[Planning ability of older and younger adults].
This investigation was aimed at the effects of ageing on planning. The investigation was carried out with young adults, middle-aged adults and elderly. Because of a suggested resemblance of problems described in elderly with impairments in patients with prefrontal cortical damage, and because the prefrontal cortex is important for planning, a test specifically sensitive to planning deficits in left prefrontal patients, the Tower of London test, was administered. On this test, the elderly solved significantly less problems in the allotted time than the two younger groups. However, the elderly did not show the same type of problem as that described in prefrontal patients. Such patients perform significantly worse on first attempts (problems correctly solved in one attempt). On this variable the elderly in our study equalled the younger adults in performance. However, they less often succeeded when the first attempt was not correct. To check whether this result could be explained simply by decreased mental and psychomotor speed and visuo-spatial function, regression functions between age and Tower score were computed, partialing out scores on tests of these more elementary abilities. A significant effect of age remained, which we tentatively interpret as an effect on planning ability.


Increased Fat Absorption From Enteral Formula Through an In-line Digestive Cartridge in Patients With Cystic Fibrosis.
Supplemental enteral nutrition (EN) is used by approximately 12% of people with cystic fibrosis (CF). The objective of this study was to evaluate the safety, tolerability, and fat absorption of a new in-line digestive cartridge (Relizorb) that hydrolyzes fat in enteral formula provided to patients with CF. Patients with CF receiving EN participated in a multicenter, randomized, double-blind, crossover trial with an open-label safety evaluation period. Plasma omega-3 fatty acid (FA) concentrations were measured and used as markers of fat absorption. Gastrointestinal symptoms were recorded to evaluate safety and tolerability. Information regarding the effect of EN on appetite and breakfast consumption was also collected. Before study entry, participants had received EN for a mean of 6.6 years at a mean volume of approximately 800 mL, yet had a mean body mass index of only 17.5 kg/m and omega-3 FA plasma concentrations were only 60% of levels found in normal healthy subjects. Compared with placebo, cartridge use resulted in a statistically significant 2.8-fold increase in plasma omega-3 FA concentrations. There were no adverse experiences associated with cartridge use, and a decrease in the frequency and severity of most symptoms of malabsorption was observed with cartridge use. Participants reported increased preservation of appetite and breakfast consumption with cartridge use compared with their pre-study regimen. Use of this in-line digestive cartridge was safe and well tolerated, and resulted in significantly increased levels of plasma omega-3 FA used with enteral formula, suggesting an overall increased fat absorption.


Tryptophan metabolism, gut microbiota, and carotid artery plaque in women with and without HIV infection.
The perturbation of tryptophan (TRP) metabolism has been linked with HIV infection and cardiovascular disease (CVD), but the interrelationship among TRP metabolites, gut microbiota, and atherosclerosis remain unclear in the context of HIV infection. We included 361 women (241 HIV+, 120 HIV-) with carotid artery plaque assessments from the Women's Interagency HIV Study, measured ten plasma TRP metabolites and profiled fecal gut microbiome. TRP metabolites-related gut bacteria were selected through the Analysis of Compositions of Microbiomes with Bias Correction method. Associations of TRP metabolites and related microbial features with plaque were examined using multivariable logistic regression. While plasma kynurenic acid (KYNA) (odds ratio [OR] = 1.93, 95% confidence interval [CI]:1.12, 3.32 per one SD increase; P = 0.02) and KYNA/TRP (OR = 1.83 [95%CI:1.08, 3.09], P = 0.02) were positively associated with plaque, indole-3-propionate (IPA) (OR = 0.62 [95%CI:0.40, 0.98], P = 0.03) and IPA/KYNA (OR = 0.51[95%CI:0.33, 0.80], P < 0.01) were inversely associated with plaque. Five gut bacterial genera and many affiliated species were positively associated with IPA (FDR-q < 0.25), including Roseburia sp., Eubacterium sp., Lachnospira sp., and Coprobacter sp.; but no bacterial genera were found to be associated with KYNA. Furthermore, an IPA-associated-bacteria score was inversely associated with plaque (OR = 0.47[95%CI:0.28, 0.79], P < 0.01). But no significant effect modification by HIV serostatus was observed in these associations. In a cohort of women living with and without HIV infection, plasma IPA levels and related gut bacteria were inversely associated with carotid artery plaque, suggesting a potential beneficial role of IPA and its gut bacterial producers in atherosclerosis and CVD.


Residents' preferences and preparation for caring for underserved populations.
Access to care by low-income persons and residents of rural and poor inner-city areas is a persistent problem, yet physicians tend to be maldistributed relative to need. The objectives were to describe preferences of resident physicians to locate in underserved areas and to assess their preparedness to provide service to low-income populations. A national survey was made of residents completing their training in eight specialties at 162 US academic health center hospitals in 1998, with 2,626 residents responding. (Of 4,832 sampled, 813 had invalid addresses or were no longer in the residency program. Among the valid sample of 4,019, the response rate was 65%.) The percentage of residents ranking public hospitals, rural areas, and poor inner-city areas as desirable employment locations and the percentage feeling prepared to provide specified services associated with indigent populations were ascertained. Logistic regressions were used to calculate adjusted percentages, controlling for sex, race/ethnicity, international medical graduate (IMG) status, plans to subspecialize, ownership of hospital, specialty, and exposure to underserved patients during residency. Only one third of residents rated public hospitals as desirable settings, although there were large variations by specialty. Desirability was not associated with having trained in a public hospital or having greater exposure to underserved populations. Only about one quarter of respondents ranked rural (26%) or poor inner-city (25%) areas as desirable. Men (29%, P <.01) and noncitizen IMGs (43%, P <.01) were more likely than others to prefer rural settings. Residents who were more likely to rate poor inner-city settings as desirable included women (28%, P =.03), noncitizen IMGs (35%, P =.01), and especially underrepresented minorities (52%, P <.01). Whereas about 90% or more of residents felt prepared to treat common clinical conditions, only 67% of residents in four primary care specialties felt prepared to counsel patients about domestic violence or to care for human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) or substance abuse patients (all 67%). Women were more likely than men to feel prepared to counsel patients about domestic violence (70% vs. 63%, P =.002) and depression (83% vs. 75%, P <.01). Underrepresented minority residents were more likely than other residents to feel prepared to counsel patients about domestic violence (P <.01) and compliance with care (P =.04). Residents with greater exposure to underserved groups were more prepared to counsel patients about domestic violence (P =.01), substance abuse (P =.01), and to treat patients with HIV/AIDS (P =.01) or with substance abuse problems (P <.01). This study demonstrates the need to expose graduate trainees to underserved populations and suggests a continuing role of minorities, women, and noncitizen physicians in caring for low-income populations.


Epigenetic mechanisms in migraine: a promising avenue?
Migraine is a disabling common brain disorder typically characterized by attacks of severe headache and associated with autonomic and neurological symptoms. Its etiology is far from resolved. This review will focus on evidence that epigenetic mechanisms play an important role in disease etiology. Epigenetics comprise both DNA methylation and post-translational modifications of the tails of histone proteins, affecting chromatin structure and gene expression. Besides playing a role in establishing cellular and developmental stage-specific regulation of gene expression, epigenetic processes are also important for programming lasting cellular responses to environmental signals. Epigenetic mechanisms may explain how non-genetic endogenous and exogenous factors such as female sex hormones, stress hormones and inflammation trigger may modulate attack frequency. Developing drugs that specifically target epigenetic mechanisms may open up exciting new avenues for the prophylactic treatment of migraine.


Analysis of situation of acute coronary syndrome based on the date of the Improving Care for Cardiovascular Disease in China-Acute Coronary Syndrome (CCC-ACS) project: single-centre observational study.
We are currently faced with an increasing burden of cardiovascular disease in China and the inadequacy of the application of guidelines in clinical practice. In the past decade, China has been strengthening the healthcare system, but it still lacked a national performance measurement system and an appropriate quality improvement strategy. Therefore, in order to improve the implementation of guideline recommendations in clinical practice, China has learnt from the successful experience of Get With The Guidelines project in 2014. Under the guidance of the Medical and Health Hospital of the National Health and Family Planning Commission, the Chinese Society of Cardiology and the American Heart Association jointly launched the Improving Care for Cardiovascular Disease in China (CCC) project. The project team provided an analysis report on the completion of key medical quality evaluation indicators of each hospital every month, supplied guidance through education, training, experience exchange and on-site investigation for problems, and certified hospitals with outstanding performance and obvious progress. The circle pattern, including evaluation, training, improvement and re-evaluation, will boost the guidelines compliance on clinical practice in China and improve the quality of medical services. This study was conducted in a centre of the Third Xiangya Hospital of Central South University. It included patients with ACS from December 2009 to December 2011 (n=225), patients with ACS in the Improving Care for Cardiovascular Disease in China-Acute Coronary Syndrome project coming from the Third Xiangya Hospital of Central South University (n=665), 12 hospitals in Hunan Province (n=4333) and 150 hospitals in China (n=63 641) from November 2014 to April 2017. It assessed the situation of drug therapy, hospitalisation day, mortality during hospitalisation, median of door-to-needle (D-to-N) time and median of door-to-balloon (D-to-B) time of patients with ST-segment elevation myocardial infarction (STEMI), the proportion of D-to-N within 30 min and D-to-B within 90 min, and the proportion of reperfusion therapy. Patients with ACS from the centre from November 2014 to April 2017 were divided into five groups (every 6 months as a group according to time). The study observed change trends in all the above-mentioned indexes. Compared with before participating in the CCC project, there were increases after participating in the CCC project in the drug usage rates of aspirin, P2Y12 inhibitor (clopidogrel or ticagrelor), β-blocker, statin and angiotensin converting enzyme inhibitor (ACEI)/angiotensin-receptor blocker (ARB). Hospitalisation day and mortality during hospitalisation were shortened. D-to-N and D-to-B times of patients with STEMI were shorter. Compared with Hunan Province and China, the drug usage rates were higher; hospitalisation day and D-to-N time were shorter; D-to-B time was longer; and the proportion of reperfusion therapy was higher. The trend of drug usage rates was on the rise. There was no significant change in the hospitalisation day and D-to-N and D-to-B times. The mortality during hospitalisation showed a downward trend. The proportion of D-to-N within 90 min and reperfusion therapy showed upward trends. Quality of care for patients with ACS improved over time in the CCC project, including taking medicine following the guidelines, increased use of reperfusion therapy and faster time to treatment. Although overall mortality has improved, we also should attach importance to high-risk patients. The influence of the CCC project, which is based on guidelines on prognosis of ACS in the centre, presents an important clinical implication that it is necessary to enhance adherence to the guidelines in the treatment of ACS.


