Reproductive aging is associated with decreased mitochondrial abundance and altered structure in murine oocytes.
To establish the phenotype of reproductive aging in our mouse model. To test the hypotheses that reproductive aging is associated with a decrease in mitochondrial abundance that could ultimately reflect dysfunction in oocytes. Breeding studies were performed in young and aged female virgin wild type C57BL6J mice to establish their reproductive phenotype by measuring time to conception, litter size, and live birth per dam. Individual oocytes were analyzed for mtDNA content. Transmission electron microscopy was used to study ultrastructure of mitochondria in oocytes. Old females were found to have significantly prolonged time to conception and fewer surviving pups in their litters. Oocytes from old mice had 2.7-fold less mtDNA compared to younger controls (p < 0.001; 95 % CI 2.1-3.5). Decrease in mitochondrial organelle abundance in old animal's oocytes was confirmed with transmission electron microscopy. Distinct morphological changes were noted in mitochondria, suggesting altered mitochondrial biogenesis in the old animals' oocytes. Reproductive aging in mice is associated with reduced reproductive competence. Aging is associated with a significant decrease in number of mitochondria in oocytes. Our data support mitochondrial organelle loss and dysfunction in oocytes as a potential etiology for reproductive senescence.


Integrating Behavioral Economic and Social Network Influences in Understanding Alcohol Misuse in a Diverse Sample of Emerging Adults.
Behavioral economic alcohol demand is a measure of motivation to consume alcohol and a robust risk factor for alcohol misuse. Social networks that are dense with alcohol are also associated with heavy drinking, but the intersection of these risk factors has not been investigated to date. This study examined these interrelationships with structural equation modeling using cross-sectional data from a diverse community sample of heavy-drinking emerging adults (N = 602). Latent variables for alcohol social network, alcohol demand, and alcohol misuse were constructed. Next, relations between the latent variables were examined, including the indirect effect of alcohol demand in the relation between alcohol social network and alcohol misuse. An alternative modeling testing the indirect effect of alcohol social network on the relation between alcohol demand and misuse was also tested. When alone in the model, social network alcohol density significantly predicted alcohol misuse. When alcohol demand was included in the model, social network alcohol density predicted alcohol demand, alcohol demand predicted alcohol misuse, and an indirect effect on alcohol misuse through alcohol demand was present. In the alternative model, an indirect effect was not present between alcohol demand and alcohol misuse by social network alcohol density. Exploratory analyses revealed significant sex, race, and college status differences. These results suggest that the influence of social network alcohol density on alcohol misuse may be, in part, through variance accounted for by alcohol reinforcing value. Longitudinal testing of this mechanistic pathway is warranted.


Piloting the Affordable Medicines Facility-malaria: what will success look like?
The Affordable Medicines Facility-malaria is an innovative financing mechanism, managed by the Global Fund to Fight AIDS, Tuberculosis and Malaria. This initiative aims to increase the use of artemisinin-based combination therapies for treating malaria. A pilot is underway in eight countries to determine whether the mechanism reduces the consumer price of these drugs and increases their availability in public and private outlets, their market share and their use. To evaluate the pilot, an analysis was done to estimate predetermined "benchmarks" of success at 1 and 2 years. The analysis used a mixed-methods approach, triangulating data from a literature review with information from 33 interviews with experts. A sensitivity analysis and other methods were used to verify the results. Benchmarks used to determine success include an increase in availability of artemisinin-based combination therapies of 40 percentage points from baseline, and an increase in their use of 10-15 percentage points from baseline at year 2. These benchmarks were based on evidence that national public health programmes aimed at increasing the use of a specific health commodity in developing countries have generally achieved only modest changes in use within a 2-year time frame. Evaluation should also take individual country contexts into account.


T1799A BRAF mutations in conjunctival melanocytic lesions.
To gain a better understanding of the molecular events leading to the development of conjunctival melanocytic lesions and conjunctival melanoma, this study was conducted to investigate the presence of T1799A BRAF oncogenic mutation in these lesions. Forty-eight surgically excised conjunctival melanocytic lesions from 48 patients were examined for the presence of the BRAF T1799A mutation. Twenty-eight lesions were conjunctival nevi, of which 20 were excised from children younger than 18 years. Fifteen lesions were conjunctival primary acquired melanosis (PAM; 11 without atypia and 4 with atypia) and five were conjunctival melanomas. To detect the BRAF T1799A mutation, both a newly developed kit (Mutector; TrimGen, Sparks, MD) and direct DNA sequence analysis of exon 15 after PCR amplification were used. The T1799A BRAF mutation was identified in 14 of 28 (50%) conjunctival nevi analyzed, but in none of the 15 conjunctival PAMs, with and without atypia. The T1799A BRAF mutation was identified in two of the five (40%) conjunctival melanomas. There was no difference in the BRAF mutation detected in conjunctival nevi in children or adults, as the BRAF mutation was detected in 50%. The results showed that conjunctival nevi, similar to skin nevi, have a high frequency of oncogenic BRAF mutations. Furthermore, the results suggest that the oncogenic event leading to BRAF mutations affect only conjunctival nevi and not conjunctival PAM. The clinical significance of these observations remains to be determined.


Chlorin-based photodynamic therapy enhances the effect of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) in bladder cancer cells.
Photodynamic therapy (PDT) is an attractive, emerging therapeutic procedure suitable for the treatment of non-muscle-invasive bladder cancer. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a death ligand that belongs to the TNF superfamily of cytokines. The ability of TRAIL to selectively induce apoptosis in cancer cells but not in normal cells promotes the development of TRAIL-based cancer therapy. However, many tumor cells are resistant to TRAIL-induced apoptosis. The purpose of the study was to overcome TRAIL-resistance in bladder cancer cells by photodynamic therapy (PDT). Three human bladder transitional cancer cell lines - T24, 647V and SW780 - were treated with TRAIL and/or chlorin-based PDT. The cytotoxicity was measured by MTT and LDH assays and apoptosis was detected using annexin V by flow cytometry. Our test confirmed that T24 and 647V bladder cancer cells are resistant to TRAIL, whereas SW780 cells are sensitive to TRAIL. Then we examined the cytotoxic and apoptotic effects of TRAIL in combination with chlorin e6-polyvinylpyrrolidone (Ce6-PVP)-mediated PDT on bladder cancer cells. We showed for the first time that pretreatment with a low dose of PDT significantly sensitizes bladder cancer cells to TRAIL-induced apoptosis. Chlorin-based PDT augments the effect of TRAIL on bladder cancer cells. PDT with Ce6-PVP photosensitizer enhances the cytotoxic and apoptotic effects of TRAIL on bladder cancer cells. The obtained results suggest that combined treatment by TRAIL and PDT may provide the basis for a new therapeutic approach to induce cell death in bladder cancer.


Physical design analysis and mainstream smoke constituent yields of the new potential reduced exposure product, Marlboro UltraSmooth.
Potential reduced exposure products (PREPs) purport to lower toxicant emissions, but without clinical and long-term health outcome data, claims for reduced harm status of PREPs depend heavily on standard machine yield smoke constituent data. Two prototypes of the new carbon-filtered PREP Marlboro UltraSmooth (MUS) were investigated using both standard (FTC/ISO) and intensive (Health Canada) machine methods to measure gas/vapor- and particulate-phase smoke constituents. Basic physical design characteristics that may influence smoke constituent yields, such as ventilation, pressure drop (resistance to draw), quantity of tobacco, and quantity and type of carbon, were measured. The possible presence of added chemical flavorant compounds was investigated using gas chromatography-mass spectroscopy. MUS prototypes were found to have several key differences in physical design compared with a conventional cigarette, including higher ventilation, lower draw resistance, and in the case of the Salt Lake City prototype, the use of vitreous carbon beads and the presence of chemical flavorants on both the beads and an embedded filter fiber. When tested under the standard regimen, gas-phase constituents of MUS prototypes were reduced compared with a conventional low-yield cigarette. However, far smaller reductions in gas-phase constituents were observed under the intensive regimen, suggesting that the carbon technology used in MUS is less effective when smoked under more intensive conditions. Particulate-phase constituents were not reduced by the carbon filter under either machine-smoking regimen. The data suggest that MUS has been designed to reduce toxic yields while preserving consumer appeal. However, MUS is less effective in reducing toxic smoke constituents when smoked under intensive conditions.


