Overexpression of transfected human ribonucleotide reductase M2 subunit in human cancer cells enhances their invasive potential.
The ribonucleotide reductase (RR) gene has been associated with malignant transformation and metastatic potential. In this report, the significance of the expression of RR mRNA and enzymatic activity to the invasive potential was examined by Boyden chamber invasion assay. Our results suggest that overexpression of RR M2 mRNA and RR enzymatic activity correlates to an increase in cell invasive potential. The drug-induced HURs clone expressed a higher level RR M2 mRNA and enzyme activity which contributes significantly to the 3-fold increase in invasive potential of the cells observed relative to the KB wild-type control. On the contrary, the HUr revertant clone decreased the RR M2 mRNA level and enzymatic activity, concomitantly decreasing their invasive potential. This phenomenon is most likely due to the return of RR to levels comparable to that of the KB wild-type cells. To confirm that this observation was not of a drug-resistance phenotype associated with multiple gene alterations, the panel of RR transfectants (M1-D transfected M1 subunit cDNA, M2-D transfected M2 subunit cDNA, X-D transfected M1/M2 cDNA) characterized in a previous study were also tested in the invasion assay. The M2-D clone expressed 6-fold higher RR M2 mRNA and RR activity and also demonstrated 6-fold higher invasive potential in vitro than either the parental or vector only transfected cell line (KB-V). The X-D clone demonstrated 3-fold higher M2 mRNA expression and revealed 4-fold higher invasive potential than control cells. The M1-D clone, in contrast, expressed a baseline level of RR M2 mRNA and higher M1 mRNA. In contrast to the X-D and M2-D cells, the invasive potential of M1-D reached an even lower level in the invasive assay than the control. These results, therefore, suggest that RR M2 overexpression plays an important role in a tumor's invasiveness.


Influence of Health Behaviors and Occupational Stress on Prediabetic State among Male Office Workers.
This study examined the influence of health behaviors and occupational stress on the prediabetic state of male office workers, and identified related risks and influencing factors. The study used a cross-sectional design and performed an integrative analysis on data from regular health checkups, health questionnaires, and a health behavior-related survey of employees of a company, using Spearman's correlation coefficients and multiple logistic regression analysis. The results showed significant relationships of prediabetic state with health behaviors and occupational stress. Among health behaviors, a diet without vegetables and fruits (Odds Ratio (OR) = 3.74, 95% Confidence Interval (CI) = 1.93⁻7.66) was associated with a high risk of prediabetic state. In the subscales on occupational stress, organizational system in the 4th quartile (OR = 4.83, 95% CI = 2.40⁻9.70) was significantly associated with an increased likelihood of prediabetic state. To identify influencing factors of prediabetic state, the multiple logistic regression was performed using regression models. The results showed that dietary habits (β = 1.20, p = 0.002), total occupational stress score (β = 1.33, p = 0.024), and organizational system (β = 1.13, p = 0.009) were significant influencing factors. The present findings indicate that active interventions are needed at workplace for the systematic and comprehensive management of health behaviors and occupational stress that influence prediabetic state of office workers.


Patterns of care for radiotherapy in vulvar cancer: a Gynecologic Cancer Intergroup study.
This study aimed to describe radiotherapeutic practice in the treatment of vulvar cancer in member study groups of the Gynecologic Cancer Intergroup (GCIG). A survey was developed and distributed to representatives of the member study groups of the GCIG, targeting the use of radiotherapy (RT) in vulvar cancer. Thirty-two surveys were returned from 12 different cooperative groups. The most common indications for neoadjuvant RT include unresectable disease or International Federation of Gynecology and Obstetrics stage >/=III. For the neoadjuvant treatment of vulvar cancer, pelvic doses were 48.2 +/- 5.0 Gy (mean +/- SD). The upper border of the pelvic field was L4/5 in 4, L5/S1 in 12, and not specified in 4. Of 21 groups that perform neoadjuvant RT, 17 use concomitant chemotherapy and 4 individualize treatment. Weekly cisplatin was the most commonly used chemotherapy. For the neoadjuvant RT treatment of the inguinal region, doses were 49.9 +/- 5.5 Gy (mean +/- SD). Sixteen of 18 groups used computed tomographic simulation for planning. After initial surgery, the most common indications for RT included positive lymph nodes or positive margins. Chemotherapy was not routinely used after surgery. Doses of RT among GCIG members are similar; however, the indications for treatment, treatment fields, and use of chemotherapy differ somewhat between groups. This is likely due to the rarity of the disease. The lack of randomized trials may contribute to the absence of a broadly accepted standard. This underscores the importance of international cooperation as in GCIG to gather more reliable data for uncommon tumors in gynecologic oncology.


Current Status of the Retrieval Rate of Retrievable Vena Cava Filters in a Tertiary Referral Center in Korea.
The purpose of this study was to review the daily practice of inferior vena cava filters (IVCFs) in a tertiary referral center in Korea and to reveal the retrieval rate and the methods for improving it. Through the electronic medical record system, a retrospective review was performed on 115 consecutive patients who underwent placement of retrievable IVCFs between February 2000 and January 2011 in Seoul National University Hospital. IVCF placement was done in 115 cases (113 patients). There were 68 men (59.1%), and the mean age was 58.5±15.5 years (range, 10-96 years). The affiliated departments were Vascular Surgery (57 cases, 49.6%), and Internal Medicine (20 cases, 17.4%). Advanced malignancy was the most commonly associated disease (n=30, 26%). The indications for IVCF placement were categorized; absolute indications in 36 cases (31.3%), relative indications in 78 cases (67.8%), and prophylactic use in 1 case (0.9%). The most common indications were thrombolysis/thrombectomy for iliocaval deep vein thrombosis (DVT) (n=55, 47.8). Of the 115 filters, 68 were retrieved (retrieval rate, 59%). The most common cause of non-retrieval was chronic high risk of venous thromboembolism in 24 patients (51%), followed by residual proximal DVT (n=7, 15%), and negligence by unknown reasons (n=6, 13%). To improve the retrieval rate, the number of follow-up losses to vascular specialists must be decreased, which can be achieved by establishment of a dedicated IVC filter clinic, implementation of a filter registry, and regular education for medical teams and patients along with their families.


Is targeted fortification of human breast milk an optimal nutrition strategy for preterm infants? An interventional study.
Fortifying human milk contributes to the prevention of postnatal growth failure in preterm infants. Because of the natural variability of human milk, targeted fortification of human milk has been advocated. However, data regarding the efficacy and safety of prolonged targeted fortification are scarce. We aimed to assess the safety of targeted fortification of human milk in preterm infants compared with standard fortification, as well as the effects on infant growth. We conducted an interventional study during hospital stay in healthy very low birth weight preterm infants who were exclusively fed human milk. Pools of human milk collected for 24 h were analysed using mid-infrared transmission spectroscopy. Targeted fortification of human milk was performed by adding macronutrients to native human milk to obtain optimal ratios of fat (4.4 g), carbohydrates (8.8 g), and protein (3 g) per 100 ml. The intervention period lasted 4-7 weeks. Weekly weight and daily growth rates were compared with those of a standardized fortification group of very low birth weight preterm infants who received standard fortified human milk (n = 10). The osmolality as well as the metabolic and gastrointestinal tolerance were monitored. Intergroup differences were evaluated using the Mann-Whitney U-test. A total of 10 preterm infants (birth weight 1223 ± 195 g; gestational age 29.1 ± 1.03 weeks) were enrolled and 118 samples of pooled milk were analysed. On average, 1.4 ± 0.1 g of protein, 2.3 ± 0.5 g of carbohydrate, and 0.3 ± 0.1 g of fat per 100 ml were added to the milk. Osmolality values after target fortification were within recommended limits (376 ± 66 mOsml/kg). Weekly weight gain (205.5 g; 95 % CI 177-233 vs 155 g; 95 % CI 132-178; p = 0.025) and daily growth rates (15.7 g/kg/day; 95 % CI 14.5-16.9 vs 12.3 g/kg/day; 95 % CI 10.7-13.9; p = 0.005) were higher in infants receiving target fortification than in infants receiving standardized fortification. The infants receiving targeted fortified milk consumed similar volumes as infants in the standardized fortification group (148 ± 4.5 vs 146 ± 4 ml/kg/day). No signs of either gastrointestinal or metabolic intolerance were observed. Target fortification appears to promote growth in very low birth weight preterm infants without any detrimental effects. Trial registration NCT02716337.


Functional inaccessibility of quiescent herpes simplex virus genomes.
Newly delivered herpes simplex virus genomes are subject to repression during the early stages of infection of human fibroblasts. This host defence strategy can limit virus replication and lead to long-term persistence of quiescent viral genomes. The viral immediate-early protein ICP0 acts to negate this negative regulation, thereby facilitating the onset of the viral replication cycle. Although few mechanistic details are available, the host repression machinery has been proposed to assemble the viral genome into a globally inaccessible configuration analogous to heterochromatin, blocking access to most or all trans-acting factors. The strongest evidence for this hypothesis is that ICP0-deficient virus is unable to reactivate quiescent viral genomes, despite its ability to undergo productive infection given a sufficiently high multiplicity of infection. However, recent studies have shown that quiescent infection induces a potent antiviral state, and that ICP0 plays a key role in disarming such host antiviral responses. These findings raise the possibility that cells containing quiescent viral genomes may be refractory to superinfection by ICP0-deficient virus, potentially providing an alternative explanation for the inability of such viruses to trigger reactivation. We therefore asked if ICP0-deficient virus is capable of replicating in cells that contain quiescent viral genomes. We found that ICP0-deficient herpes simplex virus is able to infect quiescently infected cells, leading to expression and replication of the superinfecting viral genome. Despite this productive infection, the resident quiescent viral genome was neither expressed nor replicated, unless ICP0 was provided in trans. These data document that quiescent HSV genomes fail to respond to the virally modified host transcriptional apparatus or viral DNA replication machinery provided in trans by productive HSV infection in the absence of ICP0. These results point to global repression as the basis for HSV genome quiescence, and indicate that ICP0 induces reactivation by overcoming this global barrier to the access of trans-acting factors.