Outcomes of pancreas allografts procured simultaneously with an isolated intestine allograft: single-center and national data.
Simultaneous procurement of pancreas and isolated intestine allografts from the same donor may compromise one graft or both given the shared vasculature between these two organs. This study reports the transplant outcomes for pancreas allografts procured simultaneously with an intestine graft. Separate analyses are reported from (1) 10-year US national data obtained from the United Network for Organ Sharing database including all donors from 2000 to 2010 in whom both the pancreas and isolated intestine were procured and transplanted into separate recipients and (2) local, single-center data for all donors of simultaneous pancreas and isolated intestine procurement. Scientific Registry of Transplant Recipients reference data are provided for comparison. Nationally, there were 159 simultaneous pancreas and intestine donors. The mean pancreas transplant graft and patient survival rates for this cohort were 75% and 93%, respectively. There were 34 isolated intestine allografts procured by our center during the study period. There were 14 cases (41%) in which the pancreas was transplanted. The reasons for nontransplantation of the pancreas in the other 20 donors included donor age (n=12), graft quality (n=4), and anatomic or surgical issues (n=4). For the 14 transplanted grafts, 1-year pancreas allograft survival rate was 100%. These results suggest that pancreas allografts procured simultaneously with an isolated intestine graft, although technically more challenging, do not have significantly inferior survival outcomes.


Therapeutic drug monitoring of eculizumab: Rationale for an individualized dosing schedule.
The annual cost of eculizumab maintenance therapy in paroxysmal nocturnal hemoglobinuria (PNH) and atypical hemolytic-uremic syndrome (aHUS) exceeds $300,000 per patient. A better understanding of eculizumab pharmacokinetics and subsequent individual dose adjustment could reduce this cost. We measured the trough eculizumab concentration in 9 patients with maintenance therapy (aHUS, n = 7; PNH, n = 2) and determined: 1) the intra- and inter-individual variability; 2) the influence of weight on eculizumab pharmacokinetics; and 3) the rate of elimination of eculizumab following discontinuation. A one-compartment model was developed to describe the pharmacokinetics of eculizumab and predicted complement activity by body weight. Trough eculizumab concentrations were >50 µg/mL in 9/9, >100 µg/mL in 8/9, and >300 µg/mL in 5/9 of patients. Intra-individual variability was low but eculizumab concentrations, closely correlated with patient weight (R(2) = 0.66, p = 0.034), varied broadly (55 ± 12 to 733 ± 164 µg/mL). Pharmacokinetic modeling showed that the elimination half-life varied greatly, with an increase from 7.8 d in a patient weighing 100 kg to 19.5 d in a 40 kg patient. We predicted that infusions of 1200 mg could be spaced every 4 or 6 weeks in patients weighing <90 and <70 kg, respectively. In this pilot study, the current recommended use of a fixed eculizumab dose for maintenance therapy is associated with excessively high trough concentrations in many patients. Further prospective larger studies are now required to support an individualized schedule adjusted for patient weight and based on the observed trough serum eculizumab concentration.


Elucidation of in Vitro Chlorinated Tyrosine Adducts in Blood Plasma as Selective Biomarkers of Chlorine Exposure.
Chlorine is a widely available industrial chemical and involved in a substantial number of cases of poisoning. It has also been used as a chemical warfare agent in military conflicts. To enable forensic verification, the persistent biomarkers 3-chlorotyrosine and 3,5-dichlorotyrosine in biomedical samples could be detected. An important shortfall of these biomarkers, however, is the relatively high incidence of elevated levels of chlorinated tyrosine residues in individuals with inflammatory diseases who have not been exposed to chlorine. Therefore, more reliable biomarkers are necessary to distinguish between endogenous formation and exogeneous exposure. The present study aims to develop a novel diagnostic tool for identifying site-specific chlorinated peptides as a more unambiguous indicator of exogeneous chlorine exposure. Human blood plasma was exposed in vitro to various chlorine concentrations, and the plasma proteins were subsequently digested by pronase, trypsin, or pepsin. After sample preparation, the digests were analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS) and liquid chromatography high-resolution tandem mass spectrometry (LC-HRMS/MS). In line with other studies, low levels of 3-chlorotyrosine and 3,5-dichlorotyrosine were found in blank plasma samples in this study. Therefore, 50 site-specific biomarkers were identified, which could be used as more unambiguous biomarkers for chlorine exposure. Chlorination of the peptides TY*ETTLEK, Y*KPGQTVK, Y*QQKPGQAPR, HY*EGSTVPEK, and Y*LY*EIAR could already be detected at moderate in vitro chlorine exposure levels. In addition, the latter two peptides were found to have dichlorinated fragments. Especially, Y*LY*EIAR, with a distinct chlorination pattern in the MS spectra, could potentially be used to differentiate exogeneous exposure from endogenous causes as other studies reported that this part of human serum albumin is nitrated rather than chlorinated under physiological conditions. In conclusion, trypsin digestion combined with high-resolution MS analysis of chlorinated peptides could constitute a valuable technique for the forensic verification of exposure to chlorine.


Capacitive Removal of Heavy Metal Ions from Wastewater via an Electro-Adsorption and Electro-Reaction Coupling Process.
Heavy metals widely exist in wastewater, which is a serious threat to human health or water environment. Highly efficient removal of heavy metal ions from wastewater is a major challenge to wastewater treatment. In this work, capacitive removal of heavy metal ions from wastewater via an electro-adsorption and electro-reaction coupling process was originally demonstrated. The removal efficiency of heavy metal ions in the binary-component solutions containing metal nitrate (10 mg/L) and NaCl (100 mg/L) can reach 99%. Even the removal efficiency of heavy metal ions can be close to 99% in the multi-component solution containing all the seven metal nitrates (10 mg/L for each) and 100 mg/L NaCl. Meanwhile, the electro-adsorption and electro-reaction coupling process maintained excellent regeneration ability even after 20 cycles. Furthermore, the heavy metal ions removal mechanism was proven to be the pseudocapacitive intercalation of heavy metal ions into the layered structure of the employed W18O49/graphene in the electro-adsorption and electro-reaction coupling process. This work demonstrates great potential for general applicability to wastewater treatment.


Minimal cut sets in biochemical reaction networks.
Structural studies of metabolic networks yield deeper insight into topology, functionality and capabilities of the metabolisms of different organisms. Here, we address the analysis of potential failure modes in metabolic networks whose occurrence will render the network structurally incapable of performing certain functions. Such studies will help to identify crucial parts in the network structure and to find suitable targets for repressing undesired metabolic functions. We introduce the concept of minimal cut sets for biochemical networks. A minimal cut set (MCS) is a minimal (irreducible) set of reactions in the network whose inactivation will definitely lead to a failure in certain network functions. We present an algorithm which enables the computation of the MCSs in a given network related to user-defined objective reactions. This algorithm operates on elementary modes. A number of potential applications are outlined, including network verifications, phenotype predictions, assessing structural robustness and fragility, metabolic flux analysis and target identification in drug discovery. Applications are illustrated by the MCSs in the central metabolism of Escherichia coli for growth on different substrates. Computation and analysis of MCSs is an additional feature of the FluxAnalyzer (freely available for academic users upon request, special contracts for industrial companies; see web page below). http://www.mpi-magdeburg.mpg.de/projects/fluxanalyzer


Thoracic duct cannulation of the rhesus monkey (Macaca mulatta) for lymphocyte collection: thoracic approach.
The thoracic duct of the rhesus monkey (Macaca mulatta) was cannulated with medical grade tubing through a thoracatomy at right interspace 9-10. In 42 monkeys successfully cannulated, a mean of 1.194 x 10(9) lymphocytes was collected in the 70-hour period after surgery. A second collection, 1-3 weeks later, was possible for a shorter period (up to 60 hours) in a portion of these monkeys (15) with a smaller mean yield of lymphocytes (1.076 x 10(9). A third, but still shorter, collection period of up to 48 hours was possible in only five monkeys from which a mean yield of 0.698 x 10(9) lymphocytes was collected. A temporary postoperative weight loss in the monkeys was followed by a return to normal weight several months later.


[Health education on alcohol. The necessity to abandon a purely quantitative message].
In health education, the foundations of the classic message of moderate alcohol consumption, based on the notion that there exist safe thresholds below which one can drink without danger, have become outdated. This is due to various factors: the discovery of additional routes for the oxidation of alcohol into acetaldehyde; epidemiological studies on the relationship between dose and effect that have challenged classic thresholds; and, finally, the rising emphasis on the role played by background, now better understood because of the opening of new physiological perspectives and evidence of the involvement of genetic factors. This role has led to an underlining of the notions of vulnerability and inequality in relation to alcohol. The message for health education must therefore be one of "alcohol risk", a message combining both the factor of quantity ingested and the factor of background. Without giving rise to prohibitionist attitudes, it should lead to responsible individual behavior where alcohol is concerned.


Shapes, Plasmonic Properties, and Reactivity of Magnesium Nanoparticles.
Localized surface plasmon resonances have attracted much attention due to their ability to enhance light-matter interactions and manipulate light at the subwavelength level. Recently, alternatives to the rare and expensive noble metals Ag and Au have been sought for more sustainable and large-scale plasmonic utilization. Mg supports plasmon resonances, is one of the most abundant elements in earth's crust, and is fully biocompatible, making it an attractive framework for plasmonics. This feature article first reports the hexagonal, folded, and kite-like shapes expected theoretically from a modified Wulff construction for single crystal and twinned Mg structures and describes their excellent match with experimental results. Then, the optical response of Mg nanoparticles is overviewed, highlighting Mg's ability to sustain localized surface plasmon resonances across the ultraviolet, visible, and near-infrared electromagnetic ranges. The various resonant modes of hexagons, leading to the highly localized electric field characteristic of plasmonic behavior, are presented numerically and experimentally. The evolution of these modes and the associated field from hexagons to the lower symmetry folded structures is then probed, again by matching simulations, optical, and electron spectroscopy data. Lastly, results demonstrating the opportunities and challenges related to the high chemical reactivity of Mg are discussed, including surface oxide formation and galvanic replacement as a synthetic tool for bimetallics. This Feature Article concludes with a summary of the next steps, open questions, and future directions in the field of Mg nanoplasmonics.


