The herpes simplex virus UL33 gene product is required for the assembly of full capsids.
Phenotypic analysis of the herpes simplex virus type 1 temperature-sensitive DNA-positive mutant, ts1233, revealed that the mutant had a structural defect at the nonpermissive temperature (NPT). Cells infected with ts1233 at the NPT contained large numbers of intermediate capsids, lacking dense cores but possessing some internal structure. No full capsids or enveloped virus particles were detected. In contrast to the defect in another packaging-deficient mutant ts1201, the block in the formation of dense-cored, DNA-containing capsids in ts1233-infected cells at the NPT could not be reversed by transferring the cells to the permissive temperature in the presence of a protein synthesis inhibitor. Furthermore, the capsids produced by ts1233 at the NPT had more compact internal structures than those of the gene UL26 mutant ts1201. Southern blot analysis of viral DNA in ts1233-infected cells confirmed that the mutant DNA was not encapsidated at the NPT and showed that the unpackaged DNA was not cleaved into genome-length molecules. The ts1233 mutation was mapped by marker rescue to the vicinity of genes UL32 and UL33. Sequence analysis of the DNA in this region from the mutant and two independently isolated revertants for growth revealed that ts1233 had a single base-pair change at the amino-terminal end of UL33, resulting in the substitution of an isoleucine with an asparagine. The nucleotide sequence of the revertants in this part of the genome was identical to that of wild-type virus.


Further motor unit studies in Duchenne muscular dystrophy.
Numbers of functioning motor units have been estimated in 124 muscles of boys with Duchenne dystrophy; some of the patients were studied on several occasions. In the distal muscles examined (extensor digitorum brevis, thenar, and hypothenar muscles) the losses of units were probably present at birth and did not decrease with age. In contrast, the numbers of units and of excitable muscle fibres in the soleus muscles declined significantly, especially between the ages of 9 and 12 years.


Intestinal parasites including Cryptosporidium, Cyclospora, Giardia, and Microsporidia, Entamoeba histolytica, Strongyloides, Schistosomiasis, and Echinococcus: Guidelines from the American Society of Transplantation Infectious Diseases Community of Practice.
These updated guidelines from the Infectious Diseases Community of Practice of the American Society of Transplantation review the diagnosis, prevention, and management of intestinal parasites in the pre- and post-transplant period. Intestinal parasites are prevalent in the developing regions of the world. With increasing travel to and from endemic regions, changing immigration patterns, and the expansion of transplant medicine in developing countries, they are increasingly recognized as a source of morbidity and mortality in solid-organ transplant recipients. Parasitic infections may be acquired from the donor allograft, from reactivation, or from de novo acquisition post-transplantation. Gastrointestinal multiplex assays have been developed; some of the panels include testing for Cryptosporidium, Cyclospora, Entamoeba histolytica, and Giardia, and the performance is comparable to conventional methods. A polymerase chain reaction test, not yet widely available, has also been developed to detect Strongyloides in stool samples. New recommendations have been developed to minimize the risk of Strongyloides donor-derived events. Deceased donors with epidemiological risk factors should be screened for Strongyloides and recipients treated if positive as soon as the results are available. New therapeutic agents and studies addressing the optimal treatment regimen for solid-organ transplant recipients are unmet needs.


Predator-prey interactions amongst Permo-Triassic terrestrial vertebrates as a deterministic factor influencing faunal collapse and turnover.
Unlike modern mammalian communities, terrestrial Paleozoic and Mesozoic vertebrate systems were characterized by carnivore faunas that were as diverse as their herbivore faunas. The comparatively narrow food base available to carnivores in these paleosystems raises the possibility that predator-prey interactions contributed to unstable ecosystems by driving populations to extinction. Here, we develop a model of predator-prey interactions based on diversity, abundance and body size patterns observed in the Permo-Triassic vertebrate fossil record of the Karoo Basin, South Africa. Our simulations reflect empirical evidence that despite relatively high carnivore: herbivore species ratios, herbivore abundances were sufficient for carnivores to maintain required intake levels through most of the Karoo sequence. However, high mortality rates amongst herbivore populations, even accounting for birth rates of different-sized species, are predicted for assemblages immediately preceding the end-Guadalupian and end-Permian mass extinctions, as well as in the Middle Triassic when archosaurs replaced therapsids as the dominant terrestrial fauna. These results suggest that high rates of herbivore mortality could have played an important role in biodiversity declines leading up to each of these turnover events. Such declines would have made the systems especially vulnerable to subsequent stochastic events and environmental perturbations, culminating in large-scale extinctions.


Enhanced expressions of neurodegeneration-associated factors, UPS impairment, and excess Aβ accumulation in the hippocampus of mice with persistent cerebral toxocariasis.
Toxocariasis is a worldwide zoonotic parasitic disease mainly caused by Toxocara canis. Humans can be infected by accidental ingestion of T. canis embryonated ovum-contaminated food, water, or encapsulated larvae in paratenic hosts' viscera or meat. Since humans and mice are paratenic hosts of T. canis, the wandering larvae might cause mechanical tissue damage and excretory-secretory antigens may trigger inflammatory injuries to local organs. Long-term residence of T. canis larvae in a paratenic host's brain may cause cerebral toxocariasis (CT) that contributes to cerebral damage, neuroinflammation and neuropsychiatric disorders in mice and clinical patients. Since the hippocampus has been long recognized as being responsible for learning and memory functions, parasitic invasion of this site may cause neuroinflammatory and neurodegenerative disorders. The present study intended to assess pathological changes, expressions of neurodegeneration-associated factors (NDAFs), including transforming growth factor (TGF)-β1, S100B, glial fibrillary acidic protein (GFAP), transglutaminase type 2 (TG2), claudin-5, substance P (SP) and interleukin (IL)-1β, and the ubiquitin-proteasome system (UPS) function in the hippocampus and associated cognitive behavior in ICR mice orally inoculated with a high, medium or low-dose of T. canis embryonated ova during a 20-week investigation. Results indicated although there were insignificant differences in learning and memory function between the experimental mice and uninfected control mice, possibly because the site where T. canis larvae invaded was the surrounding area but not the hippocampus per se. Nevertheless, enhanced expressions of NDAF, persistent UPS impairment and excess amyloid β (Aβ) accumulation concomitantly emerged in the experimental mice hippocampus at 8, 16 and 20 weeks post-infection. We thus postulate that progressive CT may still progress to neurodegeneration due to enhanced NDAF expressions, persistent UPS impairment and excess Aβ accumulation in the hippocampus.


Contour detection and hierarchical image segmentation.
This paper investigates two fundamental problems in computer vision: contour detection and image segmentation. We present state-of-the-art algorithms for both of these tasks. Our contour detector combines multiple local cues into a globalization framework based on spectral clustering. Our segmentation algorithm consists of generic machinery for transforming the output of any contour detector into a hierarchical region tree. In this manner, we reduce the problem of image segmentation to that of contour detection. Extensive experimental evaluation demonstrates that both our contour detection and segmentation methods significantly outperform competing algorithms. The automatically generated hierarchical segmentations can be interactively refined by user-specified annotations. Computation at multiple image resolutions provides a means of coupling our system to recognition applications.


Caffeine as a Factor Influencing the Functioning of the Human Body-Friend or Foe?
Nowadays, caffeine is one of the most commonly consumed substances, which presents in many plants and products. It has both positive and negative effects on the human body, and its activity concerns a variety of systems including the central nervous system, immune system, digestive system, respiratory system, urinary tract, etc. These effects are dependent on quantity, the type of product in which caffeine is contained, and also on the individual differences among people (sex, age, diet etc.). The main aim of this review was to collect, present, and analyze the available information including the latest discoveries on the impact of caffeine on human health and the functioning of human body systems, taking into account the role of caffeine in individual disease entities. We present both the positive and negative sides of caffeine consumption and the healing properties of this purine alkaloid in diseases such as asthma, Parkinson's disease, and others, not forgetting about the negative effects of excess caffeine (e.g., in people with hypertension, children, adolescents, and the elderly). In summary, we can conclude, however, that caffeine has a multi-directional influence on various organs of the human body, and because of its anti-oxidative properties, it was, and still is, an interesting topic for research studies including those aimed at developing new therapeutic strategies.


Alcohol, drugs, sex and violence: health risks and consequences in young British holidaymakers to the Balearics.
Substance use, risky sexual behaviour and violence are among the key youth health issues today. Whilst they are the focus of much prevention work in the UK, relatively little information is available to inform prevention in international holiday resorts, where young people can take the greatest risks with their health. We conducted a cross-sectional survey of 1033 British holidaymakers aged 16-35 in Ibiza and Majorca airports (Spain). Surveys covered frequency of alcohol and drug use at home and abroad, and drunkenness, sexual activity and experience of a range of negative experiences on holiday. Illicit drug use was more common in Ibiza. Over half (53.9%) of visitors to Ibiza reported using at least one drug on holiday (compared with 13.9% in Majorca) and over a third reported using more than one drug. However visitors to Majorca reported more frequent drunkenness and more negative outcomes (arguments, fighting, unintentional injury, illness and requiring medical attention). In both locations, frequency of substance use increased on holiday. A quarter of those travelling without a partner reported having had sex during their holiday, and of these a third did not consistently use condoms. Frequent drunkenness was associated with fighting, unintentional injury and sex with a new partner. Illicit drug use, particularly polydrug use, was associated with fighting and unprotected sex. Young holidaymakers face significant health risks on holiday. Understanding which resorts are associated with which types of health risk behaviour is critical in targeting health promotion and harm reduction measures appropriately.