Impaired nonrestricted cytolytic activity in systemic lupus erythematosus: involvement of a pathway independent of Fas, tumor necrosis factor, and extracellular ATP that is associated with little detectable perforin.
To determine the cytolytic effector pathway involved in impaired generation of nonrestricted cytolytic activity in systemic lupus erythematosus (SLE). Peripheral blood mononuclear cells (PBMC) from normal subjects and SLE patients were stimulated in vitro with anti-CD3 monoclonal antibody (MAb) and interleukin-2 to promote the generation of nonrestricted cytolytic activity. On day 13 of culture, the PBMC were assayed for cytolytic activity against Fas-Daudi cells and Fas+ Jurkat cells. The effects on cytotoxicity of calcium chelation, antagonist anti-Fas MAb, tumor necrosis factor (TNF) alpha and beta, and ATP were measured. Intracellular perforin expression was determined by intracellular staining, and perforin messenger RNA levels were determined by quantitative competitive reverse transcription-polymerase chain reaction. We demonstrated the existence of a cytolytic pathway that is independent of Fas, TNF alpha, TNF beta, and ATP, but is dependent upon extracellular calcium. Despite its calcium dependence, this pathway is associated with low-to-undetectable levels of perforin. Impaired generation of nonrestricted cytolytic activity in SLE is likely due to decreased activity of this Fas-, TNF alpha-, TNF beta-, ATP-independent pathway associated with very low levels of perforin.


Silymarin and silibinin cause G1 and G2-M cell cycle arrest via distinct circuitries in human prostate cancer PC3 cells: a comparison of flavanone silibinin with flavanolignan mixture silymarin.
Here, we assessed and compared the anticancer efficacy and associated mechanisms of silymarin and silibinin in human prostate cancer (PCA) PC3 cells; silymarin is comprised of silibinin and its other stereoisomers, including isosilybin A, isosilybin B, silydianin, silychristin and isosilychristin. Silymarin and silibinin (50-100 microg/ml) inhibited cell proliferation, induced cell death, and caused G1 and G2-M cell cycle arrest in a dose/time-dependent manner. Molecular studies showed that G1 arrest was associated with a decrease in cyclin D1, cyclin D3, cyclin E, cyclin-dependent kinase (CDK)4, CDK6 and CDK2 protein levels, and CDK2 and CDK4 kinase activity, together with an increase in CDK inhibitors (CDKIs) Kip1/p27 and Cip1/p21. Further, both agents caused cytoplasmic sequestration of cyclin D1 and CDK2, contributing to G1 arrest. The G2-M arrest by silibinin and silymarin was associated with decreased levels of cyclin B1, cyclin A, pCdc2 (Tyr15), Cdc2, and an inhibition of Cdc2 kinase activity. Both agents also decreased the levels of Cdc25B and cell division cycle 25C (Cdc25C) phosphatases with an increased phosphorylation of Cdc25C at Ser216 and its translocation from nucleus to the cytoplasm, which was accompanied by an increased binding with 14-3-3beta. Both agents also increased checkpoint kinase (Chk)2 phosphorylation at Thr68 and Ser19 sites, which is known to phosphorylate Cdc25C at Ser216 site. Chk2-specific small interfering RNA largely attenuated the silymarin and silibinin-induced G2-M arrest. An increase in the phosphorylation of histone 2AX and ataxia telangiectasia mutated was also observed. These findings indicate that silymarin and silibinin modulate G1 phase cyclins-CDKs-CDKIs for G1 arrest, and the Chk2-Cdc25C-Cdc2/cyclin B1 pathway for G2-M arrest, together with an altered subcellular localization of critical cell cycle regulators. Overall, we observed comparable effects for both silymarin and silibinin at equal concentrations by weight, suggesting that silibinin could be a major cell cycle-inhibitory component in silymarin. However, other silibinin stereoisomers present in silymarin also contribute to its efficacy, and could be of interest for future investigation.


Salmonella typhimurium proliferates and establishes a persistent infection in the intestine of Caenorhabditis elegans.
Genetic analysis of host-pathogen interactions has been hampered by the lack of genetically tractable models of such interactions. We showed previously that the human opportunistic pathogen Pseudomonas aeruginosa kills Caenorhabditis elegans, that P. aeruginosa and C. elegans genes can be identified that affect this killing, and that most of these P. aeruginosa genes are also important for mammalian pathogenesis. Here, we show that Salmonella typhimurium as well as other Salmonella enterica serovars including S. enteritidis and S. dublin can also kill C. elegans. When C. elegans is placed on a lawn of S. typhimurium, the bacteria accumulate in the lumen of the worm intestine and the nematodes die over the course of several days. This killing requires contact with live bacterial cells. The worms die with similar kinetics when placed on a lawn of S. typhimurium for a relatively short time (3-5 hours) before transfer to a lawn of E. coli. After the transfer to E. coli, a high titer of S. typhimurium persists in the C. elegans intestinal lumen for the rest of the worms' life. Furthermore, feeding for 5 hours on a 1:1000 mixture of S. typhimurium and E. coli followed by transfer to 100% E. coli, also led to death after several days. This killing correlated with an increase in the titer of S. typhimurium in the C. elegans lumen, which reached 10,000 bacteria per worm. These data indicate that, in contrast to P. aeruginosa, a small inoculum of S. typhimurium can proliferate in the C. elegans intestine and establish a persistent infection. S. typhimurium mutated in the PhoP/PhoQ signal transduction system caused significantly less killing of C. elegans.


Passive Transfer of Hepatitis B Antibodies through Intravenous Immunoglobulin in a Neonate.
Passive transfer of antibodies secondary to intravenous immunoglobulin infusion is a rare but important side effect that can lead to the wrong diagnosis and therapeutic decisions. It has never been reported in a newborn. A male newborn, vaccinated against hepatitis B and diagnosed with dilated cardiomyopathy, presented positive hepatitis B core antibodies at 12 days of life. Exclusion of hepatitis B infection was mandatory as it would be a contraindication to heart transplant. Passive transfer of antibodies was confirmed at 44 days of age, after seroreversion of hepatitis B core antibodies. Passive transfer of antibodies after intravenous immunoglobulin infusion can lead to a misleading diagnosis if not recognized. In our patient it could have been especially harmful had it prevented heart transplant. Screening for hepatitis B should be performed at least 1 month after intravenous immunoglobulin infusion.


Two-component cavity based on a regular photonic crystal nanobeam.
The authors propose and numerically examine a two-component design for an optical nanocavity. Such a nanocavity consists, first, of a photonic crystal (PC) nanobeam, in which the PC unit cell is not changed. Second, the cavity contains a fragment of some supplementary material of the size of several or several tens of PC unit cells. When the two components are combined, a defect forms in which the resonant mode can be excited. The advantages of the proposed cavity model are reported, particularly the possibility of implementing electrically pumped light sources and amplifiers and the simplification of development of nanocavities with nonlinear properties. The fabrication tolerances of the proposed nanocavity were investigated. It has been found that existing structural layer alignment technologies can be used for fabricating the suggested cavity.


Covalent organic nanosheets for bioimaging.
Covalent organic nanosheets (CONs) have attracted much attention because of their excellent physical, chemical, electronic, and optical properties. Although covalent organic nanosheets have widely been used in many applications, there are only a few CONs that have been tested for bio-medical applications. Nanometer sized triazine-based nanosheets were obtained by exfoliating their bulk counterparts in water. The obtained nanosheets were dispersible and stable in water with enhanced photoluminescence properties compared to the bulk material. The nanosheets were biocompatible and non-toxic and showed ability to stain HeLa cell nuclei without additional assistance of an external targeting agent.


Intracerebral hemorrhage in stroke patients anticoagulated with heparin.
To characterize the clinical features of patients with acute cerebral infarction who sustained intracerebral hemorrhage related to heparin anticoagulation, we describe 10 patients and review reports of 16 cases. Cardiac-source embolism was identified in seven (70%) of the 10 patients and consisted of atrial fibrillation in six of the seven. The middle cerebral artery territory was affected in nine patients (90%), with moderate-sized or large infarcts by clinical and computed tomographic criteria. The interval between stroke onset and intracerebral hemorrhage was less than 72 hours in 80% of the patients. Intracerebral hemorrhage occurred less than or equal to 24 hours after the time heparin was started in 80% of the patients. The activated partial thromboplastin time closest to the time of intracerebral hemorrhage was greater than 2 x control in seven patients. Our findings in the 10 patients are similar to those of the 16 cases previously reported and suggest that heparin-related intracerebral hemorrhage occurs early after stroke onset, usually with moderate-sized or large infarcts, and with excessive anticoagulation in some patients.


Magnitude and variability of sequential estradiol and progesterone concentrations in women using depot medroxyprogesterone acetate for contraception.
To describe the magnitude and variability of sequential serum estradiol and progesterone concentrations throughout one depot medroxyprogesterone (DMPA) injection interval. Prospective study. Family planning and women's health clinics. Thirty-one women, ages 19 to 46, using DMPA for contraception. Serum for estrogen and progesterone was collected weekly throughout one DMPA injection interval. Serum estradiol and progesterone concentrations; estradiol patterns produced from data plotted across the entire DMPA injection interval. The average daily estradiol concentrations ranged from 7.9 to 69.1 pg/mL, with a mean of 18.9 +/- 12.9 and a median of 15.4 pg/mL. Average daily progesterone concentrations ranged from 0.14 to 1.1 ng/mL, with a mean of 0.40 +/- 0.19 ng/mL and a median of 0.36 ng/mL. Two general patterns of estradiol concentrations were identified. One pattern, observed in approximately one third of the participants, reflected estradiol concentrations that were extremely low (mean, 12.7 +/- 3.6 pg/mL; median, 13.4 pg/mL) and consistently flat across the DMPA injection interval. The second pattern, seen in the remaining participants, reflected estradiol concentrations that were higher (mean, 22.2 +/- 14.9 pg/mL; median, 17.3 pg/mL) and quite variable. This study demonstrated that estradiol concentrations were lower than the 40 to 50 pg/mL reported in most studies and, for the majority of women, varied substantially across the DMPA injection interval.


