Large tree classifier with heuristic search and global training.
In the tree classifier with top-down search, a global decision is made via a series of local decisions. Although this approach gains in classification efficiency, it also gives rise to error accumulation which can be very harmful when the number of classes is very large. To overcome this difficulty, a new tree classifier with the following characteristics is proposed: 1) fuzzy logic search is used to find all ``possible correct classes,'' and some similarity measures are used to determine the ``most probable class''; 2) global training is applied to generate extended terminals in order to enhance the recognition rate; 3) both the training and search algorithms have been given a lot of flexibility, to provide tradeoffs between error and rejection rates, and between the recognition rate and speed. A computer simulation of the decision trees for the recognition of 3200 Chinese character categories yielded a very high recognition rate of 99.93 percent and a very high speed of 861 samples/s, when the program was written in a high level language and run on a large multiuser time-sharing computer.


Assessing the performance of MM/PBSA and MM/GBSA methods. 7. Entropy effects on the performance of end-point binding free energy calculation approaches.
Entropy effects play an important role in drug-target interactions, but the entropic contribution to ligand-binding affinity is often neglected by end-point binding free energy calculation methods, such as MM/GBSA and MM/PBSA, due to the expensive computational cost of normal mode analysis (NMA). Here, we systematically investigated entropy effects on the prediction power of MM/GBSA and MM/PBSA using >1500 protein-ligand systems and six representative AMBER force fields. Two computationally efficient methods, including NMA based on truncated structures and the interaction entropy approach, were used to estimate the entropic contributions to ligand-target binding free energies. In terms of the overall accuracy, we found that, for the minimized structures, in most cases the inclusion of the conformational entropies predicted by truncated NMA (enthalpynmode_min_9Å) compromises the overall accuracy of MM/GBSA and MM/PBSA compared with the enthalpies calculated based on the minimized structures (enthalpymin). However, for the MD trajectories, the binding free energies can be improved by the inclusion of the conformation entropies predicted by either truncated-NMA for a relatively high dielectric constant (εin = 4) or the interaction entropy method for εin = 1-4. In terms of reproducing the absolute binding free energies, the binding free energies estimated by including the truncated-NMA entropies based on the MD trajectories (ΔGnmode_md_9Å) give the lowest average absolute deviations against the experimental data among all the tested strategies for both MM/GBSA and MM/PBSA. Although the inclusion of the truncated NMA based on the MD trajectories (ΔGnmode_md_9Å) for a relatively high dielectric constant gave the overall best result and the lowest average absolute deviations against the experimental data (for the ff03 force field), it needs too much computational time. Alternatively, considering that the interaction entropy method does not incur any additional computational cost and can give comparable (at high dielectric constant, εin = 4) or even better (at low dielectric constant, εin = 1-2) results than the truncated-NMA entropy (ΔGnmode_md_9Å), the interaction entropy approach is recommended to estimate the entropic component for MM/GBSA and MM/PBSA based on MD trajectories, especially for a diverse dataset. Furthermore, we compared the predictions of MM/GBSA with six different AMBER force fields. The results show that the ff03 force field (ff03 for proteins and gaff with AM1-BCC charges for ligands) performs the best, but the predictions given by the tested force fields are comparable, implying that the MM/GBSA predictions are not very sensitive to force fields.


Active Life Expectancy In The Older US Population, 1982-2011: Differences Between Blacks And Whites Persisted.
Understanding long-range trends in longevity and disability is useful for projecting the likely impact of the baby-boom generation on long-term care utilization and spending. We examine changes in active life expectancy in the United States from 1982 to 2011 for white and black adults ages sixty-five and older. For whites, longevity increased, disability was postponed to older ages, the locus of care shifted from nursing facilities to community settings, and the proportion of life at older ages spent without disability increased. In contrast, for blacks, longevity increases were accompanied by smaller postponements in disability, and the percentage of remaining life spent active remained stable and well below that of whites. Older black women were especially disadvantaged in 2011 in terms of the proportion of years expected to be lived without disability. Public health measures directed at older black adults-particularly women-are needed to offset impending pressures on the long-term care delivery system as the result of population aging.


Specific differences in migratory function of myofibroblasts isolated from Crohn's disease fistulae and strictures.
Recently we found that migration of colonic lamina propria fibroblasts in Crohn's disease patients (CD-CLPF) from inflamed mucosa is significantly reduced as compared to control-CLPF. The behavior of CD-CLPFs isolated from fistulae and strictures was now investigated in detail. Initially migration assays for all CLPF cultures (CD-CLPF, fibrosis-CLPF, and fistula-CLPF) were performed in the modified 48-well Boyden chamber. Subsequently, for a migration assay more resembling the in vivo situation a 3D matrix model was developed. After seeding of cells into the 3D matrix the CLPF layer was wounded by an ERBIUM:YAG laser leading to circular cell rupture without effect on the extracellular matrix. In the modified Boyden chamber migration of fistula-CLPF was significantly reduced compared to CD-CLPF. This was correlated with a decrease in FAK-protein expression, whereas in migrating fibrosis-CLPF an increase in FAK-protein expression, -autophosphorylation and migratory potential was found. This was confirmed in the 3D matrix wounding assay: Fistula-CLPF migrated less than CD-CLPF, whereas fibrosis-CLPF migrated significantly more in the 3D matrix wounding assay. Between 1 to 36 hours incubation time fibrosis-CLPF always displayed increased migration ability as compared to CD-CLPF. In contrast, fistula-CLPF migratory potential was always below that of CD-CLPF. Myofibroblasts isolated from inflamed, fibrostenotic, or fistulized CD mucosa differ in their migratory potential both in the modified Boyden chamber as well as in a 3D matrix model. These different migratory behaviors could be an explanation for impaired or excess wound healing and subsequently for fistula and fibrosis formation.


Reducing human exposure to aflatoxin through the use of clay: a review.
Innovative sorption strategies for the detoxification of aflatoxins have been developed. NovaSil clay (NS) has been shown to prevent aflatoxicosis in a variety of animals when included in their diet. Results have shown that NS clay binds aflatoxins with high affinity and high capacity in the gastrointestinal tract, resulting in a notable reduction in the bioavailability of these toxins without interfering with the utilization of vitamins and other micronutrients. This strategy is being evaluated as a potential remedy for acute aflatoxicosis, and as a sustainable human intervention for aflatoxins via the diet. Phase I and II clinical trials confirmed the apparent safety of NS for further study in humans. A recent study in Ghanaians at high risk for aflatoxicosis has indicated that NS (at a dose level of 0.25%) is effective in decreasing biomarkers of aflatoxin exposure and does not interfere with the levels of serum vitamins A and E, and iron and zinc. In summary, enterosorption strategies/therapies based on NS clay are promising for the management of aflatoxins and as a sustainable public health intervention. The NS clay remedy is novel, inexpensive and easily disseminated. Based on the present research, aflatoxin sequestering clays should be rigorously evaluated in vitro and in vivo, and should meet the following criteria: (1) favourable thermodynamic characteristics of mycotoxin sorption, (2) tolerable levels of priority metals, dioxins/furans and other hazardous contaminants, (3) safety and efficacy in multiple animal species, (4) safety and efficacy in long-term studies, and (5) negligible interactions with vitamins, iron and zinc and other micronutrients.


Diabetes does not influence activation of coagulation, fibrinolysis or anticoagulant pathways in Gram-negative sepsis (melioidosis).
Diabetes is associated with a disturbance of the haemostatic balance and is an important risk factor for sepsis, but the influence of diabetes on the pathogenesis of sepsis remains unclear. Melioidosis ( Burkholderia pseudomallei infection) is a common cause of community-acquired sepsis in Southeast Asia and northern Australia. We sought to investigate the impact of pre-existing diabetes on the coagulation and fibrinolytic systems during sepsis caused by B.pseudomallei . We recruited a cohort of 44 patients (34 with diabetes and 10 without diabetes) with culture-proven melioidosis. Diabetes was defined as a pre-admission diagnosis of diabetes or an HbA₁c>7.8% at enrolment. Thirty healthy blood donors and 52 otherwise healthy diabetes patients served as controls. Citrated plasma was collected from all subjects; additionally in melioidosis patients follow-up specimens were collected seven and ≥ 28 days after enrolment where possible. Relative to uninfected healthy controls, diabetes per se (i.e. in the absence of infection) was characterised by a procoagulant effect. Melioidosis was associated with activation of coagulation (thrombin-antithrombin complexes (TAT), prothrombin fragment F₁+₂ and fibrinogen concentrations were elevated; PT and PTT prolonged), suppression of anti-coagulation (antithrombin, protein C, total and free protein S levels were depressed) and abnormalities of fibrinolysis (D-dimer and plasmin-antiplasmin complex [PAP] were elevated). Remarkably, none of these haemostatic alterations were influenced by pre-existing diabetes. In conclusion, although diabetes is associated with multiple abnormalities of coagulation, anticoagulation and fibrinolysis, these changes are not detectable when superimposed on the background of larger abnormalities attributable to B. pseudomallei sepsis.


The impact of single versus mixed schistosome species infections on liver, spleen and bladder morbidity within Malian children pre- and post-praziquantel treatment.
In the developing world co-infections and polyparasitism within humans appear to be the rule rather than the exception, be it any combination of inter-specific and/or inter- and intra-Genera mixed infections. Mixed infections might generate synergistic or antagonistic interactions and thereby clinically affect individuals and/or impact parasite epidemiology. The current study uniquely assesses both Schistosoma mansoni- and Schistosoma haematobium-related morbidity of the liver and the bladder as assessed by ultrasound as well as spleen and liver morbidity through clinical exams. The impact of praziquantel (PZQ) treatment on such potential inter-specific schistosome interactions and resulting morbidity using uniquely detailed longitudinal data (pre- and one year post-PZQ treatment) arising from the National Schistosomiasis Control Program in three areas of Mali: Ségou, Koulikoro and Bamako, is also evaluated. At baseline, data were collected from up to 2196 children (aged 7-14 years), 844 of which were infected with S. haematobium only, 124 with S. mansoni only and 477 with both. Follow-up data were collected from up to 1265 children. Results suggested lower liver morbidity in mixed compared to single S. mansoni infections and higher bladder morbidity in mixed compared to single S. haematobium infections. Single S. haematobium or S. mansoni infections were also associated with liver and spleen morbidity whilst only single S. haematobium infections were associated with bladder morbidity in these children (light S. haematobium infection OR: 4.3, p < 0.001 and heavy S. haematobium infection OR: 19, p < 0.001). PZQ treatment contributed to the regression of some of the forms of such morbidities. Whilst the precise biological mechanisms for these observations remain to be ascertained, the results illustrate the importance of considering mixed species infections in any analyses of parasite-induced morbidity, including that for the proposed Disability Adjusted Life Years (DALYs) revised estimates of schistosomiasis morbidity.