Computational discovery of internal micro-exons.
Very short exons, also known as micro-exons, occur in large numbers in some eukaryotic genomes. Existing annotation tools have a limited ability to recognize these short sequences, which range in length up to 25 bp. Here, we describe a computational method for the identification of micro-exons using near-perfect alignments between cDNA and genomic DNA sequences. Using this method, we detected 319 micro-exons in 4 complete genomes, of which 224 were previously unknown, human (170), the nematode Caenorhabditis elegans (4), the fruit fly Drosophila melanogaster (14), and the mustard plant Arabidopsis thaliana (36). Comparison of our computational method with popular cDNA alignment programs shows that the new algorithm is both efficient and accurate. The algorithm also aids in the discovery of micro-exon-skipping events and cross-species micro-exon conservation.


Targeted degradation of KRAS by an engineered ubiquitin ligase suppresses pancreatic cancer cell growth in vitro and in vivo.
KRAS is an attractive pancreatic ductal adenocarcinoma (PDAC) therapeutic target. E3 ligase is thought to be the component of the ubiquitin conjugation system that is directly responsible for substrate recognition. In this study, an engineered E3 ubiquitin ligase (RC-U) was generated to target the KRAS oncoprotein for ubiquitination and degradation. The engineered E3 ubiquitin ligases (RC-U) were constructed (pRC-U and lentivirus-expressing RC-U). After transfecting the pRC-U plasmid into human pancreatic cancer cells, KRAS expression levels were determined. KRAS expression was also evaluated in cells transfected with pRC-U and treated with MG-132 or cycloheximide. Interactions between RC-U and KRAS as well as whether RC-U could ubiquitinate KRAS were investigated. Extracellular signal-regulated kinase 1/2 (ERK1/2) and phosphorylated ERK 1/2 (pERK1/2) levels were examined in pancreatic cancer cells transfected with pRC-U. The effects of RC-U on pancreatic cancer cell growth were assessed. RC-U decreased KRAS protein levels. After pRC-U transfection, KRAS stability was increased in the presence of MG-132. HEK 293T cells were transfected with a mutant KRAS construct together with pRC-U and incubated with cycloheximide to inhibit new protein synthesis. The exogenous mutant KRAS oncoprotein was degraded more quickly. RC-U can bind KRAS and KRAS can be ubiquitinated by RC-U. pERK1/2 protein levels were decreased. RC-U resulted in reduced cell proliferation in vitro and in vivo. KRAS destruction by RC-U occurred through a ubiquitin-dependent, proteasome-mediated degradation pathway. RC-U inhibited pancreatic cancer cell growth in vitro and in vivo.


Comparative study of the antioxidant and reactive oxygen species scavenging properties in the extracts of the fruits of Terminalia chebula, Terminalia belerica and Emblica officinalis.
Cellular damage caused by reactive oxygen species (ROS) has been implicated in several diseases, and hence natural antioxidants have significant importance in human health. The present study was carried out to evaluate the in vitro antioxidant and reactive oxygen species scavenging activities of Terminalia chebula, Terminalia belerica and Emblica officinalis fruit extracts. The 70% methanol extracts were studied for in vitro total antioxidant activity along with phenolic and flavonoid contents and reducing power. Scavenging ability of the extracts for radicals like DPPH, hydroxyl, superoxide, nitric oxide, hydrogen peroxide, peroxynitrite, singlet oxygen, hypochlorous acid were also performed to determine the potential of the extracts. The ability of the extracts of the fruits in exhibiting their antioxative properties follow the order T. chebula >E. officinalis >T. belerica. The same order is followed in their flavonoid content, whereas in case of phenolic content it becomes E. officinalis >T. belerica >T. chebula. In the studies of free radicals' scavenging, where the activities of the plant extracts were inversely proportional to their IC50 values, T. chebula and E. officinalis were found to be taking leading role with the orders of T. chebula >E. officinalis >T. belerica for superoxide and nitric oxide, and E. officinalis >T. belerica >T. chebula for DPPH and peroxynitrite radicals. Miscellaneous results were observed in the scavenging of other radicals by the plant extracts, viz., T. chebula >T. belerica >E. officinalis for hydroxyl, T. belerica >T. chebula >E. officinalis for singlet oxygen and T. belerica >E. officinalis >T. chebula for hypochlorous acid. In a whole, the studied fruit extracts showed quite good efficacy in their antioxidant and radical scavenging abilities, compared to the standards. The evidences as can be concluded from the study of the 70% methanol extract of the fruits of Terminalia chebula, Terminalia belerica and Emblica officinalis, imposes the fact that they might be useful as potent sources of natural antioxidant.


Control of sympathetic vasomotor tone by catecholaminergic C1 neurones of the rostral ventrolateral medulla oblongata.
Increased sympathetic tone in obstructive sleep apnoea results from recurrent episodes of systemic hypoxia and hypercapnia and might be an important contributor to the development of cardiovascular disease. In this study, we re-evaluated the role of a specific population of sympathoexcitatory catecholaminergic C1 neurones of the rostral ventrolateral medulla oblongata in the control of sympathetic vasomotor tone, arterial blood pressure, and hypercapnia-evoked sympathetic and cardiovascular responses. In anaesthetized rats in vivo and perfused rat working heart brainstem preparations in situ, C1 neurones were acutely silenced by application of the insect peptide allatostatin following cell-specific targeting with a lentiviral vector to express the inhibitory Drosophila allatostatin receptor. In anaesthetized rats with denervated peripheral chemoreceptors, acute inhibition of 50% of the C1 neuronal population resulted in ∼50% reduction in renal sympathetic nerve activity and a profound fall in arterial blood pressure (by ∼25 mmHg). However, under these conditions systemic hypercapnia still evoked vigorous sympathetic activation and the slopes of the CO(2)-evoked sympathoexcitatory and cardiovascular responses were not affected by inhibition of C1 neurones. Inhibition of C1 neurones in situ resulted in a reversible fall in perfusion pressure and the amplitude of respiratory-related bursts of thoracic sympathetic nerve activity. These data confirm a fundamental physiological role of medullary catecholaminergic C1 neurones in maintaining resting sympathetic vasomotor tone and arterial blood pressure. However, C1 neurones do not appear to mediate sympathoexcitation evoked by central actions of CO(2).


Scaffolding Biomaterials for 3D Cultivated Meat: Prospects and Challenges.
Cultivating meat from stem cells rather than by raising animals is a promising solution to concerns about the negative externalities of meat production. For cultivated meat to fully mimic conventional meat's organoleptic and nutritional properties, innovations in scaffolding technology are required. Many scaffolding technologies are already developed for use in biomedical tissue engineering. However, cultivated meat production comes with a unique set of constraints related to the scale and cost of production as well as the necessary attributes of the final product, such as texture and food safety. This review discusses the properties of vertebrate skeletal muscle that will need to be replicated in a successful product and the current state of scaffolding innovation within the cultivated meat industry, highlighting promising scaffold materials and techniques that can be applied to cultivated meat development. Recommendations are provided for future research into scaffolds capable of supporting the growth of high-quality meat while minimizing production costs. Although the development of appropriate scaffolds for cultivated meat is challenging, it is also tractable and provides novel opportunities to customize meat properties.


Attachment theory, ageing and dementia: a review of the literature.
The following review draws together recent theoretical contributions and research findings highlighting the pertinence of attachment issues for older adults. Beginning with a brief overview of the principles of attachment theory, the paper proceeds to illustrate that attachment remains a key feature of relationships throughout the life cycle. Research findings suggest that past and present secure attachment relationships function as a protective resource in later life and preliminary studies highlight the continuing salience of parents as attachment figures for older adults. Particular attention is given to the significance of attachment theory in thinking about the subjective experiences of people with dementia, for whom unwilling separation and disruption of attachment bonds can be common themes. Research indicates the frequent occurrence of attachment behaviours and parent fixation amongst people with dementia living in residential care. Pre-morbid attachment style has been shown to interact with emotional and behavioural expression during the course of dementia. Clinical applications of attachment theory in dementia care are discussed and initial evidence for the effectiveness of Simulated Presence Therapy is provided. Methodological issues raised by the review are considered, together with suggestions for future research.