Phosphorylation of CYFIP2, a component of the WAVE-regulatory complex, regulates dendritic spine density and neurite outgrowth in cultured hippocampal neurons potentially by affecting the complex assembly.
Actin dynamics is a critical mechanism underlying many cellular processes in neurons. The heteropentameric WAVE-regulatory complex (WRC), consisting of WAVE, CYFIP1/2, Nap, Abi, and HSPC300, is a key regulator of actin dynamics that activates the Arp2/3 complex to initiate actin polymerization and branching. The WRC is basally inactive because of intermolecular interactions among the components, which can be modulated by bindings of phospholipids and Rac1, and phosphorylations of WAVE and Abi. However, the phosphorylation of other components of WRC and their functional significance remain largely unknown. To address this issue, we focused on CYFIP1/2, in which we found two brain-specific phosphorylation sites (S582 of CYFIP2 and T1068/T1067 of CYFIP1/2) from a publicly available phosphoproteome database. To understand their functional effects, we overexpressed wild-type, phospho-blocking, or phospho-mimetic mutants of CYFIP2 in cultured hippocampal neurons, and found that only T1067A CYFIP2 decreased the density of stubby spines. Moreover, overexpression of wild-type CYFIP2 increased neurite length, but T1067A did not exert this effect. To understand the mechanism, we modeled CYFIP2 phosphorylation in the crystal structure of WRC and found that T1067 phosphorylation could weaken the interaction between CYFIP2 and Nap1 by inducing conformational changes of CYFIP2 α-helical bundles. In the co-immunoprecipitation assay, however, wild-type, T1067A, and T1067E CYFIP2 showed similar interaction levels to Nap1, suggesting that T1067 phosphorylation alone is not sufficient to disrupt the interaction. Considering that the activation of WRC requires disassembly of the complex, our results suggest that T1067 phosphorylation, together with other factors, could contribute toward the activation process.


Quantitative proteomics reveals protein profiles underlying major transitions in aspen wood development.
Wood development is of outstanding interest both to basic research and industry due to the associated cellulose and lignin biomass production. Efforts to elucidate wood formation (which is essential for numerous aspects of both pure and applied plant science) have been made using transcriptomic analyses and/or low-resolution sampling. However, transcriptomic data do not correlate perfectly with levels of expressed proteins due to effects of post-translational modifications and variations in turnover rates. In addition, high-resolution analysis is needed to characterize key transitions. In order to identify protein profiles across the developmental region of wood formation, an in-depth and tissue specific sampling was performed. We examined protein profiles, using an ultra-performance liquid chromatography/quadrupole time of flight mass spectrometry system, in high-resolution tangential sections spanning all wood development zones in Populus tremula from undifferentiated cambium to mature phloem and xylem, including cell expansion and cell death zones. In total, we analyzed 482 sections, 20-160 μm thick, from four 47-year-old trees growing wild in Sweden. We obtained high quality expression profiles for 3,082 proteins exhibiting consistency across the replicates, considering that the trees were growing in an uncontrolled environment. A combination of Principal Component Analysis (PCA), Orthogonal Projections to Latent Structures (OPLS) modeling and an enhanced stepwise linear modeling approach identified several major transitions in global protein expression profiles, pinpointing (for example) locations of the cambial division leading to phloem and xylem cells, and secondary cell wall formation zones. We also identified key proteins and associated pathways underlying these developmental landmarks. For example, many of the lignocellulosic related proteins were upregulated in the expansion to the early developmental xylem zone, and for laccases with a rapid decrease in early xylem zones. We observed upregulation of two forms of xylem cysteine protease (Potri.002G005700.1 and Potri.005G256000.2; Pt-XCP2.1) in early xylem and their downregulation in late maturing xylem. Our data also show that Pt-KOR1.3 (Potri.003G151700.2) exhibits an expression pattern that supports the hypothesis put forward in previous studies that this is a key xyloglucanase involved in cellulose biosynthesis in primary cell walls and reduction of cellulose crystallinity in secondary walls. Our novel multivariate approach highlights important processes and provides confirmatory insights into the molecular foundations of wood development.


European myeloma network: the value of collaborative research.
The European Myeloma Network was established as a result of collaboration and cooperation between myeloma physicians and researchers. The initial impetus that brought this group together came from the Sixth Framework Programme (2002), as the group initially formed to apply for funding from this source. Although the application was unsuccessful, it provided encouragement to continue the activities, and the network was formally established in October 2003. The basic idea was to create interaction between the people working in different areas of myeloma (for example, basic science, clinical trials and patient education) and build an interface to establish standardisation of laboratory techniques such as fluorescent in-situ hybridisation (FISH) techniques for these groups. The aim of the network is to bring these groups together in some way, to deliver better quality research. Resources need to be used wisely, with priorities being set. Establishing a common databank of trials from the European Community where trial data is pooled is one such priority. This project can then extend into the future to analyse clinical, biological and genetic prognostic factors. Looking ahead to other potential benefits of the network, there may be opportunities to examine orphan treatments, and it has the potential to act as the intermediate organisation, helping pharmaceutical companies and tumour working groups in a variety of countries to establish the most appropriate clinical trials.


How altruistic organ donation may be (intrinsically) bad.
It has traditionally been assumed that organ donation must be altruistic, though the necessity of altruistic motivations has recently been questioned. Few, however, have questioned whether altruism is always a good motive. This paper considers the possibility that excessive altruism, or self-abnegation, may be intrinsically bad. How this may be so is illustrated with reference to Tom Hurka's account of the value of attitudes, which suggests that disproportionate love of one's own good-either excessive or deficient-is intrinsically bad. Whether or not we accept the details of this account, recognising that altruistic motivations may be intrinsically bad has important implications for organ procurement. One possible response is to say that we should take further measures to ensure that donors have good motives-that they are altruistic is no longer enough. An alternative is to say that, since altruistic donation need not be intrinsically good, we have less reason to object to other motivations.


Population-based estimate of the sibling recurrence risk ratio for rhegmatogenous retinal detachment.
The influence of genetic predisposition on nonsyndromic primary rhegmatogenous retinal detachment (RRD) is poorly characterized. The purpose of this study was to investigate the magnitude of genetic risk for RRD. All participants (probands) in the Scottish Retinal Detachment Study (N = 922) with known postal addresses were contacted by questionnaire to assess the personal and family history of RRD. Sibling affection status was modeled by logistic regression and generalizing estimating equations accounting for the effect of proband covariates of age, sex, spherical equivalent refraction, index birth order, and body mass index (BMI). Sibling-sibling recurrence risk ratios (λs) and parent-offspring recurrence risk ratios were calculated. Sixty-five percent of probands returned completed questionnaires. Of these, 602 families (parents, siblings, offspring), 7.8% (47) had one affected member, and 0.5% (3) had two affected members. A total of 501 sibships were included in the regression analysis. The odds ratio (OR) that a sibling would be affected, given another affected sibling, was 1.91 (95% confidence interval [CI], 1.18-3.05). With adjustment for age and sex, the OR that a sibling would be affected increased by 9.8% for each additional diopter of spherical equivalent refractive error (SER) toward myopia in the proband. The λs and the parent-offspring recurrence risk ratio of RRD were 2.1 (95% CI, 1.3-3.2) and 2.9 (95% CI, 1.9-4.2), respectively. Genetic factors are important in the etiology of myopic and nonmyopic RRD. The risk of having an affected sibling with RRD increases twofold, given that a sibling has had the condition. The sibling risk increases with the level of spherical equivalent myopia in the proband.


Optimisation of a green gas supply chain--a review.
In this review the knowledge status of and future research options on a green gas supply based on biogas production by co-digestion is explored. Applications and developments of the (bio)gas supply in The Netherlands have been considered, whereafter literature research has been done into the several stages from production of dairy cattle manure and biomass to green gas injection into the gas grid. An overview of a green gas supply chain has not been made before. In this study it is concluded that on installation level (micro-level) much practical knowledge is available and on macro-level knowledge about availability of biomass. But on meso-level (operations level of a green gas supply) very little research has been done until now. Future research should include the modeling of a green gas supply chain on an operations level, i.e. questions must be answered as where to build digesters based on availability of biomass. Such a model should also advise on technology of upgrading depending on scale factors. Future research might also give insight in the usability of mixing (partly upgraded) biogas with natural gas. The preconditions for mixing would depend on composition of the gas, the ratio of gases to be mixed and the requirements on the mixture.


Human breast adenocarcinoma MCF-7/0 cells electroporated with cytosolic class 3 aldehyde dehydrogenases obtained from tumor cells and a normal tissue exhibit differential sensitivity to mafosfamide.
The cytosolic class aldehyde dehydrogenase (ALDH-3) present in human normal tissues/secretions is apparently much less able to catalyze the oxidation aldophosphamide to carboxyphosphamide than is the ALDH-3 present in human tumor cells/tissues, suggesting that the former may be less able to protect cells from the cytotoxic action of cyclophosphamide, mafosfamide, and other oxazaphosphorines. To test this notion, relatively large and approximately equal amounts of human normal stomach mucosa ALDH-3 and catechol-induced human breast adenocarcinoma MCF-7/0 ALDH-3 were first electroporated into cells (MCF-7/0) that constitutively express only very small amounts of the enzyme. The resultant preparations were then tested for sensitivity to mafosfamide. ALDH-3 activities (NADP-dependent catalysis of benzaldehyde oxidation) were 1.7, 212, and 183 mlU/10(7) cells in sham-electroporated MCF-7/0 cells, and MCF-7/0 cells electroporated with stomach mucosa ALDH-3 and catechol-induced MCF-7/0 ALDH-3, respectively. LC90 values (concentrations of mafosfamide required to effect a 90% cell kill) were 62, 417, and >1,000 microM, respectively. The three preparations were equisensitive to phosphoramide mustard (LC90 = approximately 850 microM). Inclusion of benzaldehyde in the drug exposure medium fully restored the sensitivity of MCF-7/0 cells electroporated with either enzyme to mafosfamide. These observations support the notions that 1) cellular sensitivity to the oxazaphosphorines decreases as the cellular content of ALDH-3 increases, 2) the foregoing is the consequence of ALDH-3-catalyzed oxidation (thus detoxification) of aldophosphamide, and 3) the ALDH-3 present in at least some tumor cells/tissues is a slight variant of the ALDH-3 present in normal tissues/secretions. Furthermore, they illustrate the utility of electroporation used as a tool to determine whether a given enzyme, or even more generally, protein or other macromolecule, is a determinant of cellular sensitivity to a given cytotoxic agent.


Effect of two 12-minute culturally targeted films on intent to call 911 for stroke.
We assessed the behavioral effect of two 12-minute culturally targeted stroke films on immediately calling 911 for suspected stroke among black and Hispanic participants using a quasi-experimental pretest-posttest design. We enrolled 102 adult churchgoers (60 black and 42 Hispanic) into a single viewing of one of the 2 stroke films-a Gospel musical (English) or Telenovela (Spanish). We measured intent to immediately call 911 using the validated 28-item Stroke Action Test in English and Spanish, along with related variables, before and immediately after the intervention. Data were analyzed using repeated-measures analysis of variance. An increase in intent to call 911 was seen immediately following the single viewing. Higher self-efficacy for calling 911 was associated with intent to call 911 among Hispanic but not black participants. A composite measure of barriers to calling 911 was not associated with intent to call 911 in either group. A significant association was found between higher stroke symptom knowledge and intent to call 911 at baseline, but not immediately following the intervention. No sex associations were found; however, being older was associated with greater intent to call 911. The majority of participants would strongly recommend the films to others. One participant appropriately called 911 for a real-life stroke event. Narrative communication in the form of tailored short films may improve intent to call 911 for stroke among the black and Hispanic population.


