaging studies. We conclude that the persistence scaffold enables the identification of network elements that may support the functional integration of information across distributed brain networks.


Geriatric Syndrome Screening: Implementing an Evidence-Based Process in Hospitalized Older Adults With Cancer.
Oncology nurses are in a key position to screen patients for geriatric syndromes that can disproportionately affect older adults with cancer. Early recognition can promote informed decision-making, initiation of interventions, and tolerance of treatment. This study aimed to implement a nursing-sensitive screening process to identify types and frequency of geriatric syndromes in hospitalized older adult patients with cancer. Nurses were trained to screen for geriatric syndromes using the SPICES tool (sleep disorders, problems with eating or feeding, incontinence, confusion, evidence of falls, and skin breakdown). The electronic health record was leveraged to facilitate documentation and provide a geriatric syndrome profile for each patient. Results demonstrated ability of staff nurses to lead an evidence-based practice initiative and the challenges faced. Adherence to SPICES screening was 47%. About 80% of patients had two or more geriatric syndrome issues.


Radiofrequency ablation for T1a tumors in a solitary kidney: promising intermediate oncologic and renal function outcomes.
The experience with radiofrequency ablation (RFA) in solitary kidneys is limited in numbers and follow-up. Therefore, we report our oncologic and renal function outcomes of RFA for T1a tumors in patients with a solitary kidney. From April 2000 to August 2007, 242 patients were treated with RFA for renal cortical masses. Sixteen patients with localized tumors < or = 4 cm in a solitary kidney were identified. Clinical and radiographic data were reviewed to assess indications, complications, disease recurrence, and renal function. Twenty-one renal masses were ablated in 16 patients with a solitary kidney. The mean patient age was 66.1 years, and the mean tumor size was 2.6 cm (range, 1.1-4.0). Preoperative biopsy was diagnostic of renal cell carcinoma (RCC) in 75% of cases. At a mean follow-up of 30.7 months (range, 1.5-66.0), 14/16 (88%) patients had no radiographic evidence of disease recurrence. One patient with three masses in a solitary kidney had a local recurrence managed by salvage RFA. The other patient with a local failure also had severe baseline renal insufficiency with progression to end stage renal disease 2.5 years following RFA and underwent a radical nephrectomy. Mean glomerular filtration rate (GFR) decreased from 54.2 ml/min/1.73m2 preoperatively to 47.5 ml/min/1.73m2 at last follow-up (p = 0.015). There were no major complications, and four patients had minor complications which resolved without intervention. Radiofrequency ablation is an attractive alternative for the management of patients with T1a renal cortical tumors in a solitary kidney. Renal function appears to be adequately maintained with promising oncologic outcomes at 2.5 years.


Occupation recorded on certificates of death compared with self-report: the Atherosclerosis Risk in Communities (ARIC) Study.
Death certificates are a potential source of sociodemographic data for decedents in epidemiologic research. However, because this information is provided by the next-of-kin or other proxies, there are concerns about validity. Our objective was to assess the agreement of job titles and occupational categories derived from death certificates with that self-reported in mid and later life. Occupation was abstracted from 431 death certificates from North Carolina Atherosclerosis Risk in Communities Study participants who died between 1987 and 2001. Occupations were coded according to 1980 Bureau of Census job titles and then grouped into six 1980 census occupational categories. This information was compared with the self-reported occupation at midlife as reported at the baseline examination (1987-89). We calculated percent agreement using standard methods. Chance-adjusted agreement was assessed by kappa coefficients, with 95% confidence intervals. Agreement between death certificate and self-reported job titles was poor (32%), while 67% of occupational categories matched the two sources. Kappa coefficients ranged from 0.53 for technical/sales/administrative jobs to 0.68 for homemakers. Agreement was lower, albeit nonsignificant, for women (kappa = 0.54, 95% Confidence Interval, CI = 0.44-0.63) than men (kappa = 0.62, 95% CI = 0.54-0.69) and for African-Americans (kappa = 0.47, 95% CI = 0.34-0.61) than whites (kappa = 0.63, 95% CI = 0.57-0.69) but varied only slightly by educational attainment. While agreement between self- and death certificate reported job titles was poor, agreement between occupational categories was good. This suggests that while death certificates may not be a suitable source of occupational data where classification into specific job titles is essential, in the absence of other data, it is a reasonable source for constructing measures such as occupational SES that are based on grouped occupational data.


Residual inhibition of epidermal growth factor binding by pancreatic secretagogues and phorbol ester in rat pancreas.
Cholecystokinin-octapeptide (CCK8) inhibits 125I-labeled epidermal growth factor (EGF) cell-associated radioactivity in pancreatic acini, ostensibly as a result of its ability to mobilize cellular Ca2+. The phorbol ester tetradecanoyl phorbol acetate (TPA), a compound that activates protein kinase C, mimics the inhibitory action of CCK8. In the present study we examined the relationship between occupancy of the cholecystokinin (CCK) receptor, the subsequent inhibition of EGF binding, and the potential role of C-kinase activation in mediating this inhibition. Proglumide and dibutyryl cyclic GMP (dbGMP), two distinct competitive antagonists of CCK8, reversed the inhibitory actions of CCK8. Analysis of steady-state saturation kinetics of 125I-EGF binding indicated that CCK8 decreased the apparent affinity of the EGF receptor, mainly as a result of a marked decrease in the amount of internalized ligand. TPA also inhibited 125I-EGF internalization. Removal of CCK8 and TPA from incubation medium did not abolish their inhibitory actions. Carbachol, but not bombesin, exerted a similar residual inhibitory effect. It is suggested that in addition to acting via Ca2+, certain pancreatic secretagogues may also act through C-kinase to regulate EGF binding.


Incidence of diabetes among postmenopausal breast cancer survivors.
Evidence is emerging of an association between breast cancer and diabetes; however, it is uncertain whether diabetes incidence is increased in postmenopausal breast cancer survivors compared with women without breast cancer. The objective of this study was to determine whether postmenopausal women who develop breast cancer have a higher incidence of diabetes than those who do not develop breast cancer. We used population-based data from Ontario, Canada to compare the incidence of diabetes among women with breast cancer, aged 55 years or older, from 1996 to 2008, with that of age-matched women without breast cancer. We used Cox proportional hazard models to estimate the effect of breast cancer on the cause-specific hazard of developing diabetes overall and in the subgroup of women who received adjuvant chemotherapy. Of 24,976 breast cancer survivors and 124,880 controls, 9.7% developed diabetes over a mean follow-up of 5.8 years. The risk of diabetes among breast cancer survivors compared with women without breast cancer began to increase 2 years after diagnosis (HR 1.07 [95% CI, 1.02, 1.12]), and rose to an HR of 1.21 (95% CI, 1.09, 1.35) after 10 years. Among those who received adjuvant chemotherapy (n = 4,404), risk was highest in the first 2 years after diagnosis (HR 1.24 [95% CI 1.12, 1.38]) and then declined. We found a modest increase in the incidence of diabetes among postmenopausal breast cancer survivors that varied over time. In most women the risk began to increase 2 years after cancer diagnosis but the highest risk was in the first 2 years in those who received adjuvant therapy. Our study suggests that greater diabetes screening and prevention strategies among breast cancer survivors may be warranted.


Characterization of the main steps in first shell formation in Mytilus galloprovincialis: possible role of tyrosinase.
Bivalve biomineralization is a highly complex and organized process, involving several molecular components identified in adults and larval stages. However, information is still scarce on the ontogeny of the organic matrix before calcification occurs. In this work, first shell formation was investigated in the mussel Mytilus galloprovincialis. The time course of organic matrix and CaCO3 deposition were followed at close times post fertilization (24, 26, 29, 32, 48 h) by calcofluor and calcein staining, respectively. Both components showed an exponential trend in growth, with a delay between organic matrix and CaCO3 deposition. mRNA levels of genes involved in matrix deposition (chitin synthase; tyrosinase- TYR) and calcification (carbonic anhydrase; extrapallial protein) were quantified by qPCR at 24 and 48 hours post fertilization (hpf) with respect to eggs. All transcripts were upregulated across early development, with TYR showing highest mRNA levels from 24 hpf. TYR transcripts were closely associated with matrix deposition as shown by in situ hybridization. The involvement of tyrosinase activity was supported by data obtained with the enzyme inhibitor N-phenylthiourea. Our results underline the pivotal role of shell matrix in driving first CaCO3 deposition and the importance of tyrosinase in the formation of the first shell in M. galloprovincialis.


Probing mechanoregulation of neuronal differentiation by plasma lithography patterned elastomeric substrates.
Cells sense and interpret mechanical cues, including cell-cell and cell-substrate interactions, in the microenvironment to collectively regulate various physiological functions. Understanding the influences of these mechanical factors on cell behavior is critical for fundamental cell biology and for the development of novel strategies in regenerative medicine. Here, we demonstrate plasma lithography patterning on elastomeric substrates for elucidating the influences of mechanical cues on neuronal differentiation and neuritogenesis. The neuroblastoma cells form neuronal spheres on plasma-treated regions, which geometrically confine the cells over two weeks. The elastic modulus of the elastomer is controlled simultaneously by the crosslinker concentration. The cell-substrate mechanical interactions are also investigated by controlling the size of neuronal spheres with different cell seeding densities. These physical cues are shown to modulate with the formation of focal adhesions, neurite outgrowth, and the morphology of neuroblastoma. By systematic adjustment of these cues, along with computational biomechanical analysis, we demonstrate the interrelated mechanoregulatory effects of substrate elasticity and cell size. Taken together, our results reveal that the neuronal differentiation and neuritogenesis of neuroblastoma cells are collectively regulated via the cell-substrate mechanical interactions.


Visible-light-triggered molecular photoswitch based on reversible E/Z isomerization of a 1,2-dicyanoethene derivative.
A designed bis(dithienyl) dicyanoethene-based, strictly E/Z photoswitch (4TCE) operates through state-selective (E and Z isomer) photoactivation with visible light. The E and Z isomers of 4TCE exhibit remarkably different spectroscopic characteristics, including a large separation (70 nm) in their absorption maxima (λ(max)) and a 2.5-fold increase in molar extinction coefficient from cis to trans. The energetically stable trans form can be completely converted to the cis form within minutes when exposed to white light, whereas the reverse isomerization occurs readily upon irradiation by blue light (λ<480 nm) or completely by thermal conversion at elevated temperatures. These features together with excellent thermal stability and photostability of both isomers make this new E/Z photoswitch a promising building block for photoswitchable materials that operate without the need for UV light.