Gastric cancer: Classification, histology and application of molecular pathology.
Gastric cancer remains one of the deadly diseases with poor prognosis. New classification of gastric cancers based on histologic features, genotypes and molecular phenotypes helps better understand the characteristics of each subtype, and improve early diagnosis, prevention and treatment. The objective of this article is to review the new classification of gastric cancers and the up-to-date guidance in the application of molecular testing.


The organisation of an ophthalmological service for diabetics in a teaching hospital.
A new system of ophthalmological evaluation of diabetic patients in a teaching hospital has been devised. All the patients attending the diabetic clinic were screened by ophthalmologists. Any patient with a problem was then referred to a diabetic ophthalmology clinic where a full evaluation was done and treatment given where necessary. In an 18-month period 1,015 new ophthalmological diabetic patients were screened. Of these patients 13.6% had background retinopathy, 8.4% pre-proliferative retinopathy and 3.4% proliferative retinopathy, while 8.6% had maculopathy. These results show that even in a teaching hospital a significant percentage of patients already have pre-proliferative or even proliferative retinopathy when seen by an ophthalmologist for the first time. The importance of organising an ophthalmological service for the diabetic patient is stressed.


Interventions for acute stroke management in Africa: a systematic review of the evidence.
The past decades have witnessed a rapid evolution of research on evidence-based acute stroke care interventions worldwide. Nonetheless, the evidence-to-practice gap in acute stroke care remains variable with slow and inconsistent uptake in low-middle income countries (LMICs). This review aims to identify and compare evidence-based acute stroke management interventions with alternative care on overall patient mortality and morbidity outcomes, functional independence, and length of hospital stay across Africa. This review was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guideline. An electronic search was conducted in six databases comprising MEDLINE, Embase, Cumulative Index to Nursing and Allied Health Literature (CINAHL), Web of Science, Academic Search Complete and Cochrane Library for experimental and non-experimental studies. Eligible studies were abstracted into evidence tables and their methodological quality appraised using the Joanna Briggs Institute checklist. Data were analysed and presented narratively with reference to observed differences in patient outcomes, reporting p values and confidence intervals for any possible relationship. Initially, 1896 articles were identified and 37 fully screened. Four non-experimental studies (three cohort and one case series studies) were included in the final review. One study focused on the clinical efficacy of a stroke unit whilst the remaining three reported on thrombolytic therapy. The results demonstrated a reduction in patient deaths attributed to stroke unit care and thrombolytic therapy. Thrombolytic therapy was also associated with reductions in symptomatic intracerebral haemorrhage (SICH). However, the limited eligible studies and methodological limitations compromised definitive conclusions on the extent of and level of efficacy of evidence-based acute stroke care interventions across Africa. Evidence from this review confirms the widespread assertion of low applicability and uptake of evidence-based acute stroke care in LMICs. Despite the limited eligible studies, the overall positive patient outcomes following such interventions demonstrate the applicability and value of evidence-based acute stroke care interventions in Africa. Health policy attention is thus required to ensure widespread applicability of such interventions for improved patients' outcomes. The review findings also emphasises the need for further research to unravel the reasons for low uptake. PROSPERO CRD42016051566.


A unified framework of demographic time.
Demographic thought and practice is largely conditioned by the Lexis diagram, a two-dimensional graphical representation of the identity between age, period, and birth cohort. This relationship does not account for remaining years of life, total length of life, or time of death, whose use in demographic research is both underrepresented and incompletely situated. We describe an identity between these six demographic time measures and describe the sub-identities and diagrams that pertain to this identity. We provide an application of this framework to the measurement of late-life morbidity prevalence. We generalize these relationships to higher order identities derived from an arbitrary number of events in calendar time. Our examples are based on classic human demography, but the concepts we present can reveal patterns and relationships in any event history data, and contribute to the study of human or non-human population dynamics measured on any scale of calendar time.


Molecular characterization of localized pleural mesothelioma.
Localized pleural mesothelioma is a rare solitary circumscribed pleural tumor that is microscopically similar to diffuse malignant pleural mesothelioma. However, the molecular characteristics and nosologic relationship with its diffuse counterpart remain unknown. In a consecutive cohort of 1110 patients with pleural mesotheliomas diagnosed in 2005-2018, we identified six (0.5%) patients diagnosed with localized pleural mesotheliomas. We gathered clinical history, evaluated the histopathology, and in select cases performed karyotypic analysis and targeted next-generation sequencing. The cohort included three women and three men (median age 63; range 28-76), often presenting incidentally during radiologic evaluation for unrelated conditions. Neoadjuvant chemotherapy was administered in two patients. All tumors (median size 5.0 cm; range 2.7-13.5 cm) demonstrated gross circumscription (with microscopic invasion into lung, soft tissue, and/or rib in four cases), mesothelioma histology (four biphasic and two epithelioid types), and mesothelial immunophenotype. Of four patients with at least 6-month follow-up, three were alive (up to 8.9 years). Genomic characterization identified several subgroups: (1) BAP1 mutations with deletions of CDKN2A and NF2 in two tumors; (2) TRAF7 mutations in two tumors, including one harboring trisomies of chromosomes 3, 5, 7, and X; and (3) genomic near-haploidization, characterized by extensive loss of heterozygosity sparing chromosomes 5 and 7. Localized pleural mesotheliomas appear genetically heterogeneous and include BAP1-mutated, TRAF7-mutated, and near-haploid subgroups. While the BAP1-mutated subgroup is similar to diffuse malignant pleural mesotheliomas, the TRAF7-mutated subgroup overlaps genetically with adenomatoid tumors and well-differentiated papillary mesotheliomas, in which recurrent TRAF7 mutations have been described. Genomic near-haploidization, identified recently in a subset of diffuse malignant pleural mesotheliomas, suggests a novel mechanism in the pathogenesis of both localized pleural mesothelioma and diffuse malignant pleural mesothelioma. Our findings describe distinctive genetic features of localized pleural mesothelioma, with both similarities to and differences from diffuse malignant pleural mesothelioma.


A monocarboxylate transporter required for hepatocyte secretion of ketone bodies during fasting.
To find new genes that influence liver lipid mass, we performed a genetic screen for zebrafish mutants with hepatic steatosis, a pathological accumulation of fat. The red moon (rmn) mutant develops hepatic steatosis as maternally deposited yolk is depleted. Conversely, hepatic steatosis is suppressed in rmn mutants by adequate nutrition. Adult rmn mutants show increased liver neutral lipids and induction of hepatic lipid biosynthetic genes when fasted. Positional cloning of the rmn locus reveals a loss-of-function mutation in slc16a6a (solute carrier family 16a, member 6a), a gene that we show encodes a transporter of the major ketone body β-hydroxybutyrate. Restoring wild-type zebrafish slc16a6a expression or introducing human SLC16A6 in rmn mutant livers rescues the mutant phenotype. Radiotracer analysis confirms that loss of Slc16a6a function causes diversion of liver-trapped ketogenic precursors into triacylglycerol. Underscoring the importance of Slc16a6a to normal fasting physiology, previously fed rmn mutants are more sensitive to death by starvation than are wild-type larvae. Our unbiased, forward genetic approach has found a heretofore unrecognized critical step in fasting energy metabolism: hepatic ketone body transport. Since β-hydroxybutyrate is both a major fuel and a signaling molecule in fasting, the discovery of this transporter provides a new direction for modulating circulating levels of ketone bodies in metabolic diseases.


The effect of continuous positive airway pressure on metabolic variables in patients with obstructive sleep apnoea.
Obstructive sleep apnoea (OSA) is increasingly considered as a risk factor for metabolic disturbances, such as diabetes mellitus or dyslipidaemia. Continuous positive airway pressure (CPAP) therapy, the standard treatment for patients with OSA, may improve various metabolic variables, such as insulin sensitivity, glucose metabolism, lipids, fat distribution and adipokines. Several observational and uncontrolled clinical studies claim an improvement of these metabolic variables through the use of CPAP. However, there is only a limited number of clinical randomised controlled trials (RCTs) evaluating the effect of CPAP on metabolic variables. In this review, we summarise and discuss non-randomised studies and RCTs evaluating the effect of CPAP on metabolic variables in patients with OSA. In summary, the currently available body of evidence does not support a clinically important effect of CPAP treatment on any of the investigated metabolic variables. However, some investigators found small, but statistically significant changes in some metabolic variables, thus beneficial effects of CPAP treatment in selected patient cohorts cannot be excluded. To answer this question, more data from RCTs with well-defined study populations are warranted.


Use of failed-fertilized oocytes for diagnostic zona binding purposes after sperm binding improvement with a modified medium.
Because the availability of prophase oocytes for zona binding testing is limited, we compared sperm binding to the zona of failed-fertilized intracytoplasmic sperm injection (ICSI) and in vitro fertilization (IVF) oocytes after incubation in a standard IVF medium and a specially composed binding improvement medium. Semen samples from nine patients and nine fertile donors were separated in parallel by the standard swim-up method in both media. Subsequently, hemizona assays were performed with prophase, failed-fertilized ICSI and IVF oocytes. Sperm separation resulted in a significantly higher sperm count (P < 0.01) and progressive motility (P = 0.018) in binding improvement medium. Moreover, spermatozoa coincubated with hemizonae (prophase, failed-fertilized ICSI and IVF oocytes) in binding improvement medium bound significantly more to hemizonae than in the controls (P < 0.01). However, the hemizona index did not differ. Thus, the limited number of human zonae can be increased by the use of oocytes that failed to fertilize during ICSI or IVF. This will lead to a qualitative improvement of the diagnostic spectrum in male-factor infertility.