A structured continuum modelling framework for martensitic transformation and reorientation in shape memory materials.
Models for shape memory material behaviour can be posed in the framework of a structured continuum theory. We study such a framework in which a scalar phase fraction field and a tensor field of martensite reorientation describe the material microstructure, in the context of finite strains. Gradients of the microstructural descriptors naturally enter the formulation and offer the possibility to describe and resolve phase transformation localizations. The constitutive theory is thoroughly described by a single free energy function in conjunction with a path-dependent dissipation function. Balance laws in the form of differential equations are obtained and contain both bulk and surface terms, the latter in terms of microstreses. A natural constraint on the tensor field for martensite reorientation gives rise to reactive fields in these balance laws. Conditions ensuring objectivity as well as the relation of this framework to that provided by currently used models for shape memory alloy behaviour are discussed.


Network Flow Integer Programming to Track Elliptical Cells in Time-Lapse Sequences.
We propose a novel approach to automatically tracking elliptical cell populations in time-lapse image sequences. Given an initial segmentation, we account for partial occlusions and overlaps by generating an over-complete set of competing detection hypotheses. To this end, we fit ellipses to portions of the initial regions and build a hierarchy of ellipses, which are then treated as cell candidates. We then select temporally consistent ones by solving to optimality an integer program with only one type of flow variables. This eliminates the need for heuristics to handle missed detections due to partial occlusions and complex morphology. We demonstrate the effectiveness of our approach on a range of challenging sequences consisting of clumped cells and show that it outperforms state-of-the-art techniques.


Evaluating the consistency and specificity of neuroimaging data using meta-analysis.
Making sense of a neuroimaging literature that is growing in scope and complexity will require increasingly sophisticated tools for synthesizing findings across studies. Meta-analysis of neuroimaging studies fills a unique niche in this process: It can be used to evaluate the consistency of findings across different laboratories and task variants, and it can be used to evaluate the specificity of findings in brain regions or networks to particular task types. This review discusses examples, implementation, and considerations when choosing meta-analytic techniques. It focuses on the multilevel kernel density analysis (MKDA) framework, which has been used in recent studies to evaluate consistency and specificity of regional activation, identify distributed functional networks from patterns of co-activation, and test hypotheses about functional cortical-subcortical pathways in healthy individuals and patients with mental disorders. Several tests of consistency and specificity are described.


Sex differences in wheelchair propulsion biomechanics and mechanical efficiency in novice young able-bodied adults.
An awareness of sex differences in gait can be beneficial for detecting the early stages of gait abnormalities that may lead to pathology. The same may be true for wheelchair propulsion. The aim of this study was to determine the effect of sex on wheelchair biomechanics and mechanical efficiency in novice young able-bodied wheelchair propulsion. Thirty men and 30 women received 12 min of familiarisation training. Subsequently, they performed two 10-m propulsion tests to evaluate comfortable speed (CS). Additionally, they performed a 4-min submaximal propulsion test on a treadmill at CS, 125% and 145% of CS. Propulsion kinetics (via Smartwheel) and oxygen uptake were continuously measured in all tests and were used to determine gross mechanical efficiency (GE), net efficiency (NE) and fraction of effective force (FEF). Ratings of perceived exertion (RPE) were assessed directly after each trial. Results indicated that CS for men was faster (0.98 ± 0.24 m/s) compared to women (0.71 ± 0.18 m/s). A lower GE was found in women compared to men. Push percentage, push angle and local RPE were different across the three speeds and between men and women. NE and FEF were not different between groups. Thus, even though their CS was lower, women demonstrated a higher locally perceived exertion than men. The results suggest sex differences in propulsion characteristics and GE. These insights may aid in optimising wheelchair propulsion through proper training and advice to prevent injuries and improve performance. This is relevant in stimulating an active lifestyle for those with a disability.


Oncornavirus: isolation from a squirrel monkey (Saimiri sciureus) lung culture.
An oncornavirus isolated from a squirrel monkey (Saimiri sciureus) lung culture has a density of 1.16 to 1.17 grams per milliliter, contains 70S RNA, and has an RNA-directed DNA polymerase that prefers Mg2+ over Mn2+ in an assay in which polyribocytidylate - oligodeoxyguanylate (12-18) is used as a synthetic template. Morphologically, the virus resembles Mason-Pfizer monkey virus but is antigenically distinct from this virus. The virus grows in cells of human, chimpanzee, rhesus monkey, canine, and mink origin, but not cells of squirrel monkey origin. On the basis of its properties, the newly isolated virus can be classified as a retravirus.


Stress and food deprivation: linking physiological state to migration success in a teleost fish.
Food deprivation is a naturally occurring stressor that is thought to influence the ultimate life-history strategy of individuals. Little is known about how food deprivation interacts with other stressors to influence migration success. European populations of brown trout (Salmo trutta) exhibit partial migration, whereby a portion of the population smoltifies and migrates to the ocean, and the rest remain in their natal stream. This distinct, natural dichotomy of life-history strategies provides an excellent opportunity to explore the roles of energetic state (as affected by food deprivation) and activation of the glucocorticoid stress response in determining life-history strategy and survival of a migratory species. Using an experimental approach, the relative influences of short-term food deprivation and experimental cortisol elevation (i.e. intra-coelomic injection of cortisol suspended in cocoa butter) on migratory status, survival and growth of juvenile brown trout relative to a control were evaluated. Fewer fish migrated in both the food deprivation and cortisol treatments; however, migration of fish in cortisol and control treatments occurred at the same time while that of fish in the food deprivation treatment was delayed for approximately 1 week. A significantly greater proportion of trout in the food deprivation treatment remained in their natal stream, but unlike the cortisol treatment, there were no long-term negative effects of food deprivation on growth, relative to the control. Overall survival rates were comparable between the food deprivation and control treatments, but significantly lower for fish in the cortisol treatment. Food availability and individual energetic state appear to dictate the future life-history strategy (migrate or remain resident) of juvenile salmonids while experimental elevation of the stress hormone cortisol causes impaired growth and reduced survival of both resident and migratory individuals.


Genetic analysis of the fungus, Bremia lactucae, using restriction fragment length polymorphisms.
Restriction fragment length polymorphisms (RFLPs) were developed as genetic markers for Bremia lactucae, the biotrophic Oomycete fungus which causes lettuce downy mildew. By using 55 genomic and cDNA probes, a total of 61 RFLP loci were identified among three heterothallic isolates of B. lactucae. Of these 61 RFLP loci, 53 were heterozygous in at least one of the three strains and thus were informative for linkage analysis in at least one of two F1 crosses that were performed. Analysis of the cosegregation of these 53 RFLPs, eight avirulence loci and the mating type locus allowed the construction of a preliminary genetic linkage map consisting of 13 small linkage groups. Based on the extent of linkage detected among probes, the genome of B. lactucae can be estimated to be approximately 2000 cM. Linkage was detected between a RFLP locus and an avirulence gene, providing a potential starting point for chromosome walking to clone an avirulence gene. The high frequency of DNA polymorphism in naturally occurring isolates and the proper Mendelian segregation of loci detected by low copy number probes indicates that it will be possible to construct a detailed genetic map of B. lactucae using RFLPs as markers. The method of analysis employed here should be applicable to many other outbreeding, heterozygous species for which defined inbred lines are not available.