Frequency of COL4A3/COL4A4 mutations amongst families segregating glomerular microscopic hematuria and evidence for activation of the unfolded protein response. Focal and segmental glomerulosclerosis is a frequent development during ageing.
Familial glomerular hematuria(s) comprise a genetically heterogeneous group of conditions which include Alport Syndrome (AS) and thin basement membrane nephropathy (TBMN). Here we investigated 57 Greek-Cypriot families presenting glomerular microscopic hematuria (GMH), with or without proteinuria or chronic kidney function decline, but excluded classical AS. We specifically searched the COL4A3/A4 genes and identified 8 heterozygous mutations in 16 families (28,1%). Eight non-related families featured the founder mutation COL4A3-p.(G1334E). Renal biopsies from 8 patients showed TBMN and focal segmental glomerulosclerosis (FSGS). Ten patients (11.5%) reached end-stage kidney disease (ESKD) at ages ranging from 37-69-yo (mean 50,1-yo). Next generation sequencing of the patients who progressed to ESKD failed to reveal a second mutation in any of the COL4A3/A4/A5 genes, supporting that true heterozygosity for COL4A3/A4 mutations predisposes to CRF/ESKD. Although this could be viewed as a milder and late-onset form of autosomal dominant AS, we had no evidence of ultrastructural features or extrarenal manifestations that would justify this diagnosis. Functional studies in cultured podocytes transfected with wild type or mutant COL4A3 chains showed retention of mutant collagens and differential activation of the unfolded protein response (UPR) cascade. This signifies the potential role of the UPR cascade in modulating the final phenotype in patients with collagen IV nephropathies.


Efficient multi-gene expression in cell-free droplet microreactors.
Cell-free transcription and translation systems promise to accelerate and simplify the engineering of proteins, biological circuits and metabolic pathways. Their encapsulation on microfluidic platforms can generate millions of cell-free reactions in picoliter volume droplets. However, current methods struggle to create DNA diversity between droplets while also reaching sufficient protein expression levels. In particular, efficient multi-gene expression has remained elusive. We here demonstrate that co-encapsulation of DNA-coated beads with a defined cell-free system allows high protein expression while also supporting genetic diversity between individual droplets. We optimize DNA loading on commercially available microbeads through direct binding as well as through the sequential coupling of up to three genes via a solid-phase Golden Gate assembly or BxB1 integrase-based recombineering. Encapsulation with an off-the-shelf microfluidics device allows for single or multiple protein expression from a single DNA-coated bead per 14 pL droplet. We envision that this approach will help to scale up and parallelize the rapid prototyping of more complex biological systems.


Avoiding painful blood sampling in neonates by transcutaneous bilirubinometry.
Transcutaneous bilirubinometry, Bilicheck (kindly provided by Oxygen Care Company, Dublin) is a non-invasive, screening method for determining bilirubin levels in neonates. It has not previously been validated in Irish babies. To determine the clinical accuracy and precision of Bilicheck measurements in comparison to total serum bilirubin (TSB) measurements in Irish neonates, and to determine whether Bilicheck is a useful non-invasive screening method of avoiding blood tests in neonates. Correlation data were obtained from 53 neonates between simultaneous TSB and Bilicheck readings. Bilicheck was used to screen 100 neonates who were assessed as requiring serum bilirubin measurement. TSB was only performed on neonates whose Bilicheck was above the phototherapy line of a recognised serum bilirubin chart. Bilicheck and TSB were significantly correlated (r=0.890). Bland and Altman analysis showed that on average, Bilicheck read 30 micromol/l lower than TSB. Of 100 jaundiced neonates screened by Bilicheck, blood sampling was avoided in 70 because Bilicheck did not meet phototherapy guidelines. There is a good correlation between TSB and Bilicheck, although the latter tends to under-read by 30 micromol/l. Bilicheck may be a useful screening device to decrease the risks and discomfort associated with blood sampling in neonates.


Specific inhibition of human immunodeficiency virus type 1 replication by antisense oligonucleotides: an in vitro model for treatment.
We have developed a culture system, simulating in vivo conditions of human immunodeficiency virus type 1 (HIV-1) infection, to evaluate the long-term efficacy of antisense oligonucleotide treatment. Five oligonucleotide phosphorothioates (28-mers), complementary to different regions of HIV-1 RNA, blocked replication of the virus in a sequence-specific manner at 1 microM concentration. Variations in antiviral activity were seen among the different oligonucleotides, revealing an effect of target selection. Mismatched or random oligonucleotide phosphorothioates delayed, but did not completely inhibit, HIV-1 replication. In the case of inhibition by a splice-acceptor-site antisense oligodeoxynucleotide, a break-through phenomenon occurred after 25 days of treatment, suggesting the development of an "escape mutant." This result did not occur when the inhibitory oligodeoxynucleotides were complementary to the primary-sequence areas of the rev-responsive element and rev-1 genes. Sequential treatment of HIV-1-infected cells with a combination of different antisense oligonucleotides, each administered once, also prevented the development of escape mutants. Our results suggest that chemotherapy based on specifically targeted antisense-oligonucleotide phosphorothioates may be an effective method for reducing the viral burden in HIV-1-infected individuals at clinically achievable oligonucleotide concentrations.


Implementation of increased physical therapy intensity for improving walking after stroke: Walk 'n watch protocol for a multisite stepped-wedge cluster-randomized controlled trial.
Clinical practice guidelines support structured, progressive protocols for improving walking after stroke. Yet, practice is slow to change, evidenced by the little amount of walking activity in stroke rehabilitation units. Our recent study (n = 75) found that a structured, progressive protocol integrated with typical daily physical therapy improved walking and quality-of-life measures over usual care. Research therapists progressed the intensity of exercise by using heart rate and step counters worn by the participants with stroke during therapy. To have the greatest impact, our next step is to undertake an implementation trial to change practice across stroke units where we enable the entire unit to use the protocol as part of standard of care. What is the effect of introducing structured, progressive exercise (termed the Walk 'n Watch protocol) to the standard of care on the primary outcome of walking in adult participants with stroke over the hospital inpatient rehabilitation period? Secondary outcomes will be evaluated and include quality of life. This national, multisite clinical trial will randomize 12 sites using a stepped-wedge design where each site will be randomized to deliver Usual Care initially for 4, 8, 12, or 16 months (three sites for each duration). Then, each site will switch to the Walk 'n Watch phase for the remaining duration of a total 20-month enrolment period. Each participant will be exposed to either Usual Care or Walk 'n Watch. The trial will enroll a total of 195 participants with stroke to achieve a power of 80% with a Type I error rate of 5%, allowing for 20% dropout. Participants will be medically stable adults post-stroke and able to take five steps with a maximum physical assistance from one therapist. The Walk 'n Watch protocol focuses on completing a minimum of 30 min of weight-bearing, walking-related activities (at the physical therapists' discretion) that progressively increase in intensity informed by activity trackers measuring heart rate and step number. The primary outcome will be the change in walking endurance, measured by the 6-Minute Walk Test, from baseline (T1) to 4 weeks (T2). This change will be compared across Usual Care and Walk 'n Watch phases using a linear mixed-effects model. Additional physical, cognitive, and quality of life outcomes will be measured at T1, T2, and 12 months post-stroke (T3) by a blinded assessor. The implementation of stepped-wedge cluster-randomized trial enables the protocol to be tested under real-world conditions, involving all clinicians on the unit. It will result in all sites and all clinicians on the unit to gain expertise in protocol delivery. Hence, a deliberate outcome of the trial is facilitating changes in best practice to improve outcomes for participants with stroke in the trial and for the many participants with stroke admitted after the trial ends.


Mining a cathepsin inhibitor library for new antiparasitic drug leads.
The targeting of parasite cysteine proteases with small molecules is emerging as a possible approach to treat tropical parasitic diseases such as sleeping sickness, Chagas' disease, and malaria. The homology of parasite cysteine proteases to the human cathepsins suggests that inhibitors originally developed for the latter may be a source of promising lead compounds for the former. We describe here the screening of a unique ∼ 2,100-member cathepsin inhibitor library against five parasite cysteine proteases thought to be relevant in tropical parasitic diseases. Compounds active against parasite enzymes were subsequently screened against cultured Plasmodium falciparum, Trypanosoma brucei brucei and/or Trypanosoma cruzi parasites and evaluated for cytotoxicity to mammalian cells. The end products of this effort include the identification of sub-micromolar cell-active leads as well as the elucidation of structure-activity trends that can guide further optimization efforts.


A QTL approach in faba bean highlights the conservation of genetic control of frost tolerance among legume species.
Frost is a major abiotic stress of winter type faba beans (Vica faba L.) and has adverse effects on crop yield. Climate change, far from reducing the incidence of frost events, is making these phenomena more and more common, severe, and prolonged. Despite the important interaction that the environment has in the tolerance of faba bean to frost, this trait seems to have good levels of heritability. Several QTLs for frost tolerance have already been reported, however, a more robust identification is needed to more precisely identify the genomic regions involved in faba bean tolerance to sub-zero temperatures. Several pea (Pisum sativum L.) and barrel medic (Medicago truncatula L.) frost tolerance QTLs appear to be conserved between these two species, furthering the hypothesis that the genetic control of frost tolerance in legume species might be more generally conserved. In this work, the QTL mapping in two faba bean recombinant inbred line (RIL) populations connected by a common winter-type parent has led to the identification of five genomic regions involved in the control of frost tolerance on linkage groups I, III, IV, and V. Among them, a major and robust QTL of great interest for marker-assisted selection was identified on the lower part of the long-arm of LGI. The synteny between the faba bean frost tolerance QTLs and those previously identified in other legume species such as barrel medic, pea or soybean highlighted at least partial conservation of the genetic control of frost tolerance among different faba bean genetic pools and legume species. Four novel RILs showing high and stable levels of tolerance and the ability to recover from freezing temperatures by accumulating frost tolerance QTLs are now available for breeding programs.


