Autologous chondrocyte implantation (ACI) for the treatment of large and complex cartilage lesions of the knee.
Complex cartilage lesions of the knee including large cartilage defects, kissing lesions, and osteoarthritis (OA) represent a common problem in orthopaedic surgery and a challenging task for the orthopaedic surgeon. As there is only limited data, we performed a prospective clinical study to investigate the benefit of autologous chondrocyte implantation (ACI) for this demanding patient population. Fifty-one patients displaying at least one of the criteria were included in the present retrospective study: (1.) defect size larger than 10 cm2; (2.) multiple lesions; (3.) kissing lesions, cartilage lesions Outerbridge grade III-IV, and/or (4.) mild/moderate osteoarthritis (OA). For outcome measurements, the International Cartilage Society's International Knee Documentation Committee's (IKDC) questionnaire, as well as the Cincinnati, Tegner, Lysholm and Noyes scores were used. Radiographic evaluation for OA was done using the Kellgren score. Patient's age was 36 years (13-61), defects size 7.25 (3-17.5) cm2, previous surgical procedures 1.94 (0-8), and follow-up 30 (12-63) months. Instruments for outcome measurement indicated significant improvement in activity, working ability, and sports. Mean ICRS grade improved from 3.8 preoperatively to grade 3 postoperatively, Tegner grade 1.4 enhanced to grade 3.39. The Cincinnati score enhanced from 25.65 to 66.33, the Lysholm score from 33.26 to 64.68, the Larson score from 43.59 to 79.31, and Noyes score from 12.5 to 46.67, representing an improvement from Cincinnati grade 3.65 to grade 2.1. Lysholm grade 4 improved to grade 3.33, and Larson grade 3.96 to 2.78 (Table 1), (p < 0.001). Patients with kissing cartilage lesions had similar results as patients with single cartilage lesions. Our results suggest that ACI provides mid-term results in patients with complex cartilage lesions of the knee. If long term results will confirm our findings, ACI may be a considered as a valuable tool for the treatment of complex cartilage lesions of the knee.


Cytotoxicity of Air Pollutant 9,10-Phenanthrenequinone: Role of Reactive Oxygen Species and Redox Signaling.
Atmospheric pollution has been a principal topic recently in the scientific and political community due to its role and impact on human and ecological health. 9,10-phenanthrenequinone (9,10-PQ) is a quinone molecule found in air pollution abundantly in the diesel exhaust particles (DEP). This compound has studied extensively and has been shown to develop cytotoxic effects both in vitro and in vivo. 9, 10-PQ has been proposed to play a critical role in the development of cytotoxicity via generation of reactive oxygen species (ROS) through redox cycling. This compound also reduces expression of glutathione (GSH), which is critical in Phase II detoxification reactions. Understanding the underlying cellular mechanisms involved in cytotoxicity can allow for the development of therapeutics designed to target specific molecules significantly involved in the 9,10-PQ-induced ROS toxicity. This review highlights the developments in the understanding of the cytotoxic effects of 9, 10-PQ with special emphasis on the possible mechanisms involved.


Double resonant absorption measurement of acetylene symmetric vibrational states probed with cavity ring down spectroscopy.
A novel mid-infrared/near-infrared double resonant absorption setup for studying infrared-inactive vibrational states is presented. A strong vibrational transition in the mid-infrared region is excited using an idler beam from a singly resonant continuous-wave optical parametric oscillator, to populate an intermediate vibrational state. High output power of the optical parametric oscillator and the strength of the mid-infrared transition result in efficient population transfer to the intermediate state, which allows measuring secondary transitions from this state with a high signal-to-noise ratio. A secondary, near-infrared transition from the intermediate state is probed using cavity ring-down spectroscopy, which provides high sensitivity in this wavelength region. Due to the narrow linewidths of the excitation sources, the rovibrational lines of the secondary transition are measured with sub-Doppler resolution. The setup is used to access a previously unreported symmetric vibrational state of acetylene, ν1+ν2+ν3+ν4 (1)+ν5 (-1) in the normal mode notation. Single-photon transitions to this state from the vibrational ground state are forbidden. Ten lines of the newly measured state are observed and fitted with the linear least-squares method to extract the band parameters. The vibrational term value was measured to be at 9775.0018(45) cm(-1), the rotational parameter B was 1.162 222(37) cm(-1), and the quartic centrifugal distortion parameter D was 3.998(62) × 10(-6) cm(-1), where the numbers in the parenthesis are one-standard errors in the least significant digits.


Modulation of sensitization processes in the management of pain and the importance of descending pathways: a role for tapentadol?
Objective: This paper presents and discusses recent evidence on the pathophysiological mechanisms of pain. The role of tapentadol - an opioid characterized by an innovative mechanism of action (i.e. µ-opioid receptor [MOR] agonism and inhibition of noradrenaline [NA] reuptake [NRI]) - in the modulation of pain, and the most recent pharmacological evidence on this molecule (e.g. the µ-load concept) are also presented and commented upon.Methods: Narrative review.Results: Solid evidence has highlighted the importance of central sensitization in the transition from acute to chronic pain. In particular, the noradrenergic system holds a major role in limiting central sensitization and the progression to chronic pain. Therefore, pharmacological modulation of the noradrenergic system appears to be a well-grounded strategy for the control of chronic pain. Tapentadol is characterized by a to-date-unique mechanism of action, since it acts both as a MOR agonist and as an inhibitor of NA reuptake. The synergistic interaction of these two mechanisms allows a strong analgesic effect by acting on both ascending and descending pathways. Of note, the reduced µ-load of tapentadol limits the risk of opioid-related adverse events, such as gastrointestinal disturbances. Moreover, the NA component becomes predominant, at least, in some types of pain, with consequent specific clinical efficacy in the treatment of neuropathic and chronic pain.Conclusions: According to these characteristics, tapentadol appears suitable in the treatment of severe uncontrolled chronic pain characterized by both a nociceptive and a neuropathic component, such as osteoarthritis or back pain.


Magnetic resonance imaging of the initial active stage of equine laminitis at 4.7 T.
Equine laminitis is a severely debilitating disease. There is a poor understanding of the underlying pathophysiology, and traditional imaging modalities have limited diagnostic capacity. High field strength magnetic resonance (MR) imaging allows direct visualization of the laminae, which other modalities do not. This would prove useful both in assessment of clinical patients and in further investigation into the pathophysiology of the disease. The objective of this study was to characterize the anatomic changes within the equine foot associated with the initial active stage of laminitis. Images obtained using a 4.7 T magnet were compared with digital radiographs using histologic diagnosis as the reference standard. Objective measurements and subjective evaluation for both modalities were evaluated for the ability to predict the histologic diagnosis in horses with clinical signs of laminitis as well as in clinically normal horses and horses that were in a population at risk for developing laminitis. Signal intensity and architectural changes within the corium and laminae were readily seen at 4.7 T, and there was a strong association with the histologic diagnosis of active laminitis. Measurements obtained with MR imaging were more sensitive and specific predictors of laminitis than those obtained radiographically. Subjective evaluation with MR imaging was more sensitive than with radiography and should become more specific with greater understanding of normal anatomy.


ALDH1A isozymes are markers of human melanoma stem cells and potential therapeutic targets.
Although the concept of cancer stem cells (CSCs) is well-accepted for many tumors, the existence of such cells in human melanoma has been the subject of debate. In this study, we demonstrate the existence of human melanoma cells that fulfill the criteria for CSCs (self-renewal and differentiation) by serially xenotransplanting cells into nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice. These cells possess high aldehyde dehydrogenase (ALDH) activity with ALDH1A1 and ALDH1A3 being the predominant ALDH isozymes. ALDH-positive melanoma cells are more tumorigenic than ALDH-negative cells in both NOD/SCID mice and NSG mice. Biological analyses of the ALDH-positive melanoma cells reveal the ALDH isozymes to be key molecules regulating the function of these cells. Silencing ALDH1A by siRNA or shRNA leads to cell cycle arrest, apoptosis, decreased cell viability in vitro, and reduced tumorigenesis in vivo. ALDH-positive melanoma cells are more resistant to chemotherapeutic agents and silencing ALDH1A by siRNA sensitizes melanoma cells to drug-induced cell death. Furthermore, we, for the first time, examined the molecular signatures of ALDH-positive CSCs from patient-derived tumor specimens. The signatures of melanoma CSCs include retinoic acid (RA)-driven target genes with RA response elements and genes associated with stem cell function. These findings implicate that ALDH isozymes are not only biomarkers of CSCs but also attractive therapeutic targets for human melanoma. Further investigation of these isozymes and genes will enhance our understanding of the molecular mechanisms governing CSCs and reveal new molecular targets for therapeutic intervention of cancer.


Power optimization of wireless media systems with space-time block codes.
We present analytical and numerical solutions to the problem of power control in wireless media systems with multiple antennas. We formulate a set of optimization problems aimed at minimizing total power consumption of wireless media systems subject to a given level of QoS and an available bit rate. Our formulation takes into consideration the power consumption related to source coding, channel coding, and transmission of multiple-transmit antennas. In our study, we consider Gauss-Markov and video source models, Rayleigh fading channels along with the Bernoulli/Gilbert-Elliott loss models, and space-time block codes.


Assessing the readiness to integrate tobacco control in medical curriculum: experiences from five medical colleges in Southern India.
Making tobacco cessation a normative part of all clinical practice is the only way to substantially reduce tobacco-related deaths and the burden of tobacco-related morbidity in the short term. This study was undertaken because information on receptivity to integrate tobacco control education in the medical curriculum is extremely limited in low- and middle-income countries. From five medical colleges (two government) in southern India, 713 (men 59%) faculty and 2585 (men 48%) students participated in our cross-sectional survey. Information on self-reported tobacco use and readiness to integrate tobacco control education in the medical curriculum was collected from both the faculty and students using a pretested structured questionnaire. Multiple logistic regression analysis was done to find the associated factors. Current smoking was reported by 9.0% (95% CI 6.6-12.1) of men faculty and 13.7% (CI 11.8-15.9) by men students. Faculty who were teaching tobacco-related topics [odds ratio (OR) 2.29; 95% CI 1.65-3.20] compared to those who were not, faculty in government colleges (OR 1.69; CI 1.22-2.35) compared to those in private colleges and medical pecialists (OR 1.79; CI 1.23-2.59) compared to surgical and non-clinical specialists were more likely to be ready to integrate tobacco control education in the medical curriculum. Non-smoking students (OR 2.58; CI 2.01-3.33) compared to smokers, and women students (OR 1.80; CI 1.50-2.17) compared to men were more likely to be ready to integrate a tobacco control education in the curriculum. Faculty and students are receptive to introduce tobacco control in the medical curriculum. Government faculty, medical specialists and faculty who already teach tobacco-related topics are likely to be early introducers of this new curriculum.