Diarrhea as a cause of mortality in a mouse model of infectious colitis.
Comparative characterization of genome-wide transcriptional changes during infection can help elucidate the mechanisms underlying host susceptibility. In this study, transcriptional profiling of the mouse colon was carried out in two cognate lines of mice that differ in their response to Citrobacter rodentium infection; susceptible inbred FVB/N and resistant outbred Swiss Webster mice. Gene expression in the distal colon was determined prior to infection, and at four and nine days post-inoculation using a whole mouse genome Affymetrix array. Computational analysis identified 462 probe sets more than 2-fold differentially expressed between uninoculated resistant and susceptible mice. In response to C. rodentium infection, 5,123 probe sets were differentially expressed in one or both lines of mice. Microarray data were validated by quantitative real-time RT-PCR for 35 selected genes and were found to have a 94% concordance rate. Transcripts represented by 1,547 probe sets were differentially expressed between susceptible and resistant mice regardless of infection status, a host effect. Genes associated with transport were over-represented to a greater extent than even immune response-related genes. Electrolyte analysis revealed reduction in serum levels of chloride and sodium in susceptible animals. The results support the hypothesis that mortality in C. rodentium-infected susceptible mice is associated with impaired intestinal ion transport and development of fatal fluid loss and dehydration. These studies contribute to our understanding of the pathogenesis of C. rodentium and suggest novel strategies for the prevention and treatment of diarrhea associated with intestinal bacterial infections.


A Pilot Study into the Effects of the CB1 Cannabinoid Receptor Agonist WIN55,212-2 or the Antagonist/Inverse Agonist AM251 on Sleep in Rats.
The plant cannabinoid Δ(9)-tetrahydrocannabinol and the endocannabinoid anandamide increase the amount of sleep via a CB1 receptor mediated mechanism. Here, we explored the use of a novel electroencephalogram (EEG) recording device based on wireless EEG microchip technology (Neurologger) in freely-moving rats, and its utility in experiments of cannabinoids-induced alterations of EEG/vigilance stages. EEG was recorded through epidural electrodes placed above pre-frontal and parietal cortex (overlaying the dorsal hippocampus). As cannabinoids, we acutely administered the full synthetic CB1 receptor agonist, WIN55,212-2 (1 mg/kg), and the antagonist/inverse agonist, AM251 (2 mg/kg), either alone or together through the intraperitoneal route. WIN55,212-2 increased the total amount of NREM sleep and the length of each NREM bout, but this was unlikely due to CB1 receptor activation since it was not prevented by AM251. However, WIN55,212-2 also lowered overall EEG spectral power especially in theta and alpha frequency bands during wakefulness and NREM sleep, and this effect was reversed by AM251. The antagonist/inverse agonist caused no sleep alterations by itself and moderately increased spectral power in Theta, alpha and beta frequency bands during NREM sleep when administered on its own. Implications of endocannabinoid modulation of the sleep-wake cycle and its possible interactions with other transmitter systems are considered.


Prognostic significance of CEBPA mutations in a large cohort of younger adult patients with acute myeloid leukemia: impact of double CEBPA mutations and the interaction with FLT3 and NPM1 mutations.
To determine the clinical relevance of mutations in the CCAAT/enhancer binding protein alpha (CEBPA) gene in acute myeloid leukemia (AML) and to examine factors that might modify prognostic impact. The entire CEBPA coding sequence was screened in 1,427 young adult patients with AML, excluding acute promyelocytic leukemia, using denaturing high-performance liquid chromatography and direct sequencing. Of 107 patients (7%) with CEBPA mutations, 48 patients (45%) had one mutation (CEBPA-single), and 59 patients (55%) had two mutations (CEBPA-double). The incidence of CEBPA-double patients was similar in intermediate cytogenetic risk patients with and without a normal karyotype (6% and 5%, respectively). CEBPA-double patients had evidence of a lower coincidence with FLT3/ITDs (P = .04) and were highly unlikely to have an NPM1 mutation (P < .0001). CEBPA-double but not CEBPA-single patients had a significantly better overall survival (OS) at 8 years (34%, 31%, and 54% for CEBPA-wild-type [WT], CEBPA-single, and CEBPA-double, respectively, P = .004). This benefit was lost in the presence of a FLT3/ITD (OS for CEBPA-WT, CEBPA-single, and CEBPA-double FLT3/ITD-negative patients: 36%, 35%, 59%, respectively, P = .002; OS for CEBPA-WT, CEBPA-single, and CEBPA-double FLT3/ITD-positive patients: 26%, 21%, 14%, respectively, P = .05). There was no evidence of any additional favorable benefit for a CEBPA-single mutation in the presence of an NPM1 mutation (OS, 45%, 44%, and 56%, P = .2, for NPM1-positive/CEBPA-WT, NPM1-positive/CEBPA-single, and NPM1-negative/CEBPA-double patients, respectively). Screening for CEBPA mutations can be restricted to patients with intermediate-risk cytogenetics lacking an FLT3/ITD or NPM1 mutation. Only the presence of a CEBPA-double mutation should be used for therapy risk stratification.


Effective aortic regurgitant orifice area: description of a method based on the conservation of mass.
The natural history of aortic regurgitation is incompletely understood in part because of the lack of a simple method to estimate the defect size. A method of determining the effective regurgitant orifice area that combines Doppler catheter and Doppler echocardiographic techniques and is based on the principle of conservation of mass (the continuity equation) is described. To validate the application of the Doppler catheter system for measuring regurgitant supravalvular diastolic flow, an in vitro model of retrograde aortic flow was used. These studies indicated that measurements of supravalvular retrograde velocity with the Doppler catheter accurately reflect retrograde diastolic velocity when the aorta is less than 4.8 cm in diameter. Twenty-three patients undergoing cardiac catheterization were studied; 20 of these patients had aortic regurgitation. Retrograde supravalvular diastolic velocity was determined from a Doppler catheter positioned above the aortic valve. The effective regurgitant orifice area was calculated with use of the Doppler catheter-derived regurgitant volume and mean transvalvular diastolic velocity as determined by either catheterization or continuous wave Doppler echocardiography. The catheterization-derived regurgitant orifice area increased with the angiographic grade of as follows: 1+ (0.04 to 0.10 cm2), 2+ (0.15 to 0.49 cm2), 3+ (0.29 to 1.11 cm2) and 4+ (1.24 to 1.33 cm2). By combining Doppler catheter, echocardiographic and cardiac catheterization techniques, the effective aortic regurgitant orifice area may be estimated; this hydrodynamic area correlates with grading by supravalvular aortography. Calculation of this area provides a quantitative alternative to aortography for estimating the severity of aortic regurgitation but should be used with caution in patients with a markedly dilated aorta.


Biosynthesis of the canine zona pellucida requires the integrated participation of both oocytes and granulosa cells.
In the dog, attempts to localize the expression of zona pellucida (ZP) proteins during folliculogenesis have failed to demonstrate conclusively whether any or all of the zona proteins are synthesized in the oocyte or the granulosa cells. Probing of paraformaldehyde-fixed prepubertal canine ovarian tissue sections with a panel of fluorescently conjugated lectins localized the expression of glycoproteins during folliculogenesis. We confirm that six lectins (PSA, s-WGA, ECL, GSL-II, LEL, and STL) consistently labeled the ZP and adjacent granulosa cells of the developing follicle and that canine ZP expresses beta-gal(1,4)glcNAc, beta-gal(1,3)galNac, alpha-mannose, and terminal sialic acid residues in a developmentally specific manner. Riboprobes for canine ZPA and ZPC genes were produced and used for in situ hybridization studies of mRNA expression in canine folliculogenesis. In addition, we isolated a partial cDNA transcript from total ovarian RNA for the canine ZPB gene having a high degree of sequence identity with the felid and porcine ZPB homologues. Subsequently, the ZPA gene transcripts were localized to the cytoplasm of oocytes in primordial, primary, and early secondary follicles. We then localized expression of ZPB and ZPC gene transcripts to the granulosa cells of growing follicles, but not in squamous granulosa cells of primordial follicles or oocytes. These observations indicate that in the juvenile canine ovary, the oocyte is responsible for synthesis of the ZPA protein and directing synthesis of the ZPB and ZPC proteins by the granulosa cells and that ZP gene transcription occurs in a sequential manner during folliculogenesis.


Stress inducible proteinase inhibitor diversity in Capsicum annuum.
Wound-inducible Pin-II Proteinase inhibitors (PIs) are one of the important plant serine PIs which have been studied extensively for their structural and functional diversity and relevance in plant defense against insect pests. To explore the functional specialization of an array of Capsicum annuum (L.) proteinase inhibitor (CanPIs) genes, we studied their expression, processing and tissue-specific distribution under steady-state and induced conditions. Inductions were performed by subjecting C. annuum leaves to various treatments, namely aphid infestation or mechanical wounding followed by treatment with either oral secretion (OS) of Helicoverpa armigera or water. The elicitation treatments regulated the accumulation of CanPIs corresponding to 4-, 3-, and 2-inhibitory repeat domains (IRDs). Fourty seven different CanPI genes composed of 28 unique IRDs were identified in total along with those reported earlier. The CanPI gene pool either from uninduced or induced leaves was dominated by 3-IRD PIs and trypsin inhibitory domains. Also a major contribution by 4-IRD CanPI genes possessing trypsin and chymotrypsin inhibitor domains was specifically revealed in wounded leaves treated with OS. Wounding displayed the highest number of unique CanPIs while wounding with OS treatment resulted in the high accumulation of specifically CanPI-4, -7 and -10. Characterization of the PI protein activity through two dimensional gel electrophoresis revealed tissue and induction specific patterns. Consistent with transcript abundance, wound plus OS or water treated C. annuum leaves exhibited significantly higher PI activity and isoform diversity contributed by 3- and 4-IRD CanPIs. CanPI accumulation and activity was weakly elicited by aphid infestation yet resulted in the higher expression of CanPI-26, -41 and -43. Plants can differentially perceive various kinds of insect attacks and respond appropriately through activating plant defenses including regulation of PIs at transcriptional and post-translational levels. Based on the differentially elicited CanPI accumulation patterns, it is intriguing to speculate that generating sequence diversity in the form of multi-IRD PIs is a part of elaborative plant defense strategy to obtain a diverse pool of functional units to confine insect attack.