A radar-enabled collaborative sensor network integrating COTS technology for surveillance and tracking.
The feasibility of using Commercial Off-The-Shelf (COTS) sensor nodes is studied in a distributed network, aiming at dynamic surveillance and tracking of ground targets. Data acquisition by low-cost (<$50 US) miniature low-power radar through a wireless mote is described. We demonstrate the detection, ranging and velocity estimation, classification and tracking capabilities of the mini-radar, and compare results to simulations and manual measurements. Furthermore, we supplement the radar output with other sensor modalities, such as acoustic and vibration sensors. This method provides innovative solutions for detecting, identifying, and tracking vehicles and dismounts over a wide area in noisy conditions. This study presents a step towards distributed intelligent decision support and demonstrates effectiveness of small cheap sensors, which can complement advanced technologies in certain real-life scenarios.


The genomic region encompassing the nephropathic cystinosis gene (CTNS): complete sequencing of a 200-kb segment and discovery of a novel gene within the common cystinosis-causing deletion.
Nephropathic cystinosis is an autosomal recessive disorder caused by the defective transport of cystine out of lysosomes. Recently, the causative gene (CTNS) was identified and presumed to encode an integral membrane protein called cystinosin. Many of the disease-associated mutations in CTNS are deletions, including one >55 kb in size that represents the most common cystinosis allele encountered to date. In an effort to determine the precise genomic organization of CTNS and to gain sequence-based insight about the DNA within and flanking cystinosis-associated deletions, we mapped and sequenced the region of human chromosome 17p13 encompassing CTNS. Specifically, a bacterial artificial chromosome (BAC)-based physical map spanning CTNS was constructed by sequence-tagged site (STS)-content mapping. The resulting BAC contig provided the relative order of 43 STSs. Two overlapping BACs, which together contain all of the CTNS exons as well as extensive amounts of flanking DNA, were selected and subjected to shotgun sequencing. A total of 200,237 bp of contiguous, high-accuracy sequence was generated. Analysis of the resulting data revealed a number of interesting features about this genomic region, including the long-range organization of CTNS, insight about the breakpoints and intervening DNA associated with the common cystinosis-causing deletion, and structural information about five genes neighboring CTNS (human ortholog of rat vanilloid receptor subtype 1 gene, CARKL, TIP-1, P2X5, and HUMINAE). In particular, sequence analysis detected the presence of a novel gene (CARKL) residing within the most common cystinosis-causing deletion. This gene encodes a previously unknown protein that is predicted to function as a carbohydrate kinase. Interestingly, both CTNS and CARKL are absent in nearly half of all cystinosis patients (i.e., those homozygous for the common deletion). [The sequence data described in this paper have been submitted to the GenBank data library under accession nos. AF168787 and AF163573.]


Modulation of Dnmt3b function in vitro by interactions with Dnmt3L, Dnmt3a and Dnmt3b splice variants.
DNA methylation, an essential regulator of transcription and chromatin structure, is established and maintained by the coordinated action of three DNA methyltransferases: DNMT1, DNMT3A and DNMT3B, and the inactive accessory factor DNMT3L. Disruptions in DNMT3B function are linked to carcinogenesis and genetic disease. DNMT3B is also highly alternatively spliced in a tissue- and disease-specific manner. The impact of intra-DNMT3 interactions and alternative splicing on the function of DNMT3 family members remains unclear. In the present work, we focused on DNMT3B. Using a panel of in vitro assays, we examined the consequences of DNMT3B splicing and mutations on its ability to bind DNA, interact with itself and other DNMT3's, and methylate DNA. Our results show that, while the C-terminal catalytic domain is critical for most DNMT3B functions, parts of the N-terminal region, including the PWWP domain, are also important. Alternative splicing and domain deletions also influence DNMT3B's cellular localization. Furthermore, our data reveal the existence of extensive DNMT3B self-interactions that differentially impact on its activity. Finally, we show that catalytically inactive isoforms of DNMT3B are capable of modulating the activity of DNMT3A-DNMT3L complexes. Our studies therefore suggest that seemingly 'inactive' DNMT3B isoforms may influence genomic methylation patterns in vivo.


Waist-to-height ratio and cardiovascular risk factors in elderly individuals at high cardiovascular risk.
Several anthropometric measurements have been associated with cardiovascular disease, type-2 diabetes mellitus and other cardiovascular risk conditions, such as hypertension or metabolic syndrome. Waist-to-height-ratio has been proposed as a useful tool for assessing abdominal obesity, correcting other measurements for the height of the individual. We compared the ability of several anthropometric measurements to predict the presence of type-2 diabetes, hyperglycemia, hypertension, atherogenic dyslipidemia or metabolic syndrome. In our cross-sectional analyses we included 7447 Spanish individuals at high cardiovascular risk, men aged 55-80 years and women aged 60-80 years, from the PREDIMED study. Logistic regression models were fitted to evaluate the odds ratio of presenting each cardiovascular risk factor according to various anthropometric measures. The areas under the receiver-operating characteristic curve (AUC) were used to compare the predictive ability of these measurements. In this relatively homogeneous cohort with 48.6% of type-2 diabetic individuals, the great majority of the studied anthropometric parameters were significantly and positively associated with the cardiovascular risk factors. No association was found between BMI and body weight and diabetes mellitus. The AUCs for the waist-to-height ratio and waist circumference were significantly higher than the AUCs for BMI or weight for type-2 diabetes, hyperglycemia, atherogenic dyslipidemia and metabolic syndrome. Conversely, BMI was the strongest predictor of hypertension. We concluded that measures of abdominal obesity showed higher discriminative ability for diabetes mellitus, high fasting plasma glucose, atherogenic dyslipidemia and metabolic syndrome than BMI or weight in a large cohort of elderly Mediterranean individuals at high cardiovascular risk. No significant differences were found between the predictive abilities of waist-to-height ratio and waist circumference on the metabolic disease.


Preferential benefit of implementation of a statewide trauma system in one of two adjacent states.
Implementation of Oregon's trauma system was associated with a reduction in the risk of death for hospitalized injured patients. An alternative explanation for improved outcome, however, is favorable concurrent temporal trends, e.g., new technologies and treatments. To control for temporal trends, seriously injured hospitalized patients in Oregon and Washington were compared before either state had a trauma system (1985-1988) and when only the Oregon trauma system had been implemented (1990-1993). The study group consisted of hospitalized injured patients aged 16 to 79 years with one or more index injuries in six body regions, i.e., head, chest, spleen/liver, femur or pelvis fracture, and burns. Hospital discharge claims data were analyzed, converting International Classification of Diseases, Ninth Revision, Clinical Modification, discharge diagnosis codes to Abbreviated Injury Scale scores and Injury Severity Scores using a conversion algorithm. Multivariate logistic regression models were used to estimate the differential risk-adjusted odds of death in Oregon compared with Washington after adjustment for demographics, injury type, and injury severity. Findings indicated no difference in the risk-adjusted odds of death between Oregon and Washington while both states functioned under an ad hoc trauma system (1985-1988). A significant reduction in the risk of death, however, was noted in Oregon for patients with an index injury and an Injury Severity Score > 15 compared with Washington (adjusted odds ratio (OR) = 0.80, 95% confidence interval (CI) = 0.70-0.91) after trauma system implementation in Oregon (1990-1993). Specifically, reductions in the risk of death were demonstrated for patients with head injuries (adjusted OR = 0.70, 95% CI = 0.59-0.82) or liver/spleen injuries (adjusted OR = 0.73, 95% CI = 0.54-0.99). Assuming that the two states demonstrated similar concurrent temporal trends, the findings support the conclusion that improved outcomes among injured patients in Oregon may be attributed to the institution of a statewide trauma system.


Estrogenic-dependent glutamatergic neurotransmission from kisspeptin neurons governs feeding circuits in females.
The neuropeptides tachykinin2 (Tac2) and kisspeptin (Kiss1) in hypothalamic arcuate nucleus Kiss1 (Kiss1ARH) neurons are essential for pulsatile release of GnRH and reproduction. Since 17β-estradiol (E2) decreases Kiss1 and Tac2 mRNA expression in Kiss1ARH neurons, the role of Kiss1ARH neurons during E2-driven anorexigenic states and their coordination of POMC and NPY/AgRP feeding circuits have been largely ignored. Presently, we show that E2 augmented the excitability of Kiss1ARH neurons by amplifying Cacna1g, Hcn1 and Hcn2 mRNA expression and T-type calcium and h-currents. E2 increased Slc17a6 mRNA expression and glutamatergic synaptic input to arcuate neurons, which excited POMC and inhibited NPY/AgRP neurons via metabotropic receptors. Deleting Slc17a6 in Kiss1 neurons eliminated glutamate release and led to conditioned place preference for sucrose in E2-treated KO female mice. Therefore, the E2-driven increase in Kiss1 neuronal excitability and glutamate neurotransmission may play a key role in governing the motivational drive for palatable food in females.


Pilot programme using social network interviews to screen high-risk adolescents and young adults.
To describe a pilot programme that consisted of identifying, contacting and providing HIV testing to members of social and sexual networks of HIV-infected youth patients as a part of routine clinical care at an adolescent HIV clinic in Baltimore, Maryland, USA. Forty-nine sexually active HIV-infected adolescent patients were interviewed about their social and sexual contacts at a routine HIV clinic visit. A trained community health worker located these referred social and sexual contacts, and encouraged them to make an appointment for HIV counselling, testing and referral (CTR) services. During a period of 18 months, 26 index youths provided locating information on 53 first-generation contacts and these 53 contacts provided information on 16 second-generation contacts. A total of 32 contacts received counselling services and 25 were tested for HIV infection, yielding three new HIV-positive individuals. As a part of standard care for regular visits of HIV-infected youth patients, interviewing about their social and sexual contacts could be a viable strategy in identifying high-risk youths for HIV infection and subsequent CTR services.