Interactions of serum copper, selenium, and low density lipoprotein cholesterol in atherogenesis.
To investigate the interactions between serum copper, selenium, and low density lipoprotein cholesterol concentrations with regard to the progression of carotid atherosclerosis. Longitudinal study of a cohort of middle aged men followed up for 24 months. Epidemiological survey of the population of seven communities in eastern Finland. 126 men aged 42, 48, 54, or 60 at examination randomly selected from a population based sample of 2682 men. Increase in maximal carotid intima media thickness. The mean increase in the maximal common carotid intima media thickness after two years was greater in men with high serum copper concentrations (0.16 mm compared with 0.08 mm in men with concentrations less than 17.6 mumol/l; p = 0.010), those with low serum selenium concentrations (0.15 mm compared with 0.09 mm in men with concentrations greater than or equal to 1.40 mumol/l; p = 0.035), and those with raised serum low density lipoprotein cholesterol concentrations (0.15 mm compared with 0.08 mm in men with concentrations less than 4.0 mmol/l; p = 0.032) after adjustment for age and cigarette pack years in a three way analysis of covariance. A raised serum low density lipoprotein concentration was associated with accelerated progression of atherosclerosis only in men with higher than median serum copper concentrations (net difference 0.22 mm; p less than 0.001 for two way interaction), and this synergism was even more pronounced in men with serum selenium concentrations below the median value (net difference 0.41 mm; p = 0.042 for three way interaction). These data provide evidence of a synergistic effect of copper (a pro-oxidant), a low serum concentration of selenium (a cofactor of an enzyme that scavenges free radicals), and low density lipoprotein cholesterol concentration in atherogenesis.


Filaggrin in atopic dermatitis.
The recent identification of loss-of-function mutations in the structural protein filaggrin as a widely replicated major risk factor for eczema sheds new light on disease mechanisms in eczema, a disease that had heretofore largely been considered to have a primarily immunologic etiopathogenesis. The filaggrin gene (FLG) mutation findings are consistent with a recently proposed unifying hypothesis that offers a mechanistic understanding of eczema pathogenesis synthesizing a heritable epithelial barrier defect and resultant diminished epidermal defense mechanisms to allergens and microbes, followed by polarized T(H)2 lymphocyte responses with resultant chronic inflammation, including autoimmune mechanisms. Although compelling evidence from genetic studies on FLG implicates perturbed barrier function as a key player in the pathogenesis of eczema in many patients, much is still unknown about the sequence of biologic, physicochemical, and aberrant regulatory events that constitute the transition from an inherited barrier defect to clinical manifestations of inflammatory eczematous lesions and susceptibility to related atopic disorders. The exact contribution of FLG to the wider atopic story, factors modifying FLG expression, and the role of other barrier proteins remain to be delineated. In this review we highlight recent advances in our understanding of the FLG genetics in the cause of eczema and related complex diseases.


Part II: Functional delivery of a neurotherapeutic gene to neural stem cells using minicircle DNA and nanoparticles: Translational advantages for regenerative neurology.
Both neurotrophin-based therapy and neural stem cell (NSC)-based strategies have progressed to clinical trials for treatment of neurological diseases and injuries. Brain-derived neurotrophic factor (BDNF) in particular can confer neuroprotective and neuro-regenerative effects in preclinical studies, complementing the cell replacement benefits of NSCs. Therefore, combining both approaches by genetically-engineering NSCs to express BDNF is an attractive approach to achieve combinatorial therapy for complex neural injuries. Current genetic engineering approaches almost exclusively employ viral vectors for gene delivery to NSCs though safety and scalability pose major concerns for clinical translation and applicability. Magnetofection, a non-viral gene transfer approach deploying magnetic nanoparticles and DNA with magnetic fields offers a safe alternative but significant improvements are required to enhance its clinical application for delivery of large sized therapeutic plasmids. Here, we demonstrate for the first time the feasibility of using minicircles with magnetofection technology to safely engineer NSCs to overexpress BDNF. Primary mouse NSCs overexpressing BDNF generated increased daughter neuronal cell numbers post-differentiation, with accelerated maturation over a four-week period. Based on our findings we highlight the clinical potential of minicircle/magnetofection technology for therapeutic delivery of key neurotrophic agents.


Evaluation of blood-testis barrier integrity in terms of adhesion molecules in nonobstructive azoospermia.
Blood-testis barrier (BTB) is critical for maintaining fertility. The integrity of tight junctions (TJs) provides restricted permeability of BTB. The aim of this study was to evaluate the relationship between BTB and Sertoli cells. Testicular sperm extraction (TESE) obtained from nonobstructive azoospermia (NOA) patients was examined: Group I (spermatozoa+) and Group II (spermatozoa-). The tissues were stained with haematoxylin eosin, periodic acid-Schiff and Masson's trichrome for Johnsen's score evaluation. Apoptosis and adhesion molecules such as claudin-11, occludin and ZO-1 were assessed. In Group I, the integrity of the seminiferous tubules was intact. In Group II, some seminiferous tubule walls were lined only with Sertoli cells, had a thickening of the basement membrane, and oedema in interstitial spaces. In Group I, the seminiferous tubule consisted of a stratified columnar epithelium, claudin-11 expressions were observed as linear staining in the basal zone of the tubule, while seminiferous tubules, with low epithelium, displayed a punctate type of staining. Immunohistochemical observations were consistent with the ultrastructural findings. In Group II, high apoptosis and unstained/irregular TJ formation in claudin-11, occludin and ZO-1 were observed. In conclusion, disruption of relation between BTB and TJs may reveal inadequate spermatogenesis, which is one of the mechanisms behind azoospermia.


Spatial distribution of insecticide resistance in Caribbean populations of Aedes aegypti and its significance.
To monitor resistance to insecticides, bioassays were performed on 102 strains of the dengue vector Aedes aegypti (L.) from 16 countries ranging from Suriname in South America and through the chain of Caribbean Islands to the Bahamas, where the larvicide temephos and the adulticide malathion have been in use for 15 to 30 years. There was wide variation in the sensitivity to the larvicide in mosquito populations within and among countries. Mosquito strains in some countries such as Antigua, St. Lucia, and Tortola had consistently high resistance ratios (RR) to temephos, ranging from 5.3 to 17.7. In another group of countries--e.g., Anguilla and Curaçao--mosquitoes had mixed levels of resistance to temephos (RR = 2.5-10.6), and in a third group of countries, including St. Kitts, Barbados, Jamaica, and Suriname, mosquitoes had consistently low levels of resistance to temephos (RR = 1-4.6) (P < 0.05). On occasion significantly different levels of resistance were recorded from neighboring A. aegypti communities, which suggests there is little genetic exchange among populations. The impact of larval resistance expressed itself as reduced efficacy of temephos to kill mosquitoes when strains were treated in the laboratory or in the field in large container environments with recommended dosages. Although a sensitive strain continued to be completely controlled for up to 7 weeks, the most resistant strains had 24% survival after the first week. By week 6, 60% to 75% of all resistant strains of larvae were surviving the larval period. Responses to malathion in adult A. aegypti varied from a sensitive population in Suriname (RR = 1.3) to resistant strains in St. Vincent (RR = 4.4), Dominica (RR = 4.2), and Trinidad (RR = 4.0); however, resistance was generally not on the scale of that observed to temephos in the larval stages and had increased only slightly when compared to the levels that existed 3 to 4 years ago. Suggestions are made for a pesticide usage policy for the Caribbean region, with modifications for individual countries. This would be formulated based on each country's insecticide-resistance profile. Use of physical and biological control strategies would play a more critical role than the use of insecticides.


BAC-end sequences analysis provides first insights into coffee (Coffea canephora P.) genome composition and evolution.
Coffee is one of the world's most important agricultural commodities. Coffee belongs to the Rubiaceae family in the euasterid I clade of dicotyledonous plants, to which the Solanaceae family also belongs. Two bacterial artificial chromosome (BAC) libraries of a homozygous doubled haploid plant of Coffea canephora were constructed using two enzymes, HindIII and BstYI. A total of 134,827 high quality BAC-end sequences (BESs) were generated from the 73,728 clones of the two libraries, and 131,412 BESs were conserved for further analysis after elimination of chloroplast and mitochondrial sequences. This corresponded to almost 13 % of the estimated size of the C. canephora genome. 6.7 % of BESs contained simple sequence repeats, the most abundant (47.8 %) being mononucleotide motifs. These sequences allow the development of numerous useful marker sites. Potential transposable elements (TEs) represented 11.9 % of the full length BESs. A difference was observed between the BstYI and HindIII libraries (14.9 vs. 8.8 %). Analysis of BESs against known coding sequences of TEs indicated that 11.9 % of the genome corresponded to known repeat sequences, like for other flowering plants. The number of genes in the coffee genome was estimated at 41,973 which is probably overestimated. Comparative genome mapping revealed that microsynteny was higher between coffee and grapevine than between coffee and tomato or Arabidopsis. BESs constitute valuable resources for the first genome wide survey of coffee and provide new insights into the composition and evolution of the coffee genome.


Clinical Informatics Fellowship Programs: In Search of a Viable Financial Model: An open letter to the Centers for Medicare and Medicaid Services.
In the US, the new subspecialty of Clinical Informatics focuses on systems-level improvements in care delivery through the use of health information technology (HIT), data analytics, clinical decision support, data visualization and related tools. Clinical informatics is one of the first subspecialties in medicine open to physicians trained in any primary specialty. Clinical Informatics benefits patients and payers such as Medicare and Medicaid through its potential to reduce errors, increase safety, reduce costs, and improve care coordination and efficiency. Even though Clinical Informatics benefits patients and payers, because GME funding from the Centers for Medicare and Medicaid Services (CMS) has not grown at the same rate as training programs, the majority of the cost of training new Clinical Informaticians is currently paid by academic health science centers, which is unsustainable. To maintain the value of HIT investments by the government and health care organizations, we must train sufficient leaders in Clinical Informatics. In the best interest of patients, payers, and the US society, it is therefore critical to find viable financial models for Clinical Informatics fellowship programs. To support the development of adequate training programs in Clinical Informatics, we request that the Centers for Medicare and Medicaid Services (CMS) issue clarifying guidance that would allow accredited ACGME institutions to bill for clinical services delivered by fellows at the fellowship program site within their primary specialty.