Violence against emergency medical services personnel: A systematic review of the literature.
Violence against emergency medical services (EMS) personnel is a growing concern. The aim of this systematic review is to synthesize the current literature on violence against EMS personnel. We examined literature from 2000 to 2016. Eligibility criteria included English-language, peer-reviewed studies of EMS personnel that described violence or assaults. Sixteen searches identified 2655 studies; 25 studies from nine countries met the inclusion criteria. The evidence from this review demonstrates that violence is a common risk for EMS personnel. We identified three critical topic areas: changes in risk over time, economic impact of violence and, outcomes of risk-reduction interventions. There is a lack of peer reviewed research of interventions, with the result that current intervention programs have no reliable evidence base. EMS leaders and personnel should work together with researchers to design, implement, evaluate and publish intervention studies designed to mitigate risks of violence to EMS personnel.


Impaired mitochondrial biogenesis, defective axonal transport of mitochondria, abnormal mitochondrial dynamics and synaptic degeneration in a mouse model of Alzheimer's disease.
Increasing evidence suggests that the accumulation of amyloid beta (Aβ) in synapses and synaptic mitochondria causes synaptic mitochondrial failure and synaptic degeneration in Alzheimer's disease (AD). The purpose of this study was to better understand the effects of Aβ in mitochondrial activity and synaptic alterations in neurons from a mouse model of AD. Using primary neurons from a well-characterized Aβ precursor protein transgenic (AβPP) mouse model (Tg2576 mouse line), for the first time, we studied mitochondrial activity, including axonal transport of mitochondria, mitochondrial dynamics, morphology and function. Further, we also studied the nature of Aβ-induced synaptic alterations, and cell death in primary neurons from Tg2576 mice, and we sought to determine whether the mitochondria-targeted antioxidant SS31 could mitigate the effects of oligomeric Aβ. We found significantly decreased anterograde mitochondrial movement, increased mitochondrial fission and decreased fusion, abnormal mitochondrial and synaptic proteins and defective mitochondrial function in primary neurons from AβPP mice compared with wild-type (WT) neurons. Transmission electron microscopy revealed a large number of small mitochondria and structurally damaged mitochondria, with broken cristae in AβPP primary neurons. We also found an increased accumulation of oligomeric Aβ and increased apoptotic neuronal death in the primary neurons from the AβPP mice relative to the WT neurons. Our results revealed an accumulation of intraneuronal oligomeric Aβ, leading to mitochondrial and synaptic deficiencies, and ultimately causing neurodegeneration in AβPP cultures. However, we found that the mitochondria-targeted antioxidant SS31 restored mitochondrial transport and synaptic viability, and decreased the percentage of defective mitochondria, indicating that SS31 protects mitochondria and synapses from Aβ toxicity.


Coherent energy transport in classical nonlinear oscillators: An analogy with the Josephson effect.
By means of a simple theoretical model and numerical simulations, we demonstrate the presence of persistent energy currents in a lattice of classical nonlinear oscillators with uniform temperature and chemical potential. In analogy with the well-known Josephson effect, the currents are proportional to the sine of the phase differences between the oscillators. Our results elucidate general aspects of nonequilibrium thermodynamics and point towards a way to practically control transport phenomena in a large class of systems. We apply the model to describe the phase-controlled spin-wave current in a bilayer nanopillar.


Characterization of plasmids that encode for the K88 colonization antigen.
K88 antigen, and important virulence factor in porcine enteropathogenic Escherichia coli (EEC), can be transferred along with the ability to ferment the trisaccharide raffinose (Raf). The plasmids from a number of EEC strains that encode these two properties were isolated and characterized. In most strains the K88 and Raf genes were found on a single nonconjugative plasmid approximately 50 x 10(6) daltons in size. This plasmid core was conserved with only slight variation among the strains tested. In some transconjugants, larger conjugative plasmids were observed that were apparently recombinants between the Raf/K88 plasmid and a transfer fa(tor. Occasionally plasmids carrying only the raffinose fermentation genes arose by deletion of a deoxyribonucleic acid segment of about 20 x 10(6) daltons that included the K88 antigen gene(s).


Flying at no mechanical energy cost: disclosing the secret of wandering albatrosses.
Albatrosses do something that no other birds are able to do: fly thousands of kilometres at no mechanical cost. This is possible because they use dynamic soaring, a flight mode that enables them to gain the energy required for flying from wind. Until now, the physical mechanisms of the energy gain in terms of the energy transfer from the wind to the bird were mostly unknown. Here we show that the energy gain is achieved by a dynamic flight manoeuvre consisting of a continually repeated up-down curve with optimal adjustment to the wind. We determined the energy obtained from the wind by analysing the measured trajectories of free flying birds using a new GPS-signal tracking method yielding a high precision. Our results reveal an evolutionary adaptation to an extreme environment, and may support recent biologically inspired research on robotic aircraft that might utilize albatrosses' flight technique for engineless propulsion.


Translational homeostasis via the mRNA cap-binding protein, eIF4E.
Translational control of gene expression plays a key role in many biological processes. Consequently, the activity of the translation apparatus is under tight homeostatic control. eIF4E, the mRNA 5' cap-binding protein, facilitates cap-dependent translation and is a major target for translational control. eIF4E activity is controlled by a family of repressor proteins, termed 4E-binding proteins (4E-BPs). Here, we describe the surprising finding that despite the importance of eIF4E for translation, a drastic knockdown of eIF4E caused only minor reduction in translation. This conundrum can be explained by the finding that 4E-BP1 is degraded in eIF4E-knockdown cells. Hypophosphorylated 4E-BP1, which binds to eIF4E, is degraded, whereas hyperphosphorylated 4E-BP1 is refractory to degradation. We identified the KLHL25-CUL3 complex as the E3 ubiquitin ligase, which targets hypophosphorylated 4E-BP1. Thus, the activity of eIF4E is under homeostatic control via the regulation of the levels of its repressor protein 4E-BP1 through ubiquitination.


Efficacy of bovine milk immunoglobulin concentrate in preventing illness after Shigella flexneri challenge.
The protective efficacy of oral bovine immunoglobulin concentrates derived from colostrum against challenge with Shigella flexneri was studied in healthy adult volunteers in a randomized, double-blind fashion. Volunteers were given a product consisting of hyperimmune immunoglobulin concentrate with a high titer of anti-S. flexneri 2a lipopolysaccharide (LPS) with sodium bicarbonate or a control preparation with sodium bicarbonate three times a day for seven days. On the third day of treatment, volunteers received a challenge of 10(3) colony-forming units of S. flexneri 2a strain 2457T. None of the volunteers who received the high-titered hyperimmune product became ill, compared with 45% of volunteers who received the placebo (P less than 0.05). The duration of shedding of the challenge organism was decreased, and the active immune responses to S. flexneri LPS were less frequent and of lower magnitude in volunteers given the immunoglobulin concentrate than in those in the control group. High-titered, orally administered bovine immunoglobulin concentrate protects against shigellosis and may be useful in preventing shigellosis among travelers, military personnel, and individuals at risk during a Shigella outbreak.


Wavelet packet energy characterization of low velocity impacts and load localization by optical fiber Bragg grating sensor technique.
The low velocity impacts (LVIs) monitoring based on optical fiber Bragg grating (FBG) sensors have attracted more attention in recent years. The center wavelength migrations of FBG sensors were determined by strain and residual strain during and after LVI on composite material structure. We presented a method to discriminate the energy characters of LVI response signals related to LVI locations. By analyzing the wavelet packet energy spectra of LVI response signals monitored by FBG sensors, the sixth node's energy was found to be sensitive to LVI location. Thus, the sixth node's energies as LVI feature values, were used to predict the LVI locations by the method of support vector regression (SVR). By optimization of the SVR models' free parameters, predicting accuracy was 4.62% in the work.


Changes in the socio-demographic patterning of late adolescent health risk behaviours during the 1990s: analysis of two West of Scotland cohort studies.
Substance use and sexual risk behaviour affect young people's current and future health and wellbeing in many high-income countries. Our understanding of time-trends in adolescent health-risk behaviour is largely based on routinely collected survey data in school-aged adolescents (aged 15 years or less). Less is known about changes in these behaviours among older adolescents. We compared two cohorts from the same geographical area (West of Scotland), surveyed in 1990 and 2003, to: describe time-trends in measures of smoking, drinking, illicit drug use, early sexual initiation, number of opposite sex sexual partners and experience of pregnancy at age 18-19 years, both overall and stratified by gender and socioeconomic status (SES); and examine the effect of time-trends on the patterning of behaviours by gender and SES. Our analyses adjust for slight between-cohort age differences since age was positively associated with illicit drug use and pregnancy. Rates of drinking, illicit drug use, early sexual initiation and experience of greater numbers of sexual partners all increased significantly between 1990 and 2003, especially among females, leading to attenuation and, for early sexual initiation, elimination, of gender differences. Most rates increased to a similar extent regardless of SES. However, rates of current smoking decreased only among those from higher SES groups. In addition, increases in 'cannabis-only' were greater among higher SES groups while use of illicit drugs other than cannabis increased more in lower SES groups. Marked increases in female substance use and sexual risk behaviours have implications for the long-term health and wellbeing of young women. More effective preventive measures are needed to reduce risk behaviour uptake throughout adolescence and into early adulthood. Public health strategies should reflect both the widespread prevalence of risk behaviour in young people as well as the particular vulnerability to certain risk behaviours among those from lower SES groups.


MRP1 overexpression determines poor prognosis in prospectively treated patients with localized high-risk soft tissue sarcoma of limbs and trunk wall: an ISG/GEIS study.
Patients with localized high-risk soft tissue sarcomas (STS) of the limbs and trunk wall still have a considerable metastatic recurrence rate of more than 50%, in spite of adjuvant chemotherapy. This drug-ceiling effect of chemotherapy in sarcoma setting could be explained, at least partially, by multidrug resistance (MDR) mechanisms. The aim of this study was to ascertain whether mRNA and protein expression of ABCB1 (P-glycoprotein), ABCC1 (MRP1), and GSTA1 (glutathione S-transferase pi) was prognostic in localized high-risk STS. Immunohistochemistry and reverse transcriptase-PCR studies were performed from biopsies at the time of diagnosis. Patients of this series were prospectively enrolled into a phase III trial that compared three versus five cycles of epirubicin plus ifosfamide. The series of 102 patients found 41 events of recurrence and 37 of death with a median follow-up of 68 months. In univariate analysis, variables with a statistically significant relationship with relapse-free survival (RFS) were: MRP1 expression (5-year RFS rate of 23% in positive cases and 63% in negative cases, P = 0.029), histology (5-year RFS rate of 74% in undifferentiated pleomorphic sarcoma and 43% in synovial sarcoma, P = 0.028), and ABCC1 expression (5-year RFS rate of 33% in overexpression and 65% in downregulation, P = 0.012). Combined ABCC1/MRP1 was the only independent prognostic factor for both RFS (HR = 2.704, P = 0.005) and overall survival (HR = 2.208, P = 0.029). ABCC1/MRP1 expression shows robust prognostic relevance in patients with localized high-risk STS treated with anthracycline-based chemotherapy, which is the standard front line treatment in STS. This finding deserves attention as it points to a new targetable protein in STS.