High sensitivity ultraviolet detection based on three-dimensional graphene field effect transistors decorated with TiO2 NPs.
A three-dimensional (3D) ultraviolet (UV) photodetector was fabricated by decorating a tubular graphene field-effect transistor (GFET) with titanium dioxide (TiO2) nanoparticles (NPs). The unique tubular architecture not only provides a natural 3D optical resonant microcavity to enhance the optical field inside it, but also increases the light-matter interaction area. Strong UV absorption in the TiO2 NPs creates a number of electron-hole pairs, where the electrons are transferred to graphene, while the holes are trapped within the TiO2 NPs, leading to a strong photogating effect on the graphene channel conductance. The photoresponsivity of our 3D GFET photodetector decorated with TiO2 NPs was demonstrated up to 475.5 A W-1 at 325 nm, which is about 2 orders of magnitude higher than that of a 3D GFET photodetector without the TiO2 NP decoration (1 A W-1), and over 3 orders of magnitude higher than that of a recently reported UV photodetector based on the graphene/vertical Ga2O3 nanowire array heterojunction (0.185 A W-1). Moreover, the photoresponsivity and photoresponse speed of the device can be easily tuned by applying a small gate bias (≤3 V) and/or changing the source-drain bias. These results indicate that the photoresponsivities of graphene-based photodetectors can be significantly improved by exploiting 3D graphene structures and integrating graphene with semiconducting light harvesters simultaneously.


Drug treatment of pruritus in liver diseases.
Pruritus (itch) is a common complication of cholestatic liver diseases (CLD). It can be a distressing and debilitating symptom, causing significant impairment in quality of life. Treatment of pruritus in liver diseases can be challenging and requires specific management with early initiation and a step-wise approach using specific drugs. Clinical trials are ongoing with novel agents that demonstrate potential efficacy. Patients with cholestatic pruritus are likely to present to a variety of clinicians who would benefit from medical awareness of available treatment options. In this review, we outline the pharmaceutical agents currently used to treat cholestatic pruritus and provide the evidence base for targeted symptom control of itch in liver diseases. We also highlight recent developments in the pathophysiology of cholestatic pruritus and the emerging novel therapies.


Modified interface between pediatric ventilator and Ayre's T-piece for use in neonates.
The lowest tidal volume delivered by most of the pediatric anesthesia ventilators could be large for neonates, and may generate unacceptably high peak airway pressures. The effective tidal volume can however be reduced by connection of a small rubber bag between the ventilator and the expiratory limb of the Ayre's T-piece circuit to divide the delivered gas mixture between the added bag and the neonate's lungs. The delivered tidal volume by the ventilator can then be manipulated to achieve the desired peak inspiratory pressure; 18-20 cm H2O in neonates with normal lung compliance. The modification was found to be simple, safe and convenient.


The experience of secrecy.
The concept of secrecy calls to mind a dyadic interaction: one person hiding a secret from another during a conversation or social interaction. The current work, however, demonstrates that this aspect of secrecy is rather rare. Taking a broader view of secrecy as the intent to conceal information, which only sometimes necessitates concealment, yields a new psychology of secrecy. Ten studies demonstrate the secrets people have, what it is like to have a secret, and what about secrecy is related to lower well-being. We demonstrate that people catch themselves spontaneously thinking about their secrets-they mind-wander to them-far more frequently than they encounter social situations that require active concealment of those secrets. Moreover, independent of concealment frequency, the frequency of mind-wandering to secrets predicts lower well-being (whereas the converse was not the case). We explore the diversity of secrets people have and the harmful effects of spontaneously thinking about those secrets in both recall tasks and in longitudinal designs, analyzing more than 13,000 secrets across our participant samples, with outcomes for relationship satisfaction, authenticity, well-being, and physical health. These results demonstrate that secrecy can be studied by having people think about their secrets, and have implications for designing interventions to help people cope with secrecy. (PsycINFO Database Record


Wocania indica gen. nov., sp. nov., isolated from deep sea hydrothermal sulfide in the northwest Indian Ocean, and proposal to reclassify Flaviramulus ichthyoenteri as Wocania ichthyoenteri comb. nov.
A novel Gram-stain-negative, non-motile, strictly aerobic, rod-shaped bacterium was isolated from deep-sea hydrothermal sulfide in the northwest Indian Ocean Ridge and designated as strain IOP_32T. Strain IOP_32T could grow at 4-40 °C (optimum, 37 °C), pH 5-9 (optimum, pH 7-8) and salinity of 0-12 % NaCl (w/v; optimum, 2-3 %). The 16S rRNA gene sequence of strain IOP_32T is most similar to Bizionia fulviae EM7T, Bizionia berychis RA3-3-1T, Bizionia paragorgiae KMM 6029T and Oceanihabitans sediminis S9_10T with 95.5-95.3 % similarity. The phylogenomic analysis indicated that strain IOP_32T forms a distinct lineage with Flaviramulus ichthyoenteri Th78T within the family Flavobacteriaceae. The average nucleotide identity, average amino acid identity and percentage of conserved protein values between strain IOP_32T and the type strains of close genera were 72.3-78.5 %, 67.4-76.9 % and 56.3-61.6 %, respectively. The major cellular fatty acid was anteiso-C15 : 0. The respiratory quinone was MK-6. The polar lipids were mainly composed of phosphatidylethanolamine, an unidentified aminophospholipid and five unidentified polar lipids. Strain IOP_32T is significantly different from related genera, which is reflected by the wide adaptability to temperature and salinity levels, the composition of phospholipids and fatty acids, and the results of phylogenetic analyses. The phenotypic properties and phylogenetic data suggest that the lineage represents a novel genus and species within the family Flavobacteriaceae, for which the name Wocania indica gen. sp. nov. is proposed, with the type strain IOP_32T (=MCCC 1A14017 T=KCTC 62660 T). We also propose the reclassification of Flaviramulus ichthyoenteri as Wocania ichthyoenteri comb. nov. (Th78T=DSM 26285T=JCM 18634T=KCTC 32142T).


Effects of photoperiod on food intake, activity and metabolic rate in adult neutered male cats.
With the continued rise in feline obesity, novel weight management strategies are needed. To date, strategies aimed at altering physical activity, an important factor in weight maintenance, have been lacking. Photoperiod is known to cause physiological changes in seasonal mammals, including changes in body weight (BW) and reproductive status. Thus, our objective was to determine the effect of increased photoperiod (longer days) on voluntary physical activity levels, resting metabolic rate (RMR), food intake required to maintain BW, and fasting serum leptin and ghrelin concentrations in adult cats. Eleven healthy, adult, neutered, male domestic shorthair cats were used in a randomized crossover design study. During two 12-week periods, cats were exposed to either a short-day (SD) photoperiod of 8 h light: 16 h dark or a long-day (LD) photoperiod of 16 h light: 8 h dark. Cats were fed a commercial diet to maintain baseline BW. In addition to daily food intake and twice-weekly BW, RMR (via indirect calorimetry), body composition [via dual-energy X-ray absorptiometry (DEXA)] and physical activity (via Actical activity monitors) were measured at week 0 and 12 of each period. Fasting serum leptin and ghrelin concentrations were measured at week 0, 6 and 12 of each period. Average hourly physical activity was greater (p = 0.008) in LD vs. SD cats (3770 vs. 3129 activity counts/h), which was primarily due to increased (p < 0.001) dark period activity (1188 vs. 710 activity counts/h). This corresponded to higher (p < 0.0001) daily metabolizable energy intake (mean over 12-week period: 196 vs. 187 kcal/day), and increased (p = 0.048) RMR in LD cats (9.02 vs. 8.37 kcal/h). Body composition, serum leptin and serum ghrelin were not altered by photoperiod. More research is needed to determine potential mechanisms by which these physiological changes occurred and how they may apply to weight management strategies.


C3d Assay in Correlation With Single Antigen Bead Assay for Human Leukocyte Antigen Antibodies.
Single antigen bead assay (SAB) is a sensitive method for detecting HLA antibodies, but it does not specifically identify clinically relevant subsets. Recently, a new assay has been developed for detection of C3d bound to HLA antibody-antigen complex. We evaluated the C3d assay regarding its correlation with SAB in renal patients. A total of 138 serum samples from 109 sensitized patients were tested in parallel by SAB and C3d assay for detection of HLA class I antibodies. The relationship between C3d assay and SAB was analyzed for the numbers and median fluorescent intensity (MFI) values of the identified antibodies. Of the 138 samples, 137 were positive on SAB; of the 137 SAB-positive samples, 76 were positive on C3d assay. A total of 3748 and 685 antibodies were identified by the SAB and the C3d assay, respectively. The maximal MFI values of the SAB in the 76 samples that were C3d assay-positive were significantly higher than those of the 61 samples that were C3d assay-negative (P < .05), with the median values of 17,057 and 6066, respectively. Only 11 (0.4%) of the 2905 antibodies with MFI < 10,000 on SAB vs 501 (59.4%) of the 843 antibodies with MFI > 10,000 on SAB were identified by C3d assay with MFI > 1000. The C3d assay positivity seems to be dependent on its MFI value on SAB. Further studies are needed to ascertain the clinical significance of C3d positivity by itself.


Human dirofilariasis due to Dirofilaria repens in Ukraine, an emergent zoonosis: epidemiological report of 1465 cases.
The filarial nematode Dirofilaria repens is currently considered to be one of the most extensively spreading human and animal parasites in Europe. In Ukraine, reporting cases of dirofilariasis has been mandatory since 1975, and the disease was included in the national surveillance system for notifiable diseases. Up until December 31st 2012, a total of 1533 cases have been registered, with 1465 cases occurring within the previous 16 years. Most of the cases of dirofilariasis were registered in 6 regions: Kyiv, and the Donetsk, Zaporizhzhya, Dnipropetrovsk, Kherson and Chernihiv oblasts. In the years 1997-2002 the highest incidence rate was noted in the Kherson oblast in the south of the country (9.79 per 100 000 people), and the lowest in western Ukraine (0.07-1.68 per 100 000 people). D. repens infections were registered in all oblasts. Parasitic lesions were most often located in the head, the subconjunctival tissue and around the eyes. D. repens lesions were also found in the limbs, torso, male sexual organs, and female mammary glands. Dirofilariasis was diagnosed in persons aged from 11 months to 90 years old, most often among people between 21-40 years of age. Most patients had only one parasitic skin lesion; the majority of isolated nematodes were female. The results of our analysis point to a constant increase in D. repens dirofilariasis incidence in humans in Ukraine. Despite educational efforts, infections have become more frequent and the territory in which the disease occurs has enlarged to encompass the whole of Ukraine. Nevertheless, the Ukrainian sanitary-epidemiological services managed to achieve some measure of success, e.g. by creating a registration system for D. repens infections and establishing proper diagnostics for the disease.