Development of a Pharmacokinetic Model of Transplacental Transfer of Metformin to Predict In Vivo Fetal Exposure.
Two types of systems are used in ex vivo human placental perfusion studies to predict fetal drug exposures, that is, closed systems with recirculation of the maternal and fetal buffer and open systems using a single-pass mode without recirculation. The in vivo fetal/maternal (F:M) ratio of metformin, a cationic drug that crosses the placenta, is consistent with that reported in an open system ex vivo but not with that in a closed system. In the present study, we aimed to develop a pharmacokinetic (PK) model of transplacental transfer of metformin to predict in vivo fetal exposure to metformin and to resolve the apparent inconsistency between open and closed ex vivo systems. The developed model shows that the difference between open and closed systems is due to the difference in the time required to achieve the steady state. The model-predicted F:M ratio (approx. 0.88) is consistent with reported in vivo values [mean (95% confidence interval): 1.10 (0.69-1.51)]. The model incorporates bidirectional transport via organic cation transporter 3 (OCT3) at the basal plasma membrane, and simulations indicate that the use of trimethoprim (an OCT3 inhibitor) to prevent microbial growth in the placenta ex vivo has a negligible effect on the overall maternal-to-fetal and fetal-to-maternal clearances. The model could successfully predict in vivo fetal exposure using ex vivo human placental perfusion data from both closed and open systems. This transplacental PK modeling approach is expected to be useful for evaluating human fetal exposures to other poorly permeable compounds, besides metformin. SIGNIFICANCE STATEMENT: We developed a pharmacokinetic model of transplacental transfer of metformin, used to treat gestational diabetes mellitus, in order to predict in vivo fetal exposure and resolve the discrepancy between reported findings in open and closed ex vivo perfusion systems. The discrepancy is due to a difference in the time required to reach the steady state. The model can predict in vivo fetal exposure using data from both closed and open systems.


Observational study of treatment space in individual neonatal cot spaces.
Technology developments in neonatal intensive care units have increased the spatial requirements for clinical activities. Because the effectiveness of healthcare delivery is determined in part by the design of the physical environment and the spatial organization of work, it is appropriate to apply an evidence-based approach to architectural design. This study aimed to provide empirical evidence of the spatial requirements for an individual cot or incubator space. Observational data from 2 simulation exercises were combined with an expert review to produce a final recommendation. A validated 5-step protocol was used to collect data. Step 1 defined the clinical specialty and space. In step 2, data were collected with 28 staff members and 15 neonates to produce a simulation scenario representing the frequent and safety-critical activities. In step 3, 21 staff members participated in functional space experiments to determine the average spatial requirements. Step 4 incorporated additional data (eg, storage and circulation) to produce a spatial recommendation. Finally, the recommendation was reviewed in step 5 by a national expert clinical panel to consider alternative layouts and technology. The average space requirement for an individual neonatal intensive care unit cot (incubator) space was 13.5 m2 (or 145.3 ft2). The circulation and storage space requirements added in step 4 increased this to 18.46 m2 (or 198.7 ft2). The expert panel reviewed the recommendation and agreed that the average individual cot space (13.5 m2/[or 145.3 ft2]) would accommodate variance in working practices. Care needs to be taken when extrapolating this recommendation to multiple cot areas to maintain the minimum spatial requirement.


Economic and quality of care evaluation of dialysis service models in remote Australia: protocol for a mixed methods study.
Australia's Northern Territory (NT) has the country's highest incidence and prevalence of kidney disease. Indigenous people from remote areas suffer the heaviest disease burden. Concerns regarding cost and sustainability limit the provision of dialysis treatments in remote areas and most Indigenous people requiring dialysis relocate to urban areas. However, this dislocation of people from their family, community and support networks may prove more costly when the broader health, societal and economic consequences for the individual, family and whole of government are considered. The Dialysis Models of Care Study is a large cross organisation mixed methods study. It includes a retrospective (2000-2014) longitudinal data linkage study of two NT cohorts: Renal Cohort 1- comprising approximately 2000 adults who received dialysis and Renal Cohort 2- comprising approximately 400 children of those adults. Linkage of administrative data sets from the Australian and New Zealand Dialysis and Transplant Registry, NT Departments of Health, Housing and Education by a specialist third party (SA/NT Datalink) will enable extraction of activity, financial and outcome data. Interviews with patients, clinicians and service providers, using a snowball technique, will canvass relevant issues and assist in determining the full costs and impacts of the five most used dialysis Models of Care. The study uses a mixed methods approach to investigate the quantitative and qualitative dimensions of the full costs and outcomes associated with the choice of particular dialysis models of care for any given patient. The study includes a large data linkage component that for the first time links health, housing and education data to fully analyse and evaluate the impact on patients, their families and the broader community, resulting from the relocation of people for treatment. The study will generate a large amount of activity, financial and qualitative data that will investigate health costs less directly related to dialysis treatment, costs to government such as housing and/or education and the health, social and economic outcomes experienced by patients. This approach fills an evidence gap critical to health service planners.


BCG vaccination to prevent tuberculosis in health care workers: a decision analysis.
To perform a decision analysis to determine the optimal strategy to prevent tuberculosis (TB) in health care workers with negative tuberculin skin tests. We used a Markov model to study the occurrence of events each year and compared BCG vaccination to annual tuberculin testing plus isoniazid (INH) preventive therapy for those who become skin test positive. The outcome measures studied were the number of cases and deaths from TB and BCG and/or INH adverse reactions over 10 years. Annual tuberculin testing decreases the number of TB cases by 9% and BCG vaccination decreases the number by 49%, relative to no prevention intervention. BCG vaccination results in fewer deaths than annual tuberculin testing if the workplace incidence of Mycobacterium tuberculosis infection is greater than 0.06%, BCG vaccination effectiveness exceeds 3%, or the rate of fatal BCG adverse reactions is less than 15 times the rate reported in the literature. BCG vaccination results in less morbidity and mortality than annual tuberculin skin testing for health care workers in workplaces with documented TB transmission despite comprehensive infection control policies and procedures. Current policy on the prevention of TB among health care workers should be reconsidered.


Stronger perceptual filling-in of spatiotemporal information in the blind spot compared with artificial gaps.
Complete visual information about a scene and the objects within it is often not available to us. For example, objects may be partly occluded by other objects or have sections missing. In the retinal blind spot, there are no photoreceptors and visual input is not detected. However, owing to perceptual filling-in by the visual system we often do not perceive these gaps. There is a lack of consensus on how much of the mechanism for perceptual filling-in is similar in the case of a natural scotoma, such as the blind spot, and artificial scotomata, such as sections of the stimulus being physically removed. Part of the difficulty in assessing this relationship arises from a lack of direct comparisons between the two cases, with artificial scotomata being tested in different locations in the visual field compared with the blind spot. The peripheral location of the blind spot may explain its enhanced filling-in compared with artificial scotomata, as reported in previous studies. In the present study, we directly compared perceptual filling-in of spatiotemporal information in the blind spot and artificial gaps of the same size and eccentricity. We found stronger perceptual filling-in in the blind spot, suggesting improved filling-in for the blind spot reported in previous studies cannot be simply attributed to its peripheral location.


The p53-MDM2 network: from oscillations to apoptosis.
The p53 protein is well-known for its tumour suppressor function. The p53-MDM2 negative feedback loop constitutes the core module of a network of regulatory interactions activated under cellular stress. In normal cells, the level of p53 proteins is kept low by MDM2, i.e. MDM2 negatively regulates the activity of p53. In the case of DNA damage, the p53-mediated pathways are activated leading to cell cycle arrest and repair of the DNA. If repair is not possible due to excessive damage, the p53-mediated apoptotic pathway is activated bringing about cell death. In this paper, we give an overview of our studies on the p53-MDM2 module and the associated pathways from a systems biology perspective. We discuss a number of key predictions, related to some specific aspects of cell cycle arrest and cell death, which could be tested in experiments.


The genetics of eye disorders in the dog.
Inherited forms of eye disease are arguably the best described and best characterized of all inherited diseases in the dog, at both the clinical and molecular level and at the time of writing 29 different mutations have been documented in the scientific literature that are associated with an inherited ocular disorder in the dog. The dog has already played an important role in the identification of genes that are important for ocular development and function as well as emerging therapies for inherited blindness in humans. Similarities in disease phenotype and eye structure and function between dog and man, together with the increasingly sophisticated genetic tools that are available for the dog, mean that the dog is likely to play an ever increasing role in both our understanding of the normal functioning of the eye and in our ability to treat inherited eye disorders. This review summarises the mutations that have been associated with inherited eye disorders in the dog.


Effect of supplementing Rhodes grass hay (Chloris gayana) with Berchemia discolor or Zizyphus mucronata on the performance of growing goats in Kenya.
Twenty growing Small East African goats were used to determine the effects of feeding sun-dried leaves of the browse forages Berchemia discolor and Zizyphus mucronata as supplements to low-quality basal diet, Rhodes grass (Chloris gayana) hay, on voluntary feed intake (VFI), digestibility and growth performance. The grass hay and maize bran were used as a control. The dried leaves were then included at the rates of 15% and 30% of the dry matter intake (DMI). Berchemia discolor had the highest crude protein (CP) content of 195.5 g/kg DM, while Z. mucronata had CP content of 169.5 g/kg DM. The grass hay had the lowest CP content of 50.9 g/kg DM. The browse forages had low fibre content [Neutral detergent fibre (NDF); 257.9-369.5 g/kg DM], while the grass hay had high fibre content (NDF; 713.1 g/kg DM). Goats in the groups supplemented with either of the browse forages had higher total DMI, nitrogen (N) intake and retention and live-weight gains than those in the control diet group. The digestibility of DM and organic matter (OM) was not affected by supplementation, but the CP digestibility increased with supplementation. The use of the browse forages as supplements for goats fed on poor-quality basal diets would enhance the performance of the animals.


Pancreatic acinar cell carcinoma with extensive tumor embolism at the trunk of portal vein and pancreatic intraductal infiltration.
A 59-year-old man was admitted to the hospital with acute pancreatitis. The cause was suggested to be a pancreatic tumor based on computed tomography (CT). The pancreatic tumor was 45 mm with an extensive tumor embolism at the trunk of the portal vein and intraductal infiltration of the main pancreatic duct (MPD). The pancreatic tumor was diagnosed as acinar cell carcinoma (ACC) by endoscopic ultrasound guided fine needle aspiration. Therefore, the cause of acute pancreatitis was diagnosed to be intraductal infiltration of ACC in the MPD. Chemotherapy was initially performed because it was difficult to perform surgery due to extensive tumor embolism at the trunk of the portal vein. Degeneration and reduction of ACC and tumor embolism of the portal vein was noted on CT after chemotherapy, and extended distal pancreatectomy with portal vein reconstruction was performed. There has been no relapse for 5 years postoperative follow-up. This is an interesting and rare case because ACC with intraductal infiltration of MPD is low in frequency; most ACCs are asymptomatic when they are found, and many cases tend to have poor prognosis in spite of surgical cases.