Curcumin inhibits HIV-1 by promoting Tat protein degradation.
HIV-1 Tat is an intrinsically unfolded protein playing a pivotal role in viral replication by associating with TAR region of viral LTR. Unfolded proteins are degraded by 20S proteasome in an ubiquitin independent manner. Curcumin is known to activate 20S proteasome and promotes the degradation of intrinsically unfolded p53 tumor suppressor protein. Since HIV-1 Tat protein is largerly unfolded, we hypothesized that Tat may also be targeted through this pathway. Curcumin treated Tat transfected HEK-293T cells showed a dose and time dependent degradation of Tat protein. Contrary to this HIV-1 Gag which is a properly folded protein, remained unaffected with curcumin. Semi-quantitative RT-PCR analysis showed that curcumin treatment did not affect Tat gene transcription. Curcumin increased the rate of Tat protein degradation as shown by cycloheximide (CHX) chase assay. Degradation of the Tat protein is accomplished through proteasomal pathway as proteasomal inhibitor MG132 blocked Tat degradation. Curcumin also decreased Tat mediated LTR promoter transactivation and inhibited virus production from HIV-1 infected cells. Taken together our study reveals a novel observation that curcumin causes potent degradation of Tat which may be one of the major mechanisms behind its anti HIV activity.


The engrailed homeobox genes are required in multiple cell lineages to coordinate sequential formation of fissures and growth of the cerebellum.
The layered cortex of the cerebellum is folded along the anterior-posterior axis into lobules separated by fissures, allowing the large number of cells needed for advanced cerebellar functions to be packed into a small volume. During development, the cerebellum begins as a smooth ovoid structure with two progenitor zones, the ventricular zone and upper rhombic lip, which give rise to distinct cell types in the mature cerebellum. Initially, the cerebellar primordium is divided into five cardinal lobes, which are subsequently further subdivided by fissures. The cellular processes and genes that regulate the formation of a normal pattern of fissures are poorly understood. The engrailed genes (En1 and En2) are expressed in all cerebellar cell types and are critical for regulating formation of specific fissures. However, the cerebellar cell types that En1 and En2 act in to control growth and/or patterning of fissures has not been determined. We conditionally eliminated En2 or En1 and En2 either in both progenitor zones and their descendents or in the two complementary sets of cells derived from each progenitor zone. En2 was found to be required only transiently in the progenitor zones and their immediate descendents to regulate formation of three fissures and for general growth of the cerebellum. In contrast, En1 and En2 have overlapping functions in the cells derived from each progenitor zone in regulating formation of additional fissures and for extensive cerebellar growth. Furthermore, En1/2 function in ventricular zone-derived cells plays a more significant role in determining the timing of initiation and positioning of fissures, whereas in upper rhombic lip-derived cells the genes are more important in regulating cerebellar growth. Our studies reveal the complex manner in which the En genes control cerebellar growth and foliation in distinct cell types.


Immunology of mercury.
The heavy metal mercury is ubiquitously distributed in the environment resulting in permanent low-level exposure in human populations. Mercury can be encountered in three main chemical forms (elemental, inorganic, and organic) which can affect the immune system in different ways. In this review, we describe the effects of these various forms of mercury exposure on immune cells in humans and animals. In genetically susceptible mice or rats, subtoxic doses of mercury induce the production of highly specific autoantibodies as well as a generalized activation of the immune system. We review studies performed in this model and discuss their implications for the role of environmental chemicals in human autoimmunity.


The Fate of Bacteria in Human Digestive Fluids: A New Perspective Into the Pathogenesis of Vibrio parahaemolyticus.
Vibrio parahaemolyticus causes the most seafood-attributed gastroenteritis outbreaks worldwide and studies on its pathogenesis during passage through the human digestive fluids are limited. An in vitro continuous model system mimicking passage through saliva, gastric and intestinal fluid was used to study the survival, morphology and virulence-related gene expression of a total of sixty pathogenic, and non-pathogenic V. parahaemolyticus strains. The changes to these three characteristics for the sixty V. parahaemolyticus strains were minimal on passage through the saliva fluid. No V. parahaemolyticus strains survived passage through gastric fluid with low pH values (2.0 and 3.0) and the cells, examined microscopically, were severely damaged. However, when the pH of gastric fluid increased to 4.0, the bacterial survival rate was 54.70 ± 1.11%, and the survival rate of pathogenic strains was higher when compared to non-pathogenic strains. Even though the bactericidal effect of intestinal fluid was lower than gastric fluid, virulence-related gene expression was enhanced in the intestinal fluid. Seafood matrices can significantly raise the pH level of gastric fluid and thus aid the survival of V. parahaemolyticus through passage from human gastric acid and progression of pathogenesis in the intestinal fluid. We confirmed these phenomena in the in vitro continuous digestion model.


Generalised interrelations among mutation rates drive the genomic compliance of Chargaff's second parity rule.
Chargaff's second parity rule (PR-2), where the complementary base and k-mer contents are matching within the same strand of a double stranded DNA (dsDNA), is a phenomenon that invited many explanations. The strict compliance of nearly all nuclear dsDNA to PR-2 implies that the explanation should also be similarly adamant. In this work, we revisited the possibility of mutation rates driving PR-2 compliance. Starting from the assumption-free approach, we constructed kinetic equations for unconstrained simulations. The results were analysed for their PR-2 compliance by employing symbolic regression and machine learning techniques. We arrived to a generalised set of mutation rate interrelations in place in most species that allow for their full PR-2 compliance. Importantly, our constraints explain PR-2 in genomes out of the scope of the prior explanations based on the equilibration under mutation rates with simpler no-strand-bias constraints. We thus reinstate the role of mutation rates in PR-2 through its molecular core, now shown, under our formulation, to be tolerant to previously noted strand biases and incomplete compositional equilibration. We further investigate the time for any genome to reach PR-2, showing that it is generally earlier than the compositional equilibrium, and well within the age of life on Earth.


Chromosome-scale genome assembly of kiwifruit Actinidia eriantha with single-molecule sequencing and chromatin interaction mapping.
Kiwifruit (Actinidia spp.) is a dioecious plant with fruits containing abundant vitamin C and minerals. A handful of kiwifruit species have been domesticated, among which Actinidiaeriantha is increasingly favored in breeding owing to its superior commercial traits. Recently, elite cultivars from A. eriantha have been successfully selected and further studies on their biology and breeding potential require genomic information, which is currently unavailable. We assembled a chromosome-scale genome sequence of A. eriantha cultivar White using single-molecular sequencing and chromatin interaction map-based scaffolding. The assembly has a total size of 690.6 megabases and an N50 of 21.7 megabases. Approximately 99% of the assembly were in 29 pseudomolecules corresponding to the 29 kiwifruit chromosomes. Forty-three percent of the A. eriantha genome are repetitive sequences, and the non-repetitive part encodes 42,988 protein-coding genes, of which 39,075 have homologues from other plant species or protein domains. The divergence time between A. eriantha and its close relative Actinidia chinensis is estimated to be 3.3 million years, and after diversification, 1,727 and 1,506 gene families are expanded and contracted in A. eriantha, respectively. We provide a high-quality reference genome for kiwifruit A. eriantha. This chromosome-scale genome assembly is substantially better than 2 published kiwifruit assemblies from A. chinensis in terms of genome contiguity and completeness. The availability of the A. eriantha genome provides a valuable resource for facilitating kiwifruit breeding and studies of kiwifruit biology.


Trancranial Doppler: value in clinical practice.
The value of TCD in clinical practice is well established since it can be used to measure cerebral vasomotor reactivity and to detect and grade vasospasm (VSP) following subarachnoid haemorrhage and cerebral blood perfusion consequences of extracranial ICA stenosis or occlusion. Intracranial steno-occlusive disease can be detected more reliably by transcranial color-coded imaging (TCCI) that provides a two-dimensional imaging of parenchymal and vascular anatomy of brain too. In patients with suspected brain TCD diagnostic criteria for brain death have a sensitivity of 91 to 100% and specificity of 97 to 100% and they are particularly useful when clinical and EEG evaluations are difficult. TCD is a sensitive technique for real time detection of microembolic signals (MES) from prosthetic cardiac valves, myocardial infarction site, atrial fibrillation, aortic arch atheroma and this suggests the use of TCD for monitoring response to antithrombotic therapy. There is also a high correlation between contrast-enhanced TCD and trans-esophageal echocardiography for detecting paradoxical embolism through right-to-left cardiac or pulmonary shunts. Microembolization detected by TCD monitoring may confirm features of unstable carotid artery plaques as imaged by Duplex scanning and there is an increasing evidence that asymptomatic MES from unstable carotid plaques are an independent factor for ischemic stroke. TCD can be used as a monitoring tool during cardiac surgery and cerebrovascular operations to determine critical hemodynamic changes in cerebral arteries and to identify high-intensity transients referred to air or particulate emboli. Several research studies of the past 10 years have shown that MES may be detected by TCD during all phases of CEA and CAS and that sustained microembolism after carotid flow restoration is an indication of impending postoperative or post-procedural occlusion. Our series showed a clear difference between the number of patients with MES and the incidence rate of MES in each patient submitted to CAS (100% of cases with 35-250 MES in each case) and to CEA (74% of cases with 2-30 MES in each case). We also observed a decrease in the incidence rate of microembolic events by TCD during CAS with or without brain protection devices , 18.% and 40%, respectively. There is a statistically significant difference between the neurological deficit related to embolism during CEA (1.8% of cases) and during CAS(9 %). Furthermore DWI has shown a higher prevalence of postoperative small areas of brain ischemia due to asymptomatic embolism occurring during CAS than after carotid surgery according with a higher incidence of patients suffering from neuropsychological impairment after CAS as compared with those submitted to CEA . The use of TCD can provide new insights into pathophysiology of cerebral steno-occlusive and functional diseases, it can helps in risk stratifications of patients with cardio-embolic sources and in the choice and monitoring of medical, surgical or endovascular treatment. TCD monitoring during carotid revascularization either surgical or endovascular can alert the operator to take appropriate measures to avoid brain ischemia and provides useful data for choice and control of the different brain protection devices.