Resting-State Network Complexity and Magnitude Are Reduced in Prematurely Born Infants.
Premature birth is associated with high rates of motor and cognitive disability. Investigations have described resting-state functional magnetic resonance imaging (rs-fMRI) correlates of prematurity in older children, but comparable data in the neonatal period remain scarce. We studied 25 term-born control infants within the first week of life and 25 very preterm infants (born at gestational ages ranging from 23 to 29 weeks) without evident structural injury at term equivalent postmenstrual age. Conventional resting-state network (RSN) mapping revealed only modest differences between the term and prematurely born infants, in accordance with previous work. However, clear group differences were observed in quantitative analyses based on correlation and covariance matrices representing the functional MRI time series extracted from 31 regions of interest in 7 RSNs. In addition, the maximum likelihood dimensionality estimates of the group-averaged covariance matrices in the term and preterm infants were 5 and 3, respectively, indicating that prematurity leads to a reduction in the complexity of rs-fMRI covariance structure. These findings highlight the importance of quantitative analyses of rs-fMRI data and suggest a more sensitive method for delineating the effects of preterm birth in infants without evident structural injury.


Personalized Treatment Suggestions: The Validity and Applicability of the Risk-Prevention-Index Social in Low Back Pain Exercise Treatments.
Background: The back pain screening tool Risk-Prevention-Index Social (RPI-S) identifies the individual psychosocial risk for low back pain chronification and supports the allocation of patients at risk in additional multidisciplinary treatments. The study objectives were to evaluate (1) the prognostic validity of the RPI-S for a 6-month time frame and (2) the clinical benefit of the RPI-S. Methods: In a multicenter single-blind 3-armed randomized controlled trial, n = 660 persons (age 18-65 years) were randomly assigned to a twelve-week uni- or multidisciplinary exercise intervention or control group. Psychosocial risk was assessed by the RPI-S domain social environment (RPI-SSE) and the outcome pain by the Chronic Pain Grade Questionnaire (baseline M1, 12-weeks M4, 24-weeks M5). Prognostic validity was quantified by the root mean squared error (RMSE) within the control group. The clinical benefit of RPI-SSE was calculated by repeated measures ANOVA in intervention groups. Results: A subsample of n = 274 participants (mean = 38.0 years, SD 13.1) was analyzed, of which 30% were classified at risk in their psychosocial profile. The half-year prognostic validity was good (RMSE for disability of 9.04 at M4 and of 9.73 at M5; RMSE for pain intensity of 12.45 at M4 and of 14.49 at M5). People at risk showed significantly stronger reduction in pain disability and intensity at M4/M5, if participating in a multidisciplinary exercise treatment. Subjects at no risk showed a smaller reduction in pain disability in both interventions and no group differences for pain intensity. Regarding disability due to pain, around 41% of the sample would gain an unfitted treatment without the back pain screening. Conclusion: The RPI-SSE prognostic validity demonstrated good applicability and a clinical benefit confirmed by a clear advantage of an individualized treatment possibility.


Impact of educational interventions on adolescent attitudes and knowledge regarding vaccination: A pilot study.
Current immunisation levels in England currently fall slightly below the threshold recommended by the World Health Organization, and the three-year trend for vaccination uptake is downwards. Attitudes towards vaccination can affect future decisions on whether or not to vaccinate, and this can have significant public health implications. Interventions can impact future vaccination decisions, and these interventions can take several forms. Relatively little work has been reported on the use of vaccination interventions in young people, who form the next generation of individuals likely to make vaccination decisions. We investigated the impact of two different types of educational intervention on attitudes towards vaccination in young people in England. A cohort of young people (n = 63) was recruited via a local school. This group was divided into three sub-groups; one (n = 21) received a presentation-based intervention, one (n = 26) received an interactive simulation-based intervention, and the third (n = 16) received no intervention. Participants supplied information on (1) their attitudes towards vaccination, and (2) their information needs and views on personal choice concerning vaccination, at three time points: immediately before and after the intervention, and after six months. Neither intervention had a significant effect on participants' attitudes towards vaccination. However, the group receiving the presentation-based intervention saw a sustained uplift in confidence about information needs, which was not observed in the simulation-based intervention group. Our findings with young people are consistent with previous work on vaccination interventions aimed at adults, which have shown limited effectiveness, and which can actually reduce intention to vaccinate. Our findings on the most effective mode of delivery for the intervention should inform future discussion in the growing "games for health" domain, which proposes the use of interactive digital resources in healthcare education.


Summarising and validating test accuracy results across multiple studies for use in clinical practice.
Following a meta-analysis of test accuracy studies, the translation of summary results into clinical practice is potentially problematic. The sensitivity, specificity and positive (PPV) and negative (NPV) predictive values of a test may differ substantially from the average meta-analysis findings, because of heterogeneity. Clinicians thus need more guidance: given the meta-analysis, is a test likely to be useful in new populations, and if so, how should test results inform the probability of existing disease (for a diagnostic test) or future adverse outcome (for a prognostic test)? We propose ways to address this. Firstly, following a meta-analysis, we suggest deriving prediction intervals and probability statements about the potential accuracy of a test in a new population. Secondly, we suggest strategies on how clinicians should derive post-test probabilities (PPV and NPV) in a new population based on existing meta-analysis results and propose a cross-validation approach for examining and comparing their calibration performance. Application is made to two clinical examples. In the first example, the joint probability that both sensitivity and specificity will be >80% in a new population is just 0.19, because of a low sensitivity. However, the summary PPV of 0.97 is high and calibrates well in new populations, with a probability of 0.78 that the true PPV will be at least 0.95. In the second example, post-test probabilities calibrate better when tailored to the prevalence in the new population, with cross-validation revealing a probability of 0.97 that the observed NPV will be within 10% of the predicted NPV.


Optogenetic dissection of a behavioural module in the vertebrate spinal cord.
Locomotion relies on neural networks called central pattern generators (CPGs) that generate periodic motor commands for rhythmic movements. In vertebrates, the excitatory synaptic drive for inducing the spinal CPG can originate from either supraspinal glutamatergic inputs or from within the spinal cord. Here we identify a spinal input to the CPG that drives spontaneous locomotion using a combination of intersectional gene expression and optogenetics in zebrafish larvae. The photo-stimulation of one specific cell type was sufficient to induce a symmetrical tail beating sequence that mimics spontaneous slow forward swimming. This neuron is the Kolmer-Agduhr cell, which extends cilia into the central cerebrospinal-fluid-containing canal of the spinal cord and has an ipsilateral ascending axon that terminates in a series of consecutive segments. Genetically silencing Kolmer-Agduhr cells reduced the frequency of spontaneous free swimming, indicating that activity of Kolmer-Agduhr cells provides necessary tone for spontaneous forward swimming. Kolmer-Agduhr cells have been known for over 75 years, but their function has been mysterious. Our results reveal that during early development in zebrafish these cells provide a positive drive to the spinal CPG for spontaneous locomotion.


Single- Versus Dual-Task Functional Movement Paradigms: A Biomechanical Analysis.
Laboratory-based movement assessments are commonly performed without cognitive stimuli (ie, single-task) despite the simultaneous cognitive processing and movement (ie, dual task) demands required during sport. Cognitive loading may critically alter human movement and be an important consideration for truly assessing functional movement and understanding injury risk in the laboratory, but limited investigations exist. To comprehensively examine and compare kinematics and kinetics between single- and dual-task functional movement among healthy participants while controlling for sex. Cross-sectional study. Laboratory. Patients (or Other Participants): Forty-one healthy, physically active participants (49% female; 22.5 ± 2.1 y; 172.5 ± 11.9 cm; 71.0 ± 13.7 kg) enrolled in and completed the study. All participants completed the functional movement protocol under single- and dual-task (subtracting by 6s or 7s) conditions in a randomized order. Participants jumped forward from a 30-cm tall box and performed (1) maximum vertical jump landings and (2) dominant and (3) nondominant leg, single-leg 45° cuts after landing. The authors used mixed-model analysis of variances (α = .05) to compare peak hip, knee, and ankle joint angles (degrees) and moments (N·m/BW) in the sagittal and frontal planes, and peak vertical ground reaction force (N/BW) and vertical impulse (Ns/BW) between cognitive conditions and sex. Dual-task resulted in greater peak vertical ground reaction force compared with single-task during jump landing (mean difference = 0.06 N/BW; 95% confidence interval [CI], 0.01 to 0.12; P = .025) but less force during dominant leg cutting (mean difference = -0.08 N/BW; 95% CI, -0.14 to -0.02; P = .015). Less hip-flexion torque occurred during dual task than single task (mean difference = -0.09 N/BW; 95% CI, -0.17 to -0.02). No other outcomes were different between single and dual task (P ≥ .053). Slight, but potentially important, kinematic and kinetic differences were observed between single- and dual-task that may have implications for functional movement assessments and injury risk research. More research examining how various cognitive and movement tasks interact to alter functional movement among pathological populations is warranted before clinical implementation.


The Integrity of the TRP Domain Is Pivotal for Correct TRPV1 Channel Gating.
Transient receptor potential vanilloid subtype I (TRPV1) is a thermosensory ion channel that is also gated by chemical substances such as vanilloids. Adjacent to the channel gate, this polymodal thermoTRP channel displays a TRP domain, referred to as AD1, that plays a role in subunit association and channel gating. Previous studies have shown that swapping the AD1 in TRPV1 with the cognate from the TRPV2 channel (AD2) reduces protein expression and produces a nonfunctional chimeric channel (TRPV1-AD2). Here, we used a stepwise, sequential, cumulative site-directed mutagenesis approach, based on rebuilding the AD1 domain in the TRPV1-AD2 chimera, to unveil the minimum number of amino acids needed to restore protein expression and polymodal channel activity. Unexpectedly, we found that virtually full restitution of the AD1 sequence is required to reinstate channel expression and responses to capsaicin, temperature, and voltage. This strategy identified E692, R701, and T704 in the TRP domain as important for TRPV1 activity. Even conservative mutagenesis at these sites (E692D/R701K/T704S) impaired channel expression and abolished TRPV1 activity. However, the sole mutation of these positions in the TRPV1-AD2 chimera (D692E/K701R/S704T) was not sufficient to rescue channel gating, implying that other residues in the TRP domain are necessary to endow activity to TRPV1-AD2. A biophysical analysis of a functional chimera suggested that mutations in the TRP domain raised the energetics of channel gating by altering the coupling of stimuli sensing and pore opening. These findings indicate that inter- and/or intrasubunit interactions in the TRP domain are essential for correct TRPV1 gating.