Multiple Access Control for Cognitive Radio-Based IEEE 802.11ah Networks.
The proliferation of Internet-of-Things (IoT) technology and its reliance on the license-free Industrial, Scientific, and Medical (ISM) bands have rendered radio spectrum scarce. The IoT can nevertheless obtain great advantage from Cognitive Radio (CR) technology for efficient use of a spectrum, to be implemented in IEEE 802.11af-based primary networks. However, such networks require a geolocation database and a centralized architecture to communicate white space information on channels. On the other hand, in spectrum sensing, CR presents various challenges such as the Hidden Primary Terminal (HPT) problem. To this end, we focus on the most recently released standard, i.e., IEEE 802.11ah, in which IoT stations can first be classified into multiple groups to reduce collisions and then they can periodically access the channel. Therein, both services are similarly supported by a centralized server that requires signaling overhead to control the groups of stations. In addition, more regroupings are required over time due to the frequent variations in the number of participating stations, which leads to more overhead. In this paper, we propose a new Multiple Access Control (MAC) protocol for CR-based IEEE 802.11ah systems, called Restricted Access with Collision and Interference Resolution (RACIR). We introduce a decentralized group split algorithm that distributes the participating stations into multiple groups based on a probabilistic estimation in order to resolve collisions. Furthermore, we propose a decentralized channel access procedure that avoids the HPT problem and resolves interference with the incumbent receiver. We analyze the performance of our proposed MAC protocol in terms of normalized throughput, packet delay and energy consumption with the Markov model and analytic expressions. The results are quite promising, which makes the RACIR protocol a strong candidate for the CR-based IoT environment.


An interlaboratory study as useful tool for proficiency testing of chemical oxygen demand measurements using solid substrates and liquid samples with high suspended solid content.
In 2008, the first Proficiency Testing Scheme of Chemical Oxygen Demand (1(st)COD-PT(ADG)) was conducted to assess the results obtained for different research groups whose field work is mainly anaerobic digestion. This study was performed using four samples, two solid samples as raw materials and two solid samples to prepare high concentration suspended solid solutions. Invitations were sent to a large number of laboratories, mainly to anaerobic digestion research groups. Finally, thirty labs from sixteen countries agreed to participate, but for different reasons four participants could not send any data. In total, twenty-six results were reported to the COD-PT coordinator. This study showed the importance of continuous participation in proficiency testing (PT) schemes in order to compare the results obtained. Taking into account the lack of a general standard method and high quality certified reference materials (CRMs), the traceability of COD determination is not currently easy to check. In addition, the spread of participants' results obtained was high and pointed to the advisability of using consensus values due to their unreliability. Therefore, the theoretical oxygen demand (ThOD) values were considered as assigned values for all the samples analysed. On the other hand, in this PT the established standard deviation (ESD) has been determined by the Horwitz modified function. Participants of this 1(st)COD-PT(ADG) were asked to give a short report on the analytical method used. Although all the participants used potassium dichromate as their oxidant reagent, their experimental procedures were very different. With the purpose of comparing the results obtained, the different experimental conditions used were classified into five methods, corresponding to two main categories, open and closed reflux. The performance of laboratories was expressed by the z-score, whose value is considered satisfactory when z-score <or=+/-2. The overall analytical data evaluation showed that 64% of z-scores obtained were outside the accepted limits.


Impairment and delay of neutralizing antiviral antibody responses by virus-specific cytotoxic T cells.
After infection with some viruses that tend to persist, neutralizing antibodies are generated rather late, i.e., after 1 to 3 mo. This has been observed for HIV, Hepatitis B infection in man, or after infection with lymphocytic choriomeningitis virus (LCMV) in mice. In contrast, nonneutralizing antibodies to LCMV are generated by day 7 and reach high titers by day 10. This study attempts to evaluate reasons for late and low titered neutralizing antibody responses. After a primary infection with low doses (10(2) plaque forming units) of the poorly cytopathic LCMV-WE, neutralizing antibodies were rarely produced at detectable levels before days 60 to 120. In vivo depletion of CD8+ CTL led to a marked enhancement and acceleration of kinetics of the neutralizing antibody response. In contrast, nonneutralizing antibodies, including those specific for LCMV-GP carrying the neutralizing determinant, were detectable very early, i.e., 4 to 7 days after infection, with maximum titers usually by day 10 irrespective of the presence or absence of CTL. Mice completely lacking CD8+ T cells because of deletion by homologous recombination (CD8-/-) also exhibited neutralizing antibodies early by day 10 to 20, and by day 120 reached very high titers. The neurotropic isolate LCMV-ARMSTRONG induced low but significant titers of neutralizing antibodies relatively early (i.e., by days 7 to 10), whereas the lympho-viscerotrope mutant virus LCMV-ARMSTRONG Cl-13 did not. Early and effective CTL responses causing immunopathology (and immunosuppression) correlated with the absence of neutralizing antibodies. The discrepancy between the CTL-dependent inhibition of neutralizing versus unimpaired anti-GP ELISA responses cannot be explained by different Ag doses alone. Additionally, it may reflect different kinetics of the responses, whereby the later neutralizing responses possibly requiring IgG affinity maturation may be more susceptible to general immunosuppression; also B cells expressing neutralizing receptors, but not those with antibodies binding GP (or NP), may be actively infected, therefore they present viral peptides on class I MHC Ag and may become targets for anti-LCMV-specific CTL.


The Treatment of cardiovascular Risk in Primary care using Electronic Decision supOrt (TORPEDO) study-intervention development and protocol for a cluster randomised, controlled trial of an electronic decision support and quality improvement intervention in Australian primary healthcare.
Large gaps exist in the implementation of guideline recommendations for cardiovascular disease (CVD) risk management. Electronic decision support (EDS) systems are promising interventions to close these gaps but few have undergone clinical trial evaluation in Australia. We have developed HealthTracker, a multifaceted EDS and quality improvement intervention to improve the management of CVD risk. It is hypothesised that the use of HealthTracker over a 12-month period will result in: (1) an increased proportion of patients receiving guideline-indicated measurements of CVD risk factors and (2) an increased proportion of patients at high risk will receive guideline-indicated prescriptions for lowering their CVD risk. Sixty health services (40 general practices and 20 Aboriginal Community Controlled Health Services (ACCHSs) will be randomised in a 1:1 allocation to receive either the intervention package or continue with usual care, stratified by service type, size and participation in existing quality improvement initiatives. The intervention consists of point-of-care decision support; a risk communication interface; a clinical audit tool to assess performance on CVD-related indicators; a quality improvement component comprising peer-ranked data feedback and support to develop strategies to improve performance. The control arm will continue with usual care without access to these intervention components. Quantitative data will be derived from cross-sectional samples at baseline and end of study via automated data extraction. Detailed process and economic evaluations will also be conducted. The general practice component of the study is approved by the University of Sydney Human Research Ethics Committee (HREC) and the ACCHS component is approved by the Aboriginal Health and Medical Research Council HREC. Formal agreements with each of the participating sites have been signed. In addition to the usual scientific forums, results will be disseminated via newsletters, study websites, face-to-face feedback forums and workshops. The trial is registered with the Australian Clinical Trials Registry ACTRN 12611000478910.


Bradykinin and nerve growth factor release the capsaicin receptor from PtdIns(4,5)P2-mediated inhibition.
Tissue injury generates endogenous factors that heighten our sense of pain by increasing the response of sensory nerve endings to noxious stimuli. Bradykinin and nerve growth factor (NGF) are two such pro-algesic agents that activate G-protein-coupled (BK2) and tyrosine kinase (TrkA) receptors, respectively, to stimulate phospholipase C (PLC) signalling pathways in primary afferent neurons. How these actions produce sensitization to physical or chemical stimuli has not been elucidated at the molecular level. Here, we show that bradykinin- or NGF-mediated potentiation of thermal sensitivity in vivo requires expression of VR1, a heat-activated ion channel on sensory neurons. Diminution of plasma membrane phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2) levels through antibody sequestration or PLC-mediated hydrolysis mimics the potentiating effects of bradykinin or NGF at the cellular level. Moreover, recruitment of PLC-gamma to TrkA is essential for NGF-mediated potentiation of channel activity, and biochemical studies suggest that VR1 associates with this complex. These studies delineate a biochemical mechanism through which bradykinin and NGF produce hypersensitivity and might explain how the activation of PLC signalling systems regulates other members of the TRP channel family.


The Alcohol Marketing Landscape: Alcohol Industry Size, Structure, Strategies, and Public Health Responses.
The purpose of this study is to inform public health efforts to reduce alcohol-related harm by describing the alcohol marketing landscape. We review the size, structure, and strategies of both the U.S. national and global alcohol industries and their principal marketing activities and expenditures and provide a summary of public health responses. Primary data were obtained from advertising and alcohol industry market research firms and were supplemented by searches of peer-reviewed literature, business press, and online databases on global business and trade. Worldwide, alcohol sales totaled more than $1.5 trillion in 2017. Control of alcoholic beverage production and marketing is concentrated globally in the hands of a small number of firms. The oligopoly structure of the producing industry helps to generate high profits per dollar invested relative to other industries, which in turn fund marketing expenditures that function as barriers to entry by other firms. Advertising expenditures are high and advertising is widespread. Stakeholder marketing and corporate social responsibility campaigns assist in maintaining a policy environment conducive to extensive alcohol marketing activity. The most common regulatory response has been alcohol industry self-regulation; statutory public health responses have made little progress in recent years and have lagged behind industry innovation in digital and social marketing. Alcohol marketing is widespread globally and a structural element of the alcoholic beverage industry. Given the level of alcohol-related harm worldwide, global and regional recommendations and best practices should be used to guide policy makers in effective regulation of alcohol marketing.


Comparison of computed tomography, endosonography, and intraoperative assessment in TN staging of gastric carcinoma.
From 1986 to 1990 a prospective comparative study was undertaken to compare the relative accuracy of computed tomography, endogastric ultrasonography, and intraoperative surgical assessment in evaluating the depth of invasion (T category) and involvement of lymph nodes (N category) of patients with gastric carcinoma. One hundred and eight consecutive patients, who were treated by total gastrectomy and previously evaluated with computed tomography, endogastric ultrasonography, and intraoperative surgical assessment, entered the study. Results (T and N category) were compared with those of histopathological staging (pT and pN category). T categories were correctly staged in 43% of cases with computed tomography, 86% with endogastric ultrasonography, and 56% with intraoperative surgical assessment. Computed tomography scanning correctly staged 51% of all N1 and N2 lymph nodes compared with 74% for endogastric ultrasonography and 54% for intraoperative surgical assessment. In general, computed tomography was more accurate for advanced stages of cancer and showed a tendency to overstage the T category and understage N category of gastric tumours. By contrast, endogastric ultrasonography was equally accurate for all T categories and showed an understaging for N categories. Intraoperative surgical assessment overstaged early T stages, understaged T4 tumours, and was equally accurate for all grades of N categories. Computed tomography scanning and intraoperative surgical assessment of T and N categories were of little value in staging of gastric carcinoma. Endogastric ultrasonography is more accurate than computed tomography scanning and intraoperative surgical assessment. Therefore endogastric ultrasonography should be introduced in the preoperative assessment of patients with gastric carcinoma.