Decrease in nuclear phospholipids associated with DNA replication.
Lipid metabolism in nuclei is very active and appears involved in the transduction of signals to the genome in response to agonists acting at the plasma membrane level. However, the precise topology of nuclear lipid metabolism and the relationship between nuclear lipids and crucial events of the cell function, such as DNA replication, have not been fully elucidated. By using a recently developed cytochemical method for detecting phospholipids inside the nucleus of intact cells at the electron microscope level, we have analyzed the changes in intranuclear phospholipids in DNA-replicating versus resting cells, which are both present in the same sample of regenerating liver after partial hepatectomy. The pattern of DNA synthesis in replicating cells has been monitored by electron microscope immunocytochemistry after bromodeoxyuridine (BrdU) labeling. The data obtained, which allow a fine localization and a quantitative analysis of both DNA synthesis and phospholipid distribution, indicate a significant reduction in the phospholipids detectable inside the nucleus in all steps of the S phase. This could depend on an increased nuclear phospholipid hydrolysis, whose products should in turn activate some of the enzymes involved in the control of DNA replication.


Alteration of sarcoplasmic reticulum after denervation of chicken pectoralis muscle.
To determine the neural influence on the function of the sarcoplasmic reticulum (SR) of fast-twitch skeletal muscle, the superior pectoralis muscle of adult chicken was denervated, and the SR was isolated at 20 days post-denervation. The isolated SR was probably derived from the longitudinal SR and was relatively free of contaminants. The protein profile of the SR was quantitatively changed after denervation with an increase in the M55 and 30000-mol.wt. proteins relative to the Ca2+-ATPase. Ca2+-dependent ATPase activity and phosphoenzyme formation were lower in the denervated-muscle SR; however, the enzyme catalytic-centre activity was similar to the control value. The decrease in Ca2+-ATPase activity in denervated-muscle SR was accompanied by a lower Ca2+ accumulation so that the relationship between Ca2+ accumulation and Ca2+-dependent ATPase activity was well maintained in the SR from denervated muscle. The data imply that denervation may result in a diminution of functional Ca2+ pump sites. Evidence is presented, though, which suggests that denervation affects a single class of Ca2+-binding sites of the Ca2+-ATPase, resulting in a lower affinity for Ca2+.


Degree of host susceptibility in the initial disease outbreak influences subsequent epidemic spread.
Disease epidemics typically begin as an outbreak of a relatively small, spatially explicit population of infected individuals (focus), in which disease prevalence increases and rapidly spreads into the uninfected, at-risk population. Studies of epidemic spread typically address factors influencing disease spread through the at-risk population, but the initial outbreak may strongly influence spread of the subsequent epidemic.We initiated wheat stripe rust Puccinia striiformis f. sp. tritici epidemics to assess the influence of the focus on final disease prevalence when the degree of disease susceptibility differed between the at-risk and focus populations.When the focus/at-risk plantings consisted of partially genetic resistant and susceptible cultivars, final disease prevalence was statistically indistinguishable from epidemics produced by the focus cultivar in monoculture. In these experimental epidemics, disease prevalence was not influenced by the transition into an at-risk population that differed in disease susceptibility. Instead, the focus appeared to exert a dominant influence on the subsequent epidemic.Final disease prevalence was not consistently attributable to either the focus or the at-risk population when focus/at-risk populations were planted in a factorial set-up with a mixture (~28% susceptible and 72% resistant) and susceptible individuals. In these experimental epidemics, spatial heterogeneity in disease susceptibility within the at-risk population appeared to counter the dominant influence of the focus.Cessation of spore production from the focus (through fungicide/glyphosate application) after 1.3 generations of stripe rust spread did not reduce final disease prevalence, indicating that the focus influence on disease spread is established early in the epidemic.Synthesis and applications. Our experiments indicated that outbreak conditions can be highly influential on epidemic spread, even when disease resistance in the at-risk population is greater than that of the focus. Disease control treatments administered shortly after the initial outbreak within the focus may either prevent an epidemic from occurring or reduce its severity.


Miniature probe combining optical-resolution photoacoustic microscopy and optical coherence tomography for in vivo microcirculation study.
Photoacoustic microscopy (PAM) is sensitive to optical absorption, while optical coherence tomography (OCT) is based on optical backscattering. Combining PAM and OCT can provide complementary information about biological tissue. Here we present a combined optical-resolution PAM (ORPAM) and OCT system that is integrated through a miniature probe with an overall diameter of 2.3 mm, suitable for insertion through a standard endoscopic or laparoscopic port during minimally invasive surgery or endoscopic exam. The hybrid probe consists of a common optical path for OCT (light delivery/detection) and ORPAM (light excitation) and a 10 MHz unfocused ultrasound transducer for photoacoustic detection. The combined system yields a lateral resolution of 15 μm for both ORPAM and OCT.


[The physiopathology of retinal circulation: consequences of acute retinal vascular occlusion].
A constant oxygen supply from the retinal and choroidal vascular system is essential for the maintenance of the retinal electrical activity. Arterial or venous retinal obstruction modifies the hemodynamics in the retinal circulation and produces hypoxia in the affected area. In the case of arterial obstruction, severe hypoxia (anoxia) rapidly (in less than 45 min) produces irreversible cell damage in the inner area of the retina. With venous obstruction, the hypoxia is moderate and the retinal damage is partial. In some cases, hypoxia regresses because the formation of collateral vessels most probably provides adequate perfusion. In cases of irreversible venous obstruction, insufficient formation of collateral vessels results in the development of ischemic regions in the retina and severe hypoxia. In these cases preretinal neovascularization may develop.


Oryzisolibacter propanilivorax gen. nov., sp. nov., a propanil-degrading bacterium.
Strain EPL6T, a Gram-negative, motile, short rod was isolated from a propanil and 3,4-dichloroaniline enrichment culture produced from rice paddy soil. Based on the analyses of the 16S rRNA gene sequence, strain EPL6T was observed to be a member of the family Comamonadaceae, sharing the highest pairwise identity with type strains of the species Alicycliphilus denitrificans K601T (96.8 %) and Melaminivora alkalimesophila CY1T (96.8 %). Strain EPL6T was able to grow in a temperature range of 15-37 °C, pH 6-9 and in the presence of up to 4 % (w/v) NaCl and tested positive for catalase and oxidase reactions. The major respiratory quinone was Q8. The genomic DNA had a G+C content of 69.4±0.9 mol%. The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol, and the major fatty acid methyl esters comprised C16 : 0, C18 : 1ω7c and summed feature 3 (C16 : 1ω7c/iso-C15 : 0 2-OH). Comparison of the genome sequence of strain EPL6T and of its closest neighbours, Melaminivora alkalimesophila CY1T and Alicycliphilus denitrificans K601T, yielded values of ANI ≤84.1 % and of AAI ≤80.3 %. Therefore, the genetic, phylogenetic, phenotypic and chemotaxonomic characteristics support the classification of this organism into a new taxon. Considering the genetic divergence of strain EPL6T from the type strains of the closest species, which belong to distinct genera, we propose a new genus within the family Comamonadaceae, named Oryzisolibacter propanilivorax gen. nov., sp. nov., represented by the isolate EPL6T as the type strain of the species (=LMG 28427T=CECT 8927T).


Nebulized Heparin Attenuates Pulmonary Coagulopathy and Inflammation through Alveolar Macrophages in a Rat Model of Acute Lung Injury.
Objective Alveolar macrophages play a key role in the development and resolution of acute respiratory distress syndrome (ARDS), modulating the inflammatory response and the coagulation cascade in lungs. Anti-coagulants may be helpful in the treatment of ARDS. This study investigated the effects of nebulized heparin on the role of alveolar macrophages in limiting lung coagulation and inflammatory response in an animal model of acute lung injury (ALI). Methods Rats were randomized to four experimental groups. In three groups, ALI was induced by intratracheal instillation of lipopolysaccharide (LPS) and heparin was nebulized at constant oxygen flow: the LPS/Hep group received nebulized heparin 4 and 8 hours after injury; the Hep/LPS/Hep group received nebulized heparin 30 minutes before and 4 and 8 hours after LPS-induced injury; the LPS/Sal group received nebulized saline 4 and 8 hours after injury. The control group received only saline. Animals were exsanguinated 24 hours after LPS instillation. Lung tissue, bronchoalveolar lavage fluid (BALF) and alveolar macrophages isolated from BALF were analysed. Results LPS increased protein concentration, oedema and neutrophils in BALF as well as procoagulant and proinflammatory mediators in lung tissue and alveolar macrophages. In lung tissue, nebulized heparin attenuated ALI through decreasing procoagulant (tissue factor, thrombin-anti-thrombin complexes, fibrin degradation products) and proinflammatory (interleukin 6, tumour necrosis factor alpha) pathways. In alveolar macrophages, nebulized heparin reduced expression of procoagulant genes and the effectors of transforming growth factor beta (Smad 2, Smad 3) and nuclear factor kappa B (p-selectin, CCL-2). Pre-treatment resulted in more pronounced attenuation. Conclusion Nebulized heparin reduced pulmonary coagulopathy and inflammation without producing systemic bleeding, partly by modulating alveolar macrophages.


Tumor evolution in space: the effects of competition colonization tradeoffs on tumor invasion dynamics.
We apply competition colonization tradeoff models to tumor growth and invasion dynamics to explore the hypothesis that varying selection forces will result in predictable phenotypic differences in cells at the tumor invasive front compared to those in the core. Spatially, ecologically, and evolutionarily explicit partial differential equation models of tumor growth confirm that spatial invasion produces selection pressure for motile phenotypes. The effects of the invasive phenotype on normal adjacent tissue determine the patterns of growth and phenotype distribution. If tumor cells do not destroy their environment, colonizer and competitive phenotypes coexist with the former localized at the invasion front and the latter, to the tumor interior. If tumors cells do destroy their environment, then cell motility is strongly selected resulting in accelerated invasion speed with time. Our results suggest that the widely observed genetic heterogeneity within cancers may not be the stochastic effect of random mutations. Rather, it may be the consequence of predictable variations in environmental selection forces and corresponding phenotypic adaptations.