Human immunodeficiency virus type 1 Vpr-mediated G2 arrest requires Rad17 and Hus1 and induces nuclear BRCA1 and gamma-H2AX focus formation.
Eukaryotic cells have evolved a complex mechanism for sensing DNA damage during genome replication. Activation of this pathway prevents entry into mitosis to allow for either DNA repair or, in the event of irreparable damage, commitment to apoptosis. Under conditions of replication stress, the damage signal is initiated by the ataxia-telangiectasia-mutated and Rad3-related kinase ATR. We recently demonstrated that the human immunodeficiency virus type 1 (HIV-1) gene product viral protein R (Vpr) arrests infected cells in the G(2) phase via the activation of ATR. In the present study, we show that the activation of ATR by Vpr is analogous to activation by certain genotoxic agents, both mechanistically and in its downstream consequences. Specifically, we show a requirement for Rad17 and Hus1 to induce G(2) arrest as well as Vpr-induced phosphorylation of histone 2A variant X (H2AX) and formation of nuclear foci containing H2AX and breast cancer susceptibility protein 1. These results demonstrate that G(2) arrest mediated by the HIV-1 gene product Vpr utilizes the cellular signaling pathway whose physiological function is to recognize replication stress. These findings should contribute to a greater understanding of how HIV-1 manipulates the CD4(+)-lymphocyte cell cycle and apoptosis induction in the progressive CD4(+)-lymphocyte depletion characteristic of HIV-1 pathogenesis.


Electrolytic valving isolation of cell co-culture microenvironment with controlled cell pairing ratios.
Cancer-stromal interaction is a critical process in tumorigenesis. Conventional dish-based co-culture assays simply mix two cell types in the same dish; thus, they are deficient in controlling cell locations and precisely tracking single cell behavior from heterogeneous cell populations. Microfluidic technology can provide a good spatial-temporal control of microenvironments, but the control has been typically realized by using external pumps, making long-term cultures cumbersome and bulky. In this work, we have presented a cell-cell interaction microfluidic platform that can accurately control the co-culture microenvironment by using a novel electrolytic cell isolation scheme without using any valves or pneumatic pumps. The proposed microfluidic platform can also precisely control the number of interacting cells and pairing ratios to emulate cancer niches. More than 80% of the chambers captured the desired number of cells. The duration of cell isolation can be adjusted by electrolytic bubble generation and removal. We have verified that the electrolytic process has a negligible effect on cell viability and proliferation in our platform. To the best of our knowledge, this work is the first attempt to incorporate electrolytic bubble generation as a cell isolation method in microfluidics. For proof of feasibility, we have performed cell-cell interaction assays between prostate cancer (PC3) cells and myoblast (C2C12) cells. The preliminary results demonstrated the potential of using electrolysis for micro-environmental control during cell culture. Also, the ratio controlled cell-cell interaction assays were successfully performed which showed that the cell pairing ratios of PC3 to C2C12 affected the proliferation rate of myoblast cells due to increased secretion of growth factors from prostate cancer cells.


Plasma-Wave Terahertz Detection Mediated by Topological Insulators Surface States.
Topological insulators (TIs) represent a novel quantum state of matter, characterized by edge or surface-states, showing up on the topological character of the bulk wave functions. Allowing electrons to move along their surface, but not through their inside, they emerged as an intriguing material platform for the exploration of exotic physical phenomena, somehow resembling the graphene Dirac-cone physics, as well as for exciting applications in optoelectronics, spintronics, nanoscience, low-power electronics, and quantum computing. Investigation of topological surface states (TSS) is conventionally hindered by the fact that in most of experimental conditions the TSS properties are mixed up with those of bulk-states. Here, we activate, probe, and exploit the collective electronic excitation of TSS in the Dirac cone. By engineering Bi2Te(3-x)Sex stoichiometry, and by gating the surface of nanoscale field-effect-transistors, exploiting thin flakes of Bi2Te2.2Se0.8 or Bi2Se3, we provide the first demonstration of room-temperature terahertz (THz) detection mediated by overdamped plasma-wave oscillations on the "activated" TSS of a Bi2Te2.2Se0.8 flake. The reported detection performances allow a realistic exploitation of TSS for large-area, fast imaging, promising superb impacts on THz photonics.


The angiotensin AT2-receptor mediates inhibition of cell proliferation in coronary endothelial cells.
Angiotensin II (ANG II) is known to be a potent growth promoting factor for vascular smooth muscle cells and fibroblasts but little is known about its influence on growth in endothelial cells. We studied the effects of ANG II on endothelial growth and the role of the angiotensin receptor subtypes involved. Proliferation of rat coronary endothelial cells (CEC) and rat vascular smooth muscle cells (VSMC) was determined by [3H]thymidine incorporation, the MTT-test and by directly counting cells in a coulter counter. Angiotensin AT1- and AT2-receptors were demonstrated by binding studies and by the presence of their respective mRNA through reverse transcription polymerase chain reaction (RT-PCR). In contrast to VSMC, which in culture only express the AT1-receptor, CEC express both, AT1- and AT2-receptors simultaneously up to the third passage. Whereas ANG II stimulated growth of quiescent VSMC, an effect abolished by pretreatment with the AT1-receptor antagonist, losartan, ANG II did not induce proliferation in quiescent CEC. However, after pretreatment of quiescent endothelial cells (< passage 4) with the AT2-receptor antagonist, PD 123177, ANG II induced proliferation. This effect was reversed by additional pretreatment with losartan. ANG II significantly inhibited the proliferation of bFGF-stimulated CEC in a dose-dependent manner by maximally 50%. This effect was prevented by PD 123177 while losartan was ineffective. The AT2-receptor agonist, CGP 42112, mimicked the antiproliferative actions of ANG II, confirming the specificity of the effect. Our results show that the growth modulating actions of ANG II depend on the type of angiotensin receptor present on a given cell. In coronary endothelial cells, the antiproliferative actions of the AT2-receptor offset the growth promoting effects mediated by the AT1-receptor.


Long working hours as a risk factor for atrial fibrillation: a multi-cohort study.
Studies suggest that people who work long hours are at increased risk of stroke, but the association of long working hours with atrial fibrillation, the most common cardiac arrhythmia and a risk factor for stroke, is unknown. We examined the risk of atrial fibrillation in individuals working long hours (≥55 per week) and those working standard 35-40 h/week. In this prospective multi-cohort study from the Individual-Participant-Data Meta-analysis in Working Populations (IPD-Work) Consortium, the study population was 85 494 working men and women (mean age 43.4 years) with no recorded atrial fibrillation. Working hours were assessed at study baseline (1991-2004). Mean follow-up for incident atrial fibrillation was 10 years and cases were defined using data on electrocardiograms, hospital records, drug reimbursement registers, and death certificates. We identified 1061 new cases of atrial fibrillation (10-year cumulative incidence 12.4 per 1000). After adjustment for age, sex and socioeconomic status, individuals working long hours had a 1.4-fold increased risk of atrial fibrillation compared with those working standard hours (hazard ratio = 1.42, 95% CI = 1.13-1.80, P = 0.003). There was no significant heterogeneity between the cohort-specific effect estimates (I2 = 0%, P = 0.66) and the finding remained after excluding participants with coronary heart disease or stroke at baseline or during the follow-up (N = 2006, hazard ratio = 1.36, 95% CI = 1.05-1.76, P = 0.0180). Adjustment for potential confounding factors, such as obesity, risky alcohol use and high blood pressure, had little impact on this association. Individuals who worked long hours were more likely to develop atrial fibrillation than those working standard hours.


Changes in Provider Treatment Patterns for Dupuytren's Contracture: Analysis of Trends in Medicare Beneficiaries.
Collagenase Clostridium histolyticum (CCH) injection has been shown to be a safe and effective treatment option for Dupuytren's contracture. We hypothesize that the gaining popularity of CCH has resulted in a change in treatment patterns among providers, with increased utilization of CCH injections in the management of Dupuytren's contracture from 2012 to 2014. The Medicare Provider Utilization and Payment Data Public Use Files were used to identify all surgeons who submitted claims for surgical fasciectomy, needle aponeurotomy (NA), and CCH injection. The data were analyzed for number of providers performing the procedures, number of procedures per provider, and location of practice. From 2012 to 2014, the number of providers performing more than 10 open fasciectomies decreased from 141 to 131. In the same time, the number of providers performing more than 10 NAs increased from 63 to 70 with mean procedures per provider decreasing from 35 to 21. In contrast, the number of providers performing more than 10 CCH injections increased from 72 to 112, with mean injections per provider going from 24 to 20. The total number of injections performed increased from 1,734 to 2,220 from 2012 to 2014. The largest increase in number of injections and number of providers performing injections occurred in the South. The introduction of collagenase has changed treatment patterns with more providers treating Dupuytren's contractures with CCH injections and a statistically significant decline in the number of NA procedures per provider.


Balance of nanostructure and bimetallic interactions in Pt model fuel cell catalysts: in situ XAS and DFT study.
We have studied the effect of nanostructuring in Pt monolayer model electrocatalysts on a Rh(111) single-crystal substrate on the adsorption strength of chemisorbed species. In situ high energy resolution fluorescence detection X-ray absorption spectroscopy at the Pt L(3) edge reveals characteristic changes of the shape and intensity of the "white-line" due to chemisorption of atomic hydrogen (H(ad)) at low potentials and oxygen-containing species (O/OH(ad)) at high potentials. On a uniform, two-dimensional Pt monolayer grown by Pt evaporation in ultrahigh vacuum, we observe a significant destabilization of both H(ad) and O/OH(ad) due to strain and ligand effects induced by the underlying Rh(111) substrate. When Pt is deposited via a wet-chemical route, by contrast, three-dimensional Pt islands are formed. In this case, strain and Rh ligand effects are balanced with higher local thickness of the Pt islands as well as higher defect density, shifting H and OH adsorption energies back toward pure Pt. Using density functional theory, we calculate O adsorption energies and corresponding local ORR activities for fcc 3-fold hollow sites with various local geometries that are present in the three-dimensional Pt islands.