Focusing and extraction of light mediated by Bloch surface waves.
The control of emission from localized light sources is an objective of outstanding relevance in nanophotonics. In a recent past, a large number of metallic nanostructures has been proposed to this end, wherein plasmonic modes are exploited as energy carriers on a subwavelength scale. As an interesting alternative, we present here the use of surface modes on patterned dielectric multilayers to deliver electromagnetic power from free-space to localized volumes and vice versa. Thanks to this low-loss energy transfer, proper periodic ring structures are shown to provide a subwavelength focusing of an external radiation onto the multilayer surface. By reciprocity, the radiated power from emitters within the ring center is shown to be efficiently beamed in the free-space, with a well-controlled angular divergence. This mechanism overcomes some important limitations involved in the all-plasmonic approach, while opening new opportunities for hybrid devices in photon management applications such as optical sensing and lighting.


Self-Aligned Plasmonic Nanopores by Optically Controlled Dielectric Breakdown.
We present a novel cost-efficient method for the fabrication of high-quality self-aligned plasmonic nanopores by means of an optically controlled dielectric breakdown. Excitation of a plasmonic bowtie nanoantenna on a dielectric membrane localizes the high-voltage-driven breakdown of the membrane to the hotspot of the enhanced optical field, creating a nanopore that is automatically self-aligned to the plasmonic hotspot of the bowtie. We show that the approach provides precise control over the nanopore size and that these plasmonic nanopores can be used as single molecule DNA sensors with a performance matching that of TEM-drilled nanopores. The principle of optically controlled breakdown can also be used to fabricate nonplasmonic nanopores at a controlled position. Our novel fabrication process guarantees alignment of the nanopore with the optical hotspot of the nanoantenna, thus ensuring that pore-translocating biomolecules interact with the concentrated optical field that can be used for detection and manipulation of analytes.


One Size Does Not Fit All: The Limits of Structure-Based Models in Drug Discovery.
A major goal in computational chemistry has been to discover the set of rules that can accurately predict the binding affinity of any protein-drug complex, using only a single snapshot of its three-dimensional structure. Despite the continual development of structure-based models, predictive accuracy remains low, and the fundamental factors that inhibit the inference of all-encompassing rules have yet to be fully explored. Using statistical learning theory and information theory, here we prove that even the very best generalized structure-based model is inherently limited in its accuracy, and protein-specific models are always likely to be better. Our results refute the prevailing assumption that large data sets and advanced machine learning techniques will yield accurate, universally applicable models. We anticipate that the results will aid the development of more robust virtual screening strategies and scoring function error estimations.


Direct ex vivo analysis reveals distinct phenotypic patterns of HIV-specific CD8(+) T lymphocyte activation in response to therapeutic manipulation of virus load.
Therapeutic intervention with antiretroviral therapy (ART) enables the modulation of HIV virus load and hence provides a unique opportunity to study the consequences of varying antigen load on the phenotype of virus-specific CD8(+) T lymphocytes in a persistent human viral infection. The recent advent of tetrameric peptide / HLA class I complexes has enabled the direct phenotypic characterization of antigen-specific T cell populations ex vivo. Here, we use this technology to examine directly ex vivo the consequences of therapeutic manipulation of HIV virus load on the phenotype of HIV-specific CTL. Our observations show that: (1) distinct sequential activation patterns of CD8(+) T cells are associated with increasing virus load; (2) T cell receptor (TCR) down-regulation without apoptosis represents an early event during the generation of a T cell response in a natural infection and precedes the emergence of two distinct antigen-specific CD8(+) T cell populations which differ in TCR and CD8 expression levels. Clear differences in surface Annexin V staining were observed between these populations. The observation that CTL activation, demonstrated by TCR and CD8 down-regulation, in response to rising levels of virus load, co-segregates with apoptosis only during later stages of the response indicates that antigen-associated cell death is restricted to distinct subpopulations of CTL.


TaxaGloss - A Glossary and Translation Tool for Biodiversity Studies.
Correctly identifying organisms is key to most biological research, and is especially critical in areas of biodiversity and conservation. Yet it remains one of the greatest challenges when studying all but the few well-established model systems. The challenge is in part due to the fact that most species have yet to be described, vanishing taxonomic expertise and the relative inaccessibility of taxonomic information. Furthermore, identification keys and other taxonomic resources are based on complex, taxon-specific vocabularies used to describe important morphological characters. Using these resources is made difficult by the fact that taxonomic documentation of the world's biodiversity is an international endeavour, and keys and field guides are not always available in the practitioner's native language. To address this challenge, we have developed a publicly available on-line illustrated multilingual glossary and translation tool for technical taxonomic terms using the Symbiota Software Project biodiversity platform. Illustrations, photographs and translations have been sourced from the global community of taxonomists working with marine invertebrates and seaweeds. These can be used as single-language illustrated glossaries or to make customized translation tables. The glossary has been launched with terms and illustrations of seaweeds, tunicates, sponges, hydrozoans, sea anemones, and nemerteans, and already includes translations into seven languages for some groups. Additional translations and development of terms for more taxa are underway, but the ultimate utility of this tool depends on active participation of the international taxonomic community.


Increased mortality among individuals with chronic widespread pain relates to lifestyle factors: a prospective population-based study.
Widespread chronic pain has been related to disability and loss of quality of life, but in a few epidemiological studies also to increased mortality. The aim of this study was to further investigate the relationship between chronic pain, lifestyle factors and all cause mortality. A random sample of an adult (age 25-74) Swedish population (n = 1609) responded to a comprehensive questionnaire on pain, other symptoms, lifestyle, work and socioeconomic factors in 1988. Mortality data for this cohort between 1988 and 2002 were analysed. Survival analysis (Kaplan-Meier) and Cox proportional regression were used to study initially reported factors influencing survival. Individuals with widespread chronic pain showed an increased mortality risk (hazard ratio, HR = 1.95, CI: 1.26-3.03) compared to the group without chronic pain. Death due to cardiovascular disease accounted for the increased mortality. Adjustment for lifestyle factors eliminated the excess risk. Increased mortality among individuals with widespread chronic pain is related to factors like smoking, sleep disturbances and low physical activity. The result emphasises the importance of including lifestyle factors in a cognitive-behavioural rehabilitation process. It remains to be shown whether health promotion activities aimed at lifestyle could change mortality among individuals with chronic pain.


Inhibitory Effects of Salinomycin on Cell Survival, Colony Growth, Migration, and Invasion of Human Non-Small Cell Lung Cancer A549 and LNM35: Involvement of NAG-1.
A major challenge for oncologists and pharmacologists is to develop more potent and less toxic drugs that will decrease the tumor growth and improve the survival of lung cancer patients. Salinomycin is a polyether antibiotic used to kill gram-positive bacteria including mycobacteria, protozoans such as plasmodium falciparum, and the parasites responsible for the poultry disease coccidiosis. This old agent is now a serious anti-cancer drug candidate that selectively inhibits the growth of cancer stem cells. We investigated the impact of salinomycin on survival, colony growth, migration and invasion of the differentiated human non-small cell lung cancer lines LNM35 and A549. Salinomycin caused concentration- and time-dependent reduction in viability of LNM35 and A549 cells through a caspase 3/7-associated cell death pathway. Similarly, salinomycin (2.5-5 µM for 7 days) significantly decreased the growth of LNM35 and A549 colonies in soft agar. Metastasis is the main cause of death related to lung cancer. In this context, salinomycin induced a time- and concentration-dependent inhibition of cell migration and invasion. We also demonstrated for the first time that salinomycin induced a marked increase in the expression of the pro-apoptotic protein NAG-1 leading to the inhibition of lung cancer cell invasion but not cell survival. These findings identify salinomycin as a promising novel therapeutic agent for lung cancer.


2018 International Consensus Statement on Golf and Health to guide action by people, policymakers and the golf industry.
Scientific and public interest relating to golf and health has increased recently. Players, potential players, the golf industry and facilities, and decision makers will benefit from a better understanding of how to realise potential health benefits and minimise health issues related to golf. We outline an International Consensus on Golf and Health. A systematic literature review informed the development of a survey. Utilising modified Delphi methods, an expert panel of 25 persons including public health and golf industry leaders, took part in serial surveys providing feedback on suggested items, and proposing new items. Predefined criteria for agreement determined whether each item was included within each survey round and in the final consensus. The working group identified 79 scientifically supportable statement items from literature review and discussions. Twenty-five experts (100%) completed all three rounds of surveys, rating each item, and suggesting modifications and/or new items for inclusion in subsequent surveys. After three rounds, 83 items achieved consensus with each with >75% agreement and <10% disagreement. These items are included in the final International Consensus on Golf and Health. The final consensus presented here can inform scientific knowledge, and action plans for (1) golfers and potential golfers, (2) golf facilities and the golf industry, and (3) policy and decision makers external to golf. These outputs, if widely adopted, will contribute to an improved understanding of golf and health, and aid these groups in making evidence-informed decisions to improve health and well-being.