Maturation of parvovirus LuIII in a subcellular system. II. Isolation and characterization of nucleoprotein intermediates.
Nucleoprotein (NP) intermediates in the assembly pathway of parvovirus LuIII were isolated. These structures consist of replicating viral DNA and of preformed viral capsids into which progeny viral DNA is encapsidated concomitant with synthesis. The NP complexes sediment at between 70S and 100S. They are unstable in CsCl and dissociate in the presence of 1 M-NaCl. After fixation with glutaraldehyde, however, they accumulate at a density of 1.37 g/ml in CsCl gradients. Electron microscopy of fixed complexes revealed structures consisting of DNA threads associated with capsid-like particles. A considerable proportion of the DNA labelled during short pulses in such complexes, in a subcellular in vitro system derived from infected nuclei, can be effectively chased into 110S virions. The resulting 110S particles apparently have to undergo a sequence of morphogenic events to acquire the physicochemical properties of the mature infectious parvovirus.


Characterization of the Salmonella paratyphi C Vi polysaccharide.
The Vi capsular polysaccharide (Vi) is both a virulence factor and a protective antigen of Salmonella typhi; its pathogenic role for Salmonella paratyphi C is less well understood. We found no differences between the antigenic and immunogenic properties and the structure of the Vi from representative strains of S. paratyphi C, S. typhi, and Citrobacter freundii. There were, however, differences in both the amount produced per cell and the degree of association with the cell among the Vi from the three species of Enterobacteriaceae. S. paratyphi C produced less Vi than both the wild-type S. typhi and C. freundii did, and it showed the fastest release of Vi into the media. These findings may provide an explanation for the inability of the Vi to inhibit completely the agglutination of S. paratyphi C by anti-O sera. In an outbreak of enteric fever caused by S. paratyphi C, 66 of 78 isolates (85%) were Vi positive.


Patterns of alcohol, tobacco and cannabis use and related harm in city, regional and remote areas of Australia.
Prevalence of substance use and related harms differs across geographic locations, with prevalence increasing with remoteness. This paper builds on previous research, investigating patterns of problematic use by remoteness. Analysis of the National Drug Strategy Household Survey (NDSHS) data from 2007 by geographic location (in accordance with the Australian Standard Geographical Classification (ASGC) remoteness index). Problematic cannabis use was predicted by demographics; older males with less education were more likely to report problematic cannabis use regardless of location. Younger, less educated males in inner regional and remote areas were more likely to report risky alcohol consumption for short term harms, while older, less educated males in outer regional and remote areas were more likely to report daily drinking. People from remote areas were significantly more likely to report driving under the influence of alcohol. It is clear that men with lower levels of education were significantly more likely to report problematic alcohol and cannabis use patterns. An additional level of risk is associated with living in inner regional and remote areas, particularly in relation to risky drinking. Findings suggest a complex relationship between remoteness and substance use which requires further investigation.


Low-threshold, persistent sodium current in rat large dorsal root ganglion neurons in culture.
Dorsal root ganglion neurons from adult rats (> or = 200 g) were maintained in culture for between 1 and 3 days. Membrane currents generated by large neurons (50-75 microns apparent diameter) were recorded with the whole cell patch-clamp technique. Large neurons generated transient Na+ currents and at least two types of inward current that persisted throughout 200-ms voltage-clamp steps to +20 mV. One persistent current activated close to -35 mV (high threshold), whereas in about half of the cells another persistent current began to activate negative to -70 mV (low threshold). The high-threshold persistent current was identified as a Ca2+ current, as previously described in these neurons. The low-threshold current was reversibly suppressed either by replacing external Na+ with tetramethylammonium ions or by reducing external Na+ concentration ([Na+]) and simultaneously raising external [Ca2+]. It was blocked by tetrodotoxin (TTX) with an apparent equilibrium dissociation constant in the single nanomolar range. We conclude that the low-threshold current is a TTX-sensitive, persistent Na+ current. The persistent TTX-sensitive current contributed to steady-state membrane current from at least -70 mV to 0 mV, a wider potential range than predicted by activation-inactivation gating overlap for transient Na+ current. Because of its low threshold and fast activation kinetics, the persistent Na+ current is expected to play an important role in determining membrane excitability.


Population genetic evidence for complex evolutionary histories of four high altitude juniper species in the Qinghai-Tibetan Plateau.
Population genetics data based on multiple nuclear loci provide invaluable information to understand demographic, selective, and divergence histories of the current species. We studied nucleotide variation at 13 nuclear loci in 53 populations distributed among four closely related, but morphologically distinct juniper species of the Qinghai-Tibetan Plateau (QTP). We used a novel approach combining Approximate Bayesian Computation and a recently developed neutrality test based on the maximum frequency of derived mutations to examine the demographic and selective histories of individual species, and isolation-with-migration analyses to study the joint history of the species and detect gene flow between them. We found that (1) the four species, which diverged in response to the extensive QTP uplifts, have different demographic histories; (2) two loci, Pgi and CC0822, depart significantly from neutrality in one species and Pgi, is also marginally significant in another; and (3) shared polymorphisms are common, indicating both incomplete lineage sorting and gene flow after species divergence. In addition, the detected unidirectional gene flow provides indirect support for the theoretical prediction that introgression should mostly take place from local to invading species. Our results, together with previous studies, underscore complex evolutionary histories of plant diversification in the biodiversity-hotspot QTP.


Comparison of quantitative imaging devices and subjective optic nerve head assessment by general ophthalmologists to differentiate normal from glaucomatous eyes.
To compare the ability of subjective assessment of optic nerve head (ONH) and retinal nerve fiber layer (RNFL) by general ophthalmologists and by a glaucoma expert with objective measurements by optical coherence tomography (Stratus OCT, Carl Zeiss Meditec Inc), confocal scanning laser ophthalmoscope (HRT III; Heidelberg Engineering, Heidelberg, Germany), and scanning laser polarimetry (GDx enhanced corneal compensation; Carl Zeiss Meditec Inc, Dublin, CA) in discriminating glaucomatous and normal eyes. Sixty-one glaucomatous and 57 normal eyes of 118 subjects were included in the study. Three independent general ophthalmologists and 1 glaucoma expert evaluated ONH stereophotographs. Receiver operating characteristic curves were constructed for each imaging technique and sensitivity at fixed specificity was estimated. Comparisons of areas under these curves (aROCs) and agreement (k) were determined between stereophoto grading and best parameter from each technique. Best parameter from each technique showed larger aROC (Stratus OCT RNFL=0.92; Stratus OCT ONH vertical integrated area=0.86; Stratus OCT macular thickness=0.82; GDx enhanced corneal compensation=0.91; HRT3 global cup-to-disc area ratio=0.83; HRT3 glaucoma probability score numeric score=0.83) compared with stereophotograph grading by general ophthalmologists (0.80) in separating glaucomatous and normal eyes. Glaucoma expert stereophoto grading provided equal or larger aROC (0.92) than best parameter of each computerized imaging device. Stereophoto evaluated by a glaucoma expert showed better agreement with best parameter of each quantitative imaging technique in classifying eyes either as glaucomatous or normal compared with stereophoto grading by general ophthalmologists. The combination of subjective assessment of the optic disc by general ophthalmologists with RNFL objective parameters improved identification of glaucoma patients in a larger proportion than the combination of these objective parameters with subjective assessment of the optic disc by a glaucoma expert (29.5% vs. 19.7%, respectively). Diagnostic ability of all imaging techniques showed better performance than subjective assessment of the ONH by general ophthalmologists, but not by a glaucoma expert. Objective RNFL measurements may provide improvement in glaucoma detection when combined with subjective assessment of the optic disc by general ophthalmologists or by a glaucoma expert.


Etiologic factors in infantile asthma.
In a group of 83 infants with asthma the major etiologic factors were found to be foods, environmental inhalants and respiratory infections. Airborne pollens and molds were found to be important in a small number of patients during the latter part of infancy. IgA deficiency and general anesthesia under the age of two years played a less well defined role in the pathogenesis of infantile asthma.


Lactate dehydrogenase C is required for the protein expression of a sperm-specific isoform of lactate dehydrogenase A.
Metabolites are sensitive indicators of moment-to-moment cellular status and activity. Expecting that tissue-specific metabolic signatures unveil a unique function of the tissue, we examined metabolomes of mouse liver and testis and found that an unusual metabolite, 2-hydroxyglutarate (2-HG), was abundantly accumulated in the testis. 2-HG can exist as D- or L-enantiomer, and both enantiomers interfere with the activities of 2-oxoglutarate (2-OG)-dependent dioxygenases, such as the Jumonji family of histone demethylases. Whereas D-2-HG is produced by oncogenic mutants of isocitrate dehydrogenases (IDH) and known as an oncometabolite, L-2-HG was the major enantiomer detected in the testis, suggesting that a distinct mechanism underlies the testicular production of this metabolite. We clarified that lactate dehydrogenase C (LDHC), a testis-specific lactate dehydrogenase, is responsible for L-2-HG accumulation by generating and analysing Ldhc-deficient mice. Although the inhibitory effects of 2-HG on 2-OG-dependent dioxygenases were barely observed in the testis, the LDHA protein level was remarkably decreased in Ldhc-deficient sperm, indicating that LDHC is required for LDHA expression in the sperm. This unique functional interaction between LDH family members supports lactate dehydrogenase activity in the sperm. The severely impaired motility of Ldhc-deficient sperm suggests a substantial contribution of glycolysis to energy production for sperm motility.


Follistatin secretion is enhanced by protein, but not glucose or fat ingestion, in obese persons independently of previous gastric bypass surgery.
Follistatin is secreted from the liver and is involved in the regulation of muscle mass and insulin sensitivity via inhibition of activin A in humans. The secretion of follistatin seems to be stimulated by glucagon and inhibited by insulin, but only limited knowledge on the postprandial regulation of follistatin exists. Moreover, results on postoperative changes after Roux-en-Y gastric bypass (RYGB) are conflicting with reports of increased, unaltered, and lowered fasting concentrations of follistatin. In this study, we investigated postprandial follistatin and activin A concentrations after intake of isocaloric amounts of protein, fat, or glucose in subjects with obesity with and without previous RYGB to explore the regulation of follistatin by the individual macronutrients. Protein intake enhanced follistatin concentrations similarly in the two groups, whereas glucose and fat ingestion did not change postprandial follistatin concentrations. Concentrations of activin A were lower after protein intake compared with glucose intake in RYGB. Glucagon concentrations were also particularly enhanced by protein intake and tended to correlate with follistatin in RYGB. In conclusion, we demonstrated that protein intake, but not glucose or fat, is a strong stimulus for follistatin secretion in subjects with obesity and that this regulation is maintained after RYGB surgery.NEW & NOTEWORTHY Circulating follistatin and activin A were studied after intake of isocaloric protein, fat, or glucose drinks in subjects with obesity with and without previous Roux-en-Y gastric bypass (RYGB). Protein intake enhanced follistatin similarly in both groups, whereas glucose and fat ingestion did not change follistatin. Activin A was lower after protein compared with glucose in RYGB. The novel finding is that protein intake, but neither glucose nor fat, stimulates follistatin secretion independently of previous RYGB.