High-speed dynamic speckle interferometry: phase errors due to intensity, velocity, and speckle decorrelation.
The recently developed technique of high-speed phase-shifting speckle interferometry combined with temporal phase unwrapping allows dynamic displacement fields to be measured, even for objects containing global discontinuities such as cracks or boundaries. However, when local speckle averaging is included, small phase errors introduced at each time step are accumulated along the time axis, yielding total phase values that depend strongly on the speckle rereference rate. We present an analysis of the errors introduced in the phase evaluation by three sources: intensity errors, velocity errors, and speckle decorrelation. These errors are analyzed when they act both independently and together, for the most commonly used phase-shifting algorithms, with computer-generated speckle patterns. It is shown that, in a controlled out-of-plane geometry, errors in the unwrapped phase map that are due to speckle decorrelation rise as the time between rereferencing events is increased, whereas those due to intensity and velocity errors are reduced. It is also shown that speckle decorrelation errors are typically more important than the intensity and velocity errors. These results provide guidance as to the optimal speckle rereferencing rate in practical applications of the technique.


Repeated unilateral handgrip contractions alter functional connectivity and improve contralateral limb response times.
In humans, motor learning is underpinned by changes in sensorimotor network functional connectivity (FC). Unilateral contractions increase FC in the ipsilateral primary motor cortex (M1) and supplementary motor area (SMA); areas involved in motor planning and execution of the contralateral hand. Therefore, unilateral contractions are a promising approach to augment motor performance in the contralateral hand. In a within-participant, randomized, cross-over design, 15 right-handed adults had two magnetic resonance imaging (MRI) sessions, where functional-MRI and MR-Spectroscopic Imaging were acquired before and after repeated right-hand contractions at either 5% or 50% maximum voluntary contraction (MVC). Before and after scanning, response times (RTs) were determined in both hands. Nine minutes of 50% MVC contractions resulted in decreased handgrip force in the contracting hand, and decreased RTs and increased handgrip force in the contralateral hand. This improved motor performance in the contralateral hand was supported by significant neural changes: increased FC between SMA-SMA and increased FC between right M1 and right Orbitofrontal Cortex. At a neurochemical level, the degree of GABA decline in left M1, left and right SMA correlated with subsequent behavioural improvements in the left-hand. These results support the use of repeated handgrip contractions as a potential modality for improving motor performance in the contralateral hand.


Endoplasmic reticulum stress in the pathogenesis of fibrotic disease.
Eukaryotic cells contain an elegant protein quality control system that is crucial in maintaining cellular homeostasis; however, dysfunction of this system results in endoplasmic reticulum (ER) stress and activation of the unfolded protein response (UPR). Severe or prolonged ER stress is associated with the development of degenerative and fibrotic disorders in multiple organs, as evidenced by the identification of disease-causing mutations in epithelial-restricted genes that lead to protein misfolding or mistrafficking in familial fibrotic diseases. Emerging evidence implicates ER stress and UPR signaling in a variety of profibrotic mechanisms in individual cell types. In epithelial cells, ER stress can induce apoptosis, inflammatory signaling, and epithelial-mesenchymal transition. In other cell types, ER stress is linked to myofibroblast activation, macrophage polarization, and T cell differentiation. ER stress-targeted therapies have begun to emerge using approaches that range from global enhancement of chaperone function to selective targeting of activated ER stress sensors and other downstream mediators. As the complex regulatory mechanisms of this system are further clarified, there are opportunities to develop new disease-modifying therapeutic strategies in a wide range of chronic fibrotic diseases.


Numerical Simulations of Oscillation Modes of the Solar Convection Zone.
We use the three-dimensional hydrodynamic code of Stein & Nordlund to realistically simulate the upper layers of the solar convection zone in order to study physical characteristics of solar oscillations. Our first result is that the properties of oscillation modes in the simulation closely match the observed properties. Recent observations from the Solar and Heliospheric Observatory (SOHO)/Michelson Doppler Imager (MDI) and Global Oscillations Network Group have confirmed the asymmetry of solar oscillation line profiles, initially discovered by Duvall et al. In this Letter, we compare the line profiles in the power spectra of the Doppler velocity and continuum intensity oscillations from the SOHO/MDI observations with the simulation. We also compare the phase differences between the velocity and intensity data. We have found that the simulated line profiles are asymmetric and have the same asymmetry reversal between velocity and intensity as observed. The phase difference between the velocity and intensity signals is negative at low frequencies, and phase jumps in the vicinity of modes are also observed. Thus, our numerical model reproduces the basic observed properties of solar oscillations and allows us to study the physical properties which are not observed.


Platelet-derived growth factor receptor kinase inhibitor AG-1295 promotes osteoblast differentiation in MC3T3-E1 cells via the Erk pathway.
Previous studies have conflicting views on the effect of platelet-derived growth factor (PDGF)/PDGF receptor (PDGFR) signaling on osteogenesis. The current study investigated the effect of PDGF receptor-beta (PDGFR-β) inhibition by AG-1295 on the osteogenic differentiation of the mouse pre-osteoblastic cell line MC3T3-E1. Osteogenic differentiation was induced by treatment with β-glycerophosphate, ascorbic acid, and dexamethasone along with or absent AG-1295. Results showed that AG-1295 significantly increased alkaline phosphatase (ALP) activity and enhanced the formation of mineralized nodules in a dose-dependent manner. Furthermore, treatment with AG-1295 resulted in up-regulated mRNA expression of the osteogenic marker genes collagen type I (Col1A), runt-related transcription factor 2 (Runx2), osterix (Osx), tissue-nonspecific alkaline phosphatase (Tnap), and osteocalcin (Ocn). Consistent with its effect on osteoblast differentiation, AG-1295 also significantly suppressed the phosphorylation of Erk1/2 in MC3T3-E1 cells. In conclusion, findings suggest that blocking the PDGFR-β pathway with AG1295 markedly promotes osteoblast differentiation and matrix mineralization in mouse osteoblastic MC3T3-E1 cells and that the Erk1/2 pathway might participate in this process.


Management of menopause when estrogen cannot be used.
Estrogen deficiency, whether surgically induced or as a consequence of natural ovarian failure, has destructive effects on many organ systems. With current levels of life expectancy, untreated women may expect to spend a third of their lifetime in this state. Appropriate estrogen replacement therapy (ERT) can avert (if started promptly) or ameliorate these devastating consequences, some of which (osteoporotic fractures, increased cardiovascular morbidity) can be lethal. Nevertheless, from 10 to 20% of postmenopausal women may have significant contraindications to ERT. Treatment of symptoms and improving the quality of life is imperative, yet many physicians abjure intervention, for reasons which are not entirely clear. Recent studies of conventional intervention with sedatives or tranquilisers show results equivalent to placebo therapy. On the other hand, specific agents with demonstrated effectiveness are available for management of the major estrogen-deficiency effects, although none of them are truly adequate replacement for the effect of estrogen itself.


A rational vaccine pipeline for tuberculosis.
The development of tuberculosis (TB) vaccines is at a turning point, with the promise of new vaccines on the horizon. Over the next few years, it is possible that we will see a phase III multi-site clinical trial of at least one new TB vaccine and perhaps the introduction of a TB vaccine by the end of the decade. However, many gaps remain in our understanding of TB pathogenesis as well as the host immune responses required to provide protective immunity. A major challenge for TB vaccines is to establish a correlate of vaccine immunity which would greatly facilitate bridging studies needed to approve, license and distribute new TB vaccines in all areas endemic for TB. This will require TB vaccines that are both safe and effective in all populations. It cannot be accomplished without hard work as well as additional resources that match the ambitious goals of the TB community.


Testosterone replacement in hypogonadal men with type 2 diabetes and/or metabolic syndrome (the TIMES2 study).
This study evaluated the effects of testosterone replacement therapy (TRT) on insulin resistance, cardiovascular risk factors, and symptoms in hypogonadal men with type 2 diabetes and/or metabolic syndrome (MetS). The efficacy, safety, and tolerability of a novel transdermal 2% testosterone gel was evaluated over 12 months in 220 hypogonadal men with type 2 diabetes and/or MetS in a multicenter, prospective, randomized, double-blind, placebo-controlled study. The primary outcome was mean change from baseline in homeostasis model assessment of insulin resistance (HOMA-IR). Secondary outcomes were measures of body composition, glycemic control, lipids, and sexual function. Efficacy results focused primarily on months 0-6 (phase 1; no changes in medication allowed). Medication changes were allowed in phase 2 (months 6-12). TRT reduced HOMA-IR in the overall population by 15.2% at 6 months (P = 0.018) and 16.4% at 12 months (P = 0.006). In type 2 diabetic patients, glycemic control was significantly better in the TRT group than the placebo group at month 9 (HbA(1c): treatment difference, -0.446%; P = 0.035). Improvements in total and LDL cholesterol, lipoprotein a (Lpa), body composition, libido, and sexual function occurred in selected patient groups. There were no significant differences between groups in the frequencies of adverse events (AEs) or serious AEs. The majority of AEs (>95%) were mild or moderate. Over a 6-month period, transdermal TRT was associated with beneficial effects on insulin resistance, total and LDL-cholesterol, Lpa, and sexual health in hypogonadal men with type 2 diabetes and/or MetS.


Anesthetic equipment, facilities and services available for pediatric anesthesia in Nigeria.
Facilities and equipment are known to contribute to improved patient care and outcome. Hospitals for sub-specialized pediatric anesthetic service are routinely available worldwide. In Nigeria, such hospitals now exist. It is therefore relevant to study the facilities and equipment available for pediatric anesthetic service to measure the quality of care in a health institution. A prospective study of the anesthetic equipment, facilities and services available for pediatric anesthesia was carried out in Nigerian hospitals using anonymously administered questionnaires from October, 2008 to October, 2009, after relevant ethics approval. Ten of the 30 hospitals studied had intensive care unit facilities (33.3%) and only three of them had organized ambulatory anesthesia units for pediatric patients (10%). Facilities available for pediatric anesthetic care were service delivery oriented and not information technology based (automated anesthesia information management services - AIMS). A quarter of the hospitals studied (7) had their facilities and equipment updated recently. Also, there were no reception rooms nor post anesthetic care units specifically designed for pediatric patients. Equipment for acute pain service such as infusion pumps and patient controlled analgesia pumps (PCA pumps) and for capnography were found in only two hospitals (6.6%) at the time of the study. The anesthetic equipment used did not conform to the same standards (98%) even in the same hospital. Also, the use of disposable anesthetic equipment was low. We conclude that anesthetic services for pediatric patients should be based on the use of automated anesthesia information technology service. Anesthetic equipment should conform to the same standards and should be computerized. Disposable anesthetic equipment should also be used where applicable to minimize the incidence of nosocomial infections. This would result in improved peri-operative care in pediatric patients.