Delayed reactive distractor suppression in aging populations.
Previous studies have tended to infer that reactive control is intact in aging populations because of evidence that proactive control is impaired and that older participants appear to favor reactive control strategies. However, most of these studies did not compare reactive control in young and older participants directly. In our study, a young (18 to 21 years old) and older (60+ years old) cohort engaged in a task that assesses reactive distractor suppression where participants had to discriminate between an upright and inverted T-shape in the presence of a salient or nonsalient distractor. In previous studies using this paradigm (DiQuattro & Geng, 2011) young participants reactively used the salient distractor as an anticue and performed better (faster reaction time [RT] and higher accuracy) when it was present. It was found that older participants were not able to reactively suppress the salient distractor with a 200-ms display but were able to do so with a 600-ms display. It was concluded that the initiation of reactive suppression is delayed for older participants, but that effective suppression is possible given enough time. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Charge Inversion and Calcium Gating in Mixtures of Ions in Nanopores.
Calcium ions play important roles in many physiological processes, yet their concentration is much lower than the concentrations of potassium and sodium ions. The selectivity of calcium channels is often probed in mixtures of calcium and a monovalent salt, e.g., KCl or NaCl, prepared such that the concentration of cations is kept constant with the mole fraction of calcium varying from 0 and 1. In biological channels, even sub-mM concentration of calcium can modulate the channels' transport characteristics; this effect is often explained via the existence of high affinity Ca2+ binding sites on the channel walls. Inspired by properties of biological calcium-selective channels, we prepared a set of nanopores with tunable opening diameters that exhibited a similar response to the presence of calcium ions as biochannels. Nanopores in 15 nm thick silicon nitride films were drilled using focused ion beam and e-beam in a transmission electron microscope and subsequently rendered negatively charged through silanization. We found that nanopores with diameters smaller than 20 nm were blocked by calcium ions such that the ion currents in mixtures of KCl and CaCl2 and in CaCl2 were even ten times smaller than the ion currents in KCl solution. The ion current blockage was explained by the effect of local charge inversion where accumulated calcium ions switch the effective surface charge from negative to positive. The modulation of surface charge with calcium leads to concentration and voltage dependent local charge density and ion current. The combined experimental and modeling results provide a link between calcium ion-induced changes in surface charge properties and resulting ionic transport.


Effect of in utero exposure of iodinated intravenous contrast on neonatal thyroid function.
Administration of radioactive iodine to a pregnant woman for thyroid ablation has been shown to affect neonatal thyroid levels. Thus, there is a theoretical risk in affecting a neonate's thyroid level upon birth with prior administration of intravenous contrast containing iodine to a pregnant woman during a computed tomographic scan. However, there is no literature to support this theoretical risk. This study attempted to determine if there is any effect on a newborn's thyroid levels after exposure to this type of contrast material in utero. The medical charts of 96 pregnant women during the years 2004 through 2009 on whom computed tomographic scans were performed were reviewed. A total of 29 charts were excluded secondary to missing neonatal records, missed abortions, and intrauterine fetal demise. The rest were subdivided into those who received intravenous (IV) ± oral contrast material and those who did not. The medical records of the newborns of these patients were also reviewed. The subject group consisted of 61 pregnant women who received IV ± oral contrast and their 64 neonates. The control group consisted of 6 pregnant patients who did not receive IV contrast and their 6 neonates. Of the 64 neonates in the subject group, only one neonate was found to have a low thyroxine level with a normal thyroid stimulating hormone level. This infant was severely premature, being born at the 25th week of gestation, and developed respiratory distress syndrome and sepsis, which were potential confounding factors. All of the 6 neonates in the control group had normal thyroid levels. This study concludes that there is no significant adverse clinical risk of thyroid function abnormalities to the fetus after IV iodinated contrast material to their mothers.


Gold nanoparticles (AuNPs) impair LPS-driven immune responses by promoting a tolerogenic-like dendritic cell phenotype with altered endosomal structures.
Dendritic cells (DCs) shape immune responses by influencing T-cell activation. Thus, they are considered both an interesting model for studying nano-immune interactions and a promising target for nano-based biomedical applications. However, the accentuated ability of nanoparticles (NPs) to interact with biomolecules may have an impact on DC function that poses an unexpected risk of unbalanced immune reactions. Here, we investigated the potential effects of gold nanoparticles (AuNPs) on DC function and the consequences for effector and memory T-cell responses in the presence of the microbial inflammatory stimulus lipopolysaccharide (LPS). Overall, we found that, in the absence of LPS, none of the tested NPs induced a DC response. However, whereas 4-, 8-, and 11 nm AuNPs did not modulate LPS-dependent immune responses, 26 nm AuNPs shifted the phenotype of LPS-activated DCs toward a tolerogenic state, characterized by downregulation of CD86, IL-12 and IL-27, upregulation of ILT3, and induction of class E compartments. Moreover, this DC phenotype was less proficient in promoting Th1 activation and central memory T-cell proliferation. Taken together, these findings support the perception that AuNPs are safe under homeostatic conditions; however, particular care should be taken in patients experiencing a current infection or disorders of the immune system.


Resistance to liver fluke infection in the natural sheep host is correlated with a type-1 cytokine response.
Indonesian thin-tail (ITT) sheep can resist infection with Fasciola gigantica but not F. hepatica and presents an ideal model to investigate the mechanisms of liver fluke resistance in a natural host. This study examines the local and systemic immune responses of sheep during Fasciola infection and demonstrates that different anatomical tissues display distinct cytokine profiles consistent with liver fluke migration. The study also reveals a significant difference in the cytokine and antibody profiles of ITT sheep infected with F. gigantica compared with F. hepatica, with a higher ratio of IL-4/IFN-γ mRNA expression and specific IgG1/IgG2 antibodies strongly correlating with pathology. Interestingly, the significant type-1 cytokine profile occurred in the lymph node closest to the site of infection at a time when the effective immune response against F. gigantica liver flukes is thought to occur. When the same F. gigantica infection in the resistant ITT sheep was compared with the susceptible Merino breed, the resistant type-1 phenotype against liver fluke infection was only observed in the ITT sheep. These studies provide the first evidence to suggest that the induction of an early type-1 immune response in this natural sheep host may be responsible for the ability to resist liver fluke infection.


Effects of feeding monensin to bred heifers fed in a drylot on nutrient and energy balance.
The objective of this study was to determine if feeding monensin would improve diet digestion, energy and nitrogen balance in bred heifers receiving a limit-fed corn stalk-based diet. Sixteen pregnant Meat Animal Research Center (MARC) III composite heifers were used in a 161-d completely randomized design. Heifers were randomly assigned to one of two treatments, no monensin (CON) or 150 mg/d monensin (MON), with eight heifers in each treatment group. Heifers were limit-fed a corn stalk-based diet at 100% of MEm requirements. Effects of monensin on energy and nitrogen balance were determined via total fecal and urine collections and open-circuit respiration calorimetry. Total fecal and urine collection occurred on d 14, 42, and 161 of monensin feeding, and calorimetry measurements were made on d 0, 3, 14, 28, 42, and 161 of monensin feeding. DMI was not different (P = 0.94) for CON and MON heifers and, by design, increased (P < 0.01) from d 14 to d 161 of the trial to account for increasing fetal growth requirements. No differences (P = 0.91) in GE intake were observed between CON and MON heifers, and DE and ME intakes did not differ (P > 0.58) with monensin inclusion. DM, OM, NDF, and ADF digestion did not differ (P > 0.52) between treatments. Fecal, methane, urinary, and heat energy losses were not different (P > 0.16) for MON and CON heifers. Methane production was not different between treatments when expressed as daily liters of methane (P = 0.40); however, MON heifers produced 7% less (P = 0.03) methane per day than CON heifers when expressed as liters of methane produced on a metabolic body weight (MBW) basis. Furthermore, monensin had no effect (P = 0.36) on overall retained energy (RE). Nitrogen intake and excretion was not different (P > 0.13) between treatment groups. Results of this experiment indicate that adding monensin to limit-fed, corn stalk-based diets may not have a large effect on the energy and nitrogen balance of confined heifers.


Systemic inflammation is negatively associated with early post discharge growth following acute illness among severely malnourished children - a pilot study.
Background: Rapid growth should occur among children with severe malnutrition (SM) with medical and nutritional management. Systemic inflammation (SI) is associated with death among children with SM and is negatively associated with linear growth. However, the relationship between SI and weight gain during therapeutic feeding following acute illness is unknown. We hypothesised that growth post-hospital discharge is associated with SI among children with SM. Methods: We conducted secondary analysis of data from HIV-uninfected children with SM (n=98) who survived and were not readmitted to hospital during one year of follow-up. We examined the relationship between changes in absolute deficits in weight and mid-upper-arm circumference (MUAC) from enrolment at stabilisation to 60 days and one year later, and untargeted plasma proteome, targeted cytokines/chemokines, leptin, and soluble CD14 using multivariate regularized linear regression. Results: The mean change in absolute deficit in weight and MUAC was -0.50kg (standard deviation; SD±0.69) and -1.20cm (SD±0.89), respectively, from enrolment to 60 days later. During the same period, mean weight and MUAC gain was 3.3g/kg/day (SD±2.4) and 0.22mm/day (SD±0.2), respectively. Enrolment interleukins; IL17-alpha and IL-2, and serum amyloid P were negatively associated with weight and MUAC gain during 60 days. Lipopolysaccharide binding protein and complement component 2 were negatively associated with weight gain only. Leptin was positively associated with weight gain. Soluble CD14, beta-2 microglobulin, and macrophage inflammatory protein 1 beta were negatively associated with MUAC gain only. Glutathione peroxidase 3 was positively associated with weight and MUAC gain during one year. Conclusions: Early post-hospital discharge weight and MUAC gain were rapid and comparable to children with uncomplicated SM treated in the community. Higher concentrations of SI markers were associated with less weight and MUAC gain, suggesting inflammation negatively impacts recovery from wasting. This finding warrants further research on reducing inflammation on growth among children with SM.


Evaluation of spectrometric parameters in spectral-domain optical coherence tomography.
The parameters of the spectrometer were analyzed in order to optimize the performance of spectral-domain optical coherence tomography (SD-OCT). Because probing depth is proportional to spectral resolution, it can be increased with the choice of a higher resolution spectrometer, which implies greater pixel numbers and lower imaging speed, or by sacrificing part of the spectrum, which compromises the axial resolution and side lobes. With a dynamic range of 8 bits or more, the signal-to-noise ratio remains constant for different noise levels. The results were verified experimentally with in vivo retinal SD-OCT imaging.