Evaluation of two commercially available alternatives for HIV-1 viral load testing in resource-limited settings.
There is an urgent need for low-cost assays for HIV-1 quantitation to ensure adequate follow-up of HIV-infected patients on antiretroviral therapy (ART) in resource-limited countries. Two low-cost viral load assays are evaluated, a reverse transcriptase activity assay (ExavirLoad v2, Cavidi) and a real-time reverse transcriptase PCR assay (Generic HIV viral load, Biocentric). Both tests were compared with the ultrasensitive HIV Amplicor Monitor assay. Samples were collected in Mombasa, Kenya, from 20 HIV-1 seronegative and 150 HIV-1 seropositive individuals of whom 50 received antiretroviral treatment (ART). The ExavirLoad and the Generic HIV viral load assay were performed in a local laboratory in Mombasa, the Amplicor Monitor assay (version 1.5, Roche Diagnostics) was performed in Ghent, Belgium. ExavirLoad and Generic HIV viral load reached a sensitivity of 98.3% and 100% and a specificity of 80.0% and 90.0%, respectively. Linear regression analyses revealed good correlations between the Amplicor Monitor and the Generic HIV viral load (r=0.935, p<0.001) with high accuracy (100.1%), good precision (5.5%) and a low percent similarity coefficient of variation (5.4%). Bland-Altman analysis found 95% of the samples within clinically acceptable limits of agreement (-1.19 to 0.87logcopies/ml). Although, the ExavirLoad also showed a good linear correlation with the Amplicor Monitor (r=0.901, p<0.001), a problem with false positive results was more significant. The cost per test remains relatively high (US$ 30 for ExavirLoad and US$ 20 for the Generic HIV viral load). Hence, false positive results and the need for an expensive PCR instrument for the Generic HIV viral load assays still limit the implementation of these tests in less equipped, less experienced laboratories.


Induction of cyclooxygenase-2 by mechanical stress through a nitric oxide-regulated pathway.
Biomechanical signals play important roles in regulating the homeostasis of articular cartilage, but under abnormal conditions may be a critical factor in the onset and progression of arthritis. Prostaglandin E(2) (PGE(2)) and nitric oxide (NO), derived from the enzymes cyclo-oxygenase 2 (COX2) and NO synthase 2 (NOS2), are inflammatory mediators that modulate numerous physiological and pathophysiological processes and are potentially important pharmacological targets in osteoarthritis. The goal of this study was to determine the effect of mechanical compression on PGE(2) production in the presence of selective NOS2 and COX2 inhibitors. Articular cartilage explants harvested from 2-3-year-old pigs were subjected to intermittent compression at 0.5Hz over a range of stress magnitudes. PGE(2) and NO production into the media were determined in the presence and absence of the NOS2 inhibitor 1400W or the COX2 inhibitor NS398. COX2 protein levels were determined by immunoblot analysis. Mechanical compression significantly increased NO and PGE(2) synthesis in a manner that was dependent on the magnitude of stress. The selective COX2 inhibitor blocked compression-induced NO and PGE(2) production. Compression in the presence of 1400W further increased COX2 expression resulting in a 10-fold increase in PGE(2) production compared to uncompressed explants with 1400W and a 40-fold increase in PGE(2) compared to uncompressed explants without 1400W. Mechanical compression of articular cartilage increased COX2 and PGE(2) production through a NO-dependent pathway, and therefore pharmacological agents that target the NOS2 pathway in cartilage may have a significant influence on prostanoid production in the joint.


General practitioners' comments on video recorded consultations as an aid to understanding the doctor-patient relationship.
The aim of this study was to describe, understand and interpret physicians' experiences of consultations. Forty-six consultations were videotaped in four primary health care centres. Twelve general practitioners and 46 patients participated. Afterwards the patients and the physicians (on different occasions) commented on the recorded consultations. The comments were analysed from an inductive as well as a hypothetical-deductive model. The analyses agreed well with each other. The result demonstrated that the physicians often had difficulties understanding the patient, that they did not really know how to handle the conversation and felt that they did not really understand the context. The perceived relation to the patient was mainly related to the patients' ability to give adequate information. It is concluded that a key factor in communication is the difficulty which the physician has in understanding what the patient in fact wants-the failure to be on the same wavelength so that the message from the patient can be received.


Evidence supporting zinc as an important antioxidant for skin.
Antioxidants play a critical role in keeping skin healthy. The antioxidant benefits of vitamin C and E are well known, but the importance of the trace mineral, zinc, has been overlooked. This article reviews the evidence supporting zinc's antioxidant role in protecting against free radical-induced oxidative damage. Zinc protects against UV radiation, enhances wound healing, contributes to immune and neuropsychiatric functions, and decreases the relative risk of cancer and cardiovascular disease. All body tissues contain zinc; in skin, it is five to six times more concentrated in the epidermis than the dermis. Zinc is required for the normal growth, development and function of mammals. It is an essential element of more than 200 metalloenzymes, including the antioxidant enzyme, superoxide dismutase, and affects their conformity, stability, and activity. Zinc also is important for the proper functioning of the immune system, and for glandular, reproductive and cell health. Abundant evidence demonstrates the antioxidant role of zinc. Topical zinc, in the form of divalent zinc ions, has been reported to provide antioxidant photoprotection for skin. Two antioxidant mechanisms have been proposed for zinc: zinc ions may replace redox active molecules, such as iron and copper, at critical sites in cell membranes and proteins; alternatively, zinc ions may induce the synthesis of metallothionein, sulfhydryl-rich proteins that protect against free radicals. No matter how they work, topical zinc ions may provide an important and helpful antioxidant defense for skin.


Rotaxane rings promote oblique packing and extended lifetimes in DNA-templated molecular dye aggregates.
Molecular excitons play a central role in natural and artificial light harvesting, organic electrònics, and nanoscale computing. The structure and dynamics of molecular excitons, critical to each application, are sensitively governed by molecular packing. Deoxyribonucleic acid (DNA) templating is a powerful approach that enables controlled aggregation via sub-nanometer positioning of molecular dyes. However, finer sub-Angstrom control of dye packing is needed to tailor excitonic properties for specific applications. Here, we show that adding rotaxane rings to squaraine dyes templated with DNA promotes an elusive oblique packing arrangement with highly desirable optical properties. Specifically, dimers of these squaraine:rotaxanes exhibit an absorption spectrum with near-equal intensity excitonically split absorption bands. Theoretical analysis indicates that the transitions are mostly electronic in nature and only have similar intensities over a narrow range of packing angles. Compared with squaraine dimers, squaraine:rotaxane dimers also exhibit extended excited-state lifetimes and less structural heterogeneity. The approach proposed here may be generally useful for optimizing excitonic materials for a variety of applications ranging from solar energy conversion to quantum information science.


An automated and high-throughput-screening compatible pluripotent stem cell-based test platform for developmental and reproductive toxicity assessment of small molecule compounds.
The embryonic stem cell test (EST) represents the only validated and accepted in vitro system for the detection and classification of compounds according to their developmental and reproductive teratogenic potency. The widespread implementation of the EST, however, in particular for routine application in pharmaceutical development, has not been achieved so far. Several drawbacks still limit the high-throughput screening of potential drug candidates in this format: The long assay period, the use of non-homogeneous viability assays, the low throughput analysis of marker protein expression and the compatibility of the assay procedures to automation. We have therefore introduced several advancements into the EST workflow: A reduction of the assay period, an introduction of homogeneous viability assays, and a straightforward analysis of marker proteins by flow cytometry and high content imaging to assess the impact of small molecules on differentiation capacity. Most importantly, essential parts of the assay procedure have been adapted to lab automation in 96-well format, thus enabling the interrogation of several compounds in parallel. In addition, extensive investigations were performed to explore the predictive capacity of this next-generation EST, by testing a set of well-known embryotoxicants that encompasses the full range of chemical-inherent embryotoxic potencies possible. Due to these significant improvements, the augmented workflow provides a basis for a sensitive, more rapid, and reproducible high throughput screening compatible platform to predict in vivo developmental toxicity from in vitro data which paves the road towards application in an industrial setting. Graphical abstract •The embryonic stem cell test to predict teratogenicity was made automation-compatible. •Several key improvements to the assay procedure have been introduced to increase performance. •The workflow was adapted to human iPS cells and isogenic fibroblast donor cells.