The importance of transesophageal echocardiography in heart harvesting for cardiac transplantation.
The use of transesophageal echocardiography (TEE) during heart harvesting for transplantation can guide the heart assessment, as harvesting a marginal heart can jeopardize the cardiac transplantation. Male, 30 years old, suffered a car crash that resulted in a severe traumatic brain injury (TBI) that evolved to brain death. The patient was intubated and ventilated with a fraction of inspired oxygen of 0.6, presetting Vt 500 mL, RR 14 bpm, PEEP of 3 mm Hg, 99% O(2) saturation, and normal blood gases. He was also hypovolemic, with urine output of 9,300 mL.day(-1), sodium level of 157 mEq.L(-1), hematocrit of 27%, and BP 90/60 mm Hg maintained by infusion of norepinephrine 0.5 mcg.kg.min(-1). The patient was clinically optimized and evaluated by TEE, which showed normal size cardiac chambers, ejection fraction 66%, anatomical and functional heart valves with no changes, and foramen ovale integrity. Immediately after the confirmation of cardiac viability and clinical stabilization, the patient was taken to the operating room and the harvest began. The ischemic period lasted two hours and the heart was successfully transplanted. In most heart transplant services, the cardiac assessment is made subjectively by the surgeon who often does not have the anesthesiologist support to clinically optimize the donor. At the Instituto Nacional de Cardiologia (INC/MS), the anesthesiologist is part of the harvesting team in order to perform intraoperative TEE, evaluating objectively the harvested heart. In doing so, it provides greater chances of heart transplantation success with lower costs for the Brazilian public health system.


A Legendre-Fenchel Transform for Molecular Stretching Energies.
Single-molecular polymers can be used to analyze to what extent thermodynamics applies when the size of the system is drastically reduced. We have recently verified using molecular-dynamics simulations that isometric and isotensional stretching of a small polymer result in Helmholtz and Gibbs stretching energies, which are not related to a Legendre transform, as they are for sufficiently long polymers. This disparity has also been observed experimentally. Using molecular dynamics simulations of polyethylene-oxide, we document for the first time that the Helmholtz and Gibbs stretching energies can be related by a Legendre-Fenchel transform. This opens up a possibility to apply this transform to other systems which are small in Hill's sense.


The Lyme disease spirochete Borrelia burgdorferi induces inflammation and apoptosis in cells from dorsal root ganglia.
Lyme neuroborreliosis (LNB), caused by the spirochete Borrelia burgdorferi, affects both the peripheral and the central nervous systems. Radiculitis or nerve root inflammation, which can cause pain, sensory loss, and weakness, is the most common manifestation of peripheral LNB in humans. We previously reported that rhesus monkeys infected with B. burgdorferi develop radiculitis as well as inflammation in the dorsal root ganglia (DRG), with elevated levels of neuronal and satellite glial cell apoptosis in the DRG. We hypothesized that B. burgdorferi induces inflammatory mediators in glial and neuronal cells and that this inflammatory milieu precipitates glial and neuronal apoptosis. To model peripheral neuropathy in LNB we incubated normal rhesus DRG tissue explants with live B. burgdorferi ex vivo and identified immune mediators, producer cells, and verified the presence of B. burgdorferi in tissue sections by immunofluorescence staining and confocal microscopy. We also set up primary cultures of DRG cells from normal adult rhesus macaques and incubated the cultures with live B. burgdorferi. Culture supernatants were subjected to multiplex ELISA to detect immune mediators, while the cells were evaluated for apoptosis by the in situ TUNEL assay. A role for inflammation in mediating apoptosis was assessed by evaluating the above phenomena in the presence and absence of various concentrations of the anti-inflammatory drug dexamethasone. As Schwann cells ensheath the dorsal roots of the DRG, we evaluated the potential of live B. burgdorferi to induce inflammatory mediators in human Schwann cell (HSC) cultures. Rhesus DRG tissue explants exposed to live B. burgdorferi showed localization of CCL2 and IL-6 in sensory neurons, satellite glial cells and Schwann cells while IL-8 was seen in satellite glial cells and Schwann cells. Live B. burgdorferi induced elevated levels of IL-6, IL-8 and CCL2 in HSC and DRG cultures and apoptosis of sensory neurons. Dexamethasone reduced the levels of immune mediators and neuronal apoptosis in a dose dependent manner. In this model, B. burgdorferi induced an inflammatory response and neuronal apoptosis of DRG. These pathophysiological processes could contribute to peripheral neuropathy in LNB.


Differential Expression of OATP1B3 Mediates Unconjugated Testosterone Influx.
Castration-resistant prostate cancer (CRPC) has greater intratumoral testosterone concentrations than similar tumors from eugonadal men; simple diffusion does not account for this observation. This study was undertaken to ascertain the androgen uptake kinetics, functional, and clinical relevance of de novo expression of the steroid hormone transporter OATP1B3 (SLCO1B3). Experiments testing the cellular uptake of androgens suggest that testosterone is an excellent substrate of OATP1B3 (Km = 23.2 μmol/L; Vmax = 321.6 pmol/mg/minute), and cells expressing a doxycycline-inducible SLCO1B3 construct had greater uptake of a clinically relevant concentration of 3H-testosterone (50 nmol/L; 1.6-fold, P = 0.0027). When compared with Slco1b2 (-/-) mice, Slco1b2 (-/-)/hSLCO1B3 knockins had greater hepatic uptake (15% greater AUC, P = 0.0040) and lower plasma exposure to 3H-testosterone (17% lower AUC, P = 0.0030). Of 82 transporters genes, SLCO1B3 is the second-most differentially expressed transporter in CRPC cell lines (116-fold vs. androgen-sensitive cells), with a differentially spliced cancer-type ct-SLCO1B3 making up the majority of SLCO1B3 expression. Overexpression of SLCO1B3 in androgen-responsive cells results in 1.5- to 2-fold greater testosterone uptake, whereas siRNA knockdown of SLCO1B3 in CRPC cells did not change intracellular testosterone concentration. Primary human prostate tumors express SLCO1B3 to a greater extent than ct-SLCO1B3 (26% of total SLCO1B3 expression vs. 0.08%), suggesting that androgen uptake in these tumor cells also is greater. Non-liver tumors do not differentially express SLCO1B3.Implications: This study suggests that de novo OATP1B3 expression in prostate cancer drives greater androgen uptake and is consistent with previous observations that greater OATP1B3 activity results in the development of androgen deprivation therapy resistance and shorter overall survival. Mol Cancer Res; 15(8); 1096-105. ©2017 AACR.


Acupuncture and assisted reproductive technology.
Acupuncture is commonly undertaken during an assisted reproductive technology (ART) cycle although its role in improving live birth and pregnancy rates is unclear. To determine the effectiveness and safety of acupuncture as an adjunct to ART cycles for male and female subfertility. All reports which described randomised controlled trials of acupuncture in assisted conception were obtained through searches of the Menstrual Disorders and Subfertility Group Specialised Register, CENTRAL, Ovid MEDLINE, EMBASE, CINAHL (Cumulative Index to Nursing & Allied Health Literature), AMED , www.clinicaltrials.gov (all from inception to July 2013), National Research Register, and the Chinese clinical trial database (all to November 2012). Randomised controlled trials of acupuncture for couples who were undergoing ART, comparing acupuncture treatment alone or acupuncture with concurrent ART versus no treatment, placebo or sham acupuncture plus ART for the treatment of primary and secondary infertility. Women with medical illness that was deemed to contraindicate ART or acupuncture were excluded. Twenty randomised controlled trials were included in the review and nine were excluded. Study selection, quality assessment and data extraction were performed independently by two review authors. Meta-analysis was performed using odds ratio (OR) and 95% confidence intervals (CI). The outcome measures were live birth rate, clinical ongoing pregnancy rate, miscarriage rate, and any reported side effects of treatment. The quality of the evidence for the primary outcome (live birth) was rated using GRADE methods. This updated meta-analysis showed no evidence of overall benefit of acupuncture for improving live birth rate (LBR) regardless of whether acupuncture was performed around the time of oocyte retrieval (OR 0.87, 95% CI 0.59 to 1.29, 2 studies, n = 464, I(2) = 0%, low quality evidence) or around the day of embryo transfer (ET) (OR 1.22, 95% CI 0.87 to 1.70, 8 studies, n = 2505, I(2) = 69%, low quality evidence). There was no evidence that acupuncture had any effect on pregnancy or miscarriage rates, or had significant side effects. There is no evidence that acupuncture improves live birth or pregnancy rates in assisted conception.


High-Throughput Differentiation and Screening of a Library of Mutant Stem Cell Clones Defines New Host-Based Genes Involved in Rabies Virus Infection.
We used a genomic library of mutant murine embryonic stem cells (ESCs) and report the methodology required to simultaneously culture, differentiate, and screen more than 3,200 heterozygous mutant clones to identify host-based genes involved in both sensitivity and resistance to rabies virus infection. Established neuronal differentiation protocols were miniaturized such that many clones could be handled simultaneously, and molecular markers were used to show that the resultant cultures were pan-neuronal. Next, we used a green fluorescent protein (GFP) labeled rabies virus to develop, validate, and implement one of the first host-based, high-content, high-throughput screens for rabies virus. Undifferentiated cell and neuron cultures were infected with GFP-rabies and live imaging was used to evaluate GFP intensity at time points corresponding to initial infection/uptake and early and late replication. Furthermore, supernatants were used to evaluate viral shedding potential. After repeated testing, 63 genes involved in either sensitivity or resistance to rabies infection were identified. To further explore hits, we used a completely independent system (siRNA) to show that reduction in target gene expression leads to the observed phenotype. We validated the immune modulatory gene Unc13d and the dynein adapter gene Bbs4 by treating wild-type ESCs and primary neurons with siRNA; treated cultures were resistant to rabies infection/replication. Overall, the potential of such in vitro functional genomics screens in stem cells adds additional value to other libraries of stem cells. This technique is applicable to any bacterial or virus interactome and any cell or tissue types that can be differentiated from ESCs.