Applications and toxicity of silver nanoparticles: a recent review.
Silver nanoparticles (AgNPs) exhibit a consistent amount of flexible properties which endorse them for a larger spectrum of applications in biomedicine and related fields. Over the years, silver nanoparticles have been subjected to numerous in vitro and in vivo tests to provide information about their toxic behavior towards living tissues and organisms. Researchers showed that AgNPs have high antimicrobial efficacy against many bacteria species including Escherichia coli, Neisseria gonorrhea, Chlamydia trachomatis and also viruses. Due to their novel properties, the incorporation of silver nanoparticles into different materials like textile fibers and wound dressings can extend their utility on the biomedical field while inhibiting infections and biofilm development. Among the noble metal nanoparticles, AgNPs present a series of features like simple synthesis routes, adequate and tunable morphology, and high surface to volume ratio, intracellular delivery system, a large plasmon field area recommending them as ideal biosensors, catalysts or photo-controlled delivery systems. In bioengineering, silver nanoparticles are considered potentially ideal gene delivery systems for tissue regeneration. The remote triggered detection and release of bioactive compounds of silver nanoparticles has proved their relevance also in forensic sciences. The authors report an up to date review related to the toxicity of AgNPs and their applications in antimicrobial activity and biosensors for gene therapy.


Mid-IR hyperspectral imaging of laminar flames for 2-D scalar values.
This work presents a new emission-based measurement which permits quantification of two-dimensional scalar distributions in laminar flames. A Michelson-based Fourier-transform spectrometer coupled to a mid-infrared camera (1.5 μm to 5.5 μm) obtained 256 × 128pixel hyperspectral flame images at high spectral (δν̃ = 0.75cm(−1)) and spatial (0.52 mm) resolutions. The measurements revealed line and band emission from H2O, CO2, and CO. Measurements were collected from a well-characterized partially-premixed ethylene (C2H4) flame produced on a Hencken burner at equivalence ratios, Φ, of 0.8, 0.9, 1.1, and 1.3. After describing the instrument and novel calibration methodology, analysis of the flames is presented. A single-layer, line-by-line radiative transfer model is used to retrieve path-averaged temperature, H2O, CO2 and CO column densities from emission spectra between 2.3 μm to 5.1 μm. The radiative transfer model uses line intensities from the latest HITEMP and CDSD-4000 spectroscopic databases. For the Φ = 1.1 flame, the spectrally estimated temperature for a single pixel 10 mm above burner center was T = (2318 ± 19)K, and agrees favorably with recently reported laser absorption measurements, T = (2348 ± 115)K, and a NASA CEA equilibrium calculation, T = 2389K. Near the base of the flame, absolute concentrations can be estimated, and H2O, CO2, and CO concentrations of (12.5 ± 1.7) %, (10.1 ± 1.0) %, and (3.8 ± 0.3) %, respectively, compared favorably with the corresponding CEA values of 12.8%, 9.9% and 4.1%. Spectrally-estimated temperatures and concentrations at the other equivalence ratios were in similar agreement with measurements and equilibrium calculations. 2-D temperature and species column density maps underscore the Φ-dependent chemical composition of the flames. The reported uncertainties are 95% confidence intervals and include both statistical fit errors and the propagation of systematic calibration errors using a Monte Carlo approach. Systematic errors could warrant a factor of two increase in reported uncertainties. This work helps to establish IFTS as a valuable combustion diagnostic tool.


Pitch related to spectral edges of broadband signals.
A complex tone often evokes a pitch sensation associated with its extreme spectral components, besides the holistic pitch associated with its fundamental frequency. We studied the edge pitch created at the upper spectral edge of complexes with a low-pass spectrum by asking subjects to adjust the frequency of a sinusoidal comparison tone to the perceived pitch. Measurements were performed for different values of the fundamental frequency and of the upper frequency of the complex as well as for three different phase relations of the harmonic components. For a wide range of these parameters the subjects could adjust the comparison tone with a high accuracy, measured as the standard deviation of repeated adjustments, to a frequency close to the nominal edge frequency. The detailed dependence of the matching accuracy on temporal parameters of the harmonic complexes suggests that the perception of the edge pitch in harmonic signals is related to the temporal resolution of the hearing system. This resolution depends primarily on the time constants of basilar-membrane filters and on additional limitations due to neuronal processes.


All-trans retinoic acid in hematological malignancies, an update. GER (Gruppo Ematologico Retinoidi).
During the past ten years, the study of retinoids has undergone a total transformation. The Italian Society of Experimental Hematology decided to discuss these advances at a meeting in Florence on April 18, 1996. The material examined in the present review includes articles and abstracts published in journals covered by the Science Citation Index and Medline. In addition, all the authors of the present article have been actively working in the field of retinoids and have contributed several papers. Summaries of their oral presentations at the Florence meeting are reported in the Appendix to this review article. One of the most important advances has been the elucidation of new molecular mechanisms of control of gene expression by retinoids. A number of new retinoids have been synthesized by chemists, some of which are being screened for potential clinical use, and a few have already had a tremendous impact on clinical practice. The most important achievements have been obtained in acute promyelocytic leukemia. In 1988 a Chinese group working in Shanghai showed that using all-trans retinoic acid (ATRA) alone 94% of acute promyelocytic leukemic patients obtained complete remission through differentiation of the leukemic clone. This result transformed a dream into reality and allowed researchers to move from laboratory experience to clinical applications of this differentiating therapy. Expanding the spectrum of hematological malignancies that may respond to ATRA remains a challenge; however, several results show some activity of retinoids alone or in combination with other drugs in juvenile chronic myeloid leukemia (CML), myelodysplastic syndrome, cutaneous T-cell lymphoma and CML. Particularly interesting are the studies that explored the potential clinical synergism of ATRA-based combination therapies with growth factors, other differentiating agents such as vitamin D3, immunomodulators like interferons, or chemotherapeutic agents, in particular Ara-C, all of which show promising in vitro effects when used in combination with retinoids.


Increased horizontal viewing zone angle of a hologram by resolution redistribution of a spatial light modulator.
The narrow viewing zone angle is one of the problems associated with electronic holography. We propose a technique that enables the ratio of horizontal and vertical resolutions of a spatial light modulator (SLM) to be altered. This technique increases the horizontal resolution of a SLM several times, so that the horizontal viewing zone angle is also increased several times. A SLM illuminated by a slanted point light source array is imaged by a 4f imaging system in which a horizontal slit is located on the Fourier plane. We show that the horizontal resolution was increased four times and that the horizontal viewing zone angle was increased approximately four times.


Just do it! How performing an action enhances remembering in transient global amnesia.
Transient global amnesia (TGA) is a clinical syndrome characterized by the sudden onset of a massive episodic memory deficit that spares other cognitive functions. As such, it provides a unique human amnesia model for testing the enactment effect (i.e., better memory for performed actions than for verbally encoded sentences). Our main aim was to test whether the enactment effect is preserved in TGA patients, both to have a better understanding and to test the robustness of this effect in a massive amnesia. Object-action pairs were encoded under four conditions: verbal, experimenter-performed, and two enacted conditions (self-performed and self-performed with choice). We tested object-action pair retrieval using cued recall (CR) and recognition tasks, and source memory using a free recall task. We also assessed binding, executive functions, short-term memory, episodic memory, anxiety and mood. We run correlations to control for their putative effects on memory for action. Data were collected from 24 patients, 16 of whom were examined during the acute phase and eight the day-after, as well as from 18 healthy controls. The memory performances of the patients in the acute phase improved for both (i) the CR score, between the verbal, experimenter-performed and self-performed with choice conditions, and (ii) the total recognition score, between the verbal condition and the two enacted conditions. Correlations were found between self-performed task (SPT) enhancement and both the binding and anxiety. In spite of their severely impaired episodic memory, patients with TGA benefit from the enactment effect. These results are discussed in relation to the role of motor components and episodic integration in memory for actions. We suggest that enactment effect can be used in clinical practice and rehabilitation, possible even for patients with a massive memory impairment.


Justifying the More Restrictive Alternative: Ethical Justifications for One Health AMR Policies Rely on Empirical Evidence.
Global consumption of antibiotics has accelerated the evolution of bacterial antimicrobial resistance. Yet, the risks from increasing bacterial antimicrobial resistance are not restricted to human populations: transmission of antimicrobial resistant bacteria occurs between humans, farms, the environment and other reservoirs. Policies that take a 'One Health' approach deal with this cross-reservoir spread, but are often more restrictive concerning human actions than policies that focus on a single reservoir. As such, the burden of justification lies with these more restrictive policies. We argue that an ethical justification for preferring One Health policies over less restrictive alternatives relies on empirical evidence as well as theory. The ethical justification for these policies is based on two arguments: (i) comparatively greater effectiveness, and (ii) comparatively better tracking of moral responsibility. Yet the empirical assumptions on which these claims rest are limited by existing empirical knowledge. Using livestock farming as an example, we suggest that scientific research into characterising antimicrobial resistance and linking practices to outcomes ought to be guided (at least in part) by the imperative to supply the context-specific data needed to ethically justify preferring a One Health policy over less restrictive alternatives.