Forecasting the future of cardiovascular disease in the United States: a policy statement from the American Heart Association.
Cardiovascular disease (CVD) is the leading cause of death in the United States and is responsible for 17% of national health expenditures. As the population ages, these costs are expected to increase substantially. To prepare for future cardiovascular care needs, the American Heart Association developed methodology to project future costs of care for hypertension, coronary heart disease, heart failure, stroke, and all other CVD from 2010 to 2030. This methodology avoided double counting of costs for patients with multiple cardiovascular conditions. By 2030, 40.5% of the US population is projected to have some form of CVD. Between 2010 and 2030, real (2008$) total direct medical costs of CVD are projected to triple, from $273 billion to $818 billion. Real indirect costs (due to lost productivity) for all CVD are estimated to increase from $172 billion in 2010 to $276 billion in 2030, an increase of 61%. These findings indicate CVD prevalence and costs are projected to increase substantially. Effective prevention strategies are needed if we are to limit the growing burden of CVD.


The development and evaluation of a five-language multi-perspective standardised measure: clinical decision-making involvement and satisfaction (CDIS).
The aim of this study was to develop and evaluate a brief quantitative five-language measure of involvement and satisfaction in clinical decision-making (CDIS) - with versions for patients (CDIS-P) and staff (CDIS-S) - for use in mental health services. An English CDIS was developed by reviewing existing measures, focus groups, semistructured interviews and piloting. Translations into Danish, German, Hungarian and Italian followed the International Society for Pharmacoeconomics and Outcomes Research (ISPOR) Task Force principles of good practice for translation and cultural adaptation. Psychometricevaluation involved testing the measure in secondary mental health services in Aalborg, Debrecen, London, Naples, Ulm and Zurich. After appraising 14 measures, the Control Preference Scale and Satisfaction With Decision-making English-language scales were modified and evaluated in interviews (n = 9), focus groups (n = 22) and piloting (n = 16). Translations were validated through focus groups (n = 38) and piloting (n = 61). A total of 443 service users and 403 paired staff completed CDIS. The Satisfaction sub-scale had internal consistency of 0.89 (0.86-0.89 after item-level deletion) for staff and 0.90 (0.87-0.90) for service users, both continuous and categorical (utility) versions were associated with symptomatology and both staff-rated and service userrated therapeutic alliance (showing convergent validity), and not with social disability (showing divergent validity), and satisfaction predicted staff-rated (OR 2.43, 95%CI 1.54- 3.83 continuous, OR 5.77, 95%CI 1.90-17.53 utility) and service user-rated (OR 2.21, 95%CI 1.51-3.23 continuous, OR 3.13, 95%CI 1.10-8.94 utility) decision implementation two months later. The Involvement sub-scale had appropriate distribution and no floor or ceiling effects, was associated with stage of recovery, functioning and quality of life (staff only) (showing convergent validity), and not with symptomatology or social disability (showing divergent validity), and staff-rated passive involvement by the service user predicted implementation (OR 3.55, 95%CI 1.53-8.24). Relationships remained after adjusting for clustering by staff. CDIS demonstrates adequate internal consistency, no evidence of item redundancy, appropriate distribution, and face, content, convergent, divergent and predictive validity. It can be recommended for research and clinical use. CDIS-P and CDIS-S in all 3 five languages can be downloaded at http://www.cedar-net.eu/instruments. ISRCTN75841675.


Clinical study protocol for the ARCH project - computational modeling for improvement of outcome after vascular access creation.
Despite clinical guidelines and the possibility of diagnostic vascular imaging, creation and maintenance of a vascular access (VA) remains problematic: avoiding short- and long-term VA dysfunction is challenging. Although prognostic factors for VA dysfunction have been identified in previous studies, their potential interplay at a systemic level is disregarded. Consideration of multiple prognostic patient specific factors and their complex interaction using dedicated computational modeling tools might improve outcome after VA creation by enabling a better selection of VA configuration. These computational modeling tools are developed and validated in the ARCH project: a joint initiative of four medical centers and three industrial partners (FP7-ICT-224390). This paper reports the rationale behind computational modeling and presents the clinical study protocol designed for calibrating and validating these modeling tools. The clinical study is based on the pre-operative collection of structural and functional data at a vascular level, as well as a VA functional evaluation during the follow-up period. The strategy adopted to perform the study and for data collection is also described here.


Programmed death-1 (PD-1) expression in cervical intraepithelial neoplasia and its relationship with recurrence after conization.
Impaired local cellular immunity contributes to persistent human papillomavirus (HPV) infection and development of cervical intraepithelial neoplasia (CIN). Programmed death-1 (PD-1) and its ligands PD-ligand-1 (L1) and PD-L2 are negative regulators of T cell activity in various cancers, but few studies exist. The aim of this study was to determine the clinicopathologic and immunologic parameters (PD-1, PD-L1, and PD-L2) related to the persistence/recurrence of CIN after conization. Medical records of 652 patients diagnosed with CIN and underwent conization were reviewed. The associations between clinicopathologic parameters (e.g., age, parity, initial HPV load, etc.) and persistence/recurrence of CIN were analyzed. Expression of PD-1, PD-L1, and PD-L2 was assessed on 100 conization specimens by immunohistochemistry (IHC) in women matched for propensity-score (50 with persistence/recurrence and 50 without). Initial HPV load (>1,000 relative light unit) and positive margin were shown to be significantly associated with CIN persistence/recurrence (p=0.012 and p<0.001, respectively). Multivariate analysis showed that margin status was an independent predictor of persistence/recurrence (hazard ratio=8.86; 95% confidence interval=1.67-16.81; p<0.001). On IHC analysis, none of the patients expressed PD-L1. PD-1+ T cells were observed in 25 of 100 patients. Also, PD-1+ T cells were significantly correlated with increasing grade of CIN (p=0.031). In addition, patients with persistence/recurrence had increased expression of PD-1 compared with those without (36% vs. 14%, respectively; p=0.020). Although PD-L2 expression did not differ between 2 groups, it was significantly higher in patients with high-grade CIN compared to low-grade (34.7% vs. 12%, respectively; p=0.041). Positive surgical margin and expression of PD-1+ T cells were associated with CIN persistence/recurrence after conization.


Genotype announcement to Japanese smokers who attended a health checkup examination.
Genotype announcement may be one of the effective methods to induce smoking cessation, but the studies are limited throughout the world. Subjects were smokers who attended a health checkup examination provided by a local government in Hokkaido, Japan, 2003. Those who agreed to know their genotypes were informed of the genotypes of glutathione S-transferease (GST) M1 present/null, GSTT1 present/null, and NAD(P)H:quinone oxidoreductase 1 (NQO1) C609T (Pro187Ser). Out of 143 smokers (92 males and 51 females), 101 individuals participated in the present study. A postal questionnaire one year after the genotype announcement found that 8 persons (6 males and 2 females) of 41 respondents had quitted smoking. Two of 8 quitters stated that they had quitted smoking due to the announcement. There were none who regretted the genotype tests. Although the cessation rate, 7.9% (8/101) at least, was not marked, no harmful effects were observed among the respondents.


The role of MDM2 and MDM4 in breast cancer development and prevention.
The major cause of death from breast cancer is not the primary tumour, but relapsing, drug-resistant, metastatic disease. Identifying factors that contribute to aggressive cancer offers important leads for therapy. Inherent defence against carcinogens depends on the individual molecular make-up of each person. Important molecular determinants of these responses are under the control of the mouse double minute (MDM) family: comprised of the proteins MDM2 and MDM4. In normal, healthy adult cells, the MDM family functions to critically regulate measured, cellular responses to stress and subsequent recovery. Proper function of the MDM family is vital for normal breast development, but also for preserving genomic fidelity. The MDM family members are best characterized for their negative regulation of the major tumour suppressor p53 to modulate stress responses. Their impact on other cellular regulators is emerging. Inappropriately elevated protein levels of the MDM family are highly associated with an increased risk of cancer incidence. Exploration of the MDM family members as cancer therapeutic targets is relevant for designing tailored anti-cancer treatments, but successful approaches must strategically consider the impact on both the target cancer and adjacent healthy cells and tissues. This review focuses on recent findings pertaining to the role of the MDM family in normal and malignant breast cells.


Coding of Visual, Auditory, Rule, and Response Information in the Brain: 10 Years of Multivoxel Pattern Analysis.
How is the processing of task information organized in the brain? Many views of brain function emphasize modularity, with different regions specialized for processing different types of information. However, recent accounts also highlight flexibility, pointing especially to the highly consistent pattern of frontoparietal activation across many tasks. Although early insights from functional imaging were based on overall activation levels during different cognitive operations, in the last decade many researchers have used multivoxel pattern analyses to interrogate the representational content of activations, mapping out the brain regions that make particular stimulus, rule, or response distinctions. Here, we drew on 100 searchlight decoding analyses from 57 published papers to characterize the information coded in different brain networks. The outcome was highly structured. Visual, auditory, and motor networks predominantly (but not exclusively) coded visual, auditory, and motor information, respectively. By contrast, the frontoparietal multiple-demand network was characterized by domain generality, coding visual, auditory, motor, and rule information. The contribution of the default mode network and voxels elsewhere was minor. The data suggest a balanced picture of brain organization in which sensory and motor networks are relatively specialized for information in their own domain, whereas a specific frontoparietal network acts as a domain-general "core" with the capacity to code many different aspects of a task.