Atherosclerotic lesions in genetically modified mice quantified in vivo by non-invasive high-resolution magnetic resonance microscopy.
We have previously shown that magnetic resonance microscopy (MRM) accurately quantifies atherosclerosis in Apolipoprotein E deficient (ApoE(-/-)) mice aged 36-84 weeks. The present study tests MRM in the quantification of aortic atherosclerosis over a broader range of lesion severity. Younger mice with less advanced disease were imaged in order to evaluate sensitivity, specificity and maximum practical resolution of MRM. Nineteen mice underwent in vivo MRM. Wall area measurements by MRM and light microscopy (LM) (n=43) were highly correlated (r=0.85, slope=0.88, P<0.0001). Wall areas by MRM ranged from 0.114 to 0.934 (median, 0.334) mm(2). A threshold of 0.35 mm(2), for the upper limit of normal, gave MRM positive predictive value (PPV) for detecting abnormally thickened arteries=89.5% and negative predictive value (NPV)=75%, referred to LM. Lesion shape assessed by LM and MRM were also well correlated (r=0.72, P<0.001). Increased wall area in atherosclerosis was found by MRM (P=0.01) and LM (P<0.0001) to be accommodated entirely by 'positive remodeling', confirming the importance of determining plaque size directly. MRM accurately quantifies mouse aortic atherosclerosis and will enhance studies in this important animal model.


Probabilistic inference of transcription factor concentrations and gene-specific regulatory activities.
Quantitative estimation of the regulatory relationship between transcription factors and genes is a fundamental stepping stone when trying to develop models of cellular processes. Recent experimental high-throughput techniques, such as Chromatin Immunoprecipitation (ChIP) provide important information about the architecture of the regulatory networks in the cell. However, it is very difficult to measure the concentration levels of transcription factor proteins and determine their regulatory effect on gene transcription. It is therefore an important computational challenge to infer these quantities using gene expression data and network architecture data. We develop a probabilistic state space model that allows genome-wide inference of both transcription factor protein concentrations and their effect on the transcription rates of each target gene from microarray data. We use variational inference techniques to learn the model parameters and perform posterior inference of protein concentrations and regulatory strengths. The probabilistic nature of the model also means that we can associate credibility intervals to our estimates, as well as providing a tool to detect which binding events lead to significant regulation. We demonstrate our model on artificial data and on two yeast datasets in which the network structure has previously been obtained using ChIP data. Predictions from our model are consistent with the underlying biology and offer novel quantitative insights into the regulatory structure of the yeast cell. MATLAB code is available from http://umber.sbs.man.ac.uk/resources/puma


Hemolytic anemias associated with deficient or dysfunctional spectrin.
Elliptocytes from patients with hereditary elliptocytosis (HE) form elliptical ghosts and membrane skeletons. The composition of HE skeletons is quantitatively normal; however, in some but not all kindreds the major membrane skeletal protein, spectrin, is abnormally heat-sensitive, presumably due to a molecular defect which diminishes its conformational stability. Red cells from four mutants of the common house mouse (Mus musculus) with severe, recessive hemolytic anemias show marked membrane budding, fragmentation, and spherocytosis, which suggest membrane instability. Ghosts spontaneously vesiculate and are spectrin-deficient. The amount of spectrin varies from none to one-half the normal amount and correlates with the clinical severity of the four mutations. The cause of this deficiency remains to be determined. These mutants prove that spectrin is a critical determinant of membrane structural integrity and provide a unique opportunity to test, in intact red cells, putative functions of spectrin. Spectrin extracted from ghosts at low ionic strength is heterogeneous. At physiologic ionic strengths part (46 +/- 5%) is polymerized (P-spectrin) and complexed with actin, and part remains as nonpolymerized spectrin dimers and tetramers (NP-spectrin). We postulate that these are native membrane species which exist in a metabolically controlled equilibrium in vivo and that the proportion of these species regulates membrane shape, strength, and flexibility.


The oncogenic role of miR-155 in breast cancer.
miR-155 is an oncogenic miRNA with well described roles in leukemia. However, additional roles of miR-155 in breast cancer progression have recently been described. A thorough literature search was conducted to review all published data to date, examining the role of miR-155 in breast cancer. Data on all validated miR-155 target genes was collated to identify biologic pathways relevant to miR-155 and breast cancer progression. Publications describing the clinical relevance, functional characterization, and regulation of expression of miR-155 in the context of breast cancer are reviewed. A total of 147 validated miR-155 target genes were identified from the literature. Pathway analysis of these genes identified likely roles in apoptosis, differentiation, angiogenesis, proliferation, and epithelial-mesenchymal transition. The large number of validated miR-155 targets presented here provide many avenues of interest as to the clinical potential of miR-155. Further investigation of these target genes will be required to elucidate the specific mechanisms and functions of miR-155 in breast cancer. This is the first review examining the role of miR-155 in breast cancer progression. The collated data of target genes and biologic pathways of miR-155 identified in this review suggest new avenues of research for this oncogenic miRNA.


Visual perceptual learning in human object recognition areas: a repetition priming study using high-density electrical mapping.
It is often the case that only partial or degraded views of an object are available to an observer, and yet in many of these cases, object recognition is accomplished with surprising ease. The perceptual filling-in or "closure" that makes this possible has been linked to a group of object recognition areas in the human brain, the lateral occipital (LO) complex, and has been shown to have a specific electrophysiological correlate, the N(cl) component of the event related potential. Perceptual closure presumably occurs because repeated and varied exposure to different classes of objects has caused the brain to undergo "perceptual learning," which promotes a robust mnemonic representation, accessible under partial information circumstances. The present study examined the impact of perceptual learning on closure-related brain processes. Fragmented pictures of common objects were presented, such that information content was incrementally increased until just enough information was present to permit closure and object recognition. Periodic repetition of a subset of these picture sequences was used to induce repetition priming due to perceptual learning. This priming has an electrophysiological signature that is putatively generated in the LO complex, but significantly precedes the electrophysiological correlate of closure. The temporal progression of priming- and closure-related activity in the LO complex supports the view that sensory processing entails multiple reentrant stages of activity within processing modules of the visual hierarchy. That the earliest priming-related activity occurs over LO complex, suggests that the sensory trace itself may reside in these object recognition areas.


[Evaluating signal intensity of movement-impaired myocardial segments in MR delayed images after administration of Gd-DTPA. Correlation of regional increase in contraction after revascularization].
The aim of the study was to correlate delayed contrast enhancement of dysfunctional regions of the myocardium after injection of Gd-DTPA with the improvement of regional contractility 3 months after revascularization. Eleven patients with coronary artery disease and wall motion abnormalities underwent MR imaging before and 3 months after revascularization therapy (PTCA or CABG). Short-axis images were acquired using a cine gradient echo sequence. After revascularization, a representative slice was analyzed 14 +/- 1 min after injection of 0.1 mmol/kg Gd-DTPA using a T1-weighted turbo fast low-angle shot sequence. Improved systolic wall thickening 3 months after revascularization served as criterion of viability and was correlated with delayed contrast enhancement patterns. After revascularization, 6 patients showed complete recovery of the dysfunctional area, 1 patient partial recovery, and 4 patients remained unchanged. All 4 patients with persisting wall motion abnormalities, 1/6 patients with recovery and the patient with partial recovery revealed a delayed enhancement. Three months after revascularization, delayed enhancement of dysfunctional myocardium is evident in patients without regional wall motion improvement, while the lack of delayed enhancement correlates with improvement of regional contractility.


Enhancing care for hospitalized older adults with cognitive impairment: a randomized controlled trial.
Approximately 40% of hospitalized older adults have cognitive impairment (CI) and are more prone to hospital-acquired complications. The Institute of Medicine suggests using health information technology to improve the overall safety and quality of the health care system. Evaluate the efficacy of a clinical decision support system (CDSS) to improve the quality of care for hospitalized older adults with CI. A randomized controlled clinical trial. A public hospital in Indianapolis. A total of 998 hospitalized older adults were screened for CI, and 424 patients (225 intervention, 199 control) with CI were enrolled in the trial with a mean age of 74.8, 59% African Americans, and 68% female. A CDSS alerts the physicians of the presence of CI, recommends early referral into a geriatric consult, and suggests discontinuation of the use of Foley catheterization, physical restraints, and anticholinergic drugs. Orders of a geriatric consult and discontinuation orders of Foley catheterization, physical restraints, or anticholinergic drugs. Using intent-to-treat analyses, there were no differences between the intervention and the control groups in geriatric consult orders (56% vs 49%, P = 0.21); discontinuation orders for Foley catheterization (61.7% vs 64.6%, P = 0.86); physical restraints (4.8% vs 0%, P = 0.86), or anticholinergic drugs (48.9% vs 31.2%, P = 0.11). A simple screening program for CI followed by a CDSS did not change physician prescribing behaviors or improve the process of care for hospitalized older adults with CI.