Narrowing of the dorsal vein complex technique during laparoscopic radical prostatectomy: a simple trick to simplify the control of venous plexus.
The control of the Dorsal Venous Complex (DVC) is crucial to the recovery of urinary continence during Laparoscopic Radical Prostatectomy (LRP). The size of DVC may affect the venous control. We developed a trick to simplify the suturing of the DVC. Forty-seven patients with localized prostate cancer were divided in two groups: group 1 (n = 24) underwent LRP with a conventional ligature of DVC, and in group 2 (n = 23) the venous control was done with "Narrowing" of DVC technique (N-DVC). Our technique involves maintaining pressure on a metallic urethral sound inserted into the urethra, just at the time of ligature. The width of DVC in group 2 was measured before and after applying the technique. The numbers of attempts to place the stitch adequately were recorded and compared in both groups. The demographic dates, perioperative dates and results were compared retrospectively. Operative time, estimated blood loss, prostate weight, positive surgical margins rates and potency results showed no significant differences between the groups. The immediate 1-month, and 3-month continence rates were significantly greater in group 2 (30.4% vs. 12.5%, P = .048; 73.9% vs. 50%, P = .037, respectively). For all patients in group 2, width of DVC decreased and the ligation stitch was effective at the first attempt. In 37.5% of patients in group 1, the controlling of the DVC was obtained in more than one attempt. The N-DVC simplifies the control of DVC during LRP and may contribute to the early recovery of continence.


Spontaneous oscillation and fluid-structure interaction of cilia.
The exact mechanism to orchestrate the action of hundreds of dynein motor proteins to generate wave-like ciliary beating remains puzzling and has fascinated many scientists. We present a 3D model of a cilium and the simulation of its beating in a fluid environment. The model cilium obeys a simple geometric constraint that arises naturally from the microscopic structure of a real cilium. This constraint allows us to determine the whole 3D structure at any instant in terms of the configuration of a single space curve. The tensions of active links, which model the dynein motor proteins, follow a postulated dynamical law, and together with the passive elasticity of microtubules, this dynamical law is responsible for the ciliary motions. In particular, our postulated tension dynamics lead to the instability of a symmetrical steady state, in which the cilium is straight and its active links are under equal tensions. The result of this instability is a stable, wave-like, limit cycle oscillation. We have also investigated the fluid-structure interaction of cilia using the immersed boundary (IB) method. In this setting, we see not only coordination within a single cilium but also, coordinated motion, in which multiple cilia in an array organize their beating to pump fluid, in particular by breaking phase synchronization.


The Effect of Educational Program Based on BASNEF Model for Eye Care in Non-insulin Dependent Diabetic Patients.
The purpose of this study was to determine the effects of educational program based on the BASNEF (Belief, Attitude, Subjective Norm, and Enabling Factors) Model on eye care among patients with insulin independent diabetes mellitus in Shiraz City, Fars Province, Iran. We enrolled 100 patients with non-insulin independent diabetes mellitus (Type II) fulfilling the inclusion criteria of this experimental study. The participants were randomly divided into two groups, one experimental, and one control group. All groups completed the questionnaires based on the BASNEF Model, a checklist related to patient's practice including patients' HbA1c and FBS levels, the pre-test results of an ophthalmologist's eyes examination and the results of three months follow-up. The experimental group participated in eight educational sessions during the interventional. The data analysis used including chi square-test, t-test, and ANOVA. The knowledge and all BASNEF Model components were significantly increased in the experimental group compared to the control group after intervention. In addition, behavioral eye care, such as physical activities, regular taking medicine, having eye examination, FBS checking, having appropriate diet, HbA1c level , and fasting blood sugar levels improved significantly among the experimental group compared to the control group. Applying the BASNEF Model is very effective for developing an educational program for diabetic patients, in order to control their blood sugar and enhancing behavioral eye care. Besides such programs, follow up education on controlling and monitoring is highly recommended.


Pharmacology and legal status of cannabidiol.
Cannabidiol (CBD) is the second most abundant cannabinoid present in Cannabis sativa L. It is not associated with psychotropic activity and is capable to mitigate the psychotomimetic effects produced by tetrahydrocannabinol (THC). The latest cannabis decriminalization policies and the high applicability in therapeutic and technologic-industrial fields, have determined an exponential marketing growth of foods, cosmetics and in particularly medicinal products containing CBD, which are easily available for consumers. Most importantly, on 2018 United States Food and Drug Administration approved CBD oral solution with the trade name of Epidiolex® for the treatment of two rare and severe forms of epilepsy, "Lennox-Gastaut syndrome" and "Dravet syndrome", in pediatric patients. The aim of this review was to focus on pharmacology and on legal status of CBD, to highlight the lack of harmonization of international regulatory laws over the marketing authorization of CBD-based products.


Plasmid integration, amplification and cytogenetics in CHO cells: questions and comments.
This paper addresses seven questions with respect to the transfer, integration, amplification and genetic stability of recombinant DNA in mammalian genomes. Most of these questions were raised with those issues in mind which are frequently discussed in the context of the manufacture of biologicals of therapeutic value on the basis of recombinant cell lines. For the most part, a high degree of ignorance has to be acknowledged and only very limited fragments of information are available. The reason for this ignorance is, as will become clear from this article, the sheer size and complexity of the mammalian genome and the inadequacy of presently available tools to unravel this complexity.


Effect of diethyl ether and ethanol as an oxygenated additive on Calophyllum inophyllum biodiesel in CI engine.
The present experimental investigation is conducted to examine the working characteristics of compression ignition (CI) engine using oxygenated additives such as diethyl ether and ethanol to the blends of Calophyllum inophyllum biodiesel. Experiments are conducted on a water-cooled single-cylinder constant speed DI (direct injection) diesel engine under same operating circumstances. The results indicates that an enhancement of brake thermal efficiency is up to 3.7% and 6.2% with an addition of ethanol and DEE additive in biodiesel blended fuels respectively at maximum load condition. This is attributable to the existence of higher oxygen value and volatile nature of the additives. The combustion characteristics like in-cylinder pressure and net heat release rate of DEE blended fuels are 1.1-5.2% and 0.4-2.7% higher than other blends respectively. This is mainly due to higher cetane number and high volatile nature of DEE. This also results in higher NOX emissions. But 2-9% drop in nitrous oxide emission is noticed for biodiesel with ethanol blends compared to all other blends. This is as a result of superior latent heat of vaporization and lesser cetane number of ethanol which results in lower cylinder pressure about 0.4-2.6% and this end up with more unburned HC and CO emissions. Energy and exergy studies help to analyze the result of ethanol and DEE additive along with blends of biodiesel. The exergy efficiencies of DEE blended fuels enhance up to 2.3-6% compared to other blends at all load conditions. This is attributable to the superior combustion characteristics of DEE.


Assessing the targeting and fate of cathepsin k antibody-modified nanoparticles in a rat abdominal aortic aneurysm model.
Abdominal aortic aneurysms (AAAs), a prototypic proteolytic cardiovascular disorder, are localized expansions of the aortal wall. Chronically upregulated and overexpressed proteases irreversibly degrade and disrupt the elastic matrix, which provides stretch and recoil properties to the aortal wall. Adult vascular smooth muscle cells are inherently unable to produce sufficient elastin to form new elastic fibers to naturally repair the aortal wall and the AAA continues to grow until fatal rupture. Surgical intervention is reserved for AAAs with a high risk of rupture, but there is currently no treatment for small, still growing AAAs. We have previously developed matrix regenerative PEG-PLGA nanoparticles (NPs) with pro-elastogenic and anti-proteolytic properties that act synergistically with a released therapeutic. However, strategies are required to effectively deliver these NPs to the disease site to avail of these benefits. We have identified cathepsin K, a protease overexpressed in AAA tissue, as a potential substrate for antibody based active targeting. We sought to assess the safety and biocompatibility of NPs with anti-cathepsin K antibodies conjugated to the NP surface (cat K Ab-NPs) and then assess their biodistribution and retention in both the targeted aorta and non-target organs in a rat AAA model. In this work, we show that cat K Ab-NPs can selectively target the aneurysmal aorta in a rat AAA model. However, there is unwanted NP uptake and retention in non-target organs that can be addressed in future work. Still, cathepsin K is a viable target for active delivery of NPs in an AAA model. STATEMENT OF SIGNIFICANCE: We have previously developed elastic matrix regenerative polymer nanoparticles (NPs), but require strategies to efficiently target the disease site. Antibodies against cathepsin K, an overexpressed protease in abdominal aortic aneurysms, have been conjugated to the NP surface to act as a targeting moiety. In this work, we assessed NP safety and in vivo biodistribution in an aneurysmal rat model and demonstrated positive targeting and retention for up to 2 weeks within the aortal wall.


Neuromechanical factors involved in the formation and propulsion of fecal pellets in the guinea-pig colon.
The neuromechanical processes involved in the formation and propulsion of fecal pellets remain incompletely understood. We analyzed motor patterns in isolated segments of the guinea-pig proximal and distal colon, using video imaging, during oral infusion of liquid, viscous material, or solid pellets. Colonic migrating motor complexes (CMMCs) in the proximal colon divided liquid or natural semisolid contents into elongated shallow boluses. At the colonic flexure these boluses were formed into shorter, pellet-shaped boluses. In the non-distended distal colon, spontaneous CMMCs produced small dilations. Both high- and low-viscosity infusions evoked a distinct motor pattern that produced pellet-shaped boluses. These were propelled at speeds proportional to their surface area. Solid pellets were propelled at a speed that increased with diameter, to a maximum that matched the diameter of natural pellets. Pellet speed was reduced by increasing resistive load. Tetrodotoxin blocked all propulsion. Hexamethonium blocked normal motor patterns, leaving irregular propagating contractions, indicating the existence of neural pathways that did not require nicotinic transmission. Colonic migrating motor complexes are responsible for the slow propulsion of the soft fecal content in the proximal colon, while the formation of pellets at the colonic flexure involves a content-dependent mechanism in combination with content-independent spontaneous CMMCs. Bolus size and consistency affects propulsion speed suggesting that propulsion is not a simple reflex but rather a more complex process involving an adaptable neuromechanical loop.