Biochemical properties and PCR performance of a family B DNA polymerase from hyperthermophilic Euryarchaeon Thermococcus peptonophilus.
The Thermococcus peptonophilus (Tpe) DNA polymerase gene was expressed under the control of the T7lac promoter on pET-22b(+) in Escherichia coli BL21-CodonPlus(DE3)-RIL in order to fully elucidate its biochemical properties and evaluate its feasibility in polymerase chain reaction (PCR) application. The expressed enzyme was then purified by heat treatment followed by two steps of column chromatography after which optimum pH and temperature of the enzyme were evaluated to be 7.0 and 75 degrees C, respectively. The optimal buffer for PCR with Tpe DNA polymerase consisted of 50 mM Tris-HCl (pH 8.0), 2 mM MgCl(2), 80 mM KCl, and 0.02% Triton X-100. Tpe DNA polymerase revealed a 3.6-fold higher fidelity (3.37 x 10(-6)) than Taq DNA polymerase (12.13 x 10(-6)) and performed significantly more efficiently in PCR amplification than both Taq and Pfu DNA polymerases. Ratios of 31:1 of Taq to Tpe DNA polymerases allowed PCR amplification of targets up to 15 kb in length with a 2.2-fold higher fidelity than Taq DNA polymerase. The results of the PCR experiments indicate that Tpe DNA polymerase may provide a higher fidelity DNA amplification in a shorter reaction time.


UVB irradiation regulates ERK1/2- and p53-dependent thrombomodulin expression in human keratinocytes.
Thrombomodulin (TM) is highly expressed in endothelial cells and acts as a natural anticoagulation factor to maintain circulation homeostasis. TM is an interesting molecule with many physiological functions, including anti-inflammation, anti-thrombosis, and carcinogenesis inhibition. TM can also be detected on the spinous layer of epidermal keratinocytes. However, the role of epidermal TM is still under investigation. In this study, we investigated keratinocyte TM expression and regulation in response to sub-cytotoxic ultraviolet B (UVB) irradiation. Oxidative stress was assessed with DCF and the results revealed that UVB irradiation significantly and dose-dependently augmented reactive oxygen species (ROS) production in HaCaT cells. In addition, low-dose UVB irradiation decreased TM mRNA and protein levels. Blocking ROS production and ERK activation prevented UVB-induced TM down-regulation. The nuclear p53 accumulation and TM promoter binding was observed within 3 h after UVB exposure. Small interfering RNA-mediated p53 knockdown disrupted the UVB-mediated TM protein down-regulation. Our study demonstrates that UVB irradiation results in ROS accumulation and ERK activation, which causes the nuclear p53 accumulation and TM promoter binding to inhibit TM expression. This study provides novel evidence demonstrating that p53 serves as a key regulator of keratinocyte TM expression.


Intraspecific variations of Dekkera/Brettanomyces bruxellensis genome studied by capillary electrophoresis separation of the intron splice site profiles.
In enology, "Brett" character refers to the wine spoilage caused by the yeast Dekkera/Brettanomyces bruxellensis and its production of volatile phenolic off-flavours. However, the spoilage potential of this yeast is strain-dependent. Therefore, a rapid and reliable recognition at the strain level is a key point to avoid serious economic losses. The present work provides an operative tool to assess the genetic intraspecific variation in this species through the use of introns as molecular targets. Firstly, the available partial D./B. bruxellensis genome sequence was investigated in order to build primers annealing to introns 5' splice site sequence (ISS). This analysis allowed the detection of a non-random vocabulary flanking the site and, exploiting this feature, the creation of specific probes for strain discrimination. Secondly, the separation of the intron splice site PCR fragments was obtained throughout the set up of a capillary electrophoresis protocol, giving a 94% repeatability threshold in our experimental conditions. The comparison of results obtained with ISS-PCR/CE versus the ones performed by mtDNA RFLP revealed that the former protocol is more discriminating and allowed a reliable identification at strain level. Actually sixty D./B. bruxellensis isolates were recognised as unique strains, showing a level of similarity below 79% and confirming the high genetic polymorphism existing within the species. Two main clusters were grouped at similarity levels of about 46% and 47%, respectively, showing a poor correlation with the geographic area of isolation. Moreover, from the evolutionary point of view, the proposed technique could determine the frequency of the genome rearrangements that can occur in D./B. bruxellesis populations.


The effect of being human and the basis of grammatical word order: Insights from novel communication systems and young sign languages.
This study identifies a central factor that gives rise to the different word orders found in the world's languages. In the last decade, a new window on this long-standing question has been provided by data from young sign languages and invented gesture systems. Previous work has assumed that word order in both invented gesture systems and young sign languages is driven by the need to encode the semantic/syntactic roles of the verb's arguments. Based on the responses of six groups of participants, three groups of hearing participants who invented a gestural system on the spot, and three groups of signers of relatively young sign languages, we identify a major factor in determining word order in the production of utterances in novel and young communication systems, not suggested by previous accounts, namely the salience of the arguments in terms of their human/animacy properties: human arguments are introduced before inanimate arguments ('human first'). This conclusion is based on the difference in word order patterns found between responses to depicted simple events that vary as to whether both subject and object are human or whether the subject is human and the object inanimate. We argue that these differential patterns can be accounted for uniformly by the 'human first' principle. Our analysis accounts for the prevalence of SOV order in clauses with an inanimate object in all groups (replicating results of previous separate studies of deaf signers and hearing gesturers) and the prevalence of both SOV and OSV in clauses with a human object elicited from the three groups of participants who have the least interference from another linguistic system (nonliterate deaf signers who have had little or no exposure to another language). It also provides an explanation for the basic status of SOV order suggested by other studies, as well as the scarcity of the OSV order in languages of the world, despite its appearance in novel communication systems. The broadest implication of this study is that the basic cognitive distinction between humans and inanimate entities is a crucial factor in setting the wheels of word ordering in motion.


Lycopene, sulforaphane, quercetin, and curcumin applied together show improved antiproliferative potential in colon cancer cells in vitro.
Lycopene, sulforaphane, quercetin, and curcumin, ingredients of daily diet, show significant anticancer and chemopreventive potential; however, no data are available showing thorough evaluation of jointly used phytochemicals on cancer cell proliferation. Here, we compare anticancer potential of mentioned substances applied separately or in combination (as MIX) by measuring mitochondrial activity (MTT test), DNA synthesis (BrdU test) and lactate dehydrogenase release (LDH test) in colon epithelial (CCD841 CoTr), and colon cancer (HT-29, LS174T) cells. Additive inhibitory effect of simultaneously used phytochemicals on cancer cells proliferation has been shown. In epithelial cells, tested combination effectively inhibited mitochondrial activity, but not DNA synthesis. LDH test revealed cytotoxicity of tested mixture against cancer cells without negative effect on normal cells. Furthermore, we demonstrated that MIX enhances antiproliferative effect of common cytostatics: 5-fluorouracil and cisplatin. Presented data suggest chemopreventive potential of the proposed combination of natural substances and their usefulness as adjuvant strategy during chemotherapy. PRACTICAL APPLICATIONS: Colorectal cancer is one of the most common causes of cancer death worldwide. Since its development and progression is strongly correlated with dietary habits, healthy diet as well as supplementation with proved anticancer agents seems to be reasonable strategy of colon cancer prevention and treatment. In the present study, we have focused on four natural compounds abundantly found in daily diet i.e., lycopene, sulforaphane, quercetin, and curcumin, with well established anticancer potential. Their individual and collective impact both on normal colon epithelium cells and colon cancer cells viability, growth, and proliferation was examined. Furthermore, activity of the substances combined as MIX to influence antiproliferative potential of commonly used in colon cancer treatment cytostatics, 5-fluorouracil, and cisplatin was verified. Proposed in the study combination of phytochemicals with experimentally proven antiproliferative activity may propose an effective strategy for prevention and treatment of colon cancer.


A New Figure of Merit for Organic Solar Cells with Transport-limited Photocurrents.
Compared to their inorganic counterparts, organic semiconductors suffer from relatively low charge carrier mobilities. Therefore, expressions derived for inorganic solar cells to correlate characteristic performance parameters to material properties are prone to fail when applied to organic devices. This is especially true for the classical Shockley-equation commonly used to describe current-voltage (JV)-curves, as it assumes a high electrical conductivity of the charge transporting material. Here, an analytical expression for the JV-curves of organic solar cells is derived based on a previously published analytical model. This expression, bearing a similar functional dependence as the Shockley-equation, delivers a new figure of merit α to express the balance between free charge recombination and extraction in low mobility photoactive materials. This figure of merit is shown to determine critical device parameters such as the apparent series resistance and the fill factor.


Sorting of mannose 6-phosphate receptors and lysosomal membrane proteins in endocytic vesicles.
The intracellular distributions of the cation-independent mannose 6-phosphate receptor (MPR) and a 120-kD lysosomal membrane glycoprotein (lgp120) were studied in rat hepatoma cells. Using quantitative immunogold cytochemistry we found 10% of the cell's MPR located at the cell surface. In contrast, lgp120 was not detectable at the plasma membrane. Intracellularly, MPR mainly occurred in the trans-Golgi reticulum (TGR) and endosomes. lgp120, on the other hand, was confined to endosomes and lysosomes. MPR was present in both endosomal tubules and vacuoles, whereas lgp120 was confined to the endosomal vacuoles. In cells incubated for 5-60 min with the endocytic tracer cationized ferritin, four categories of endocytic vacuoles could be discerned, i.e., vacuoles designated MPR+/lgp120-, MPR+/lgp120+, MPR-/lgp120+, and vacuoles nonimmunolabeled for MPR and lgp120. Tracer first reached MPR+/lgp120-, then MPR+/lgp120+, and finally MPR-/lgp120+ vacuoles, which are assumed to represent lysosomes. To study the kinetics of appearance of endocytic tracers in MPR-and/or lgp120-containing pools in greater detail, cells were allowed to endocytose horse-radish peroxidase (HRP) for 5-90 min. The reduction in detectability of MPR and lgp120 antigenicity on Western blots, due to treatment of cell homogenates with 3'3-diaminobenzidine, was followed in time. We found that HRP reached the entire accessible pool of MPR almost immediately after internalization of the tracer, while prolonged periods of time were required for HRP to maximally access lgp120. The combined data suggest that MPR+/lgp120+ vacuoles are endocytic vacuoles, intermediate between MPR+/lgp120-endosomes and MPR-/lgp120+ lysosomes, and represent the site where MPR is sorted from lgp120 destined for lysosomes. We propose that MPR is sorted from lgp120 by selective lateral distribution of the receptor into the tubules of this compartment, resulting in the retention of lgp120 in the vacuoles and the net transport of lgp120 to lysosomes.