Gene expression profiling in murine autoimmune arthritis during the initiation and progression of joint inflammation.
We present here an extensive study of differential gene expression in the initiation, acute and chronic phases of murine autoimmune arthritis with the use of high-density oligonucleotide arrays interrogating the entire mouse genome. Arthritis was induced in severe combined immunodeficient mice by using adoptive transfer of lymphocytes from proteoglycan-immunized arthritic BALB/c mice. In this unique system only proteoglycan-specific lymphocytes are transferred from arthritic mice into syngeneic immunodeficient recipients that lack adaptive immunity but have intact innate immunity on an identical (BALB/c) genetic background.Differential gene expression in response to donor lymphocytes that migrated into the joint can therefore be monitored in a precisely timed manner, even before the onset of inflammation. The initiation phase of adoptively transferred disease (several days before the onset of joint swelling) was characterized by differential expression of 37 genes, mostly related to chemokines, interferon-gamma and tumor necrosis factor-alpha signaling, and T cell functions. These were designated early arthritis 'signature' genes because they could distinguish between the naive and the pre-arthritic state. Acute joint inflammation was characterized by at least twofold overexpression of 256 genes and the downregulation of 21 genes, whereas in chronic arthritis a total of 418 genes with an equal proportion of upregulated and downregulated transcripts were expressed differentially. Hierarchical clustering and functional classification of inflammation-related and arthritis-related genes indicated that the most common biological activities were represented by genes encoding interleukins, chemokine receptors and ligands, and by those involved in antigen recognition and processing.


The effect of sodium fluorescein on anterior eye surface measurements.
During image acquisition, certain topographers require the addition of sodium fluorescein (NaFl) dye to the tear film. This study investigates the effect of NaFl dye on corneal topography and tear surface quality. The E300 corneal topographer (Medmont International Pty Ltd., Victoria, Australia) was used to measure ocular surface topography and quality of 57 eyes of 57 healthy individuals without dry eye symptoms, age 35.1 ± 15.2 years (mean ± standard deviation) ranging between 19 and 65 years. The mean of three simulated keratometry values, a variety of corneal shape descriptors, and Tear Film Surface Quality (TFSQ) were measured under three different conditions; without NaFl (baseline), with the addition of a single dose NaFl, and using a double dose of NaFl. Compared to baseline, the Inferior-Superior (IS) index decreased significantly after a single dose (P = 0.034) or double dose of NaFl (P = 0.030). The corneal surface was significantly more regular without NaFl (P = 0.003) or one insertion of NaFl (P = 0.024) when compared to two doses of NaFl. There was no association with age, or dry eye signs or symptoms on the variance observed in any of the indices between baseline, intervention I, and intervention II (P > 0.05). Agreement between corneal surface indices reduced following the addition of NaFl. In comparison to measurements taken without an ocular dye, one dose of NaFl resulted in increased reliability and consistency in corneal topography measurements using the E300 topographer, but 2 doses decreased reliability and consistency. Practitioners ought to be aware that tear film surface regularity and inferior-superior corneal power changed significantly following the addition of NaFl in those with healthy corneas. Its effect in diseased corneas is unknown.


M. tuberculosis persistence, latency, and drug tolerance.
The success of Mycobacterium tuberculosis as a pathogen is largely attributable to its ability to persist in host tissues, where drugs that are rapidly bactericidal in vitro require prolonged administration to achieve comparable effects. Latency is a frequent outcome of untreated or incompletely treated M. tuberculosis infection, creating a long-standing reservoir of future disease and contagion. Although the interactions between the bacterium and its host that result in chronic or latent infection are still largely undefined, recent years have seen a resurgence of interest and research activity in this area. Here we review some of the classic studies that have led to our current understanding of M. tuberculosis persistence, and discuss the varied approaches that are now being brought to bear on this important problem.


Early Home Activities and Oral Language Skills in Middle Childhood: A Quantile Analysis.
Oral language development is a key outcome of elementary school, and it is important to identify factors that predict it most effectively. Commonly researchers use ordinary least squares regression with conclusions restricted to average performance conditional on relevant covariates. Quantile regression offers a more sophisticated alternative. Using data of 17,687 children from the United Kingdom's Millennium Cohort Study, we compared ordinary least squares and quantile models with language development (verbal similarities) at 11 years as the outcome. Gender had more of an effect at the top of the distribution, whereas poverty, early language, and reading to the child had a greater effect at the bottom. The picture for TV watching was more mixed. The results are discussed in terms of the provision of universal and targeted interventions.


Altered susceptibility to in vitro oxidation of LDL in LDL complexes and LDL aggregates.
Low density lipoprotein (LDL) is known to form complexes with polysulfated compounds, like heparin, dextran sulfate (DS), and chondroitin sulfate. In particular, chondroitin 6-sulfate (C6S)-rich proteoglycans of the arterial intima can associate with LDL, resulting in accumulation of LDL in atherosclerotic lesions. Besides LDL complex formation, LDL self-aggregation has been recently suggested to play a role in atherogenesis. Oxidative modification of LDL has also been implicated as a factor in the generation of the atherosclerotic plaque. Assuming that LDL self-aggregation may alter the molecule's susceptibility to oxidative modification, we have studied the sensitivity of LDL in LDL a aggregates as well as in insoluble and soluble LDL-C6S, LDL-heparin, and LDL-DS complexes to in vitro oxidation by cooper ions. Complexing the LDL with C6S and heparin resulted in an increased susceptibility of LDL to in vitro oxidation, whereas the oxidation of LDL complexed with DS was unaffected. In great contrast to the oxidation of LDL in LDL complexes, the in vitro oxidation of LDL in LDL aggregates (self-aggregation by denaturation) was strongly reduced. The results suggest that complex or aggregate formation may alter the susceptibility of the lipoprotein to oxidative modification and finally its metabolic fate or biological activity.


Putative active site template model for cytochrome P4502C9 (tolbutamide hydroxylase).
Binding of substrates to the active site of cytochrome P450 enzymes largely relies on hydrophobic interactions. However, other binding interactions can take place giving the enzyme high regioselectivity and even stereoselectivity. For instance, within the major human cytochrome P450s involved in drug metabolism, cytochrome P4502D6 (CYP2D6) relies on an ion-pair interaction as a major binding factor. There are now a number of substrates reported that have routes of metabolism ascribed specifically to cytochrome P4502C9 (CYP2C9), the isoform mainly responsible for tolbutamide hydroxylation. Although chemically diverse, these substrates have the capability to be hydrogen bond donors (or acceptors). The substrate specificity has been rationalized in terms of a hydrogen bond donor/acceptor model and, by use of molecular modeling, an active site template model for CYP2C9 has been generated. The substrates modeled were phenytoin, warfarin, ibuprofen, naproxen, diclofenac, delta 1-tetrahydrocannabinol, 58C80, and tolbutamide. In addition to the substrates, the potent, selective inhibitor sulfaphenazole was also included in the modeling. An initial hydrogen bond donor site (N2) was identified on phenytoin, the most rigid of the substrates. Corresponding hydrogen bond donation sites were then identified on all of the molecules studied. Using molecular modeling, the site of metabolism and the hydrogen bond donation sites of the molecules were then overlaid on phenytoin to produce the putative active site model. The resultant model is described by a, the distance between the site of metabolism (Y), and the hydrogen bond donor heteroatom (X) and C, the angle between this and the hydrogen bond. The mean dimensions (+/- SD) for the nine substrates and one inhibitor (a = 6.7 +/- 1.0 A, C = 133 +/- 21 degrees) illustrate the degree of overlap achieved.


Contradictory shear stress distribution prevents restenosis after provisional stenting for bifurcation lesions.
Endothelial shear stress is one of the local hemodynamic factors suspected in the development of coronary atherosclerosis in bifurcation lesions. In patients with provisional stenting, the endothelial shear stress (SS) distribution is unknown. The aim of this study was to investigate the magnitude and distribution of the SS of coronary bifurcation lesions stenting by the provisional approach. Ten consecutive patients were included in this study. Quantitative coronary analysis, flow study, and three-dimensional computational analysis with the aid of the commercial software CD STAR-CCM+ were done before and after the provisional stenting procedure and also 8 months later. Clinical and angiographic follow-up were available in all patients. No patient had a side branch (SB) stent. At the 8-month follow-up, no major adverse cardiac event (MACE) occurred. There was also no clinical and angiographic restenosis. Before PCI, the distal main vessel (MV)-lateral, and the SB-lateral subsegments had relative nonsignificant lower SS value (4.08 +/- 2.78 Pa and 4.35 +/- 5.04 Pa, respectively) when compared to other segments. After 8-month follow-up, sustained decreased SS value was shown in the distal MV-lateral segment (4.08 +/- 2.78-1.68 +/- 1.65 Pa), when compared with significantly increased SS value in the SB-lateral subsegment 4.35 +/- 5.04-16.50 +/- 40.45 Pa). The explanation is that after stenting in the MV, the flow was redistributed immediately after percutaneous coronary intervention (PCI) and reversed back to its original 8 months later. However, the growth of the fibrous tissue causing in-stent restenosis (ISR) is prohibited by sirolimus on the stent struts. In contrast, in a branch opened up by plain old balloon angioplasty (POBA), the flow did not change much, the flow could even be worse because it is shifted to the MV after the cross-sectional area of the MV improved by stenting. However, thanks to POBA, there is increased fibrous tissue formation, enough to increase the SS and prevent further accumulation of cell and cholesterol needed for more restenosis. In the provisional approach, low endothelial SS correlated with no restenosis for patients who underwent stenting of the MV, while a contradictory combination of high SS and no restenosis was seen in the SB after only POBA. The mechanism of prevention of restenosis in the SB is by increasing the SS while in the MV, the mechanism of prevention of ISR is secondary to sirolimus on the stents struts.