A series of 146 peripheral non-neural sheath nerve tumors: 30-year experience at Louisiana State University Health Sciences Center.
This is a retrospective review of 146 surgically treated benign and malignant peripheral non-neural sheath tumors (PNNSTs). Tumor classifications with patient numbers, locations of benign PNNSTs, and surgical techniques and adjunctive treatments are presented. The results of a literature review regarding tumor frequencies are presented. One hundred forty-six patients with 111 benign and 35 malignant PNNSTs were treated between 1969 and 1999 at the Louisiana State University Health Sciences Center (LSUHSC). The benign tumors included 33 ganglion cysts, 16 cases of localized hypertrophic neuropathy, 12 lipomas, 12 tumors of vascular origin, and 11 desmoid tumors. There were four each of lipofibrohamartomas, myositis ossificans, osteochondromas, and ganglioneuromas; two each of meningiomas, cystic hygromas, myoblastoma or granular cell tumors, triton tumors, and lymphangiomas; and one epidermoid cyst. The locations of benign PNNSTs were the following: 33 in the brachial plexus region, 39 in an upper extremity, one in the pelvic plexus, and 38 in a lower extremity. The malignant PNNSTs included 35 surgically treated carcinomas, 15 of which originated in the breast and nine in the lung. There were two melanomas metastatic to nerve and one tumor each that had metastasized from the bladder, rectum, skin, head and neck, and thyroid, and from a primary Ewing sarcoma. There was a single lymphoma that had metastasized to the radial nerve and one chordoma and one osteosarcoma, each of which had metastasized to the brachial plexus. There were more benign PNNSTs than malignant ones. Benign tumors were relatively equally distributed in the brachial plexus region and upper and lower extremities, with the exception of the pelvic plexus, which had only one tumor.


Indicators of acute bacterial meningitis in children at a rural Kenyan district hospital.
Acute bacterial meningitis remains an important cause of death and neurologic sequelae in African children. The clinical features of meningitis are often nonspecific and in this setting may overlap with those of malaria. Early diagnosis and appropriate antibiotic treatment are perhaps the most important steps in management, but published data suggest that fewer than half of the cases of childhood meningitis are identified at first assessment in hospitals in this region. The objective of this study was to identify clinical indicators of acute bacterial meningitis by examining components of the World Health Organization Integrated Management of Childhood Illness (IMCI) referral criteria for meningitis (lethargy, unconsciousness, inability to feed, stiff neck, or seizures) and other symptoms and signs. Kilifi District Hospital, serving approximately 200,000 people in a rural, malaria-endemic area of the Kenyan coast, was studied. A Kenya Medical Research Institute research center is located at the hospital. All pediatric admissions aged > or =60 days between June 2001 and July 2002 were eligible. A total of 91 (2.0%) of 4582 admissions had meningitis, including 77 (4.0%) of 1929 of those who met the IMCI referral criteria for meningitis at admission (sensitivity: 85%; specificity: 59%). Independent indicators of the presence of meningitis were a bulging fontanel, neck stiffness, cyanosis, impaired consciousness, partial seizures, and seizures outside the febrile convulsions age range. One or more of these indicators was present in 895 (19%) of admissions, 72 (8.0%) of whom had meningitis (sensitivity: 79%; specificity: 80%). Independent indicators of the absence of meningitis were the absence of a history of fever, a history of diarrhea, and a positive malaria slide. The area under the receiver operating characteristic curve for a set of simple screening rules based on the positive indicators identified was 0.88 (95% confidence interval: 0.85-0.92). The presence of > or =1 of a bulging fontanel, neck stiffness, cyanosis, impaired consciousness, partial seizures, and seizures outside the febrile convulsions age range is a clear indication for lumbar puncture and/or presumptive treatment. However, careful observation and reassessment may be the only practical way to identify one fifth of meningitis cases in this setting.


Presence of C1-inhibitor polymers in a subset of patients suffering from hereditary angioedema.
Hereditary angioedema (HAE) is a potentially life-threatening disease caused by mutations in the gene encoding the serine protease inhibitor (serpin) C1 inhibitor (C1-inh). The mutations cause decreased functional plasma levels of C1-inh, which triggers unpredictable recurrent edema attacks. Subjects suffering from HAE have been classified in type I patients with decreased functional and antigenic levels of C1-inh, and type II patients with decreased functional but normal antigenic C1-inh levels. However, a few reports have demonstrated that some mutations cause C1-inh polymerization in vitro, and it is speculated that C1-inh polymers may exist in patient plasma, challenging the current classification of HAE patients. To investigate the presence of C1-inh polymers in patient plasma samples, we developed an immunological method, where monoclonal antibodies produced against polymerized C1-inh were applied in native PAGE western blotting. Using this approach we analyzed genuine plasma samples from 31 Danish HAE families, and found that plasma samples from three genotypically distinct HAE type I families (classified upon C1-inh plasma concentrations) contained C1-inh polymers. Identical C1-inh polymerization phenotypes were observed in four affected family members from one of these families. Genotyping of the families revealed that the polymerogenic mutations of two families were located in proximity to the reactive center loop insertion site in C1-inh (p.Ile271Thr and p.Ser258_Pro260del),and one mutation affected helix C (p.Thr167Asn). In conclusion, we demonstrate that C1-inh polymers are present in the plasma of a subgroup of HAE type I patients.


Novel implications of Lingo-1 and its signaling partners in schizophrenia.
Myelination and neurite outgrowth both occur during brain development, and their disturbance has been previously been implicated in the pathophysiology of schizophrenia. Leucine-rich repeat and immunoglobulin domain-containing protein (Lingo-1) is a potent negative regulator of axonal myelination and neurite extension. As co-factors of Lingo-1 signaling (Nogo receptor (NgR), With No Lysine (K) (WNK1) and Myelin transcription factor 1 (Myt1)) have been implicated in the genetics of schizophrenia, we explored for the first time the role of Lingo-1 signaling pathways in this disorder. Lingo-1 protein, together with its co-receptor and co-factor proteins NgR, tumor necrosis factor (TNF) receptor orphan Y (TROY), p75, WNK1 and Myt1, have never been explored in the pathogenesis of schizophrenia. We examined protein levels of Lingo-1, NgR, TROY, p75, WNK1, Myt1 and myelin basic protein (MBP) (as a marker of myelination) within the post-mortem dorsolateral prefrontal cortex (DLPFC) (37 schizophrenia patients versus 37 matched controls) and hippocampus (Cornu Ammonis, CA1 and CA3) (20 schizophrenia patients versus 20 matched controls from the same cohort). Both of these brain regions are highly disrupted in the schizophrenia pathophysiology. There were significant increases in Lingo-1 (P<0.001) and Myt1 (P=0.023) and a reduction in NgR (P<0.001) in the DLPFC in schizophrenia subjects compared with controls. There were also increases in both TROY (P=0.001) and WNK1 (P=0.011) in the CA1 of schizophrenia subjects and, in contrast to the DLPFC, there was an increase in NgR (P=0.006) in the CA3 of schizophrenia subjects compared with controls. No significant difference was reported for MBP levels (P>0.05) between the schizophrenia and control groups in the three tested regions. This is the first time that a study has shown altered Lingo-1 signaling in the schizophrenia brain. Our novel findings may present a direct application for the use of a Lingo-1 antagonist to complement current and future schizophrenia therapies.


Cholinergic modulation of midbrain dopaminergic systems.
Dopamine neurons in the midbrain respond to behavioral events and environmental stimuli. Their different patterns of activation in turn modulate the activity of forebrain regions and modulate the expression of selective behavioral responses. However, their activity is closely dependent on the cholinergic systems in the brainstem. Ascending cholinergic projections from the pedunculopontine and laterodorsal tegmental nuclei target dopaminergic neurons in the substantia nigra compacta and ventral tegmental area following a topographical gradient. These projections, by means of the activation of acetylcholine receptors, influence the firing of dopamine neurons and therefore their responsiveness, ultimately affecting the release of dopamine in their forebrain targets. Brainstem cholinergic neurons are thus in a position to critically influence the activity of dopaminergic neurons in the midbrain, and thereby have a critical role in the expression of behavior.


Sexual experience enhances cognitive flexibility and dendritic spine density in the medial prefrontal cortex.
The medial prefrontal cortex is important for cognitive flexibility, a capability that is affected by environmental conditions and specific experiences. Aversive experience, such as chronic restraint stress, is known to impair performance on a task of cognitive flexibility, specifically attentional set-shifting, in rats. Concomitant with this performance decrement, chronic stress reduces the number of dendritic spines on pyramidal neurons in the medial prefrontal cortex. No previous studies have examined whether a rewarding experience, namely mating, affects cognitive flexibility and dendritic spines in the medial prefrontal cortex of male rats. To test this possibility, we exposed adult male rats to sexual receptive females once daily for one week, assessed attentional set-shifting performance, and then analyzed their brains for changes in dendritic spines. We found that sexual experience improved performance on extradimensional set-shifting, which is known to require the medial prefrontal cortex. Additionally, we observed increased dendritic spine density on apical and basal dendrites of pyramidal neurons in the medial prefrontal cortex, but not the orbitofrontal cortex, after sexual experience. We also found that sexual experience enhanced dendritic spine density on granule neurons of the dentate gyrus. The ventral hippocampus sends a direct projection to the medial prefrontal cortex, raising the possibility that experience-dependent changes in the hippocampus are necessary for alterations in medial prefrontal cortex structure and function. As a first attempt at investigating this, we inactivated the ventral hippocampus with the GABA agonist muscimol, after each daily bout of sexual experience to observe whether the beneficial effects on cognitive flexibility were abolished. Contrary to our hypothesis, blocking hippocampal activity after sexual experience had no impact on enhanced cognitive flexibility. Taken together, these findings indicate that sexual experience enhances medial prefrontal cortex dendritic spine density and cognitive flexibility but that these effects may not require continual input from the hippocampus.