Engrailed-2 (EN2): a tumor specific urinary biomarker for the early diagnosis of prostate cancer.
Prostate cancer (PC) is the second most common cause of cancer related death in men. A number of key limitations with prostate specific antigen (PSA), currently the standard detection test, has justified evaluation of new biomarkers. We have assessed the diagnostic potential of Engrailed-2 (EN2) protein, a homeodomain-containing transcription factor expressed in PC cell lines and secreted into the urine by PC in men. EN2 expression in PC cell lines and prostate cancer tissue was determined by semi-quantative RT-PCR and immunohistochemistry. First pass urine [without prior digital rectal examination (DRE)] was collected from men presenting with urinary symptoms (referred to exclude/confirm the presence of prostate cancer) and from controls. EN2 protein was measured by ELISA in urine from men with PC (n = 82) and controls (n = 102). EN2 was expressed and secreted by PC cell lines and PC tissue but not by normal prostate tissue or stroma. The presence of EN2 in urine was highly predictive of PC, with a sensitivity of 66% and a specificity of 88.2%, without requirement for DRE. There was no correlation with PSA levels. These results were confirmed independently by a second academic center. Urinary EN2 is a highly specific and sensitive candidate biomarker of prostate cancer. A larger multicenter study to further evaluate the diagnostic potential of EN2 is justified.


Dynamic changes in short- and long-term bacterial composition following fecal microbiota transplantation for recurrent Clostridium difficile infection.
Fecal microbiota transplantation (FMT) is an effective treatment for recurrent Clostridium difficile infection (CDI) that often fails standard antibiotic therapy. Despite its widespread recent use, however, little is known about the stability of the fecal microbiota following FMT. Here we report on short- and long-term changes and provide kinetic visualization of fecal microbiota composition in patients with multiply recurrent CDI that were refractory to antibiotic therapy and treated using FMT. Fecal samples were collected from four patients before and up to 151 days after FMT, with daily collections until 28 days and weekly collections until 84 days post-FMT. The composition of fecal bacteria was characterized using high throughput 16S rRNA gene sequence analysis, compared to microbiota across body sites in the Human Microbiome Project (HMP) database, and visualized in a movie-like, kinetic format. FMT resulted in rapid normalization of bacterial fecal sample composition from a markedly dysbiotic state to one representative of normal fecal microbiota. While the microbiome appeared most similar to the donor implant material 1 day post-FMT, the composition diverged variably at later time points. The donor microbiota composition also varied over time. However, both post-FMT and donor samples remained within the larger cloud of fecal microbiota characterized as healthy by the HMP. Dynamic behavior is an intrinsic property of normal fecal microbiota and should be accounted for in comparing microbial communities among normal individuals and those with disease states. This also suggests that more frequent sample analyses are needed in order to properly assess success of FMT procedures.


Hamiltonian approach for optimization of phase-sensitive double-pumped parametric amplifiers.
In this work we applied a Hamiltonian formalism to simplify the equations of non-degenerate nonlinear four-wave mixing to the one-degree-of-freedom Hamiltonian equations with a three-parameter Hamiltonian. Thereby, a problem of signal amplification in a phase-sensitive double-pumped parametric fiber amplifier with pump depletion was reduced to a geometrical study of the phase portraits of the one-degree-of-freedom Hamiltonian system. For a symmetric case of equal pump powers and equal signal and idler powers at the fiber input, it has been shown that the theoretical maximum gain occurs on the extremal trajectories. However, to reduce the nonlinear interaction of waves, we proposed to choose the separatrix as the optimal trajectory on the phase plane. Analytical expressions were found for the maximum amplification, as well as the length of optical fiber and the relative phase of interacting waves allowing this amplification. Using the proposed approach, we optimized of the phase-sensitive parametric amplifier. As a result, the optimal parameters of the phase-sensitive amplifier were found and the maximum possible signal amplification was realized in a broad range of signal wavelengths.


APOBEC-1 deletion enhances cisplatin-induced acute kidney injury.
Cisplatin (CP) induces acute kidney injury (AKI) whereby proximal tubules undergo regulated necrosis. Repair is almost complete after a single dose. We now demonstrate a role for Apolipoprotein B mRNA editing enzyme, catalytic polypeptide 1 (Apobec-1) that is prominently expressed at the interface between acute and chronic kidney injury (CKD), in the recovery from AKI. Apobec-1 knockout (KO) mice exhibited greater mortality than in wild type (WT) and more severe AKI in both CP- and unilateral ischemia reperfusion (IR) with nephrectomy. Specifically, plasma creatinine (pCr) 2.6 ± 0.70 mg/dL for KO, n = 10 and 0.16 ± 0.02 for WT, n = 6, p < 0.0001 in CP model and 1.34 ± 0.22 mg/dL vs 0.75 ± 0.06, n = 5, p < 0.05 in IR model. The kidneys of Apobec-1 KO mice showed increased necrosis, increased expression of KIM-1, NGAL, RIPK1, ASCL4 and increased lipid accumulation compared to WT kidneys (p < 0.01). Neutrophils and activated T cells were both increased, while macrophages were reduced in kidneys of Apobec-1 KO animals. Overexpression of Apobec-1 in mouse proximal tubule cells protected against CP-induced cytotoxicity. These findings suggest that Apobec-1 mediates critical pro-survival responses to renal injury and increasing Apobec-1 expression could be an effective strategy to mitigate AKI.


New developments in the InterPro database.
InterPro is an integrated resource for protein families, domains and functional sites, which integrates the following protein signature databases: PROSITE, PRINTS, ProDom, Pfam, SMART, TIGRFAMs, PIRSF, SUPERFAMILY, Gene3D and PANTHER. The latter two new member databases have been integrated since the last publication in this journal. There have been several new developments in InterPro, including an additional reading field, new database links, extensions to the web interface and additional match XML files. InterPro has always provided matches to UniProtKB proteins on the website and in the match XML file on the FTP site. Additional matches to proteins in UniParc (UniProt archive) are now available for download in the new match XML files only. The latest InterPro release (13.0) contains more than 13 000 entries, covering over 78% of all proteins in UniProtKB. The database is available for text- and sequence-based searches via a webserver (http://www.ebi.ac.uk/interpro), and for download by anonymous FTP (ftp://ftp.ebi.ac.uk/pub/databases/interpro). The InterProScan search tool is now also available via a web service at http://www.ebi.ac.uk/Tools/webservices/WSInterProScan.html.


Successful management of recalcitrant infection related to total hip replacement using pedicled rectus femoris or vastus lateralis muscle flaps.
Deep prosthetic joint infection remains an uncommon but serious complication of total hip replacement. We reviewed 24 patients with recalcitrant hip wounds following infected total hip replacement treated with either pedicled rectus femoris or vastus lateralis muscle flaps between 1998 and 2009. The mean age of the patients was 67.4 years (42 to 86) with ten men and 14 women. There had been a mean of four (1 to 8) previous attempts to close the wound. A total of 20 rectus femoris and five vastus lateralis flaps were used, with one of each type of flap failing and requiring further reconstruction. All patients had positive microbiology. At a mean follow-up of 47 months (9 to 128), 22 patients had a healed wound and two had a persistent sinus. The prosthesis had been retained in five patients. In the remainder it had been removed, and subsequently re-implanted in nine patients. Six patients continued to take antibiotics at final follow-up. This series demonstrates the effectiveness of pedicled muscle flaps in healing these infected wounds. The high number of previous debridements suggests that these flaps could have been used earlier.


A metagenomic assessment of the bacteria associated with Lucilia sericata and Lucilia cuprina (Diptera: Calliphoridae).
Lucilia Robineau-Desvoidy (Diptera: Calliphoridae) is a blow fly genus of forensic, medical, veterinary, and agricultural importance. This genus is also famous because of its beneficial uses in maggot debridement therapy (MDT). Although the genus is of considerable economic importance, our knowledge about microbes associated with these flies and how these bacteria are horizontally and trans-generationally transmitted is limited. In this study, we characterized bacteria associated with different life stages of Lucilia sericata (Meigen) and Lucilia cuprina (Wiedemann) and in the salivary gland of L. sericata by using 16S rDNA 454 pyrosequencing. Bacteria associated with the salivary gland of L. sericata were also characterized using light and transmission electron microscopy (TEM). Results from this study suggest that the majority of bacteria associated with these flies belong to phyla Proteobacteria, Firmicutes, and Bacteroidetes, and most bacteria are maintained intragenerationally, with a considerable degree of turnover from generation to generation. In both species, second-generation eggs exhibited the highest bacterial phylum diversity (20 % genetic distance) than other life stages. The Lucilia sister species shared the majority of their classified genera. Of the shared bacterial genera, Providencia, Ignatzschineria, Lactobacillus, Lactococcus, Vagococcus, Morganella, and Myroides were present at relatively high abundances. Lactobacillus, Proteus, Diaphorobacter, and Morganella were the dominant bacterial genera associated with a survey of the salivary gland of L. sericata. TEM analysis showed a sparse distribution of both Gram-positive and Gram-negative bacteria in the salivary gland of L. sericata. There was more evidence for horizontal transmission of bacteria than there was for trans-generational inheritance. Several pathogenic genera were either amplified or reduced by the larval feeding on decomposing liver as a resource. Overall, this study provides information on bacterial communities associated with different life stages of Lucilia and their horizontal and trans-generational transmission, which may help in the development of better vector-borne disease management and MDT methods.


The ESC-EORP Chronic Ischaemic Cardiovascular Disease Long Term (CICD LT) registry.
The European Society of Cardiology (ESC) EURObservational Research Programme (EORP) Chronic Ischaemic Cardiovascular Disease Long Term (CICD LT) registry aims to study the clinical profile, treatment modalities, and outcomes of patients diagnosed with CICD in a contemporary environment in order to assess whether these patients at high cardiovascular (CV) risk are treated according to ESC guidelines on prevention or on stable coronary disease and to determine mid- and long-term outcomes and their determinants in this population. Nine thousand one hundred and seventy-four patients over 18 years with documented CICD defined by a history acute coronary syndrome with/without ST elevation, previous coronary revascularization, or stable coronary artery disease were enrolled between 1 May 2015 and 31 July 2018. Individual patient data on clinical profile, biology, and treatment modalities were collected across 154 centres from 20 ESC countries. Two years of follow-up is scheduled in order to determine the following clinical outcomes: all-cause and CV death, all-cause and CV hospitalizations, changes in medications, and quality of life using the EuroQol5D-5L score. The CICD LT is an international registry of care and outcomes of patients hospitalized with CICD which will provide insights into the contemporary profile and management of patients with this common disease.