WDM compatible and electrically tunable SPE-OCDMA system based on the temporal self-imaging effect.
A coding/decoding setup for a spectral phase encoding optical code-division multiple access (SPE-OCDMA) system has been developed. The proposal is based on the temporal self-imaging effect and the use of an easily tunable electro-optic phase modulator to achieve line-by-line coding of the transmitted signal, thus assuring compatibility with WDM techniques. Modulation of the code is performed at the same rate as the data, avoiding the use of high-bandwidth electro-optic modulators. As proof of concept of the technique, experimental results are presented for a back-to-back coder/decoder setup transmitting a 10 GHz unmodulated optical pulse train within an 80 GHz optical window and using 8-chip Hadamard codes.


Cost-effectiveness of the fixed-dose combination tiotropium/olodaterol versus tiotropium monotherapy or a fixed-dose combination of long-acting β2-agonist/inhaled corticosteroid for COPD in Finland, Sweden and the Netherlands: a model-based study.
Chronic obstructive pulmonary disease (COPD) guidelines advocate treatment with combinations of long-acting bronchodilators for patients with COPD who have persistent symptoms or continue to have exacerbations while using a single bronchodilator. This study assessed the cost-utility of the fixed dose combination of the bronchodilators tiotropium and olodaterol versus two comparators, tiotropium monotherapy and long-acting β2 agonist/inhaled corticosteroid (LABA/ICS) combinations, in three European countries: Finland, Sweden and the Netherlands. A previously published COPD patient-level discrete event simulation model was updated with most recent evidence to estimate lifetime quality-adjusted life years (QALYs) and costs for COPD patients receiving either tiotropium/olodaterol, tiotropium monotherapy or LABA/ICS. Treatment efficacy covered impact on trough forced expiratory volume in 1 s (FEV1), total and severe exacerbations and pneumonias. The unit costs of medication, maintenance treatment, exacerbations and pneumonias were obtained for each country. The country-specific analyses adhered to the Finnish, Swedish and Dutch pharmacoeconomic guidelines, respectively. Treatment with tiotropium/olodaterol gained QALYs ranging from 0.09 (Finland and Sweden) to 0.11 (the Netherlands) versus tiotropium and 0.23 (Finland and Sweden) to 0.28 (the Netherlands) versus LABA/ICS. The Finnish payer's incremental cost-effectiveness ratio (ICER) of tiotropium/olodaterol was €11 000/QALY versus tiotropium and dominant versus LABA/ICS. The Swedish ICERs were €6200/QALY and dominant, respectively (societal perspective). The Dutch ICERs were €14 400 and €9200, respectively (societal perspective). The probability that tiotropium/olodaterol was cost-effective compared with tiotropium at the country-specific (unofficial) threshold values for the maximum willingness to pay for a QALY was 84% for Finland, 98% for Sweden and 99% for the Netherlands. Compared with LABA/ICS, this probability was 100% for all three countries. Based on the simulations, tiotropium/olodaterol is a cost-effective treatment option versus tiotropium or LABA/ICS in all three countries. In both Finland and Sweden, tiotropium/olodaterol is more effective and cost saving (ie, dominant) in comparison with LABA/ICS.


Cigarette smoke-related hydroquinone induces filamentous actin reorganization and heat shock protein 27 phosphorylation through p38 and extracellular signal-regulated kinase 1/2 in retinal pigment epithelium: implications for age-related macular degeneration.
Retinal pigment epithelium (RPE)-derived membranous debris named blebs, may accumulate and contribute to sub-RPE deposit formation, which is the earliest sign of age-related macular degeneration (AMD). Oxidative injury to the RPE might play a significant role in AMD. However, the underlying mechanisms are unknown. We previously reported that hydroquinone (HQ), a major pro-oxidant in cigarette smoke, foodstuff, and atmospheric pollutants, induces actin rearrangement and membrane blebbing in RPE cells as well as sub-RPE deposits in mice. Here, we show for the first time that phosphorylated Heat shock protein 27 (Hsp27), a key regulator of actin filaments dynamics, is up-regulated in RPE from patients with AMD. Also, HQ-induced nonlethal oxidative injury led to Hsp27mRNA up-regulation, dimer formation, and Hsp27 phosphorylation in ARPE-19 cells. Furthermore, we found that a cross talk between p38 and extracellular signal-regulated kinase (ERK) mediates HQ-induced Hsp27 phosphorylation and actin aggregate formation, revealing ERK as a novel upstream mediator of Hsp27 phosphorylation. Finally, we demonstrated that Hsp25, p38, and ERK phosphorylation are increased in aging C57BL/6 mice chronically exposed to HQ, whereas Hsp25 expression is decreased. Our data suggest that phosphorylated Hsp27 might be a key mediator in AMD and HQ-induced oxidative injury to the RPE, which may provide helpful insights into the early cellular events associated with actin reorganization and bleb formation involved in sub-RPE deposits formation relevant to the pathogenesis of AMD.


Oxygen vacancy migration/diffusion induced synaptic plasticity in a single titanate nanobelt.
Neuromorphic computational systems that emulate biological synapses in the human brain are fundamental in the development of artificial intelligence protocols beyond the standard von Neumann architecture. Such systems require new types of building blocks, such as memristors that access a quasi-continuous and wide range of conductive states, which is still an obstacle for the realization of high-efficiency and large-capacity learning in neuromorphoric simulation. Here, we introduce hydrogen and sodium titanate nanobelts, the intermediate products of hydrothermal synthesis of TiO2 nanobelts, to emulate the synaptic behavior. Devices incorporating a single titanate nanobelt demonstrate robust and reliable synaptic functions, including excitatory postsynaptic current, paired pulse facilitation, short term plasticity, potentiation and depression, as well as learning-forgetting behavior. In particular, the gradual modulation of conductive states in the single nanobelt device can be achieved by a large number of identical pulses. The mechanism for synaptic functionality of the titanate nanobelt device is attributed to the competition between an electric field driven migration of oxygen vacancies and a thermally induced spontaneous diffusion. These results provide insight into the potential use of titanate nanobelts in synaptic applications requiring continuously addressable states coupled with high processing efficiency.


A Place-based Assessment of Flash Flood Hazard and Vulnerability in the Contiguous United States.
Flash flood is among the most catastrophic natural hazards which causes disruption in the environment and societies. Flash flood is mainly initiated by intense rainfall, and due to its rapid onset (within six hours of rainfall), taking action for effective response is challenging. Building resilience to flash floods require understanding of the socio-economic characteristics of the societies and their vulnerability to these extreme events. This study provides a comprehensive assessment of socio-economic vulnerability to flash floods and investigates the main characteristics of flash flood hazard, i.e. frequency, duration, severity, and magnitude. A socio-economic vulnerability index is developed at the county level across the Contiguous United States (CONUS). For this purpose, an ensemble of social and economic variables from the US Census and the Bureau of Economic Analysis were analyzed. Then, the coincidence of socio-economic vulnerability and flash flood hazard were investigated to identify the critical and non-critical regions. Results show that the southwest U.S. experienced severe flash flooding with high magnitude, whereas the Northern Great Plains experience lower severity and frequency. Critical counties (high-vulnerable-hotspot) are mostly located in the southern and southwestern parts of the U.S. The majority of counties in the Northern Great Plains indicate a non-critical status.


An Injectable Hydrogel as Bone Graft Material with Added Antimicrobial Properties.
Currently, the technique which provides the best chances for a successful bone graft, is the use of bone tissue from the same patient receiving it (autograft); the main limitations are the limited availability and the risks involved in removing living bone tissue, for example, explant site pain and morbidity. Allografts and xenografts may overcome these limitations; however, they increase the risk of rejection. For all these reasons the development of an artificial bone graft material is particularly important and hydrogels are a promising alternative for bone regeneration. Gels were prepared using 1,4-butanediol diacrylate as crosslinker and alpha tricalciumphosphate; ZnCl2 and SrCl2 were added to the aqueous phase. MTT results demonstrated that the addition of strontium had a beneficial effect on the osteoblast cells density on hydrogels, and zinc instead did not increase osteoblast proliferation. The amount of calcium produced by the osteoblast cells quantified through the Alizarin Red protocol revealed that both strontium and zinc positively influenced the formation of calcium; furthermore, their effect was synergistic. Rheology properties were used to mechanically characterize the hydrogels and especially the influence of crosslinker's concentration on them, showing the hydrogels presented had extremely good mechanical properties. Furthermore, the antimicrobial activity of strontium and zinc in the hydrogels against methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis was determined.


Amyotrophic lateral sclerosis: analysis of ALS cases in a predominantly admixed population of Ecuador.
Recent studies suggest amyotrophic lateral sclerosis (ALS) prevalence, incidence, and age of onset are heterogeneous across populations. These include studies from South America (SA) where lower prevalence, earlier onset, and reduced survival time of ALS are reported. However, the scarcity of epidemiological and clinical data confounds effective comparison. To investigate ALS incidence in the predominantly admixed population of Ecuador, we analyzed patient data. We analyzed case data from two major hospitals. To confirm diagnosis, we evaluated clinical and EMG examinations in a cohort of patients. For 2000-2012, we found 116 patients with ALS diagnosis in the two hospitals. Crude incidence was 0.2-0.6 per 100,000. Median age of onset was 54.3 (+ 15.06 SD). Clinical re-evaluation found misdiagnosis in three cases in the cohort. In conclusion, ALS incidence in the Ecuadorian hospital population is in accord with rates reported in recent studies for other admixed populations, and lower than that in the United States and Europe. Our study found that appropriate EMG administration and interpretation for the purposes of supporting a diagnosis of ALS with current consensus guidelines prevent adequate use of this test as an essential tool in the evaluation and diagnosis of ALS. Training for required standardization in Ecuador is recommended.