Probing the Catalytic Mechanism and Inhibition of SAMHD1 Using the Differential Properties of Rp- and Sp-dNTPαS Diastereomers.
SAMHD1 is a fundamental regulator of cellular dNTPs that catalyzes their hydrolysis into 2'-deoxynucleoside and triphosphate, restricting the replication of viruses, including HIV-1, in CD4+ myeloid lineage and resting T-cells. SAMHD1 mutations are associated with the autoimmune disease Aicardi-Goutières syndrome (AGS) and certain cancers. More recently, SAMHD1 has been linked to anticancer drug resistance and the suppression of the interferon response to cytosolic nucleic acids after DNA damage. Here, we probe dNTP hydrolysis and inhibition of SAMHD1 using the Rp and Sp diastereomers of dNTPαS nucleotides. Our biochemical and enzymological data show that the α-phosphorothioate substitution in Sp-dNTPαS but not Rp-dNTPαS diastereomers prevents Mg2+ ion coordination at both the allosteric and catalytic sites, rendering SAMHD1 unable to form stable, catalytically active homotetramers or hydrolyze substrate dNTPs at the catalytic site. Furthermore, we find that Sp-dNTPαS diastereomers competitively inhibit dNTP hydrolysis, while Rp-dNTPαS nucleotides stabilize tetramerization and are hydrolyzed with similar kinetic parameters to cognate dNTPs. For the first time, we present a cocrystal structure of SAMHD1 with a substrate, Rp-dGTPαS, in which an Fe-Mg-bridging water species is poised for nucleophilic attack on the Pα. We conclude that it is the incompatibility of Mg2+, a hard Lewis acid, and the α-phosphorothioate thiol, a soft Lewis base, that prevents the Sp-dNTPαS nucleotides coordinating in a catalytically productive conformation. On the basis of these data, we present a model for SAMHD1 stereospecific hydrolysis of Rp-dNTPαS nucleotides and for a mode of competitive inhibition by Sp-dNTPαS nucleotides that competes with formation of the enzyme-substrate complex.


Performance characteristics of Parkour practitioners: Who are the traceurs?
Parkour is a modern physical activity that consists of using the environment, mostly urban, as a playground of obstacles. The aims of this study were (i) to investigate age, anthropometric and training characteristics of Parkour practitioners, called 'traceurs' and (ii) to assess jump performances and muscular characteristics of traceurs, compared to those of gymnasts and power athletes. The mean age of the population of traceurs studied (n = 130) was 19.4 ± 4.3 years, women represented 12.4% of the total field and mean training volume was 8.1 ± 0.5 hours/week. Vertical and long jump performances were analysed on smaller samples of participants (four groups, n = 15 per group); and eccentric (-90° s(-1), -30° s(-1)), concentric (30° s(-1), 90° s(-1)) and isometric knee extensors torques were evaluated by means of an isokinetic dynamometer. Traceurs showed greater (P < .01) drop jump performance (64.9 ± 1.5 cm) than gymnasts (60.9 ± 1.1 cm) and greater (P < .001) counter movement jump with arms (59.2 ± 1.5 cm) than power athletes (53.0 ± 1.4 cm). Standing long jump performances were greater (P < .05) for traceurs (282.7 ± 5.2 cm) compared to other athletes (gymnasts: 273.9 ± 7.3 cm; power athletes: 261.3 ± 6.7 cm). Eccentric knee extension torques were greater (P < .05) for traceurs compared to other athletes. This study revealed that Parkour training induces major development of jump and muscular skills. The use of such training has several practical applications as it provides a better resistance to high eccentric load and helps reinforce musculoskeletal structures.


Social representations and interface layout: A new way of enhancing persuasive technology applied to organ donation.
Although campaigns promoting organ donation have proved their effectiveness, increasing the number of people who explicitly agree to become donors is still difficult. Based on the social marketing notion of persuasive technology, we reasoned that it was timely to focus on the design of this persuasive technology and to analyze its contribution in particularly challenging contexts such as organ donation. More specifically, the originality of the present study lay in the way we linked the field of persuasive technology to the theory of social representations, and combined them with an analysis of the ergonomic aspects of interface layout. This study had two complementary goals. The first was to determine whether the sociocognitive salience of the central elements of social representations (i.e., the most frequent and important themes related to the subject-here, organ donation-for individuals), can be used to achieve persuasive outcomes. The second was to determine whether interface layout, in terms of information location and background characteristics (color and contrast), can strengthen the persuasive impact. University students (N > 200) were exposed to a computer screen displaying a message involving either central or peripheral elements of the social representations of organ donation (status), placed either in the middle or on one side of the screen (location), and shown against either a white or a blue background (background). Eye-tracking data were recorded, in addition to self-reported data. In line with the elaboration likelihood model, results showed that participants who were exposed to central (vs. peripheral) elements of the social representations of organ donation followed the central route in processing information. Moreover, they had stronger attitudes, and more of them stated that they were actual organ donors. Importantly, however, at least for some variables, these status-related effects were not independent of the interface layout. More specifically, the persuasive impact of the central elements was enhanced when the information was displayed in the middle (vs. the side) of the screen and when it was displayed on a white (vs. blue) background. We discuss the theoretical and practical issues raised by these results.


Estimating the effects of immigration status on mental health care utilizations in the United States.
Immigration status is a likely deterrent of mental health care utilization in the United States. Using the Medical Expenditure Panel Survey and National Health Interview survey from 2002 to 2006, multivariable logistic regressions were used to estimate the effects of immigration status on mental health care utilization among patients with depression or anxiety disorders. Multivariate regressions showed that immigrants were significantly less likely to take any prescription drugs, but not significantly less likely to have any physician visits compared to US-born citizens. Results also showed that improving immigrants' health care access and health insurance coverage could potentially reduce disparities between US-born citizens and immigrants by 14-29% and 9-28% respectively. Policy makers should focus on expanding the availability of regular sources of health care and immigrant health coverage to reduce disparities on mental health care utilization. Targeted interventions should also focus on addressing immigrants' language barriers, and providing culturally appropriate services.


HIV-1 and HIV-2 reverse transcriptases: different mechanisms of resistance to nucleoside reverse transcriptase inhibitors.
As anti-HIV therapy becomes more widely available in developing nations, it is clear that drug resistance will continue to be a major problem. The related viruses HIV-1 and HIV-2 share many of the same resistance pathways to nucleoside reverse transcriptase inhibitors (NRTIs). However, clinical data suggest that while HIV-1 reverse transcriptase (RT) usually uses an ATP-dependent excision pathway to develop resistance to the nucleoside analog zidovudine (AZT), HIV-2 RT does not appear to use this pathway. We previously described data that suggested that wild-type (WT) HIV-2 RT has a much lower ability to excise AZT monophosphate (AZTMP) than does WT HIV-1 RT and suggested that this is the reason that HIV-2 RT more readily adopts an exclusion pathway against AZT triphosphate (AZTTP), while HIV-1 RT is better able to exploit the ATP-dependent pyrophosphorolysis mechanism. However, we have now done additional experiments, which show that while HIV-1 RT can adopt either an exclusion- or excision-based resistance mechanism against AZT, HIV-2 RT can use only the exclusion mechanism. All of our attempts to make HIV-2 RT excision competent did not produce an AZT-resistant RT but instead yielded RTs that were less able to polymerize than the WT. This suggests that the exclusion pathway is the only pathway available to HIV-2.


Chromatin condensation during apoptosis is accompanied by degradation of lamin A+B, without enhanced activation of cdc2 kinase.
Chromatin condensation paralleled by DNA fragmentation is one of the most important criteria which are used to identify apoptotic cells. However, comparable changes are also observed in interphase nuclei which have been treated with cell extracts from mitotic cells. In this respect it is known that in mitosis, the lamina structure is broken down as a result of lamin solubilization and it is possible that a similar process is happening in apoptotic cells. The experiments described in this study have used confluent cultures of an embryonic fibroblast cell line which can be induced to undergo either apoptosis at low serum conditions or mitosis. Solubilization of lamin A+B was analyzed by immunoblotting and indirect immunofluorescence. These studies showed that in mitotic cells lamina breakdown is accompanied by lamin solubilization. In apoptotic cells, a small amount of lamin is solubilized before the onset of apoptosis, thereafter, chromatin condensation is accompanied by degradation of lamin A+B to a 46-kD fragment. Analysis of cellular lysates by probing blots with anti-PSTAIR followed by anti-phosphotyrosine showed that in contrast to mitosis, dephosphorylation on tyrosine residues did not occur in apoptotic cells. At all timepoints after the onset of apoptosis there was no significant increase in the activation of p34cdc2 as determined in the histone H1 kinase assay. Coinduction of apoptosis and mitosis after release of cells from aphidicolin block showed that apoptosis could be induced in parallel with S-phase. The sudden breakdown of chromatin structure may be the result of detachment of the chromatin loops from their anchorage at the nuclear matrix, as bands of 50 kbp and corresponding multimers were detectable by field inversion gel electrophoresis (FIGE). In apoptotic cells all of the DNA was fragmented, but only 14% of the DNA was smaller than 50 kbp. DNA strand breaks were detected at the periphery of the condensed chromatin by in situ tailing (ISTAIL). Chromatin condensation during apoptosis appears to be due to a rapid proteolysis of nuclear matrix proteins which does not involve the p34cdc2 kinase.


Assessment of high resolution melting analysis as a potential SNP genotyping technique in forensic casework.
High resolution melting (HRM) analysis is a simple, cost effective, closed tube SNP genotyping technique with high throughput potential. The effectiveness of HRM for forensic SNP genotyping was assessed with five commercially available HRM kits evaluated on the ViiA™ 7 Real Time PCR instrument. Four kits performed satisfactorily against forensically relevant criteria. One was further assessed to determine the sensitivity, reproducibility, and accuracy of HRM SNP genotyping. The manufacturer's protocol using 0.5 ng input DNA and 45 PCR cycles produced accurate and reproducible results for 17 of the 19 SNPs examined. Problematic SNPs had GC rich flanking regions which introduced additional melting domains into the melting curve (rs1800407) or included homozygotes that were difficult to distinguish reliably (rs16891982; a G to C SNP). A proof of concept multiplexing experiment revealed that multiplexing a small number of SNPs may be possible after further investigation. HRM enables genotyping of a number of SNPs in a large number of samples without extensive optimization. However, it requires more genomic DNA as template in comparison to SNaPshot®. Furthermore, suitably modifying pre-existing forensic intelligence SNP panels for HRM analysis may pose difficulties due to the properties of some SNPs.


Incidence and variance of foot and ankle injuries in elite college football players.
We conducted a study on the risk for foot and ankle injuries in college football players on the basis of injury type and player position. In February 2006, we evaluated 320 intercollegiate football players at the National Football League Combine. All pathologic conditions and surgical procedures of the foot and ankle were recorded, and data were analyzed by player position to detect any trends. Seventy-two percent (n = 231) of the players had a history of foot and ankle injuries, with a total of 287 foot and ankle injuries (1.24 injuries/player injured). The most common injuries were lateral ankle sprain (n = 115), syndesmotic sprain (50), metatarsophalangeal dislocation/turf toe (36), and fibular fracture (25). Foot and ankle injuries were most common in kickers/punters (100% incidence), special teams (100%), running backs (83%), wide receivers (83%), and offensive linemen (80%). Lateral ankle sprains, the most common injuries, were treated surgically only 2.6% of the time. Offensive linemen were most likely to have had syndesmotic sprains (32%), and quarterbacks had the highest incidence of fibular fractures (16%). Foot and ankle injuries are common in collegiate football players, affecting 72% of players. Thirteen percent underwent surgical treatment. Trends are seen in the types of injuries for the different player positions.