Transcriptomic Profiling of Dromedary Camels Immunised with a MERS Vaccine Candidate.
Middle East Respiratory Syndrome coronavirus (MERS-CoV) infects dromedary camels and zoonotically infects humans, causing a respiratory disease with severe pneumonia and death. With no approved antiviral or vaccine interventions for MERS, vaccines are being developed for camels to prevent virus transmission into humans. We have previously developed a chimpanzee adenoviral vector-based vaccine for MERS-CoV (ChAdOx1 MERS) and reported its strong humoral immunogenicity in dromedary camels. Here, we looked back at total RNA isolated from whole blood of three immunised dromedaries pre and post-vaccination during the first day; and performed RNA sequencing and bioinformatic analysis in order to shed light on the molecular immune responses following a ChAdOx1 MERS vaccination. Our finding shows that a number of transcripts were differentially regulated as an effect of the vaccination, including genes that are involved in innate and adaptive immunity, such as type I and II interferon responses. The camel Bcl-3 and Bcl-6 transcripts were significantly upregulated, indicating a strong activation of Tfh cell, B cell, and NF-κB pathways. In conclusion, this study gives an overall view of the first changes in the immune transcriptome of dromedaries after vaccination; it supports the potency of ChAdOx1 MERS as a potential camel vaccine to block transmission and prevent new human cases and outbreaks.


Motivations for and barriers to choosing an interventional cardiology career path: results from the EAPCI Women Committee worldwide survey.
Very few women become interventional cardiologists, although a substantial proportion of cardiologists and the majority of medical students are women. In accordance with the EAPCI Women Committee mission of attaining gender equality at the professional level, a worldwide survey was recently conducted aiming to understand better the motivations and the barriers for women in selecting interventional cardiology (IC) as a career path. A total of 1,787 individuals (60.7% women) responded to the survey. Women compared to men were less frequently married (women vs. men, 57.0% vs. 79.8%, p<0.001) and more frequently childless (46.6% vs. 20.5%, p<0.002). The most prevalent reason for choosing IC was passion (83.3% vs. 76.1%, p=0.12), while those for not choosing were, sequentially, lack of opportunity (29.0% vs. 45.7%), radiation concerns (19.9% vs. 11.6%) and preference (16.2% vs. 29.5%), p<0.001. According to 652 men replying to why, in their opinion, women do not choose IC, on-calls and long working hours were the most frequent reasons (35.3%). Several barriers preclude women from choosing IC, including lack of opportunity, concerns regarding radiation exposure and the prejudices of their male colleagues. This highlights the need to develop new strategies for future training, education, and support of women in order to choose IC.


Isokinetic profile of wrist and forearm strength in elite female junior tennis players.
In tennis, injuries to the elbow and wrist occur secondary to the repetitive nature of play and are seen at increasingly young ages. Isokinetic testing can be used to determine muscular strength levels, but dominant/non-dominant and agonist/antagonist relations are needed for meaningful interpretation of the results. To determine whether there are laterality differences in wrist extension/flexion (E/F) and forearm supination/pronation (S/P) strength in elite female tennis players. 32 elite female tennis players (age 12 to 16 years) with no history of upper extremity injury underwent bilateral isokinetic testing using a Cybex 6000 dynamometer. Peak torque and single repetition work values for wrist E/F and forearm S/P were measured at speeds of 90 degrees/s and 210 degrees/s, with random determination of the starting extremity. Repeated measures analysis of variance was used to determine differences between extremities for peak torque and single repetition work values. Significantly greater (p<0.01) dominant arm wrist E/F and forearm pronation strength was measured at both testing speeds. Significantly less (p<0.01) dominant side forearm supination strength was measured at both testing speeds. Greater dominant arm wrist E/F and forearm pronation strength is common and normal in young elite level female tennis players. These strength relations indicate sport specific muscular adaptations in the dominant tennis playing extremity. The results of this study can guide clinicians who work with young athletes from this population. Restoring greater dominant side wrist and forearm strength is indicated after an injury to the dominant upper extremity in such players.


Remote automated multi-generational growth and observation of an animal in low Earth orbit.
The ultimate survival of humanity is dependent upon colonization of other planetary bodies. Key challenges to such habitation are (patho)physiologic changes induced by known, and unknown, factors associated with long-duration and distance space exploration. However, we currently lack biological models for detecting and studying these changes. Here, we use a remote automated culture system to successfully grow an animal in low Earth orbit for six months. Our observations, over 12 generations, demonstrate that the multi-cellular soil worm Caenorhabditis elegans develops from egg to adulthood and produces progeny with identical timings in space as on the Earth. Additionally, these animals display normal rates of movement when fully fed, comparable declines in movement when starved, and appropriate growth arrest upon starvation and recovery upon re-feeding. These observations establish C. elegans as a biological model that can be used to detect changes in animal growth, development, reproduction and behaviour in response to environmental conditions during long-duration spaceflight. This experimental system is ready to be incorporated on future, unmanned interplanetary missions and could be used to study cost-effectively the effects of such missions on these biological processes and the efficacy of new life support systems and radiation shielding technologies.


Expansion of nurse anesthesia educational programs: where are the barriers?
CRNAs have been a major contributor to direct patient anesthesia care for more than 100 years. All indicators suggest a greater need for nurse anesthetist services in the future. Our ability to meet this increased demand will depend on wise decisions in accreditation and professional agendas and support of all CRNAs in this critical issue. As stated by AANA Past President Linda Williams, CRNA, JD, students represent only 10% of our membership but they represent 100% of our future. Let's unite all forces to secure a future in which CRNAs will be indispensable healthcare providers throughout the 21st century.


Soluble TNF receptor production by activated T lymphocytes: differential effects of acute and chronic exposure to TNF.
Soluble tumour necrosis factor receptors (sTNF-R) are up-regulated at sites of chronic inflammation such as rheumatoid synovial joints. The p75 sTNF-R is the more abundant, suggesting an important role for this TNF inhibitor in regulating TNF bioactivity in vivo. As the precise cellular source of these soluble receptors is not known, we investigated the production and regulation of sTNF-R by T lymphocytes, an abundant cell type in inflammatory infiltrates, which upon activation express high levels of p75 surface receptors. Using panels of T-cell lines and clones expressing high levels of p75 TNF-R, we found that p75 sTNF-R production upon stimulation is a feature common to all subsets of T cells, including cells of the CD4-CD8- double negative phenotype expressing either alpha beta or gamma delta T-cell receptors (TCR). In contrast, levels of p55 sTNF-R were only detected when T cells were stimulated at higher densities and by potent mitogens such as phorbol 12-myristate 13-acetate (PMA). Detailed kinetic analyses revealed that the production of p75 sTNF-R was biphasic, the first phase was activation dependent, occurring in the absence of detectable TNF, while the second phase of p75 sTNF-R production was regulated by cytokines such as TNF. Unlike short-term exposure to TNF which enhances sTNF-R production in vitro and in vivo, prolonged exposure of T lymphocytes to TNF suppressed p75 sTNF-R (but not p55 sTNF-R) production in a dose- and time-dependent fashion. These results suggest that in patients with chronic inflammatory disease, which are exposed to augmented levels of bioactive TNF for prolonged periods, the production of p75 sTNF-R may be impaired.


Medications that reduce emergency hospital admissions: an overview of systematic reviews and prioritisation of treatments.
Rates of emergency hospitalisations are increasing in many countries, leading to disruption in the quality of care and increases in cost. Therefore, identifying strategies to reduce emergency admission rates is a key priority. There have been large-scale evidence reviews to address this issue; however, there have been no reviews of medication therapies, which have the potential to reduce the use of emergency health-care services. The objectives of this study were to review systematically the evidence to identify medications that affect emergency hospital admissions and prioritise therapies for quality measurement and improvement. This was a systematic review of systematic reviews. We searched MEDLINE, PubMed, the Cochrane Database of Systematic Reviews & Database of Abstracts of Reviews of Effects, Google Scholar and the websites of ten major funding agencies and health charities, using broad search criteria. We included systematic reviews of randomised controlled trials that examined the effect of any medication on emergency hospital admissions among adults. We assessed the quality of reviews using AMSTAR. To prioritise therapies, we assessed the quality of trial evidence underpinning meta-analysed effect estimates and cross-referenced the evidence with clinical guidelines. We identified 140 systematic reviews, which included 1968 unique randomised controlled trials and 925,364 patients. Reviews contained 100 medications tested in 47 populations. We identified high-to moderate-quality evidence for 28 medications that reduced admissions. Of these medications, 11 were supported by clinical guidelines in the United States, the United Kingdom and Europe. These 11 therapies were for patients with heart failure (angiotensin-converting-enzyme inhibitors, angiotensin II receptor blockers, aldosterone receptor antagonists and digoxin), stable coronary artery disease (intensive statin therapy), asthma exacerbations (early inhaled corticosteroids in the emergency department and anticholinergics), chronic obstructive pulmonary disease (long-acting muscarinic antagonists and long-acting beta-2 adrenoceptor agonists) and schizophrenia (second-generation antipsychotics and depot/maintenance antipsychotics). We identified 11 medications supported by strong evidence and clinical guidelines that could be considered in quality monitoring and improvement strategies to help reduce emergency hospital admission rates. The findings are relevant to health systems with a large burden of chronic disease and those managing increasing pressures on acute health-care services.