Phylogenetic analysis of beak and feather disease virus across a host ring-species complex.
Pathogens have been hypothesized to play a major role in host diversity and speciation. Susceptibility of hybrid hosts to pathogens is thought to be a common phenomenon that could promote host population divergence and subsequently speciation. However, few studies have tested for pathogen infection across animal hybrid zones while testing for codivergence of the pathogens in the hybridizing host complex. Over 8 y, we studied natural infection by a rapidly evolving single-strand DNA virus, beak and feather diseases virus (BFDV), which infects parrots, exploiting a host-ring species complex (Platycercus elegans) in Australia. We found that host subspecies and their hybrids varied strikingly in both BFDV prevalence and load: both hybrid and phenotypically intermediate subspecies had lower prevalence and load compared with parental subspecies, while controlling for host age, sex, longitude and latitude, as well as temporal effects. We sequenced viral isolates throughout the range, which revealed patterns of genomic variation analogous to Mayr's ring-species hypothesis, to our knowledge for the first time in any host-pathogen system. Viral phylogeny, geographic location, intraspecific host density, and parrot community diversity and composition did not explain the differences in BFDV prevalence or load between subpopulations. Overall, our analyses suggest that functional host responses to infection, or force of infection, differ between subspecies and hybrids. Our findings highlight the role of host hybridization and clines in altering host-pathogen interactions, dynamics that can have important implications for models of speciation with gene flow, and offer insights into how pathogens may adapt to diverging host populations.


The role of body dissatisfaction and bingeing in the self-esteem of women with type II diabetes.
This study investigated the impact of body dissatisfaction and binge eating on self-esteem in women with Type II diabetes. The relationship of body dissatisfaction and bingeing to perceived blood glucose control was also assessed. Questionnaires were completed by a total of 215 women: 125 women with Type II diabetes and 90 comparison women, who were roughly matched for age, education, and ethnicity. When actual weight (BMI) was statistically controlled, there was no difference between the groups in body dissatisfaction or bingeing behavior. The women with diabetes, however, had significantly lower self-esteem. Further, bingeing made a significant contribution to their self-esteem, in contrast to the women without diabetes. For the women with diabetes, body dissatisfaction and bingeing were also related to perceived blood glucose control, although only bingeing remained significant when both variables were entered into the regression equation. It was concluded that diabetes broadens the domains of body dissatisfaction which are related to self-esteem.


The changing pattern and determinants of declining consanguinity in Jordan during 1990-2012.
Consanguinity is a deep rooted cultural trait in Jordan. To examine the patterns and determinants of declining rates of consanguineous marriage in Jordan during 1990-2012 in the context of the changing pattern of socio-economic and demographic conditions. The data come from the 1990 and 2012 Jordan Population and Family Health Surveys (JPFHSs). A total of 6461 women in 1990 and 11,352 women in 2012 were successfully interviewed. Descriptive and multivariate statistical techniques were used for data analysis. Consanguinity was found to be widely practiced (35% in 2012) until recent times in Jordan. However, there has been a secular declining trend over the last few decades as the practice of consanguinity has declined from 56% in 1990 to 35% in 2012. Increasing age at marriage and female education, higher level of education of husbands, declining family size, increasing rate of urbanisation and female employment, exposure to mass media and higher economic status appeared as significant predictors of declining consanguinity in Jordan. The findings of this study support Goode's hypothesis of a decrease of consanguinity with modernisation. Although consanguinity is a deeply rooted cultural trend in Jordan, it is gradually losing ground due to modernisation and socio-demographic transition of the country.


Does memory improve with age? CD85j (ILT-2/LIR-1) expression on CD8 T cells correlates with 'memory inflation' in human cytomegalovirus infection.
CMV-specific memory CD8(+) T cells accumulate over time to reach high frequencies amongst peripheral blood lymphocytes - a phenomenon termed 'memory inflation'. Using tetramer staining on samples from a large number of subjects and multivariate regression analysis, we were able to relate this to the phenotype of CD8(+) T cells. We made the following observations: (i) CD85j (ILT-2/LIR-1) was highly expressed alongside CD57 - an established effector memory marker - on CMV-specific CD8(+) T cells; (ii) on CD8(+) T cells as a whole, with increasing age, CD57 and CD85j (ILT-2/LIR-1) expression increased whereas CCR7 expression decreased, indicating increasing maturation of the total CD8(+) T-cell compartment with age; (iii) unit increases in the percentage of CMV-specific CD8(+) T cells expressing CD57 and CD85j (ILT-2/LIR-1) were associated with incremental expansion of these T-cell populations; (iv) CMV seropositivity is associated with a marked effect on the overall phenotype of CD8(+) T cells (at any given age, CMV seropositivity is associated with an 18.7% increase in CD85j (ILT-2/LIR-1) expression); and (v) from our observations we estimated from this an apparent 'ageing effect' of CMV on CD8(+) T cells of 35.4 years. The data presented are consistent with a predictable, unidirectional and linear model of virus specific T-cell differentiation and maturation.


CSEQ-SIMULATOR: A DATA SIMULATOR FOR CLIP-SEQ EXPERIMENTS.
CLIP-Seq protocols such as PAR-CLIP, HITS-CLIP or iCLIP allow a genome-wide analysis of protein-RNA interactions. For the processing of the resulting short read data, various tools are utilized. Some of these tools were specifically developed for CLIP-Seq data, whereas others were designed for the analysis of RNA-Seq data. To this date, however, it has not been assessed which of the available tools are most appropriate for the analysis of CLIP-Seq data. This is because an experimental gold standard dataset on which methods can be accessed and compared, is still not available. To address this lack of a gold-standard dataset, we here present Cseq-Simulator, a simulator for PAR-CLIP, HITS-CLIP and iCLIP-data. This simulator can be applied to generate realistic datasets that can serve as surrogates for experimental gold standard dataset. In this work, we also show how Cseq-Simulator can be used to perform a comparison of steps of typical CLIP-Seq analysis pipelines, such as the read alignment or the peak calling. These comparisons show which tools are useful in different settings and also allow identifying pitfalls in the data analysis.


Potential of marine natural products against drug-resistant bacterial infections.
Natural products have been a rich source of compounds with structural and chemical diversity for drug discovery. However, antibiotic resistance in bacteria has been reported for nearly every antibiotic once it is used in clinical practice. In the past decade, pharmaceutical companies have reduced their natural product discovery projects because of challenges, such as high costs, low return rates, and high rediscovery rates. The largely unexplored marine environment harbours substantial diversity and is a large resource to discover novel compounds with novel modes of action, which is essential for the treatment of drug-resistant bacterial infections. In this Review, we report compounds derived from marine sources that have shown in-vivo and in-vitro efficacy against drug-resistant bacteria. Analysis of the physicochemical properties of these marine natural products with activity against drug-resistant bacteria showed that 60% of the compounds have oral bioavailability potential. Their overall distribution pattern of drug characteristics agrees with the observation that marketed antibacterial drugs have a polar distribution, with a lower median calculated logP. The aim of this Review is to summarise the diversity of these marine natural products, with a special focus on analysis of drug bioavailability. Such biologically active compounds, with high degrees of bioavailability, have the potential to be developed as effective drugs against infectious diseases.


Distribution and phylogenetic significance of the 71-kb inversion in the plastid genome in Funariidae (Bryophyta).
The recent assembly of the complete sequence of the plastid genome of the model taxon Physcomitrella patens (Funariaceae, Bryophyta) revealed that a 71-kb fragment, encompassing much of the large single copy region, is inverted. This inversion of 57% of the genome is the largest rearrangement detected in the plastid genomes of plants to date. Although initially considered diagnostic of Physcomitrella patens, the inversion was recently shown to characterize the plastid genome of two species from related genera within Funariaceae, but was lacking in another member of Funariidae. The phylogenetic significance of the inversion has remained ambiguous. Exemplars of all families included in Funariidae were surveyed. DNA sequences spanning the inversion break ends were amplified, using primers that anneal to genes on either side of the putative end points of the inversion. Primer combinations were designed to yield a product for either the inverted or the non-inverted architecture. The survey reveals that exemplars of eight genera of Funariaceae, the sole species of Disceliaceae and three generic representatives of Encalyptales all share the 71-kb inversion in the large single copy of the plastid genome. By contrast, the plastid genome of Gigaspermaceae (Funariales) is characterized by a gene order congruent with that described for other mosses, liverworts and hornworts, and hence it does not possess this inversion. The phylogenetic distribution of the inversion in the gene order supports a hypothesis only weakly supported by inferences from sequence data whereby Funariales are paraphyletic, with Funariaceae and Disceliaceae sharing a common ancestor with Encalyptales, and Gigaspermaceae sister to this combined clade. To reflect these relationships, Gigaspermaceae are excluded from Funariales and accommodated in their own order, Gigaspermales order nov., within Funariideae.


Enhanced Cell Response to Zirconia Surface Immobilized with Type I Collagen.
Zirconia (ZrO2) dental implants provide good biocompatibility, have good corrosion resistance, and have a color that is similar to that of natural teeth. Unfortunately, ZrO2 is a bioinert material and therefore achieves osseointegration difficultly. In this study, we sought to enhance osseointegration by producing rough ZrO2 surfaces that contain hydroxyl groups (designated ZSA) through the use of sandblasting in conjunction with alkaline treatment. We immobilized type I collagen on ZSA surfaces using the natural cross-linker, procyanidin. Our results further showed that surfaces produced in ZSA-P/C featured more and steadier type I collagen than surfaces produced in ZSA-C. The ZSA-P/C also presented superior cell responses in terms of adhesion, proliferation, and mineralization of human bone marrow mesenchymal stem cells. The enhanced cell responses in the ZSA-P/C were induced through the prolonged activation of focal adhesion kinase, AKT (the phosphoinositide 3-kinase pathway), and p38 (the mitogen-activated protein kinase pathway). The simple and novel approach to immobilize type I collagen on roughened ZrO2 surfaces presented in this article can likely benefit dental implant applications.


Levels, distribution and air-soil exchange fluxes of polychlorinated biphenyls (PCBs) in the environment of Punjab Province, Pakistan.
An initial survey of the concentrations of polychlorinated biphenyl (PCB) compounds in air and soils across industrial and agricultural areas of Punjab Province, Pakistan, was conducted from January to March 2011. The total concentration of all PCBs (<Sigma>31 PCBs) ranged from 34 to 389pgm(-3) in air and from 7 to 45ngg(-1) dry weight in soils, where both ranges were similar to the average ranges in other areas of the world. PCBs were elevated across industrial regions near urban and industrial sources. Consistently low air concentrations of PCBs at the agricultural sites suggest that they are less widespread or uniformly distributed in the Pakistani atmosphere. The calculated air and soil fugacity fraction values indicated that soils are a potential secondary source of PCBs in agricultural areas, whereas they are in equilibrium or atmospheric deposition in industrial and urban areas. TEQ concentrations of dioxin-like PCBs for soil samples met the Canadian standard. However, local authorities should address the human health threats from urban and industrial soils in Punjab Province, Pakistan.