Transcripts of maize RbcS genes accumulate differentially in C3 and C4 tissues.
RbcS genes exist as multigene families in most plant species examined. In this paper, we report an investigation into the expression patterns of two maize RbcS genes, designated in this report as RbcS1 and RbcS2. We present the sequence of RbcS2 and show that the structure of the gene has several features in common with other monocot RbcS genes. To determine whether RbcS1 and RbcS2 fulfil different functional roles with respect to the C3 and C4 carbon fixation pathways, we have investigated the expression patterns of the two genes in different maize tissue types. Transcripts of both genes are found at high levels specifically in bundle-sheath cells of maize seedling leaves, indicating that both genes are expressed in the C4-type pattern. However, we show that RbcS1 transcripts are relatively more abundant than RbcS2 transcripts in C3 tissues such as husk leaves. These results are discussed with respect to the evolution of C4 carbon fixation and the mechanisms required for the cell-specific expression of RbcS genes.


Ciliary proteins specify the cell inflammatory response by tuning NFκB signalling, independently of primary cilia.
Complex inflammatory signalling cascades define the response to tissue injury but also control development and homeostasis, limiting the potential for these pathways to be targeted therapeutically. Primary cilia are subcellular regulators of cellular signalling, controlling how signalling is organized, encoded and, in some instances, driving or influencing pathogenesis. Our previous research revealed that disruption of ciliary intraflagellar transport (IFT), altered the cell response to IL-1β, supporting a putative link emerging between cilia and inflammation. Here, we show that IFT88 depletion affects specific cytokine-regulated behaviours, changing cytosolic NFκB translocation dynamics but leaving MAPK signalling unaffected. RNA-seq analysis indicates that IFT88 regulates one third of the genome-wide targets, including the pro-inflammatory genes Nos2, Il6 and Tnf Through microscopy, we find altered NFκB dynamics are independent of assembly of a ciliary axoneme. Indeed, depletion of IFT88 inhibits inflammatory responses in the non-ciliated macrophage. We propose that ciliary proteins, including IFT88, KIF3A, TTBK2 and NPHP4, act outside of the ciliary axoneme to tune cytoplasmic NFκB signalling and specify the downstream cell response. This is thus a non-canonical function for ciliary proteins in shaping cellular inflammation.This article has an associated First Person interview with the first author of the paper.


Mitochondrial dynamics in diabetic cardiomyopathy.
Cardiac function is energetically demanding, reliant on efficient well-coupled mitochondria to generate adenosine triphosphate and fulfill the cardiac demand. Predictably then, mitochondrial dysfunction is associated with cardiac pathologies, often related to metabolic disease, most commonly diabetes. Diabetic cardiomyopathy (DCM), characterized by decreased left ventricular function, arises independently of coronary artery disease and atherosclerosis. Dysregulation of Ca(2+) handling, metabolic changes, and oxidative stress are observed in DCM, abnormalities reflected in alterations in mitochondrial energetics. Cardiac tissue from DCM patients also presents with altered mitochondrial morphology, suggesting a possible role of mitochondrial dynamics in its pathological progression. Abnormal mitochondrial morphology is associated with pathologies across diverse tissues, suggesting that this highly regulated process is essential for proper cell maintenance and physiological homeostasis. Highly structured cardiac myofibers were hypothesized to limit alterations in mitochondrial morphology; however, recent work has identified morphological changes in cardiac tissue, specifically in DCM. Mitochondrial dysfunction has been reported independently from observations of altered mitochondrial morphology in DCM. The temporal relationship and causative nature between functional and morphological changes of mitochondria in the establishment/progression of DCM is unclear. Altered mitochondrial energetics and morphology are not only causal for but also consequential to reactive oxygen species production, hence exacerbating oxidative damage through reciprocal amplification, which is integral to the progression of DCM. Therefore, targeting mitochondria for DCM will require better mechanistic characterization of morphological distortion and bioenergetic dysfunction.


Encapsulation of dye molecules and nanoparticles in hollow organogel fibers of a nonchiral polyurethane model compound.
We present the case of a nonchiral organogelator that forms hollow fibers and encapsulates silver nanoparticles (SNP) and a dye molecule. The biscarbamate molecule (a model compound for polyurethanes), which has two hydrogen-bonding motifs symmetrically attached to n-dodecyl side chains (C(12)), gels benzonitrile with hollow fibrillar morphology. The C(12) molecules form sheets that eventually wrap into hollow fibers to form the gel network. Herein, two-component gels were prepared with C(12) as one component and SNP, phthalocyanine (Pc), or perylene (Pe) as the other. Microscopic analysis and partial melting experiments confirmed the inclusion of the silver nanoparticles and phthalocyanine into the hollow fibers. On the other hand, Pe molecules tend to form crystals at the outer surface of the C(12) fibers, which results in a significant increase in the width of the gel fibers. This difference in the behavior of Pc and Pe molecules were accounted for by their crystal geometry and significantly different crystal growth rate compared with that of C(12) fiber formation in the gels. Pc crystallizes in a needle shape that facilitates occlusion in the gel fibers, whereas Pe forms large platelets. X-ray diffraction and spectroscopic analysis of the two-component gels along with their neat components confirmed that there was no change in the packing behavior of the Pc and Pe molecules in the gels. Therefore, these are examples of two-component physical gels in which the Pc crystals are occluded within the hollow fibers of C(12) by physical mixing of the components without the aid of any inter-molecular interactions between the different components. We have thus shown that lumen-loaded gel fibers with nanoparticles and dye molecules can be prepared by the two-component gel route, provided that the above growth rate, shape, and size conditions are satisfied.


Functional and genomic analyses of alpha-solenoid proteins.
Alpha-solenoids are flexible protein structural domains formed by ensembles of alpha-helical repeats (Armadillo and HEAT repeats among others). While homology can be used to detect many of these repeats, some alpha-solenoids have very little sequence homology to proteins of known structure and we expect that many remain undetected. We previously developed a method for detection of alpha-helical repeats based on a neural network trained on a dataset of protein structures. Here we improved the detection algorithm and updated the training dataset using recently solved structures of alpha-solenoids. Unexpectedly, we identified occurrences of alpha-solenoids in solved protein structures that escaped attention, for example within the core of the catalytic subunit of PI3KC. Our results expand the current set of known alpha-solenoids. Application of our tool to the protein universe allowed us to detect their significant enrichment in proteins interacting with many proteins, confirming that alpha-solenoids are generally involved in protein-protein interactions. We then studied the taxonomic distribution of alpha-solenoids to discuss an evolutionary scenario for the emergence of this type of domain, speculating that alpha-solenoids have emerged in multiple taxa in independent events by convergent evolution. We observe a higher rate of alpha-solenoids in eukaryotic genomes and in some prokaryotic families, such as Cyanobacteria and Planctomycetes, which could be associated to increased cellular complexity. The method is available at http://cbdm.mdc-berlin.de/~ard2/.


CpG-containing oligonucleotides are efficient adjuvants for induction of protective antiviral immune responses with T-cell peptide vaccines.
Synthetic nonmethylated oligonucleotides containing CpG dinucleotides (CpG-ODNs) have been shown to exhibit immunostimulatory activity. CpG-ODNs have the capacity to directly activate B cells, macrophages, and dendritic cells, and we show here that this is reflected by cell surface binding of oligonucleotides to these cell subsets. However, T cells are not directly activated by CpG-ODNs, which correlates with the failure to bind to the T-cell surface. Efficient competition for CpG-induced B-cell activation by non-CpG-containing oligonucleotides suggests that oligonucleotides might bind to an as yet undefined sequence-nonspecific receptor prior to cellular activation. Induction of protective T-cell responses against challenge infection with lymphocytic choriomeningitis virus (LCMV) or with recombinant vaccinia virus expressing the LCMV glycoprotein was achieved by immunizing mice with the immunodominant major histocompatibility complex class I-binding LCMV glycoprotein-derived peptide gp33 together with CpG-ODNs. In these experiments, B cells, potentially serving as CpG-ODN-activated antigen-presenting cells (APCs), were not required for induction of protective immunity since CpG-ODN-gp33-immunized B-cell-deficient mice were equally protected against challenge infection with both viruses. This finding suggested that macrophages and/or dendritic cells were sufficiently activated in vivo by CpG-ODNs to serve as potent APCs for the induction of naive T cells. Furthermore, treatment with CpG-ODN alone induced protection against infection with Listeria monocytogenes via antigen-independent activation of macrophages. These data suggest that CpG activation of macrophages and dendritic cells may provide a critical step in CpG-ODN adjuvant activity.


Attenuation of the development of spontaneous hypertension in rats by chronic central administration of captopril.
Captopril infused into the lateral ventricle (ICV) of adult spontaneously hypertensive rats (SHR) decreases blood pressure. The current study was designed to explore the effects of brain converting-enzyme inhibition in young animals before the development of established hypertension and to characterize changes induced by captopril in a variety of pressor systems that might be responsible for the development of hypertension in this strain. Captopril (1.25 micrograms/0.5 microliter/hr) was infused into male SHR starting at 7 weeks of age. Four weeks later systolic blood pressure was only 157 +/- 3.3 compared to 181 +/- 3.9 mm Hg in vehicle-infused controls, and the pressor effect of ICV-injected angiotensin I was attenuated by 50%. When the same dose of captopril was infused intravenously, hypertension progressed as in vehicle-treated rats. Serum angiotensin-converting enzyme activity (SACE) and plasma arginine vasopressin (AVP) concentration were significantly higher (p less than 0.001 and 0.05, respectively), in the ICV captopril group than in the ICV vehicle group, while plasma aldosterone concentration and renin activity, fluid intake, urine volume, and urinary sodium excretion were similar in the two groups. Peripheral sympathetic nervous system activity assessed in the resting state was not altered by captopril treatment. In addition, AVP content of the telencephalon, diencephalon, mesencephalon, and pons medulla were not altered by ICV captopril. Renin activity was elevated in the telencephalon of ICV captopril-treated animals but unaltered in the other brain regions examined. These data demonstrate that ICV administration of captopril attenuates the development of hypertension in young SHR by mechanisms apparently independent of altered fluid and sodium balance and the sympathoadrenal system. The effect on blood pressure occurs in the absence of changes in renin activity or AVP content of plasma or those brain regions most often associated with blood pressure control.