Laser interstitial thermal therapy for newly diagnosed and recurrent glioblastoma.
OBJECTIVE Glioblastoma (GBM) is the most common and deadly malignant primary brain tumor. Better surgical therapies are needed for newly diagnosed GBMs that are difficult to resect and for GBMs that recur despite standard therapies. The authors reviewed their institutional experience of using laser interstitial thermal therapy (LITT) for the treatment of newly diagnosed or recurrent GBMs. METHODS This study reports on the pre-LITT characteristics and post-LITT outcomes of 8 patients with newly diagnosed GBMs and 13 patients with recurrent GBM who underwent LITT. RESULTS Compared with the group with recurrent GBMs, the patients with newly diagnosed GBMs who underwent LITT tended to be older (60.8 vs 48.9 years), harbored larger tumors (22.4 vs 14.6 cm3), and a greater proportion had IDH wild-type GBMs. In the newly diagnosed GBM group, the median progression-free survival and the median survival after the procedure were 2 months and 8 months, respectively, and no patient demonstrated radiographic shrinkage of the tumor on follow-up imaging. In the 13 patients with recurrent GBM, 5 demonstrated a response to LITT, with radiographic shrinkage of the tumor following ablation. The median progression-free survival was 5 months, and the median survival was greater than 7 months. CONCLUSIONS In carefully selected patients with recurrent GBM, LITT may be an effective alternative to surgery as a salvage treatment. Its role in the treatment of newly diagnosed unresectable GBMs is not established yet and requires further study.


The mere presence of a goalkeeper affects the accuracy of penalty kicks.
The keeper-independent strategy, in which a football penalty kicker selects a target location in advance and ignores the goalkeeper's actions during the run-up, has been suggested to be the preferable strategy for taking a penalty kick. The current in-field experiment investigated the question of whether the goalkeeper can indeed be ignored. Ten intermediate-level football players were instructed to adopt a goalkeeper-independent strategy and to perform penalty kicks directed at one of two targets located in the upper corners of the goal under three conditions: without a goalkeeper, in the presence of a goalkeeper (who tried to save the ball), and in the presence of a goalkeeper who was informed by the penalty kickers where they intended to direct the ball. The mere presence of a goalkeeper impaired shot accuracy. The shots were more centralised, that is, biased toward the goalkeeper. The effects were enhanced for the condition in which the penalty kicker knew the goalkeeper was knowledgeable about ball direction. The findings were consistent with the response activation model that holds that aiming at a target can be biased toward salient visual non-targets. The implications for adopting and practising goalkeeper-independent strategies are discussed.


Inhibition of interleukin-1 responsiveness by type II receptor gene transfer: a surface "receptor" with anti-interleukin-1 function.
The hypothesis that the type II receptor (RII) acts as a decoy for interleukin-1 (IL-1) was tested by gene transfer in cells expressing only the type I receptor (8387 fibroblasts). RII-transfected cells showed defective responsiveness to IL-1 in terms of NFkappaB activation, cytokine gene expression and production. Blocking monoclonal antibodies against RII restored the capacity of RII-transfected cells to respond to IL-1 beta. Hence defective IL-1 responsiveness of RII-transfected cells requires surface expression of the molecule. RII-transfected cells showed normal responsiveness to TNF, which shares functional properties and elements in the signal transduction pathway with IL-1. Cells transfected with a deletion mutant of RII missing 26 of 29 amino acids of the cytoplasmic portion of the molecule showed impaired responsiveness to IL-2. Cells transfected with full-length or the cytoplasmic deletion mutant of RII released copious amounts of RII in the supernatant. However, transfected cells showed defective responsiveness to brief exposure to IL-1, in the absence of measurable released RII. These results indicate that impairment of the responsiveness to IL-1 following RII gene transfer was dependent upon surface expression of the molecule, specific for IL-1 and unaffected by truncation of the cytoplasmic portion. Thus, the type II "receptor" is a decoy surface molecule, regulated by antiinflammatory signals, whose only known function is to capture and block IL-1.


A tutorial on individual participant data meta-analysis using Bayesian multilevel modeling to estimate alcohol intervention effects across heterogeneous studies.
This paper provides a tutorial companion for the methodological approach implemented in Huh et al. (2015) that overcame two major challenges for individual participant data (IPD) meta-analysis. Specifically, we show how to validly combine data from heterogeneous studies with varying numbers of treatment arms, and how to analyze highly-skewed count outcomes with many zeroes (e.g., alcohol and substance use outcomes) to estimate overall effect sizes. These issues have important implications for the feasibility, applicability, and interpretation of IPD meta-analysis but have received little attention thus far in the applied research literature. We present a Bayesian multilevel modeling approach for combining multi-arm trials (i.e., those with two or more treatment groups) in a distribution-appropriate IPD analysis. Illustrative data come from Project INTEGRATE, an IPD meta-analysis study of brief motivational interventions to reduce excessive alcohol use and related harm among college students. Our approach preserves the original random allocation within studies, combines within-study estimates across all studies, overcomes between-study heterogeneity in trial design (i.e., number of treatment arms) and/or study-level missing data, and derives two related treatment outcomes in a multivariate IPD meta-analysis. This methodological approach is a favorable alternative to collapsing or excluding intervention groups within multi-arm trials, making it possible to directly compare multiple treatment arms in a one-step IPD meta-analysis. To facilitate application of the method, we provide annotated computer code in R along with the example data used in this tutorial.


Purification and molecular cloning of Plx1, a Cdc25-regulatory kinase from Xenopus egg extracts.
Cdc2, the cyclin-dependent kinase that controls mitosis, is negatively regulated by phosphorylation on its threonine-14 and tyrosine-15 residues. Cdc25, the phosphatase that dephosphorylates both of these residues, undergoes activation and phosphorylation by multiple kinases at mitosis. Plx1, a kinase that associates with and phosphorylates the amino-terminal domain of Cdc25, was purified extensively from Xenopus egg extracts. Cloning of its complementary DNA revealed that Plx1 is related to the Polo family of protein kinases. Recombinant Plx1 phosphorylated Cdc25 and stimulated its activity in a purified system. Cdc25 phosphorylated by Plx1 reacted strongly with MPM-2, a monoclonal antibody to mitotic phosphoproteins. These studies indicate that Plx1 may participate in control of mitotic progression.


Targeted microbubbles for imaging tumor angiogenesis: assessment of whole-body biodistribution with dynamic micro-PET in mice.
To evaluate in vivo whole-body biodistribution of microbubbles (MBs) targeted to tumor angiogenesis-related vascular endothelial growth factor (VEGF) receptor 2 (VEGFR2) by using dynamic micro-positron emission tomography (PET) in living mice. Animal protocols were approved by the Institutional Administrative Panel on Laboratory Animal Care. Lipid-shell perfluorocarbon-filled MBs, targeted to VEGFR2 via anti-VEGFR2 antibodies, were radiolabeled by conjugating the radiofluorination agent N-succinimidyl-4-[(18)F]fluorobenzoate (SFB) to the anti-VEGFR2 antibodies. These MBs were then injected intravenously into nude mice (n = 4) bearing angiosarcomas, and the whole-body biodistribution of these probes was assessed for 60 minutes by using dynamic micro-PET. Results were compared with ex vivo gamma counting (n = 6) and immunofluorescence staining (n = 6). Control studies in angiosarcoma-bearing mice were performed with injection of the radiolabeled antibodies alone (n = 3) or free SFB (n = 3). A mixed-effects regression of MB accumulation on fixed effects of time and tissue type (tumor or muscle) and random effect of animal was performed. VEGFR2-targeted MBs rapidly cleared from the blood circulation (50% blood clearance after approximately 3.5 minutes) and accumulated in the liver (mean, 33.4% injected dose [ID]/g +/- 13.7 [standard deviation] at 60 minutes) and spleen (mean, 9.3% ID/g +/- 6.5 at 60 minutes) on the basis of micro-PET imaging. These findings were confirmed with ex vivo gamma counting. Uptake of targeted MBs was significantly higher (P < .0001) in tumor than in adjacent skeletal muscle tissue. Immunofluorescence staining demonstrated accumulation of the targeted MBs within hepatic Kupffer cells and splenic macrophages. Biodistribution of the radiolabeled antibodies and free SFB differed from the distribution of the targeted MBs. Dynamic micro-PET allows assessment of in vivo biodistribution of VEGFR2-targeted MBs.


Association between Serum Gamma-Glutamyltransferase and Prevalence of Metabolic Syndrome Using Data from the Korean Genome and Epidemiology Study.
The aim of this study was to determine whether there is a positive correlation between gamma-glutamyltransferase (GGT) levels and the prevalence of metabolic syndrome and whether GGT can be used as an easily checkable metabolic index using data from the large-scale Korean Genome and Epidemiology Study (KoGES). We obtained data of 211,725 participants of the KoGES. The collected data included age, sex, height, weight, waist circumference, and various biochemical characteristics, including serum GGT levels. The data of study participants who ingested more than 40 g/day of alcohol and who were diagnosed with metabolic syndrome at baseline was excluded. We analyzed the prevalence of metabolic syndrome according to GGT quartiles in both genders. The GGT level was significantly higher in subjects with metabolic syndrome compared to normal subjects (37.92±48.20 mg/dL vs. 25.62±33.56 mg/dL). The prevalence of metabolic syndrome showed a stepwise increase with GGT quartiles in both male and female subjects. Compared to the lowest GGT quartile, the odds ratio was 1.534 (95% confidence interval [CI], 1.432 to 1.643), 1.939 (95% CI, 1.811 to 2.076), and 2.754 (95% CI, 2.572 to 2.948) in men and 1.155 (95% CI, 1.094 to 1.218), 1.528 (95% CI, 1.451 to 1.609), and 2.022 (95% CI, 1.921 to 2.218) in women with increasing GGT quartile. The cutoff value of GGT predicting risk of metabolic syndrome was 27 IU/L in men and 17 IU/L in women. We suggested that GGT could be an easily checkable marker for the prediction of metabolic syndrome.


Diffraction-based overlay metrology using angular-multiplexed acquisition of dark-field digital holograms.
In semiconductor device manufacturing, optical overlay metrology measures pattern placement between two layers in a chip with sub-nm precision. Continuous improvements in overlay metrology are needed to keep up with shrinking device dimensions in modern chips. We present first overlay metrology results using a novel off-axis dark-field digital holographic microscopy concept that acquires multiple holograms in parallel by angular multiplexing. We show that this concept reduces the impact of source intensity fluctuations on the noise in the measured overlay. With our setup we achieved an overlay reproducibility of 0.13 nm and measurements on overlay targets with known programmed overlay values showed good linearity of R2= 0.9993. Our data show potential for significant improvement and that digital holographic microscopy is a promising technique for future overlay metrology tools.