Persistent Viral Reservoirs in Lymphoid Tissues in SIV-Infected Rhesus Macaques of Chinese-Origin on Suppressive Antiretroviral Therapy.
Understanding HIV latent reservoirs in tissues is essential for the development of new strategies targeting these sites for eradication. Here, we assessed the size of latent reservoirs and the source of residual viruses in multiple lymphoid tissues of SIV-infected and fully suppressed rhesus macaques of Chinese-origin (cRMs). Eight cRMs were infected with SIVmac251 and treated with tenofovir and emtricitabine daily for 24 weeks initiated 4 weeks post-infection. Four of the eight animals reached sustained full viral suppression with undetectable viremia. The levels of cell-associated SIV DNA varied in peripheral blood mononuclear cells (PBMCs) and multiple lymphoid tissues, but with higher levels in the mesenteric lymph nodes (MesLNs). The levels of cell-associated SIV RNA also varied in different tissues. The higher frequency of viral RNA detection in the MesLNs was also observed by in situ hybridization. Consistently, the infection unit per million cells (IUPM) in the MesLNs was higher than in PBMCs and other tested lymphoid tissues by quantitative viral outgrowth assay (QVOA). Furthermore, env gp120 from tissue SIV RNA was amplified by single genome amplification. Phylogenetic analysis revealed diverse variants from tissues parallel to the viral inoculum in all viral suppressed animals. These results demonstrate that the latency and viral reservoirs in the lymphoid tissues still exist in aviremic macaques under full suppressive therapy. Moreover, the size of viral latent reservoirs differs in various lymphoid tissues with a relatively larger size in the MesLNs.


Robustness and evolvability: a paradox resolved.
Understanding the relationship between robustness and evolvability is key to understand how living things can withstand mutations, while producing ample variation that leads to evolutionary innovations. Mutational robustness and evolvability, a system's ability to produce heritable variation, harbour a paradoxical tension. On one hand, high robustness implies low production of heritable phenotypic variation. On the other hand, both experimental and computational analyses of neutral networks indicate that robustness enhances evolvability. I here resolve this tension using RNA genotypes and their secondary structure phenotypes as a study system. To resolve the tension, one must distinguish between robustness of a genotype and a phenotype. I confirm that genotype (sequence) robustness and evolvability share an antagonistic relationship. In stark contrast, phenotype (structure) robustness promotes structure evolvability. A consequence is that finite populations of sequences with a robust phenotype can access large amounts of phenotypic variation while spreading through a neutral network. Population-level processes and phenotypes rather than individual sequences are key to understand the relationship between robustness and evolvability. My observations may apply to other genetic systems where many connected genotypes produce the same phenotypes.


Sex differences in correlates of steady state and pulsatile component of blood pressure.
1. The aim of the present study was to evaluate whether metabolic factors are linked to the steady component and the pulsatile component of blood pressure, evaluated as mean arterial pressure and pulse pressure respectively, in a sex-specific manner. 2. A cohort of 299 subjects (152 males, 147 females; 25-80 years of age) was studied. Patients presenting congestive heart failure, coronary insufficiency, severe valvular heart disease, neurological accident in the last 6 months, renal or respiratory failure, cancer, diabetes mellitus or acute infectious disease were excluded. None of the women was taking oral contraceptives or oestrogen supplementation. All cardioactive drugs were withdrawn at least 2 weeks before the subjects entered the study. 3. Men presented higher mean arterial pressure (120 +/- 15 compared with 115 +/- 16 mmHg, P < 0.01) and lower pulse pressure values (63 +/- 16 compared with 67 +/- 18 mmHg, P < 0.05) than women. In men, no significant relation between mean arterial pressure and the tested variables was detected; multiple regression analysis demonstrated that age contributed independently to the model for pulse pressure with a multiple r2 of 0.10 (P < 0.01). In women, body mass index contributed independently to the model for mean arterial pressure, with a multiple of 0.12 (P < 0.005); age and, to a lesser extent, body mass index, glycaemia and triglyceridaemia persisted as independent determinants of pulse pressure at the multiple regression analysis, with a multiple r2 of 0.20 (P < 0.001). 4. Our findings suggest that metabolic risk factors are associated differently with pulse pressure and mean arterial pressure values in the two sexes.


Time Is Not Always the Matter: An Instance of Encapsulating Peritoneal Sclerosis Developing in a Patient on Peritoneal Dialysis for a Short Term.
Encapsulating peritoneal sclerosis (EPS) is an infrequent but serious complication that is observed mostly in patients on long-term peritoneal dialysis (PD). However it can occur after short-term PD, in association with "second hit" risk factors such as peritonitis, acute cessation of PD, or kidney transplantation with the use of calcineurin inhibitors.In our case, a young woman with second-hit risk factors presented with clinical and abdominal computed tomography findings consistent with EPS after short-term PD. She was treated conservatively with nutritional support and was discharged in improved and stable clinical status.In general, the diagnosis of EPS requires clinical findings of bowel obstruction combined with typical computed tomography imaging features. However, the clinical manifestations can be very vague, and the diagnosis is often unclear. A recent study categorized EPS into 4 clinical stages, from pre-EPS to chronic ileus, with associated management from conservative treatment to surgical intervention.In association with second-hit risk factors, EPS can occur after short-term PD. Severity is variable, and the outcome is often devastating. Timely recognition and expert management of EPS can change the outcome very favorably.


Microwave-assisted one-pot synthesis of new ionic iridium complexes of [Ir(bzq)2(N^N)]+A- type and their selected electroluminescent properties.
Iridium C,N-cyclometalated complexes with an ionic structure are considered to be promising candidates for application in host/guest solid-state phosphorescent single-layer devices because the employment of such dopants offers the possibility of reducing their concentration in organic matrices as well as allows obtaining organic light emitting devices (OLEDs) with interesting emission parameters. We report herein a methodology enabling the synthesis of cyclometalated ionic iridium(iii) complexes of the type [Ir(C^N)2(N^N)]+A- according to a three-component one-pot strategy involving the acceleration of the reaction via microwave irradiation. The developed protocol allowed efficient synthesis of a series of new cationic iridium(iii) coordination derivatives, which were isolated and spectroscopically characterized, while the structures of two of them were determined by the X-ray method. Moreover, the iridium(iii) derivatives were subjected to the cyclic voltammetry studies in order to determine the energies of the HOMO and LUMO levels as well as to estimate their electrochemical properties and to predict some electronic properties. Additionally, the ONIOM calculation scheme that was used to predict HOMO-LUMO gaps for the studied Ir(iii) complexes showed a good correlation between the experimental and calculated values. In order to determine the influence of the structure and nature of the ancillary ligand on the location of the maximum emission band, the photophysical properties of the synthesized iridium complexes were characterized. Finally, the selected compounds were used as emitters for the construction of polymer light emitting diodes (PLEDs) based on a poly(N-vinylcarbazole)/2-(4-tert-butylphenyl)-5-(4-biphenyl)-1,3,4-oxadiazole (PVK/PBD) matrix. The highest luminance, above 10 000 cd m-2, was recorded for the device containing only 1.0 wt% of [Ir(bzq)2(1,10-phenanthroline)]+PF6- in the PVK/PBD. The fabricated PLEDs exhibit current efficiency in the range of 1.0 to 2.2 cd A-1.


Nearest Neighbor Networks: clustering expression data based on gene neighborhoods.
The availability of microarrays measuring thousands of genes simultaneously across hundreds of biological conditions represents an opportunity to understand both individual biological pathways and the integrated workings of the cell. However, translating this amount of data into biological insight remains a daunting task. An important initial step in the analysis of microarray data is clustering of genes with similar behavior. A number of classical techniques are commonly used to perform this task, particularly hierarchical and K-means clustering, and many novel approaches have been suggested recently. While these approaches are useful, they are not without drawbacks; these methods can find clusters in purely random data, and even clusters enriched for biological functions can be skewed towards a small number of processes (e.g. ribosomes). We developed Nearest Neighbor Networks (NNN), a graph-based algorithm to generate clusters of genes with similar expression profiles. This method produces clusters based on overlapping cliques within an interaction network generated from mutual nearest neighborhoods. This focus on nearest neighbors rather than on absolute distance measures allows us to capture clusters with high connectivity even when they are spatially separated, and requiring mutual nearest neighbors allows genes with no sufficiently similar partners to remain unclustered. We compared the clusters generated by NNN with those generated by eight other clustering methods. NNN was particularly successful at generating functionally coherent clusters with high precision, and these clusters generally represented a much broader selection of biological processes than those recovered by other methods. The Nearest Neighbor Networks algorithm is a valuable clustering method that effectively groups genes that are likely to be functionally related. It is particularly attractive due to its simplicity, its success in the analysis of large datasets, and its ability to span a wide range of biological functions with high precision.


Parasomnias: an updated review.
Parasomnias are abnormal behaviors emanating from or associated with sleep. Sleepwalking and related disorders result from an incomplete dissociation of wakefulness from nonrapid eye movement (NREM) sleep. Conditions that provoke repeated cortical arousals, or promote sleep inertia lead to NREM parasomnias by impairing normal arousal mechanisms. Changes in the cyclic alternating pattern, a biomarker of arousal instability in NREM sleep, are noted in sleepwalking disorders. Sleep-related eating disorder (SRED) is characterized by a disruption of the nocturnal fast with episodes of feeding after an arousal from sleep. SRED is often associated with the use of sedative-hypnotic medications; in particular, the widely prescribed benzodiazepine receptor agonists. Recently, compelling evidence suggests that nocturnal eating may in some cases be a nonmotor manifestation of Restless Legs Syndrome (RLS). rapid eye movement (REM) Sleep Behavior Disorder (RBD) is characterized by a loss of REM paralysis leading to potentially injurious dream enactment. The loss of atonia in RBD often predates the development of Parkinson's disease and other disorders of synuclein pathology. Parasomnia behaviors are related to an activation (in NREM parasomnias) or a disinhibition (in RBD) of central pattern generators (CPGs). Initial management should focus on decreasing the potential for sleep-related injury followed by treating comorbid sleep disorders. Clonazepam and melatonin appear to be effective therapies in RBD, whereas paroxetine has been reported effective in some cases of sleep terrors. At this point, pharmacotherapy for other parasomnias is less certain, and further investigations are necessary.


Autophagy Supports Breast Cancer Stem Cell Maintenance by Regulating IL6 Secretion.
Autophagy is a mechanism by which cells degrade cellular material to provide nutrients and energy for survival during stress. The autophagy is thought to be a critical process for cancer stem cell (CSC) or tumor-initiating cell maintenance but the mechanisms by which autophagy supports survival of CSCs remain poorly understood. In this study, inhibition of autophagy by knockdown of ATG7 or BECN1 modified the CD44(+)/CD24(low/-) population in breast cancer cells by regulating CD24 and IL6 secretion. In a breast cancer cell line that is independent of autophagy for survival, autophagy inhibition increased IL6 secretion to the media. On the other hand, in an autophagy-dependent cell line, autophagy inhibition decreased IL6 secretion, cell survival, and mammosphere formation. In these cells, IL6 treatment or conditioned media from autophagy-competent cells rescued the deficiency in mammosphere formation induced by autophagy inhibition. These results reveal that autophagy regulates breast CSC maintenance in autophagy-dependent breast cancer cells by modulating IL6 secretion implicating autophagy as a potential therapeutic target in breast cancer. Modulation of autophagy in breast cancer has different and even opposite effects, indicating the need for a selection strategy when trying to manipulate autophagy in the context of cancer therapy.