Percutaneously placed inferior vena cava filter for the prevention of pulmonary emboli: a plea for early insertion.
After surgery or trauma, thrombosis of a lower-extremity vein can lead to potentially fatal pulmonary embolization. Although anticoagulation has traditionally been the preferred method of preventing emboli, this approach is often ineffective or contraindicated. In such cases, partial or complete interruption of the inferior vena cava with a specially designed filter can prevent emboli from reaching the pulmonary circulation. The author reviews the history of vena caval interruption and describes his own experience with this technique, which has the potential for becoming the treatment of choice in the prevention of pulmonary emboli.


PREVALENCE OF LAWSONIA INTRACELLULARIS INFECTION IN NONHUMAN PRIMATES AND PEST RODENTS IN A ZOOLOGICAL COLLECTION.
In 2016 and 2017, Lawsonia intracellularis was isolated from several pileated gibbons (Hylobates pileatus) presenting with diarrhea in Mulhouse Zoo (eastern France). To this day, infection with this bacterium has rarely been described in nonhuman primates (NHP) in captivity or in the wild and there are no data about the prevalence or transmission of the disease. This study focuses on finding the prevalence of this infection amongst Mulhouse Zoo's NHP collection and trying to identify a source of contamination responsible for this epizooty. Forty-eight real-time PCR were conducted on feces from all NHP species in the zoo and on small mammals trapped in the NHP housing structures. No NHP was experiencing symptoms at the time of the study, however test results showed that Lawsonia intracellularis can be found in 61.76% (21/34) of the group total (n = 34) and the prevalence even increases to 92.3% (12/13) in the Lemuriform infraorder (n = 13). In small mammals (n = 14), prevalence of the bacterium is 57.17% (8/14) including 77.78% in rodents (7/9). The results of this study show that several NHP species are healthy carriers and some species of small mammals can be considered as a potential source of contamination. Because of the difficulty encountered trying to isolate the bacterium, it is plausible that infections caused by Lawsonia intracellularis have been underdiagnosed to this day, and that it could be an emerging disease in Europe. Therefore, using real-time PCR to search for this bacterium seems essential in case of diarrhea occurring in nonhuman primates. Moreover, even though further studies on contamination sources need to be conducted, the issue of the presence of rodents in NHP housing structures has to be taken very seriously and tackled with the utmost care.


Structure of the Macrobrachium rosenbergii nodavirus: A new genus within the Nodaviridae?
Macrobrachium rosenbergii nodavirus (MrNV) is a pathogen of freshwater prawns that poses a threat to food security and causes significant economic losses in the aquaculture industries of many developing nations. A detailed understanding of the MrNV virion structure will inform the development of strategies to control outbreaks. The MrNV capsid has also been engineered to display heterologous antigens, and thus knowledge of its atomic resolution structure will benefit efforts to develop tools based on this platform. Here, we present an atomic-resolution model of the MrNV capsid protein (CP), calculated by cryogenic electron microscopy (cryoEM) of MrNV virus-like particles (VLPs) produced in insect cells, and three-dimensional (3D) image reconstruction at 3.3 Å resolution. CryoEM of MrNV virions purified from infected freshwater prawn post-larvae yielded a 6.6 Å resolution structure, confirming the biological relevance of the VLP structure. Our data revealed that unlike other known nodavirus structures, which have been shown to assemble capsids having trimeric spikes, MrNV assembles a T = 3 capsid with dimeric spikes. We also found a number of surprising similarities between the MrNV capsid structure and that of the Tombusviridae: 1) an extensive network of N-terminal arms (NTAs) lines the capsid interior, forming long-range interactions to lace together asymmetric units; 2) the capsid shell is stabilised by 3 pairs of Ca2+ ions in each asymmetric unit; 3) the protruding spike domain exhibits a very similar fold to that seen in the spikes of the tombusviruses. These structural similarities raise questions concerning the taxonomic classification of MrNV.


Biomimetic Architectured Graphene Aerogel with Exceptional Strength and Resilience.
Materials combining lightweight, robust mechanical performances, and multifunctionality are highly desirable for engineering applications. Graphene aerogels have emerged as attractive candidates. Despite recent progresses, the bottleneck remains how to simultaneously achieve both strength and resilience. While multiscale architecture designs may offer a possible route, the difficulty lies in the lack of design guidelines and how to experimentally achieve the necessary structure control over multiple length scales. The latter is even more challenging when manufacturing scalability is taken into account. The Thalia dealbata stem is a naturally porous material that is lightweight, strong, and resilient, owing to its architecture with three-dimensional (3D) interconnected lamellar layers. Inspired by such, we assemble graphene oxide (GO) sheets into a similar architecture using a bidirectional freezing technique. Subsequent freeze-drying and thermal reduction results in graphene aerogels with highly tunable 3D architectures, consequently an unusual combination of strength and resilience. With their additional electrical conductivity, these graphene aerogels are potentially useful for mechanically switchable electronics. Beyond such, our study establishes bidirectional freezing as a general method to achieve multiscale architectural control in a scalable manner that can be extended to many other material systems.


Bone Marrow Stimulation for Osteochondral Lesions of the Talus: Are Clinical Outcomes Maintained 10 Years Later?
Arthroscopic bone marrow stimulation (BMS) is considered the first-line treatment for osteochondral lesions of the talus (OLTs). However, the long-term stability of the clinical success of BMS remains unclear. To investigate the long-term clinical outcomes among patients who underwent BMS for OLT and to identify prognostic factors for the need for revision surgery. Case series; Level of evidence, 4. A retrospective analysis was performed on 202 ankles (189 patients) that were treated with BMS for OLT and had a minimum follow-up of 10 years. The visual analog scale for pain, American Orthopaedic Foot & Ankle Society ankle-hindfoot score, and the Foot and Ankle Outcome Score (FAOS) were assessed by repeated measures analysis of variance. Prognostic factors associated with revision surgery were evaluated with Cox proportional hazard regression models and log-rank tests. The mean lesion size was 105.32 mm2 (range, 19.75-322.79); 42 ankles (20.8%) had large lesions (≥150 mm2). The mean visual analog scale for pain improved from 7.11 ± 1.73 (mean ± SD) preoperatively to 1.44 ± 1.52, 1.46 ± 1.57, and 1.99 ± 1.67 at 1, 3 to 6, and ≥10 years, respectively, after BMS (P < .001). The mean ankle-hindfoot score also improved, from 58.22 ± 13.57 preoperatively to 86.88 ± 10.61, 86.17 ± 10.23, and 82.76 ± 11.65 at 1, 3 to 6, and ≥10 years after BMS (P < .001). The FAOS at the final follow-up was 82.97 ± 13.95 for pain, 81.81 ± 14.64 for symptoms, 83.49 ± 11.04 for activities of daily living, 79.34 ± 11.61 for sports, and 78.71 ± 12.42 for quality of life. Twelve ankles underwent revision surgery after a mean 53.5 months. Significant prognostic factors associated with revision surgery were the size of the lesion (preoperative magnetic resonance imaging measurement ≥150 mm2; P = .014) and obesity (body mass index ≥25; P = .009). BMS for OLT yields satisfactory clinical outcomes at a mean follow-up of 13.9 years. The success of the surgery may depend on the lesion size and body mass index of the patient.


Biventricular repair for double outlet right ventricle with non-committed ventricular septal defect.
Outcomes of biventricular repair for patients with double outlet right ventricle and non-committed ventricular septal defect (DORVncVSD) are not well defined. We aim to report our experience with biventricular repair of this anomaly in proposing an original surgical management that simplifies the anatomical correction. From January 2005 to December 2013, 75 consecutive patients with DORVncVSD who had undergone biventricular repair in our institution were retrospectively included. The patients were divided into 2 groups: 40 patients in Group A had the ventricular septal defect rerouted to the aorta, and 35 patients in Group B had the ventricular septal defect rerouted to the pulmonary artery. Concomitant tricuspid procedures, conal resection and ventricular septal defect enlargement were used to favour intracardiac tunnel geometry. Five types of biventricular repair and 16 concomitant procedures were performed. Mean age at biventricular repair was 2.2 ± 2.0 years. There were 6 (8.0%) early deaths and 4 (6.1%) early intracardiac baffle obstructions. During the 4.1 ± 4.0 years follow-up, 3 (4.3%) late deaths occurred with an 87.1% estimated overall survival at 5 years (early deaths included). Six late-onset intracardiac tunnel obstructions were noted and three of them required reoperation. Comparing the 2 groups, Group A patients have more late-onset (6 in Group A vs 0 in Group B, P = 0.026) and overall tunnel obstructions (10 in Group A vs 0 in Group B, P = 0.001). Concomitant tricuspid procedures significantly reduced intracardiac obstruction (0 in 16 vs 10 in 24, P = 0.003) without development of any tricuspid regurgitation and stenosis. Using appropriate intracardiac tunnel establishment strategy and techniques, biventricular repair of DORVncVSD is feasible with encouraging outcomes. Concomitant tricuspid procedures can reduce intracardiac tunnel geometry without increase of mortality and morbidity.


Analysis of 109 Japanese children's lip and nose shapes using 3-dimensional digitizer.
We assessed lip and nose shapes, which played an important role in performance evaluations, before and after cleft lip and nose surgery. We used a noncontact-type semiconductor laser 3-dimensional measurement system on normal Japanese children to obtain 3-dimensional images of noses and lips, which were accurate enough to measure facial shapes. We could rotate these images on the computer, therefore we measured the following points: the distance between the peaks of the Cupid's bow and the width of the labial fissure (frontal view), and the width of the nose and the nasal tip protrusion (basal view). Lip and nose shapes were evaluated for each gender. Additionally, nasolabial angles (NLA) were measured on the lateral views of faces. We classified the morphology of the philtral columns into four types; (1) triangular type, (2) parallel type, (3) concave type, and (4) flat type. We also classified nostril shapes into four types: (1) teardrop type, (2) heart shaped type, (3) round type, and (4) triangular type. We calculated the average of the 3-dimensional coordinate values for each landmark, and created standard facial models of normal Japanese children. Moreover, we invented a new morphological evaluation method before and after cleft lip and nose surgery, using the 3-dimensional data converting and editing software. The method was more feasible to evaluate the assessment cleft lip and nose surgery, by quantifying the surface areas of right and left nostrils and the surface areas of upper and lower vermilions, even now by measuring with the eyes and comparing them.