An experimental study of the relationship between thoughts and eating behaviour in bulimia nervosa.
This study tested the hypothesis that cognitive disturbance has a causal role in the maintenance of disturbed eating in bulimia nervosa. Thoughts about eating, weight and shape were activated in one group of patients with bulimia nervosa (the experimental group) but not in another (the control group). There was an increase in negative self-statements in the experimental group following the experimental manipulation. Food consumption was then measured in a taste test. As predicted, the experimental group ate less in the short term than the control group. Contrary to expectations, they did not subsequently report more objective bulimic episodes as a consequence of this decreased food intake. They did, however, report significantly fewer subjective bulimic episodes.


Prevalence and correlates of microalbuminuria in Yemeni children with sickle cell disease.
Microalbuminuria (MA) has been recognized as a sensitive marker of early glomerular injury and a predictor of kidney dysfunction in patients with sickle cell disease (SCD). Limited data are available about MA in SCD children in the Arab countries and none from Yemen. The aim of this study is to determine the prevalence and correlates of MA among 101 children aged 1-16 years, with SCD at their steady state. Children were recruited during their routine health-care visits to the pediatric outpatient clinic in Al-Sadaqa general teaching hospital, Aden, Yemen, between September 2014 and February 2015. A random spot urine sample for each child was screened for MA using Micral-Test strips method. Data on clinical history, anthropometry, blood pressure (BP), and laboratory investigations were obtained. The overall prevalence of MA in this sample was 30.7%, with male predominance (80.6%) (P <0.05). The mean age of children with MA was 7.5 ± 3.2 years, and 10% of them were under five years of age. MA was correlated to both hemoglobin and hematocrit levels, which found to have protective effect against MA (Odds ratio = 0.17 and 0.59, respectively, P <0.05). No correlations were found between MA with age, height, weight, body mass index, recurrent clinical events (painful crises, blood transfusions, and hospitalizations), or fetal hemoglobin levels. BP measurements for all individuals were within the normal ranges, but systolic and diastolic BP were significantly higher in those with MA than without. This study demonstrated a high prevalence of MA in Yemeni children with SCD, and affecting young children as early as 2.5 years of age. Screening for MA as one of the early renal injury markers in children with SCD may help in the prevention of permanent loss of renal function and subsequent renal insufficiency in adulthood.


Characterizing specimen induced aberrations for high NA adaptive optical microscopy.
Aberrations are known to severely compromise image quality in optical microscopy, especially when high numerical aperture (NA) lenses are used in confocal fluorescence microscopy (CFM) and two-photon microscopy (TPM). The method of adaptive optics may correct aberrations and restore diffraction limited operation. So far the problem of aberrations that occur in the imaging of biological specimens has not been quantified. However, this information is essential for the design of adaptive optics systems. We have therefore built an interferometer incorporating high NA objective lenses to measure the aberrations introduced by biological specimens. The measured wavefronts were decomposed into their Zernike mode content in order both to classify and quantify the aberrations. We calculated the potential benefit of correcting different numbers of Zernike modes using different NAs in an adaptive CFM by comparing the signal levels before and after correction. The results indicate that adaptive correction of low order Zernike modes can provide significant benefit for many specimens. The results also show that quantitative fluorescence microscopy may be strongly affected by specimen induced aberrations in non-adaptive systems.


Requirement of Smad4 from Ocular Surface Ectoderm for Retinal Development.
Microphthalmia is characterized by abnormally small eyes and usually retinal dysplasia, accounting for up to 11% of the blindness in children. Right now there is no effective treatment for the disease, and the underlying mechanisms, especially how retinal dysplasia develops from microphthalmia and whether it depends on the signals from lens ectoderm are still unclear. Mutations in genes of the TGF-β superfamily have been noted in patients with microphthalmia. Using conditional knockout mice, here we address the question that whether ocular surface ectoderm-derived Smad4 modulates retinal development. We found that loss of Smad4 specifically on surface lens ectoderm leads to microphthalmia and dysplasia of retina. Retinal dysplasia in the knockout mice is caused by the delayed or failed differentiation and apoptosis of retinal cells. Microarray analyses revealed that members of Hedgehog and Wnt signaling pathways are affected in the knockout retinas, suggesting that ocular surface ectoderm-derived Smad4 can regulate Hedgehog and Wnt signaling in the retina. Our studies suggest that defective of ocular surface ectoderm may affect retinal development.


Probing driving forces in aerolysin and α-hemolysin biological nanopores: electrophoresis versus electroosmosis.
The transport of macromolecules through nanopores is involved in many biological functions and is today at the basis of promising technological applications. Nevertheless the interpretation of the dynamics of the macromolecule/nanopore interaction is still misunderstood and under debate. At the nanoscale, inside biomimetic channels under an external applied voltage, electrophoresis, which is the electric force acting on electrically charged molecules, and electroosmotic flow (EOF), which is the fluid transport associated with ions, contribute to the direction and magnitude of the molecular transport. In order to decipher the contribution of the electrophoresis and electroosmotic flow, we explored the interaction of small, rigid, neutral molecules (cyclodextrins) and flexible, non-ionic polymers (poly(ethylene glycol), PEG) that can coordinate cations under appropriate experimental conditions, with two biological nanopores: aerolysin (AeL) and α-hemolysin (aHL). We performed experiments using two electrolytes with different ionic hydration (KCl and LiCl). Regardless of the nature of the nanopore and of the electrolyte, cyclodextrins behaved as neutral analytes. The dominant driving force was attributed to EOF, acting in the direction of the anion flow and stronger in LiCl than in KCl. The same qualitative behaviour was observed for PEGs in LiCl. In contrast, in KCl, PEGs behaved as positively charged polyelectrolytes through both AeL and aHL. Our results are in agreement with theoretical predictions about the injection of polymers inside a confined geometry (ESI). We believe our results to be of significant importance for better control of the dynamics of analytes of different nature through biological nanopores.


Structure and dynamic properties of a glucocorticoid receptor-induced chromatin transition.
Activation of the mouse mammary tumor virus (MMTV) promoter by the glucocorticoid receptor (GR) is associated with a chromatin structural transition in the B nucleosome region of the viral long terminal repeat (LTR). Recent evidence indicates that this transition extends upstream of the B nucleosome, encompassing a region larger than a single nucleosome (G. Fragoso, W. D. Pennie, S. John, and G. L. Hager, Mol. Cell. Biol. 18:3633-3644). We have reconstituted MMTV LTR DNA into a polynucleosome array using Drosophila embryo extracts. We show binding of purified GR to specific GR elements within a large, multinucleosome array and describe a GR-induced nucleoprotein transition that is dependent on ATP and a HeLa nuclear extract. Previously uncharacterized GR binding sites in the upstream C nucleosome region are involved in the extended region of chromatin remodeling. We also show that GR-dependent chromatin remodeling is a multistep process; in the absence of ATP, GR binds to multiple sites on the chromatin array and prevents restriction enzyme access to recognition sites. Upon addition of ATP, GR induces remodeling and a large increase in access to enzymes sites within the transition region. These findings suggest a dynamic model in which GR first binds to chromatin after ligand activation, recruits a remodeling activity, and is then lost from the template. This model is consistent with the recent description of a "hit-and-run" mechanism for GR action in living cells (J. G. McNally, W. G. Müller, D. Walker, and G. L. Hager, Science 287:1262-1264, 2000).