Ecological causes of multilevel covariance between size and first-year survival in a wild bird population.
Estimates of selection in natural populations are frequent but our understanding of ecological causes of selection, and causes of variation in the direction, strength and form of selection is limited. Here, we apply a multilevel framework to partition effects of great tit fledging mass on first-year survival to hierarchical levels and quantify their ecological dependence using a data set spanning 51 years. We show that estimates of the effect of fledging mass on first-year survival decline threefold from year- to brood- to individual level, so that estimates of selection depend strongly on the level at which they are calculated. We identify variables related to summer and winter food availability as underlying higher-level effects of fledging mass on first-year survival and show experimentally that brood-level effects originate early in development. Further, we show that predation and conspecific density modulate individual-level effects of fledging mass on first-year survival. These analyses demonstrate how correlations between traits, fitness and environment influence estimates of selection and show how partitioning trait effects between levels of selection and environmental factors is a promising approach to identify potential agents of selection.


Development of joint engagement in young deaf and hearing children: effects of chronological age and language skills.
To evaluate joint engagement (JE) in age-matched children with and without hearing and its relationship to oral language skills. Participants were 180 children with severe-to-profound hearing loss prior to cochlear implant surgery, and 96 age-matched children with normal hearing; all parents were hearing. JE was evaluated in a 10-minute videotaped free play task with parents. Engagement states ranged from the lowest (unengaged) to the highest level (symbol-infused coordinated). Standardized language measures were administered. Multivariate analyses were conducted between the groups, stratified by chronological and language age. Children who were deaf (Deaf) spent less time in total symbol-infused JE than children with normal hearing (NH) across all ages. The majority of the Deaf group (83%) fell in the lowest language age group, in comparison to 35% of the NH group, and spent significantly less time in symbol-infused JE than hearing children. These delays were also observed in the Deaf group, who fell into the 18-36 month language age. No children in the Deaf group had achieved a language age of > 36 months. Young children with and without hearing had different developmental trajectories of JE, which were related to oral language skills.


Characteristics of Carotid Artery Structure and Mechanical Function and Their Relationships with Aortopathy in Patients with Bicuspid Aortic Valves.
Patients with a bicuspid aortic valve (BAV) often have proximal aortic dilatation and systemic vascular dysfunction. We hypothesized that BAV patients would have different carotid artery structural and functional characteristics compared to tricuspid aortic valve (TAV) patients. In 28 patients with surgically confirmed BAV and 27 patients with TAV, intima media thickness (IMT), number of plaques, fractional area change (FAC), global circumferential strain (GCS), and standard deviation of CS (SD-CS) in both common carotid arteries were assessed using duplex ultrasound and velocity vector imaging (VVI). Patients with BAV were younger and had less co-morbidity, but showed a significantly larger ascending aorta (43.3 ± 7.5 vs. 37.0 ± 6.2 mm, p < 0.001) and a higher prevalence of aortopathy (61 vs. 30%, p = 0.021) than those with TAV. BAV patients showed a significantly lower IMT and fewer plaques. Although FAC and GCS were not significantly different between the two groups, they tended to be lower in the BAV group when each group was divided into three subgroups according to age. There was a significant age-dependent increase in IMT and decreases in FAC and GCS in the TAV group (p = 0.005, p = 0.001, p = 0.002, respectively), but this phenomenon was not evident in the BAV group (p = 0.074, p = 0.248, p = 0.394, respectively). BAV patients with aortopathy showed a higher SD-CS than those without aortopathy (p = 0.040), reflecting disordered mechanical function. In conclusion, BAV patients have different carotid artery structure and function compared with TAV patients, suggesting intrinsic vascular abnormalities that are less affected by established cardiovascular risk factors and more strongly related to the presence of aortopathy.


Intrinsic Differential Scanning Fluorimetry for Fast and Easy Identification of Adeno-Associated Virus Serotypes.
Recombinant adeno-associated virus (AAV) vectors have evolved as the most promising technology for gene therapy due to their good safety profile, high transduction efficacy, and long-term gene expression in non-dividing cells. AAV-based gene therapy holds great promise for treating genetic disorders like inherited blindness, muscular atrophy, or bleeding disorders. Multiple naturally occurring and engineered AAV serotypes exist, which differ in capsid sequence and as a consequence in cellular tropism. Individual AAV capsids differ in thermal stability and have a characteristic melting temperature (Tm), which enables serotype-specific discrimination of AAV vectors. Differential scanning fluorimetry (DSF) combined with a dye-like SYPRO Orange (SO-DSF), which binds to hydrophobic regions of unfolded proteins, has been successfully applied to determine the Tm of AAV capsids. Here, we present DSF measurement of intrinsic fluorescence signal (iDSF) as a simple alternative method for determination of AAV capsid Tm. The study demonstrates that DSF measurement of intrinsic fluorescence signal is a simple, accurate, and rapid alternative to SO-DSF, which enables characterization of AAV capsid stability with excellent precision and without the need of SO or any other dye.


Motion tracking for medical imaging: a nonvisible structured light tracking approach.
We present a system for head motion tracking in 3D brain imaging. The system is based on facial surface reconstruction and tracking using a structured light (SL) scanning principle. The system is designed to fit into narrow 3D medical scanner geometries limiting the field of view. It is tested in a clinical setting on the high resolution research tomograph (HRRT), Siemens PET scanner with a head phantom and volunteers. The SL system is compared to a commercial optical tracking system, the Polaris Vicra system, from NDI based on translatory and rotary ground truth motions of the head phantom. The accuracy of the systems was similar, with root mean square (rms) errors of 0.09 degrees for ±20 degrees axial rotations, and rms errors of 0.24 mm for ± 25 mm translations. Tests were made using (1) a light emitting diode (LED) based miniaturized video projector, the Pico projector from Texas Instruments, and (2) a customized version of this projector replacing a visible light LED with a 850 nm near infrared LED. The latter system does not provide additional discomfort by visible light projection into the patient's eyes. The main advantage over existing head motion tracking devices, including the Polaris Vicra system, is that it is not necessary to place markers on the patient. This provides a simpler workflow and eliminates uncertainties related to marker attachment and stability. We show proof of concept of a marker less tracking system especially designed for clinical use with promising results.


Anxiety and Depressive Symptoms as Predictors of All-Cause Mortality among People with Insulin-Naïve Type 2 Diabetes: 17-Year Follow-Up of the Second Nord-Trøndelag Health Survey (HUNT2), Norway.
To examine whether elevated anxiety and/or depressive symptoms are related to all-cause mortality in people with Type 2 diabetes, not using insulin. 948 participants in the community-wide Nord-Trøndelag Health Survey conducted during 1995-97 completed the Hospital Anxiety and Depression Scale with subscales of anxiety (HADS-A) and depression (HADS-D). Elevated symptoms were defined as HADS-A or HADS-D ≥8. Participants with type 2 diabetes, not using insulin, were followed until November 21, 2012 or death. Cox regression analyses were used to estimate associations between baseline elevated anxiety symptoms, elevated depressive symptoms and mortality, adjusting for sociodemographic factors, HbA1c, cardiovascular disease and microvascular complications. At baseline, 8% (n = 77/948) reported elevated anxiety symptoms, 9% (n = 87/948) elevated depressive symptoms and 10% (n = 93/948) reported both. After a mean follow-up of 12 years (SD 5.1, range 0-17), 541 participants (57%) had died. Participants with elevated anxiety symptoms only had a decreased mortality risk (unadjusted HR 0.66, 95% CI 0.46-0.96). Adjustment for HbA1c attenuated this relation (HR 0.73, 95% CI 0.50-1.07). Those with elevated depression symptoms alone had an increased mortality risk (fully adjusted model HR 1.39, 95% CI 1.05-1.84). Having both elevated anxiety and depressive symptoms was not associated with increased mortality risk (adjusted HR 1.30, 95% CI 0.96-1.74). Elevated depressive symptoms were associated with excess mortality risk in people with Type 2 diabetes not using insulin. No significant association with mortality was found among people with elevated anxiety symptoms. Having both elevated anxiety and depressive symptoms was not associated with mortality. The hypothesis that elevated levels of anxiety symptoms leads to behavior that counteracts the adverse health effects of Type 2 diabetes needs further investigation.


Evidence of a genetic basis of Morgagni-Stewart-Morel syndrome. A case report of identical twins.
We report two 71-year-old female monozygotic twins presenting with advanced hyperostosis frontalis interna, obesity, shortness and cognitive impairment. They both have suffered from generalized seizures since their early adulthood. Moreover, the patients showed some additional conditions only occurring in one individual or the other such as migraine, marked recurrent depressive disorder or polyarthrosis. The symptoms common to both twins appear to correspond to the Morgagni-Stewart-Morel syndrome and indicate a genetic basis of this disorder as these features occur in genetically identical patients.