Diagnostic utility of ERG immunostaining in dermatofibroma.
Dermatofibroma/fibrous histiocytoma (DF/FH) is a common cutaneous mesenchymal neoplasm exhibiting benign biological behaviour. However, the immunohistochemical utility of erythroblast transformation-specific-related gene (ERG) for diagnosing DF remains unknown. The authors reviewed the immunohistochemical status of ERG in different subtypes of DF and in its differential diagnoses. Overall, 97 cases of ordinary DF/FH, 6 cases of aneurysmal FH, 10 cases of cellular FH, 5 cases of angiomatoid FH, 2 cases of epithelioid FH, 64 cases of dermatofibrosarcoma protuberans (DFSP) and 52 cases of fibrous scar were retrieved. As the other histological types of cutaneous neoplasms, 6 cases of myxofibrosarcoma, 4 cases of undifferentiated pleomorphic sarcoma, 11 cases of atypical fibroxanthoma, 19 cases of malignant melanoma, 20 cases of nevocellular nevus, 20 cases of neurofibroma, 19 cases of schwannoma, 8 cases of angioleiomyoma and 1 case of pilar leiomyoma were included. Immunohistochemical positivity for ERG was demonstrated in 87 of 97 cases (89.6%) of ordinary DF/FH, 7 of 10 cases (70%) of cellular FH, 3 of 6 cases (50%) of aneurysmal FH, 1 of 5 cases (20%) of angiomatoid FH and 1 of 52 cases (0.1%) of fibrous scar. All cases of DFSP, epithelioid FH and other types of cutaneous neoplasms included in the current investigation were negative for ERG. The intensity of ERG immunohistochemical staining in spindle-shaped cells appeared weaker than that in endothelial cells. DF/FH was frequently positive for ERG immunostaining. ERG immunostaining may thus be useful to distinguish DF/FH from DFSP.


Suspected carotid artery stenosis: cost-effectiveness of CT angiography in work-up of patients with recent TIA or minor ischemic stroke.
To assess the effectiveness and cost-effectiveness of state-of-the-art noninvasive diagnostic imaging strategies in patients with a transient ischemic attack (TIA) or minor stroke who are suspected of having carotid artery stenosis (CAS). All prospectively evaluated patients provided informed consent, and the local ethics committee approved this study. Diagnostic performance, treatment, long-term events, quality of life, and costs resulting from strategies employing duplex ultrasonography (US), computed tomographic (CT) angiography, contrast material-enhanced magnetic resonance (MR) angiography, and combinations of these modalities were modeled in a decision tree and Markov model. Data sources included a prospective diagnostic cohort study, a meta-analysis, and a review of the literature. Outcomes were costs, quality-adjusted life-years (QALYs), incremental cost-effectiveness ratios, and net health benefits (QALY-equivalents), with a willingness-to-pay threshold of euro 50,000 per QALY and a societal perspective. The strategy with the highest net health benefit was considered the most cost effective. Extensive one-way, two-way, and probabilistic sensitivity analyses to explore the effect of varying parameter values were performed. The reference case analysis assumed that patients underwent surgery 2-4 weeks after the first symptoms, and the effect of earlier intervention was explored. The reference case analysis showed that duplex US combined with CT angiography and surgery for 70%-99% stenoses was the most cost-effective strategy, with a net health benefit of 13.587 and 15.542 QALY-equivalents in men and women, respectively. In men, the CT angiography strategy with a 70%-99% cutoff yielded slightly more QALYs, at an incremental cost of euro 71,419 per QALY, compared with duplex US combined with CT angiography. In patients with a high-risk profile, in patients with a high prior probability of disease, and when patients could be treated within 2 weeks after the first symptoms, the CT angiography strategy with surgery for 50%-99% stenoses was the most cost-effective strategy. In diagnosing CAS, duplex US should be the initial test, and, if its results are positive, CT angiography should be performed; patients with 70%-99% stenoses should then undergo carotid endarterectomy. In patients with a high-risk profile, a high probability of CAS, or who can undergo surgery without delay, immediate CT angiography and surgery for 50%-99% stenoses is indicated.


FDM 3D Printed Composites for Bone Tissue Engineering Based on Plasticized Poly(3-hydroxybutyrate)/poly(d,l-lactide) Blends.
Tissue engineering is a current trend in the regenerative medicine putting pressure on scientists to develop highly functional materials and methods for scaffolds' preparation. In this paper, the calibrated filaments for Fused Deposition Modeling (FDM) based on plasticized poly(3-hydroxybutyrate)/poly(d,l-lactide) 70/30 blend modified with tricalcium phosphate bioceramics were prepared. Two different plasticizers, Citroflex (n-Butyryl tri-n-hexyl citrate) and Syncroflex (oligomeric adipate ester), both used in the amount of 12 wt%, were compared. The printing parameters for these materials were optimized and the printability was evaluated by recently published warping test. The samples were studied with respect to their thermal and mechanical properties, followed by biological in vitro tests including proliferation, viability, and osteogenic differentiation of human mesenchymal stem cells. According to the results from differential scanning calorimetry and tensile measurements, the Citroflex-based plasticizer showed very good softening effect at the expense of worse printability and unsatisfactory performance during biological testing. On the other hand, the samples with Syncroflex demonstrated lower warping tendency compared to commercial polylactide filament with the warping coefficient one third lower. Moreover, the Syncroflex-based samples exhibited the non-cytotoxicity and promising biocompatibility.


Autophagy-independent senescence and genome instability driven by targeted telomere dysfunction.
Telomere dysfunction plays a complex role in tumorigenesis. While dysfunctional telomeres can block the proliferation of incipient cancer clones by inducing replicative senescence, fusion of dysfunctional telomeres can drive genome instability and oncogenic genomic rearrangements. Therefore, it is important to define the regulatory pathways that guide these opposing effects. Recent work has shown that the autophagy pathway regulates both senescence and genome instability in various contexts. Here, we apply models of acute telomere dysfunction to determine whether autophagy modulates the resulting genome instability and senescence responses. While telomere dysfunction rapidly induces autophagic flux in human fibroblast cell lines, inhibition of the autophagy pathway does not have a significant impact upon the transition to senescence, in contrast to what has previously been reported for oncogene-induced senescence. Our results suggest that this difference may be explained by disparities in the development of the senescence-associated secretory phenotype. We also show that chromosome fusions induced by telomere dysfunction are comparable in autophagy-proficient and autophagy-deficient cells. Altogether, our results highlight the complexity of the senescence-autophagy interface and indicate that autophagy induction is unlikely to play a significant role in telomere dysfunction-driven senescence and chromosome fusions.


Geographical and geological origin of natural graphite heavily influence the electrical and electrochemical properties of chemically modified graphenes.
Natural graphite is an important precursor for the production of chemically modified graphenes in bulk quantities for electrochemical applications. These natural graphites have varying fundamental properties due to the different geological processes and environments at their points of origin, which are expected to affect their chemical reactivity and hence the properties of the derived graphene materials. Four different natural graphites with known geographical and geological origins were exposed to a modified Hummers oxidation method and the resulting graphite oxides were studied. The graphite oxides were shown to have different extents of oxidation and types of oxygen groups, which directly influenced their electrochemical properties. These differences were propagated further in the subsequent chemical reduction of the graphite oxides, and the reduced graphene oxides exhibited significantly different reduction efficiencies and electrical conductivities. These findings show that the choice of natural graphite of known origin is important to synthesize chemically modified graphenes with a desired set of properties.


Insights on the Regeneration Potential of Müller Glia in the Mammalian Retina.
Müller glia, the major glial cell types in the retina, maintain retinal homeostasis and provide structural support to retinal photoreceptors. They also possess regenerative potential that might be used for retinal repair in response to injury or disease. In teleost fish (such as zebrafish), the Müller glia response to injury involves reprogramming events that result in a population of proliferative neural progenitors that can regenerate the injured retina. Recent studies have revealed several important mechanisms for the regenerative capacity of Müller glia in fish, which may shed more light on the mechanisms of Müller glia reprogramming and regeneration in mammals. Mammalian Müller glia can adopt stem cell characteristics, and in response to special conditions, be persuaded to proliferate and regenerate, although their native regeneration potential is limited. In this review, we consider the work to date revealing the regenerative potential of the mammalian Müller glia and discuss whether they are a potential source for cell regeneration therapy in humans.


Tailoring charge density and hydrogen bonding of imidazolium copolymers for efficient gene delivery.
Conventional free radical polymerization with subsequent postpolymerization modification afforded imidazolium copolymers with controlled charge density and side chain hydroxyl number. Novel imidazolium-containing copolymers where each permanent cation contained one or two adjacent hydroxyls allowed precise structure-transfection efficiency studies. The degree of polymerization was identical for all copolymers to eliminate the influence of molecular weight on transfection efficiency. DNA binding, cytotoxicity, and in vitro gene transfection in African green monkey COS-7 cells revealed structure-property-transfection relationships for the copolymers. DNA gel shift assays indicated that higher charge densities and hydroxyl concentrations increased DNA binding. As the charge density of the copolymers increased, toxicity of the copolymers also increased; however, as hydroxyl concentration increased, cytotoxicity remained constant. Changing both charge density and hydroxyl levels in a systematic fashion revealed a dramatic influence on transfection efficiency. Dynamic light scattering of the polyplexes, which were composed of copolymer concentrations required for the highest luciferase expression, showed an intermediate DNA-copolymer binding affinity. Our studies supported the conclusion that cationic copolymer binding affinity significantly impacts overall transfection efficiency of DNA delivery vehicles, and the incorporation of hydroxyl sites offers a less toxic and effective alternative to more conventional highly charged copolymers.


Monodisperse colloidal gallium nanoparticles: synthesis, low temperature crystallization, surface plasmon resonance and Li-ion storage.
We report a facile colloidal synthesis of gallium (Ga) nanoparticles with the mean size tunable in the range of 12-46 nm and with excellent size distribution as small as 7-8%. When stored under ambient conditions, Ga nanoparticles remain stable for months due to the formation of native and passivating Ga-oxide layer (2-3 nm). The mechanism of Ga nanoparticles formation is elucidated using nuclear magnetic resonance spectroscopy and with molecular dynamics simulations. Size-dependent crystallization and melting of Ga nanoparticles in the temperature range of 98-298 K are studied with X-ray powder diffraction, specific heat measurements, transmission electron microscopy, and X-ray absorption spectroscopy. The results point to delta (δ)-Ga polymorph as a single low-temperature phase, while phase transition is characterized by the large hysteresis and by the large undercooling of crystallization and melting points down to 140-145 and 240-250 K, respectively. We have observed size-tunable plasmon resonance in the ultraviolet and visible spectral regions. We also report stable operation of Ga nanoparticles as anode material for Li-ion batteries with storage capacities of 600 mAh g(-1), 50% higher than those achieved for bulk Ga under identical testing conditions.