Myocardial metabolism during increased work states in the porcine left ventricle in vivo.
It is not known whether myocardial energy requirements can be increased to the degree that they exceed myocardial O2 availability in the absence of abnormalities of coronary blood flow or coronary reserve. To determine whether this form of "demand ischemia" occurs, 10 swine were subjected to pressure overload induced by aortic constriction, inotropic and chronotropic stimulation by dobutamine, and the combination of these interventions. In an additional 9 animals, intravenous adenosine was administered during the combination of constriction and dobutamine to determine whether further increases in coronary flow could be achieved and if they would attenuate the metabolic changes. Left ventricular anterior wall transmural blood flow was measured by radioactive microspheres. Energy phosphates were assessed by 31P magnetic resonance spectroscopy using the Fourier series window technique to increase the proportion of signal derived from the subendocardium. Myocardial lactate release was quantified independent of net lactate uptake using an isotopic tracer technique. The three interventions produced 39% to 195% increases in myocardial O2 uptake from control measurements. The phosphocreatine to ATP ratio (PCr/ATP), uncorrected for partial saturation, fell significantly, from 1.39 +/- 0.10 at control conditions to 1.25 +/- 0.10 with dobutamine alone and 1.15 +/- 0.08 with dobutamine plus constriction (P < .05 for both). Myocardial lactate release rose from 0.21 +/- 0.03 mumol.g-1.min-1 at control conditions to 0.45 +/- 0.05 and 0.59 +/- 0.10 mumol.g-1.min-1, respectively (P < .05 for both), with these two interventions. Although transmurally averaged left ventricular blood flow rose from 0.97 +/- 0.09 mL.g-1.min-1 at control conditions to 3.25 +/- 0.47 mL.g-1.min-1 (P < .001) and subendocardial blood flow increased from 1.02 +/- 0.09 to 2.92 +/- 0.45 mL.g-1.min-1 (P < .001) at the highest of the three increased work states, the subendocardial to subepicardial flow ratio declined progressively from 1.13 +/- 0.08 to 0.87 +/- 0.04 (P < .05). With a further increase in aortic constriction, myocardial O2 uptake and subepicardial blood flow rose, whereas subendocardial blood flow did not change, and there was a further decline in PCr/ATP and a rise in lactate release. Although adenosine increased the average myocardial blood flow during high work state from 3.79 +/- 0.91 to 6.29 +/- 1.08 mL.g-1.min-1 (P < .001), the further rise in subendocardial flow from 3.08 +/- 0.62 to 3.78 +/- 0.68 mL.g-1.min-1 was not significant, nor were the accompanying changes in PCr/ATP or lactate metabolism.(ABSTRACT TRUNCATED AT 400 WORDS)


Risk factors for readmission in schizophrenia patients following involuntary admission.
Individuals with schizophrenia who are involuntarily admitted may have poorer prognosis, including higher readmission rates, than those voluntarily admitted. However, little is known about the risk factors for readmission in those schizophrenia patients who are involuntarily admitted. We aim to explore the risk factors for readmission in this population. We enrolled 138 schizophrenia patients with involuntary admission from July 2008 to June 2013 and followed those patients for readmission outcomes at 3 months and at 1 year. The one-year and 3-months readmission rates were 33.3% and 15.2%, respectively. Unmarried status (adjusted odds ratio (aOR) = 6.28, 95% CI: 1.48-26.62), previous history of involuntary admission (aOR = 4.08, 95% CI: 1.19-14.02), longer involuntary admission days (aOR = 1.04, 95% CI: 1.01-1.07) and shorter total admission days (aOR = 1.03, 95% CI: 1.01-1.05) were associated with increased risk for 1-year readmission. Younger age (aOR = 1.10, 95% CI 1.02-1.18) was associated with increased risk for 3-months readmission. Unmarried status, prior history of involuntary admission, longer involuntary admission days and shorter total admission days were associated with increased risk for 1-year readmission. Healthcare providers may need to focus on patients with these risk factors to reduce subsequent readmissions.


Congenital hypothyroidism: assessment with ultrasound.
To determine the clinical utility of cervical ultrasound in patients suspected of having congenital hypothyroidism. Thirty-seven patients with suspected congenital hypothyroidism underwent ultrasound and scintigraphic evaluation of the thyroid anatomy, morphology, and function. The ultrasound findings and laboratory data were compared with the standard-of-reference scintigraphic findings and laboratory data for diagnosing specific causes in those patients, and prognosis was correlated with the ultrasound findings. Ultrasound was not reliable for detecting ectopia (n = 8) or differentiating ectopia from aplasia (n = 1). Ultrasound showed ectopia in six (four in the mouth floor and two in the tongue base) of eight cases (75% sensitivity). Ultrasound did not show one ectopia in the floor of mouth because its echogenicity was similar to that of surrounding tissues. A second ectopia, in the hypopharynx, was missed because of hindrance of the laryngeal air. Radioactive iodine uptake and scintigraphy was required for the patients with enlarged glands in the normal place to differentiate dyshormonogenesis from other categories. Specific causes were diagnosed correctly with ultrasound findings and laboratory data alone in all of the 20 patients who had hemiaplasia or small or normal-size glands in the normal location. Incidences of heterogeneity and hypoechogenicity of the thyroid gland in patients with prolonged clinical course (whose replacement therapy or follow-up extended for more than 1 year) were significantly higher than those in patients with short clinical course. Ultra-sound can obviate the need for scintigraphy in more than half (54%) of patients with possible congenital hypothyroidism. Ultrasound has a potential to predict prognosis of these patients.


Acidic residues in the nucleotide-binding site of the bacteriophage T7 DNA primase.
DNA primases catalyze the synthesis of oligoribonucleotides to initiate lagging strand DNA synthesis during DNA replication. Like other prokaryotic homologs, the primase domain of the gene 4 helicase-primase of bacteriophage T7 contains a zinc motif and a catalytic core. Upon recognition of the sequence, 5'-GTC-3' by the zinc motif, the catalytic site condenses the cognate nucleotides to produce a primer. The TOPRIM domain in the catalytic site contains several charged residues presumably involved in catalysis. Each of eight acidic residues in this region was replaced with alanine, and the properties of the altered primases were examined. Six of the eight residues (Glu-157, Glu-159, Asp-161, Asp-207, Asp-209, and Asp-237) are essential in that altered gene 4 proteins containing these mutations cannot complement T7 phage lacking gene 4 for T7 growth. These six altered gene 4 proteins can neither synthesize primers de novo nor extend an oligoribonucleotide. Despite the inability to catalyze phosphodiester bond formation, the altered proteins recognize the sequence 5'-GTC-3' in the template and deliver preformed primer to T7 DNA polymerase. The alterations in the TOPRIM domain result in the loss of binding affinity for ATP as measured by surface plasmon resonance assay together with ATP-agarose affinity chromatography.


Risk factors for breast cancer, including occupational exposures.
The knowledge on the etiology of breast cancer has advanced substantially in recent years, and several etiological factors are now firmly established. However, very few new discoveries have been made in relation to occupational risk factors. The International Agency for Research on Cancer has evaluated over 900 different exposures or agents to-date to determine whether they are carcinogenic to humans. These evaluations are published as a series of Monographs (www.iarc.fr). For breast cancer the following substances have been classified as "carcinogenic to humans" (Group 1): alcoholic beverages, exposure to diethylstilbestrol, estrogen-progestogen contraceptives, estrogen-progestogen hormone replacement therapy and exposure to X-radiation and gamma-radiation (in special populations such as atomic bomb survivors, medical patients, and in-utero exposure). Ethylene oxide is also classified as a Group 1 carcinogen, although the evidence for carcinogenicity in epidemiologic studies, and specifically for the human breast, is limited. The classification "probably carcinogenic to humans" (Group 2A) includes estrogen hormone replacement therapy, tobacco smoking, and shift work involving circadian disruption, including work as a flight attendant. If the association between shift work and breast cancer, the most common female cancer, is confirmed, shift work could become the leading cause of occupational cancer in women.


Whole exome and targeted deep sequencing identify genome-wide allelic loss and frequent SETDB1 mutations in malignant pleural mesotheliomas.
Malignant pleural mesothelioma (MPM), a rare malignancy with a poor prognosis, is mainly caused by exposure to asbestos or other organic fibers, but the underlying genetic mechanism is not fully understood. Genetic alterations and causes for multiple primary cancer development including MPM are unknown. We used whole exome sequencing to identify somatic mutations in a patient with MPM and two additional primary cancers who had no evidence of venous, arterial, lymphovascular, or perineural invasion indicating dissemination of a primary lung cancer to the pleura. We found that the MPM had R282W, a key TP53 mutation, and genome-wide allelic loss or loss of heterozygosity, a distinct genomic alteration not previously described in MPM. We identified frequent inactivating SETDB1 mutations in this patient and in 68 additional MPM patients (mutation frequency: 10%, 7/69) by targeted deep sequencing. Our observations suggest the possibility of a new genetic mechanism in the development of either MPM or multiple primary cancers. The frequent SETDB1 inactivating mutations suggest there could be new diagnostic or therapeutic options for MPM.