A head and neck cancer tumor response-specific gene signature for cisplatin, 5-fluorouracil induction chemotherapy fails with added taxanes.
It is a major clinical challenge to predict which patients, with advanced stage head and neck squamous cell carcinoma, will not exhibit a reduction in tumor size following induction chemotherapy in order to avoid toxic effects of ineffective chemotherapy and delays for instituting other therapeutic options. Further, it is of interest to know to what extent a gene signature, which identifies patients with tumors that will not respond to a particular induction chemotherapy, is applicable when additional chemotherapeutic agents are added to the regimen. To identify genes that predict tumor resistance to induction with cisplatin/5-fluorouracil (PF) or PF and a taxane, we analyzed patient tumor biopsies with whole genome microarrays and quantitative reverse transcriptase-PCR (TLDA) cards. A leave one out cross-validation procedure allowed evaluation of the prediction tool. A ten-gene microarray signature correctly classified 12/13 responders and 7/10 non-responders to PF (92% specificity, 82.6% accuracy). TLDA analysis (using the same classifier) of the patients correctly classified 12/12 responders and 8/10 non-responders (100% specificity, 90.9% accuracy). Further, TLDA analysis correctly predicted the response of 5 new patients and, overall, 12/12 responders and 13/15 non-responders (100% specificity, 92.6% accuracy). The protein products of the genes constituting the signature physically associate with 27 other proteins, involved in regulating gene expression, constituting an interaction network. In contrast, TLDA-based prediction (with the same gene signature) of responses to induction with PF and either of two taxanes was poor (0% specificity, 25% accuracy and 33.3% specificity, 25% accuracy). Successful transfer of the microarray-based gene signature to an independent, PCR-based technology suggests that TLDA-based signatures could be a useful hospital-based technology for determining therapeutic options. Although highly specific for tumor responses to PF induction, the gene signature is unsuccessful when taxanes are added. The results illustrate the subtlety in developing "personalized medicine".


Inhibition of the CXCL12/CXCR4-axis as preventive therapy for radiation-induced pulmonary fibrosis.
A devastating late injury caused by radiation is pulmonary fibrosis. This risk may limit the volume of irradiation and compromise potentially curative therapy. Therefore, development of a therapy to prevent this toxicity can be of great benefit for this patient population. Activation of the chemokine receptor CXCR4 by its ligand stromal cell-derived factor 1 (SDF-1/CXCL12) may be important in the development of radiation-induced pulmonary fibrosis. Here, we tested whether MSX-122, a novel small molecule and partial CXCR4 antagonist, can block development of this fibrotic process. The radiation-induced lung fibrosis model used was C57BL/6 mice irradiated to the entire thorax or right hemithorax to 20 Gy. Our parabiotic model involved joining a transgenic C57BL/6 mouse expressing GFP with a wild-type mouse that was subsequently irradiated to assess for migration of GFP+ bone marrow-derived progenitor cells to the irradiated lung. CXCL12 levels in the bronchoalveolar lavage fluid (BALF) and serum after irradiation were determined by ELISA. CXCR4 and CXCL12 mRNA in the irradiated lung was determined by RNase protection assay. Irradiated mice were treated daily with AMD3100, an established CXCR4 antagonist; MSX-122; and their corresponding vehicles to determine impact of drug treatment on fibrosis development. Fibrosis was assessed by serial CTs and histology. After irradiation, CXCL12 levels increased in BALF and serum with a corresponding rise in CXCR4 mRNA within irradiated lungs consistent with recruitment of a CXCR4+ cell population. Using our parabiotic model, we demonstrated recruitment of CXCR4+ bone marrow-derived mesenchymal stem cells, identified based on marker expression, to irradiated lungs. Finally, irradiated mice that received MSX-122 had significant reductions in development of pulmonary fibrosis while AMD3100 did not significantly suppress this fibrotic process. CXCR4 inhibition by drugs such as MSX-122 may alleviate potential radiation-induced lung injury, presenting future therapeutic opportunities for patients requiring chest irradiation.


Dyspnea assessment and adverse events during sputum induction in COPD.
The inhalation of normal or hypertonic saline during sputum induction (SI) may act as an indirect bronchoconstrictive stimulus leading to dyspnea and lung function deterioration. Our aim was to assess dyspnea and adverse events in COPD patients who undergo SI following a safety protocol. Sputum was induced by normal and hypertonic (4.5%) saline solution in 65 patients with COPD of varying severity. In order to minimize saline-induced bronchoconstriction a protocol based on the European Respiratory Society sputum induction Task group report was followed. Dyspnea change was scored using the Borg scale and lung function was assessed by spirometry and oximetry. Borg score changes [median(IQR) 1.5(0-2)] were observed during SI in 40 subjects; 16 patients required temporary discontinuation of the procedure due to dyspnea-general discomfort and 2 did not complete the session due to dyspnea-wheezing. The change in Borg dyspnea score was significantly correlated with oxygen saturation and heart rate changes and with discontinuation of the procedure due to undesired symptoms. 19 subjects presented an hyperresponsive reaction (decline>20% from baseline FEV1). No significant correlation between Borg changes and FEV1 decline was found. Patients with advanced COPD presented significantly greater Borg and oxygen saturation changes than patients with less severe disease (p = 0.02 and p = 0.001, respectively). Baseline FEV1, oxygen saturation and 6MWT demonstrated significant diagnostic values in distinguishing subjects who develop an adverse physiologic reaction during the procedure. COPD patients undergoing SI following a safety protocol do not experience major adverse events. Dyspnea and oxygen desaturation is more likely to occur in patients with disease in advanced stages, leading to short discontinuation or less frequently to termination of the procedure. Baseline FEV1, oxygen saturation and 6MWT may have a prognostic value for the development of these adverse events and might be useful to be evaluated in advance.


Human immunodeficiency virus associated thrombotic thrombocytopenic purpura, a clinical conundrum.
HIV complicates the diagnostic and therapeutic approaches to idiopathic thrombotic thrombocytopenic purpura (TTP), prompting debate in the literature regarding the benefit of plasma exchange versus simple plasma infusion. Herein we present a case of HIV-TTP, initially treated conservatively with plasma infusion but because of progressive neurologic decline, required urgent plasma exchange for resolution of hematologic derangements and neurologic sequelae. Based on the available literature, there appears to be a spectrum of HIV-associated TTP disorders. Patients with advanced HIV disease and opportunistic infections who present with thrombotic microangiopathy tend to respond to simple plasma infusion, while patients with less progressive HIV disease tend to behave like those with idiopathic TTP, requiring plasma exchange rather than simple plasma infusion. This article illustrates that in patients with HIV-TTP who do not respond to plasma infusion, early escalation to plasma exchange may help avoid life-threatening complications such as seizures and even death.


Design, synthesis and biological evaluation of 3',4',5'-trimethoxy flavonoid benzimidazole derivatives as potential anti-tumor agents.
A series of 3',4',5'-trimethoxy flavonoids with benzimidazole linked by different chain alkanes have been designed and synthesized. The potential activity of these compounds as anti-tumor agents was evaluated by cytotoxicity assay in MGC-803 (human gastric cancer), MCF-7 (human breast cancer), HepG-2 (human hepatoma) and MFC (mouse gastric cancer) tumor cell lines. Among them, compound 15 7-(3-(2-chloro-1H-benzo[d]imidazol-1-yl)propoxy)-2-(3,4,5-trimethoxyphenyl)-4H-chromen-4-one displayed the most potent antiproliferative activity, with IC50 values of 20.47 ± 2.07, 43.42 ± 3.56, 35.45 ± 2.03 μM and 23.47 ± 3.59 μM, respectively. The flow cytometry (FCM) results showed that compound 15 caused the cell cycle to be arrested in G1 phase and induced apoptosis of MFC cells in a dose-dependent manner. In addition, compound 15 exhibited a significant inhibitory effect on tumor growth in vivo. All the results outlined the great potential of compound 15 for further exploitation as anti-tumor agent.


Metabolic disturbances of the vitamin A pathway in human diaphragmatic hernia.
Congenital diaphragmatic hernia (CDH) is a common life-threatening congenital anomaly resulting in high rates of perinatal death and neonatal respiratory distress. Some of the nonisolated forms are related to single-gene mutations or genomic rearrangements, but the genetics of the isolated forms (60% of cases) still remains a challenging issue. Retinoid signaling (RA) is critical for both diaphragm and lung development, and it has been hypothesized that subtle disruptions of this pathway could contribute to isolated CDH etiology. Here we used time series of normal and CDH lungs in humans, in nitrofen-exposed rats, and in surgically induced hernia in rabbits to perform a systematic transcriptional analysis of the RA pathway key components. The results point to CRPBP2, CY26B1, and ALDH1A2 as deregulated RA signaling genes in human CDH. Furthermore, the expression profile comparisons suggest that ALDH1A2 overexpression is not a primary event, but rather a consequence of the CDH-induced lung injury. Taken together, these data show that RA signaling disruption is part of CDH pathogenesis, and also that dysregulation of this pathway should be considered organ specifically.


Hereditary diffuse gastric cancer due to a previously undescribed CDH1 splice site mutation.
Our patient was a 52-year-old man who was diagnosed with signet ring cell gastric adenocarcinoma. An extensive family history of gastric cancer raised suspicion for hereditary diffuse gastric cancer. Sequencing of the patient's CDH1 gene revealed a novel point mutation in a strictly conserved splice site within intron 6, c.833-2 A > G. This mutation was predicted to result in loss of function due to defective RNA splicing. To characterize the pathogenic mechanism of this mutation, we amplified the patient's CDH1 gene products by reverse transcriptase polymerase chain reaction. Primers flanking the region of the mutation detected 3 distinct transcripts. In addition to the wild-type product, a larger product consistent with activation of a cryptic splice site within intron 6 and a smaller product shown to result from exon 7 skipping were detected. In summary, we have identified a novel CDH1 mutation in a large hereditary diffuse gastric cancer kindred and identified its pathogenic mechanism.