Characterization of a monoclonal antibody recognizing an epitope designated as canine leukocyte-associated antigen.
An IgG1 monoclonal antibody (mAb), designated as 15F1.5, was generated against surface determinants of a dog peripheral blood-derived PHA-induced IL-2-dependent T cell line. It reacted with 65-80% of peripheral blood mononuclear cells (PBMCs), 90-95% of polymorphonuclear cells (PMNs), 65-70% of thymocytes, 85-95% of Thy-1 positive cells and 85-95% of IL-2-dependent T lymphoid cells in flow cytometry. It was nonreactive with peripheral blood red cells and platelets. It immunoprecipitated 95 and 150 Kd proteins derived from detergent solubilized lymphocyte membranes. Indirect immunofluorescent and immunoperoxidase staining of frozen tissue sections demonstrated positive reactivity to cells in lymphoid but not nonlymphoid tissues. The 15F1.5 antibody was not directly mitogenic for PBMC's. It caused significant decrease (P < or = 0.05) in the lymphoproliferative response to T-dependent B cell mitogens, such as pokeweed mitogen (PWM) and staphage lysate (SPL), without significant effects on responses to the T cell mitogens, phytohemagglutinin (PHA), and concanavalin A (Con A). The mixed lymphocyte culture (MLC) response and both the proliferative and effector arms of the cell-mediated cytotoxicity reactions (CMC) were inhibited in a dose-dependent manner. The mAb also inhibited the auto- and allolymphoproliferative reactivity of mixed lymphocyte kidney or islet cell cultures (MLKC and MLIC), and the adhesion of T lymphoblasts and PMA-treated PMNs to endothelial cells. In vivo administration of the 15F1.5 (20 mg/day for 5 days) caused an immediate and prolonged reduction in MLC responses, associated with cell binding of the mAb to PBMC and epitope modulation during the course of treatment, as indicated by flow cytometry. These results suggest that 15F1.5 is an immunomodulating antibody reacting with canine LFA-1. Thus, this mAb would be useful in studying the role of LFA-1/ICAM-1 in graft rejection as well as other inflammatory responses. It would also allow the use of an animal model to investigate the immunoregulatory effects of in vivo administration of anti-CD11/CD18 antibodies in organ/tissue transplants.


Pernicious effect of physical cutoffs in fractal analysis.
Fractal scaling appears ubiquitous, but the typical extension of the scaling range observed is just one to two decades. A recent study has shown that an apparent fractal scaling spanning a similar range can emerge from the randomness in dilute sets. We show that this occurs also in most kinds of nonfractal sets irrespective of defining the fractal dimension by box counting, minimal covering, the Minkowski sausage, Walker's ruler, or the correlation dimension. We trace this to the presence of physical cutoffs, which induce smooth changes in the scaling, and a bias over a couple of decades around some characteristic length. The latter affects also the practical measure of fractality of truly fractal objects. A defensive strategy against artifacts and bias consists in carefully identifying the cutoffs and a quick-and-dirty thumb rule requires to observe fractal scaling over at least three decades.


Stepping stones: Principal career paths and school outcomes.
More than one out of every five principals leaves their school each year. In some cases, these career changes are driven by the choices of district leadership. In other cases, principals initiate the move, often demonstrating preferences to work in schools with higher achieving students from more advantaged socioeconomic backgrounds. Principals often use schools with many poor or low-achieving students as stepping stones to what they view as more desirable assignments. We use longitudinal data from one large urban school district to study the relationship between principal turnover and school outcomes. We find that principal turnover is, on average, detrimental to school performance. Frequent turnover of school leadership results in lower teacher retention and lower student achievement gains. Leadership changes are particularly harmful for high poverty schools, low-achieving schools, and schools with many inexperienced teachers. These schools not only suffer from high rates of principal turnover but are also unable to attract experienced successors. The negative effect of leadership changes can be mitigated when vacancies are filled by individuals with prior experience leading other schools. However, the majority of new principals in high poverty and low-performing schools lack prior leadership experience and leave when more attractive positions become available in other schools.


Vaccines targeting tumor blood vessel antigens promote CD8(+) T cell-dependent tumor eradication or dormancy in HLA-A2 transgenic mice.
We have recently shown that effective cytokine gene therapy of solid tumors in HLA-A2 transgenic (HHD) mice lacking murine MHC class I molecule expression results in the generation of HLA-A2-restricted CD8(+) T effector cells selectively recognizing tumor blood vessel-associated pericytes and/or vascular endothelial cells. Using an HHD model in which HLA-A2(neg) tumor (MC38 colon carcinoma or B16 melanoma) cells are not recognized by the CD8(+) T cell repertoire, we now show that vaccines on the basis of tumor-associated blood vessel Ags (TBVA) elicit protective Tc1-dependent immunity capable of mediating tumor regression or extending overall survival. Vaccine efficacy was not observed if (HLA-A2(neg)) wild-type C57BL/6 mice were instead used as recipient animals. In the HHD model, effective vaccination resulted in profound infiltration of tumor lesions by CD8(+) (but not CD4(+)) T cells, in a coordinate reduction of CD31(+) blood vessels in the tumor microenvironment, and in the "spreading" of CD8(+) T cell responses to alternate TBVA that were not intrinsic to the vaccine. Protective Tc1-mediated immunity was durable and directly recognized pericytes and/or vascular endothelial cells flow-sorted from tumor tissue but not from tumor-uninvolved normal kidneys harvested from these same animals. Strikingly, the depletion of CD8(+), but not CD4(+), T cells at late time points after effective therapy frequently resulted in the recurrence of disease at the site of the regressed primary lesion. This suggests that the vaccine-induced anti-TBVA T cell repertoire can mediate the clinically preferred outcomes of either effectively eradicating tumors or policing a state of (occult) tumor dormancy.


Correlation of estrophilin content of primary mammary cancer to eventual endocrine treatment.
The estrogen receptor protein content of recurrent breast cancer correlates well with the clinical response of hormonal manipulation. This predictive value of the ER of the primary tumor obtained at the time of mastectomy has not yet been proven. If this predictive capability should hold for primary tumors and eventual endocrine treatment, the ramifications are obvious: endocrine therapy could be offered to patients on a rational basis and adjuvant therapy could be considered on a plausible biochemical basis. This report details our observations as to the ER content of the primary tumor and the eventual result of endocrine therapy. Thirty-seven patients whose tumor ER was determined from the primary tumor eventually underwent some form of endocrine therapy for recurrent disease. Fifteen of the primary tumors had significant ER content and 22 possessed insignificant amounts. Only one of the 22 patients with insignificant ER content was benefited by endocrine treatment. Those patients whose tumors contained low amounts of ER experienced recurrence of their disease more rapidly than did those with higher ER content. There was no correlation of age, cell type of tumor or metastatic site with the ER content of the tumor. There is a positive correlation between response to chemotherapy and ER content of tumor. Measurement of the estrogen receptor protein content of the primary breast tumor is a reliable method for choosing patients for eventual endocrine therapy. Those patients whose tumors contain insignificant estrophilin are not candidates for such attempts at palliation.


Epidemic of Potato virus Y and Cucumber mosaic virus in Henan Province Tobacco.
Flue-cured tobacco is an important crop in Henan Province, China. During the 2000 growing season, many tobacco plants showed various degrees of mottling, mosaic, vein clearing, or vein necrosis in most of the counties. Some plants even died at an early stage of growth. A survey was conducted in May-June in several tobacco-growing counties, and the incidence of symptomatic plants in individual fields ranged from 10 to 85%. The most widely planted tobacco varieties, NC89, K326, and K346, were highly susceptible. Symptomatic plants were collected from Jiaxian and Xiangcheng counties and samples were tested by enzyme-linked immunosorbent assay for Tobacco mosaic virus (TMV), Cucumber mosaic virus (CMV), Potato virus Y (PVY), and Potato virus X (PVX). Of 65 samples tested, 21 were positive for only PVY, 16 positive for only CMV, one each was positive for only TMV or PVX. Nineteen samples were doubly infected with various combinations of these viruses and six were infected with combinations of three viruses. The causal agent(s) in the remaining sample could not be determined. In total, CMV was detected in 40 samples, PVY in 38, PVX in 10, and TMV in 7 samples. TMV and CMV used to be the most important viruses and PVY occurred only rarely. But PVY has become prevalent in Henan and in neighboring Shandong province (2). CMV and TMV were reported to be the most prevalent viruses in Shanxi (1) and Fujian Provinces (3). Because resistant varieties are not available, and mixed infections are more common, the results presented here explain why huge damage is occurring in tobacco crops in recent years. Some varieties are partially resistant to TMV and CMV but the varieties commonly grown are highly susceptible to PVY. Therefore, breeding for resistance to viruses, especially to PVY, is urgent to control the occurrence of tobacco viral diseases. References: (1) J. L. Cheng et al. Acta Tabacaria Sin. 4:43, 1998. (2) J. B. Wang et al. Chinese Tobacco Sci. 1:26, 1998. (3) L. H. Xie et al. Acta Tabacaria Sin. 2:25, 1994.


Purification and basic biochemical characterization of 19 recombinant plant peroxidase isoenzymes produced in Pichia pastoris.
The plant enzyme horseradish peroxidase (HRP) is used in several important industrial and medical applications, of which especially biosensors and diagnostic kits describe an emerging field. Although there is an increasing demand for high amounts of pure enzyme preparations, HRP is still isolated from the plant as a mixture of different isoenzymes with different biochemical properties. Based on a recent next generation sequencing approach of the horseradish transcriptome, we produced 19 individual HRP isoenzymes recombinantly in the yeast Pichia pastoris. After optimizing a previously reported 2-step purification strategy for the recombinant isoenzyme HRP C1A by substituting an unfavorable size exclusion chromatography step with an anion exchange step using a monolithic column, we purified the 19 HRP isoenzymes with varying success. Subsequent basic biochemical characterization revealed differences in catalytic activity, substrate specificity and thermal stability of the purified HRP preparations. The preparations of the isoenzymes HRP A2A and HRP A2B were found to be highly interesting candidates for future applications in diagnostic kits with increased sensitivity.


Late Endobronchial Pulmonary Metastasis in a Patient with Breast Cancer.
The lung is a common location for malignant metastases. However, endobronchial metastases from nonpulmonary neoplasms are rare. Metastatic breast cancer usually occurs in 2-3 years of the disease course. A 65-year-old woman visited our hospital for the evaluation of dry cough. The patient had a history of breast cancer, which was treated with modified radical mastectomy and axillary dissection 10 years ago; she was then treated with aromatase inhibitor for 5 years. Chest X-ray revealed right hilum enlargement. Thorax computed tomography revealed a 35-mm diameter mass that was localized in the right hilum. Fiberoptic bronchoscopy was performed, and an endobronchial lesion was observed in the right main lobe carina. Pathological evaluation revealed that the mass was a metastasis of the invasive ductal carcinoma of the breast. Weekly paklitaxel chemotherapy was initiated because of the symptomatic disease. We reported the case of a patient with breast cancer who had an endobronchial metastasis. Her disease-free interval was 10 years. This case indicates that a long-term follow-up of breast cancer is necessary, and biopsies must be performed to make a final diagnosis when any suspicious hilum enlargement is observed.