Microbial copper reduction method to scavenge anthropogenic radioiodine.
Unexpected reactor accidents and radioisotope production and consumption have led to a continuous increase in the global-scale contamination of radionuclides. In particular, anthropogenic radioiodine has become critical due to its highly volatile mobilization and recycling in global environments, resulting in widespread, negative impact on nature. We report a novel biostimulant method to effectively scavenge radioiodine that exhibits remarkable selectivity for the highly difficult-to-capture radioiodine of >500-fold over other anions, even under circumneutral pH. We discovered a useful mechanism by which microbially reducible copper (i.e., Cu(2+) to Cu(+)) acts as a strong binder for iodide-iodide anions to form a crystalline halide salt of CuI that is highly insoluble in wastewater. The biocatalytic crystallization of radioiodine is a promising way to remove radioiodine in a great capacity with robust growth momentum, further ensuring its long-term stability through nuclear I(-) fixation via microcrystal formation.


Gold nanoparticle surface deposition induced quantum efficiency enhancement for Si-multi-pixel photon counters.
With recent development of nanotechnology, novel devices with nanostructures arise to improve the performance of photodetectors. Here, we demonstrated that by surface decoration with gold nanoparticles on the active area, the quantum detection efficiency of a multi-pixel photon counter was increased due to surface plasmon resonance enhancement. The deposited gold nano-particles actually brought about almost the same enhancement factor for any photon-number fields. As a result, the photon-number-resolving capability of the multi-pixel photon counter was well reserved with the gold nano-particle deposition induced efficiency augment. This result provides guidance to the development of the high-efficiency photon-number-resolving detectors.


Bear bile: dilemma of traditional medicinal use and animal protection.
Bear bile has been used in Traditional Chinese Medicine (TCM) for thousands of years. Modern investigations showed that it has a wide range of pharmacological actions with little toxicological side effect and the pure compounds have been used for curing hepatic and biliary disorders for decades. However, extensive consumption of bear bile made bears endangered species. In the 1980's, bear farming was established in China to extract bear bile from living bears with "Free-dripping Fistula Technique". Bear farming is extremely inhumane and many bears died of illness such as chronic infections and liver cancer. Efforts are now given by non-governmental organizations, mass media and Chinese government to end bear farming ultimately. At the same time, systematic research has to be done to find an alternative for bear bile. In this review, we focused on the literature, laboratory and clinical results related to bear bile and its substitutes or alternative in English and Chinese databases. We examined the substitutes or alternative of bear bile from three aspects: pure compounds derived from bear bile, biles from other animals and herbs from TCM. We then discussed the strategy for stopping the trading of bear bile and issues of bear bile related to potential alternative candidates, existing problems in alternative research and work to be done in the future.


Self-induced oscillations in an optomechanical system driven by bolometric backaction.
We have explored the nonlinear dynamics of an optomechanical system consisting of an illuminated Fabry-Perot cavity, one of whose end mirrors is attached to a vibrating cantilever. The backaction induced by the bolometric light force produces negative damping such that the system enters a regime of nonlinear oscillations. We study the ensuing attractor diagram describing the nonlinear dynamics. A theory is presented that yields quantitative agreement with experimental results. This includes the observation of a regime where two mechanical modes of the cantilever are excited simultaneously.


Atmospheric optical measurements in the vicinity of crater lake, Oregon. Part 2.
This paper, Part 2, presents additional atmospheric optical data, comparable to the data presented in Part 1, but for a different type of day, and for four descents instead of two. All descents were before local apparent noon. Simultaneous spectral irradiance data were recorded at the surface of Crater Lake. Data presented are altitude profiles of heading of aircraft, temperature, relative humidity, equilibrium luminance, horizontal path function, attenuation length, nadir luminance, upwelling and downwelling illuminances and their ratios, reflectance calculated from nadir luminance, atmospheric beam transmittances for vertical path of sight as a function of solar zenith angle, and downwelling illuminance as a function of solar zenith angle.


Gene expression profile of the rat eye iridocorneal angle: NEIBank expressed sequence tag analysis.
To characterize gene expression pattern in the combined tissues of the rat iridocorneal angle by expressed sequence tag (EST) analysis, as part of the NEIBank project. RNA was extracted from dissected tissues of the rat iridocorneal angle (iris, ciliary body, trabecular meshwork, and Schlemm's canal) and used to construct unamplified, non-normalized cDNA libraries in the pSPORT1 vector. Approximately 5000 clones were sequenced from the 5'-end. Clones were clustered and identified using the GRIST software, a procedure based on BLAST comparisons. Complete sequences of several novel cDNAs showing eye-preferred expression patterns were obtained. The expression patterns of several genes have been investigated by Northern blot and in situ hybridization, as well as by RT-PCR. After analysis and removal of non-mRNA sequences, 2195 independent clusters, potentially representing individual eye angle-expressed clones were obtained. The expression profile of the combined rat eye angle tissues was more similar to that of the human iris than to human trabecular meshwork. Several cDNAs encoding transcription factors essential for normal eye development and function including Pax-6, Six3, c-Maf, Maf1, Sox-4, Foxc1, Rx, and Ldb2 were present among sequenced clones. A number of tested cDNAs showed eye-preferred expression patterns. Myocilin, which is abundant in human eye angle tissues, was not observed in the rat collection; however, transcripts for three other olfactomedin-domain proteins were seen. Latrotoxin receptor (CL1AA) and optimedin were shown to be expressed in the iris and ciliary body, as well as in the ganglion and inner nuclear cell layers of the retina, whereas the rat orthologue of the human HNOEL-iso gene was expressed in the iris and sclera and less actively in the trabecular meshwork, retina, and optic nerve. The iridocorneal libraries are a good source of novel uncharacterized genes and molecular markers for the tissues of the eye angle. Although myocilin is not abundantly expressed in rat eye angle, other olfactomedin-containing genes are expressed there and may play important roles in normal eye function and disease.


Restricted daily consumption of a highly palatable food (chocolate Ensure(R)) alters striatal enkephalin gene expression.
Brain opioid peptide systems are known to play an important role in motivation, emotion, attachment behaviour, the response to stress and pain, and the control of food intake. Opioid peptides within the ventral striatum are thought to play a key role in the latter function, regulating the affective response to highly palatable, energy-dense foods such as those containing fat and sugar. It has been shown previously that stimulation of mu opiate receptors within the ventral striatum increases intake of palatable food. In the present study, we examined enkephalin peptide gene expression within the striatum in rats that had been given restricted daily access to an energy-dense, palatable liquid food, chocolate Ensure(R). Rats maintained on an ad libitum diet of rat chow and water were given 3-h access to Ensure(R) daily for two weeks. One day following the end of this period, preproenkephalin gene expression was measured with quantitative in situ hybridization. Compared with control animals, rats that had been exposed to Ensure(R) had significantly reduced enkephalin gene expression in several striatal regions including the ventral striatum (nucleus accumbens), a finding that was confirmed in a different group with Northern blot analysis. Rats fed this regimen of Ensure(R) did not differ in weight from controls. In contrast to chronic Ensure(R), acute ingestion of Ensure(R) did not appear to affect enkephalin peptide gene expression. These results suggest that repeated consumption of a highly rewarding, energy-dense food induces neuroadaptations in cognitive-motivational circuits.


Proton pump inhibitors: Risks of long-term use.
Proton pump inhibitors are among the most commonly prescribed classes of drugs, and their use is increasing, in particular for long-term treatment, often being over-prescribed and used for inappropriate conditions. In recent years, considerable attention has been directed towards a wide range of adverse effects, and even when a potential underlying biological mechanism is plausible, the clinical evidence of the adverse effect is often weak. Several long-term side effects have been investigated ranging from interaction with other drugs, increased risk of infection, reduced intestinal absorption of vitamins and minerals, and more recently kidney damage and dementia. The most recent literature regarding these adverse effects and their association with long-term proton pump inhibitor treatment is reviewed, and the mechanisms through which these possible complications might develop are discussed.


Passive sampling of perfluorinated chemicals in water: flow rate effects on chemical uptake.
A recently developed modified polar organic chemical integrative sampler (POCIS) provides a means for monitoring perfluorinated chemicals (PFCs) in water. However, changes in external flow rates may alter POCIS sampling behaviour and consequently affect estimated water concentrations of analytes. In this work, uptake kinetics of selected PFCs, over 15 days, were investigated. A flow-through channel system was employed with spiked river water at flow rates between 0.02 and 0.34 m s(-1). PFC sampling rates (Rs) (0.09-0.29 L d(-1) depending on analyte and flow rate) increased from the lowest to highest flow rate employed for some PFCs (MW ≤ 464) but not for others (MW ≥ 500). Rs's for some of these smaller PFCs were increasingly less sensitive to flow rate as this increased within the range investigated. This device shows promise as a sampling tool to support monitoring efforts for PFCs in a range of flow rate conditions.


Lanadelumab for the treatment of hereditary angioedema.
Introduction: Hereditary angioedema due to C1 inhibitor deficiency (C1-INH-HAE) is a rare yet still probably underdiagnosed clinical condition. Recurrent episodes of subcutaneous and sub-mucosal swelling may involve the skin, the gastrointestinal tract or even the upper airways, exposing the patients to the risk of death. With the aim of improving patients' quality of life, the therapeutic scenario has expanded over the years.Areas covered: The focus of the present review is lanadelumab, a fully human, κ-light-chain, monoclonal immunoglobulin G1 against plasma kallikrein, currently approved for long-term prophylaxis of C1-INH-HAE attacks in the USA and Canada and designated as an orphan drug by the European Medicines Agency.Expert opinion: Lanadelumab is able to inhibit plasma kallikrein with high selectivity and affinity. The subsequent phases of drug development and the ongoing open-label trial have proven its safety and efficacy. It overcomes some of the limitations of other drugs available for long-term prophylaxis, given the easy route of administration, the simple administration schedule and the possibility to tailor the treatment to each patient. Further studies are needed to test its efficacy also in other types of angioedema for which a central role of plasma kallikrein is envisaged.


Modelling the effects of transcranial alternating current stimulation on the neural encoding of speech in noise.
Transcranial alternating current stimulation (tACS) can non-invasively modulate neuronal activity in the cerebral cortex, in particular at the frequency of the applied stimulation. Such modulation can matter for speech processing, since the latter involves the tracking of slow amplitude fluctuations in speech by cortical activity. tACS with a current signal that follows the envelope of a speech stimulus has indeed been found to influence the cortical tracking and to modulate the comprehension of the speech in background noise. However, how exactly tACS influences the speech-related cortical activity, and how it causes the observed effects on speech comprehension, remains poorly understood. A computational model for cortical speech processing in a biophysically plausible spiking neural network has recently been proposed. Here we extended the model to investigate the effects of different types of stimulation waveforms, similar to those previously applied in experimental studies, on the processing of speech in noise. We assessed in particular how well speech could be decoded from the neural network activity when paired with the exogenous stimulation. We found that, in the absence of current stimulation, the speech-in-noise decoding accuracy was comparable to the comprehension of speech in background noise of human listeners. We further found that current stimulation could alter the speech decoding accuracy by a few percent, comparable to the effects of tACS on speech-in-noise comprehension. Our simulations further allowed us to identify the parameters for the stimulation waveforms that yielded the largest enhancement of speech-in-noise encoding. Our model thereby provides insight into the potential neural mechanisms by which weak alternating current stimulation may influence speech comprehension and allows to screen a large range of stimulation waveforms for their effect on speech processing.


Factors contributing to inter- and intra-annual variation in condition of cod Gadus morhua in the Barents Sea.
1. Relative body condition (the quantity of stored energy) is an important tool in understanding demographic variation and the ability of a population to respond to environmental stressors, varying food availability and competition. 2. A high-resolution database was used to examine causes of variation in the condition of north-east Arctic cod (Gadus morhua L.) for the period 1967-2004, over annual and monthly timescales. Community dynamics and climate variation were also tested as potential causes. 3. Temperature was shown to have a positive impact on condition at both inter- and intra-annual timescales. Inter-annually, temperature may affect stock distribution, in particular its overlap with the capelin stock. At shorter timescales it is likely that temperature directly affects the metabolism of the cod. 4. Intra-annually, the quantity of capelin in cod stomachs positively affected cod condition in the current and the preceding month for all lengths of cod. This indicated a time lag between a change in food consumption and a subsequent change in condition, or 'latency'. 5. Our study has shown that variation in temperature is a vital determinant of changes in condition, both at inter- and intra-annual timescales. Furthermore, the principle of latency has been demonstrated at the population level. Indirect effects of competition for energy-rich resources have been shown to have a negative effect on condition. This study supplements our knowledge of the implications for condition of changes in climate and in potential food resources.


The role of the amygdala in atypical gaze on emotional faces in autism spectrum disorders.
Reduced focus toward the eyes is a characteristic of atypical gaze on emotional faces in autism spectrum disorders (ASD). Along with the atypical gaze, aberrant amygdala activity during face processing compared with neurotypically developed (NT) participants has been repeatedly reported in ASD. It remains unclear whether the previously reported dysfunctional amygdalar response patterns in ASD support an active avoidance of direct eye contact or rather a lack of social attention. Using a recently introduced emotion classification task, we investigated eye movements and changes in blood oxygen level-dependent (BOLD) signal in the amygdala with a 3T MRI scanner in 16 autistic and 17 control adult human participants. By modulating the initial fixation position on faces, we investigated changes triggered by the eyes compared with the mouth. Between-group interaction effects revealed different patterns of gaze and amygdalar BOLD changes in ASD and NT: Individuals with ASD gazed more often away from than toward the eyes, compared with the NT group, which showed the reversed tendency. An interaction contrast of group and initial fixation position further yielded a significant cluster of amygdala activity. Extracted parameter estimates showed greater response to eyes fixation in ASD, whereas the NT group showed an increase for mouth fixation. The differing patterns of amygdala activity in combination with differing patterns of gaze behavior between groups triggered by direct eye contact and mouth fixation, suggest a dysfunctional profile of the amygdala in ASD involving an interplay of both eye-avoidance processing and reduced orientation.


Segregational Drift and the Interplay between Plasmid Copy Number and Evolvability.
The ubiquity of plasmids in all prokaryotic phyla and habitats and their ability to transfer between cells marks them as prominent constituents of prokaryotic genomes. Many plasmids are found in their host cell in multiple copies. This leads to an increased mutational supply of plasmid-encoded genes and genetically heterogeneous plasmid genomes. Nonetheless, the segregation of plasmid copies into daughter cells during cell division is considered to occur in the absence of selection on the plasmid alleles. We investigate the implications of random genetic drift of multicopy plasmids during cell division-termed here "segregational drift"-to plasmid evolution. Performing experimental evolution of low- and high-copy non-mobile plasmids in Escherichia coli, we find that the evolutionary rate of multicopy plasmids does not reflect the increased mutational supply expected according to their copy number. In addition, simulated evolution of multicopy plasmid alleles demonstrates that segregational drift leads to increased loss frequency and extended fixation time of plasmid mutations in comparison to haploid chromosomes. Furthermore, an examination of the experimentally evolved hosts reveals a significant impact of the plasmid type on the host chromosome evolution. Our study demonstrates that segregational drift of multicopy plasmids interferes with the retention and fixation of novel plasmid variants. Depending on the selection pressure on newly emerging variants, plasmid genomes may evolve slower than haploid chromosomes, regardless of their higher mutational supply. We suggest that plasmid copy number is an important determinant of plasmid evolvability due to the manifestation of segregational drift.


Surgical bleeding after pre-operative unfractionated heparin and low molecular weight heparin for coronary bypass surgery.
Since the impairment of platelet function may cause excess peri-operative bleeding, pre-operative discontinuation of aspirin and heparin bridging are common for cardiac surgery. We evaluated the impact of pre-operative administration of enoxaparin and unfractionated heparin (UFH) on coagulation parameters and peri-operative bleeding in patients undergoing elective coronary artery bypass grafting (CABG) surgery after discontinuation of aspirin. Forty-three patients with three-vessel coronary artery disease undergoing elective CABG surgery discontinued aspirin and were randomized to receive either UFH 180 UI/Kg x 2/day s.c. or enoxaparin 100 UI/Kg x 2/day s.c. until 12 h before surgery (median pre-operative treatment 8 days, range 6-12 days). Surgery was performed as usual with UFH. Neither UFH nor any low molecular weight heparin was given in the immediate post-operative period. The effects of UFH and enoxaparin were monitored by the activated partial thromboplastin time (aPTT) and the Enox-test (sensitive to factor Xa inhibition) using a Rapidpoint Coagulation Analyzer. aPTT and factor Xa activity were also measured by standard methods. Peri-operative bleeding and the nadirs of hemoglobin concentration, hematocrit and platelet count were monitored post-operatively. Patients in the two groups were similar for number of bypasses, on-pump time, total surgery time, and time from the last heparin administration. Coagulation parameters increased significantly and similarly at 30 min and 6 h with both treatments, but returned within the normal range at 12 h. Hemoglobin, hematocrit and platelet counts significantly decreased to the same extent after CABG and re-normalized at the same time. Transfusional requirements of blood and plasma units were similar in the two groups. From the kinetics of coagulation parameters and the evaluation of bleeding, enoxaparin is a safe alternative to UFH as a bridging therapy to CABG after discontinuation of aspirin.


Sexual risk among orphaned adolescents: is country-level HIV prevalence an important factor?
Previous studies from sub-Saharan Africa have found that orphans experience increased sexual risk compared to non-orphans. We developed a theoretical framework for the investigation of determinants of HIV risk and used it to generate specific hypotheses regarding the effect of country-level HIV prevalence on the sexual risk experience of orphans. We expected that countries with high HIV prevalence would experience a higher prevalence of orphanhood. We further hypothesised that orphans in countries with high HIV prevalence would experience increased sexual risk, compared to non-orphans, due to pressure on the extended family network, which is primarily responsible for the care of orphans in sub-Saharan Africa, resulting in poorer standards of care and guidance. We used hierarchical logistic regression models to investigate this hypothesis using cross-sectional, Demographic and Health Survey data from 10 sub-Saharan African countries. We found that countries with high HIV prevalence did indeed have higher prevalence of orphanhood. We also found that, amongst female adolescents, maternal and double orphans were significantly more likely to have started sex than non-orphans in countries with high HIV prevalence but were not at increased risk in low HIV prevalence countries. This effect of country-level HIV prevalence on the sexual risk of orphans was not explained by household level factors such as wealth, overcrowding or age of the household head. The same pattern of risk was not observed for male adolescents - male orphans were not more likely to have started sex than non-orphans. This suggests that orphaned adolescent women are an important target group for HIV prevention and that efforts should be made to integrate prevention messages into existing support programmes for orphans and vulnerable children.


Drug release into hydrogel-based subcutaneous surrogates studied by UV imaging.
Upon subcutaneous administration, the distribution of drug between the delivery vehicle and the biological tissue critically affects the absorption of drug substances. Utilization of physical models resembling the native tissues appears promising for obtaining a detailed understanding of the performance of drug delivery systems based on in vitro experiments. The objective of this study was to evaluate a UV imaging-based method for real-time characterization of the release and transport of piroxicam in hydrogel-based subcutaneous tissue mimics/surrogates. Piroxicam partitioning from medium chain triglyceride (MCT) into 0.5% (w/v) agarose or 25% (w/v) F127-based hydrogels was investigated by monitoring the concentration profiles of the drug in the gels. The effect of pH on piroxicam distribution and diffusion coefficients was studied. For both hydrogel systems, the diffusion of piroxicam in the gels was not affected significantly by the pH change from 4.0 to 7.4 but a considerable change in the oil-gel distribution coefficients was found (24 and 34 times less at pH 7.4 as compared those observed at pH 4.0 for F127 and agarose gels, respectively). In addition, the release and transport processes of piroxicam upon the injection of aqueous or MCT solutions into an agarose-based hydrogel were investigated by UV imaging. The spatial distribution of piroxicam around the injection site in the gel matrix was monitored in real-time. The disappearance profiles of piroxicam from the injected aqueous solution were obtained. This study shows that the UV imaging methodology has considerable potential for characterizing transport properties in hydrogels, including monitoring the real-time spatial concentration distribution in vitro after administration by injection.