Transmission of multidrug-resistant tuberculosis.
To compare the Mycobacterium tuberculosis isolates of adult index cases with multidrug-resistant (MDR) tuberculosis to the isolates obtained from their child contacts. A 4-year prospective study in the Western Cape Province of South Africa. We evaluated 149 child contacts of 80 adult MDR pulmonary tuberculosis cases. This report includes those cases where a culture for M. tuberculosis was obtained from both the adult source case and the child contact. Isolates were compared by drug susceptibility pattern and restriction fragment length polymorphism analysis. Six adult-child pairs with cultures for M. tuberculosis were identified. Two children had contact with more than one adult tuberculosis case. One child received previous isoniazid prophylaxis. Drug susceptibility pattern and restriction fragment length polymorphism analysis were identical for five adult-child pairs. One child, with no other known source case, had a strain different from that of the identified source case, but the MDR M. tuberculosis strain with which he was infected was prevalent in the community in which he resided. All children responded well to treatment. This study confirms that most of the childhood contacts of adults with MDR tuberculosis are likely to be infected by these MDR source cases despite their exposure to other drug-susceptible adults with tuberculosis in some instances. Child contacts of adults with MDR tuberculosis should be treated according to the drug susceptibility patterns of the likely source cases' M. tuberculosis strains unless their own strain's susceptibility testing indicates otherwise. Contact tracing remains of fundamental importance in identifying children at risk.


Prevalence of diabetes mellitus in pregnancy among Yup'ik Eskimos, 1987-1988.
To evaluate the prevalence of diabetes mellitus in pregnancy in Yup'ik Eskimos. A retrospective review of consecutive birth-log data and medical records was conducted. Records were taken from the primary care and tertiary referral facilities in Alaska Area Native Health Service. Some 630 consecutive deliveries were reviewed from 1 March 1987 to 29 February 1988, with 25 excluded for ancestry other than Alaska Native. Another 605 Alaska Native patient charts were reviewed, with 545 Alaska Native patients screened for diabetes in pregnancy. The study population had a mean age of 25.6 yr, mean gravidity of 3.4, mean parity of 1.9, and mean birth weight of 3567 +/- 493 g. Patients were screened with a 50-g glucose oral load with a plasma glucose 1 h later. 156 of 605 (25.7%) patients, with a screen > or = 7.8 mM received a 100-g OGTT. Of 605, 35 (5.8%) patients met O'Sullivan criteria, and 2 of 605 (0.3%) patients met WHO criteria for previous diabetes mellitus, for a total 37 of 605 (6.7% [corrected]) women with diabetes in pregnancy. The subjects who met O'Sullivan criteria had statistically greater mean age (29.9 yr), gravidity (4.9), parity (2.9), and birth weight of their infants (3678 +/- 389 g), compared with women with a screen < 7.8 mM. The prevalence of diabetes in pregnancy among Yup'ik Eskimos is twice the rate for the U.S. for all races, despite the Yup'ik having the lowest rate of diabetes mellitus among Alaska Natives. This may represent a large number of undiagnosed patients with impaired glucose tolerance, and may reflect the wide-spread dietary and life-style changes that have occurred in the Yup'ik in the last 30 yr. The Yup'ik present a unique opportunity to apply prevention techniques in a population with an emerging problem with glucose tolerance.


Dorsal anterior cingulate-brainstem ensemble as a reinforcement meta-learner.
Optimal decision-making is based on integrating information from several dimensions of decisional space (e.g., reward expectation, cost estimation, effort exertion). Despite considerable empirical and theoretical efforts, the computational and neural bases of such multidimensional integration have remained largely elusive. Here we propose that the current theoretical stalemate may be broken by considering the computational properties of a cortical-subcortical circuit involving the dorsal anterior cingulate cortex (dACC) and the brainstem neuromodulatory nuclei: ventral tegmental area (VTA) and locus coeruleus (LC). From this perspective, the dACC optimizes decisions about stimuli and actions, and using the same computational machinery, it also modulates cortical functions (meta-learning), via neuromodulatory control (VTA and LC). We implemented this theory in a novel neuro-computational model-the Reinforcement Meta Learner (RML). We outline how the RML captures critical empirical findings from an unprecedented range of theoretical domains, and parsimoniously integrates various previous proposals on dACC functioning.


Sex, clinical presentation, and outcome in patients with acute coronary syndromes. Global Use of Strategies to Open Occluded Coronary Arteries in Acute Coronary Syndromes IIb Investigators.
Studies have reported that women with acute myocardial infarction have in-hospital and long-term outcomes that are worse than those of men. To assess sex-based differences in presentation and outcome, we examined data from the Global Use of Strategies to Open Occluded Coronary Arteries in Acute Coronary Syndromes IIb study, which enrolled 12,142 patients (3662 women and 8480 men) with acute coronary syndromes, including infarction with ST-segment elevation, infarction with no ST-segment elevation, and unstable angina. Overall, the women were older than the men, and had significantly higher rates of diabetes, hypertension, and prior congestive heart failure. They had significantly lower rates of prior myocardial infarction and were less likely ever to have smoked. A smaller percentage of women than men had infarction with ST elevation (27.2 percent vs. 37.0 percent, P<0.001), and of the patients who presented with no ST elevation (those with myocardial infarction or unstable angina), fewer women than men had myocardial infarction (36.6 percent vs. 47.6 percent, P<0.001). Women had more complications than men during hospitalization and a higher mortality rate at 30 days (6.0 percent vs. 4.0 percent, P<0.001) but had similar rates of reinfarction at 30 days after presentation. However, there was a significant interaction between sex and the type of coronary syndrome at presentation (P=0.001). After stratification according to coronary syndrome and adjustment for base-line variables, there was a nonsignificant trend toward an increased risk of death or reinfarction among women as compared with men only in the group with infarction and ST elevation (odds ratio, 1.27; 95 percent confidence interval, 0.98 to 1.63; P=0.07). Among patients with unstable angina, female sex was associated with an independent protective effect (odds ratio for infarction or death, 0.65; 95 percent confidence interval, 0.49 to 0.87; P=0.003). Women and men with acute coronary syndromes had different clinical profiles, presentation, and outcomes. These differences could not be entirely accounted for by differences in base-line characteristics and may reflect pathophysiologic and anatomical differences between men and women.


The Fas counterattack: Fas-mediated T cell killing by colon cancer cells expressing Fas ligand.
Tumors escape immunological rejection by a diversity of mechanisms. In this report, we demonstrate that the colon cancer cell SW620 expresses functional Fas ligand (FasL), the triggering agent of Fas receptor (FasR)-mediated apoptosis within the immune system. FasL mRNA and cell surface FasL were detected in SW620 cells using reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemical staining, respectively. We show that SW620 kills Jurkat T cells in a Fas-mediated manner. FasR-specific antisense oligonucleotide treatment, which transiently inhibited FasR expression, completely protected Jurkat cells from killing by SW620. FasL-specific antisense oligonucleotide treatment of SW620 inhibited its Jurkat-killing activity. FasL has recently been established as a mediator of immune privilege in mouse retina and testis. Our finding that colon cancer cells express functional FasL suggests it may play an analogous role in bestowing immune privilege on human tumors. HT29 and SW620 colon cancer cells were found to express FasR mRNA and cell surface FasR using RT-PCR and immunofluorescence flow cytometry, respectively. However, neither of these cells underwent apoptosis after treatment by the anti-FasR agonistic monoclonal antibody CH11. Our results therefore suggest a Fas counterattack model for immune escape in colon cancer, whereby the cancer cells resist Fas-mediated T cell cytotoxicity but express functional FasL, an apoptotic death signal to which activated T cells are inherently sensitive.


Gene- and pathway-based association tests for multiple traits with GWAS summary statistics.
To identify novel genetic variants associated with complex traits and to shed new insights on underlying biology, in addition to the most popular single SNP-single trait association analysis, it would be useful to explore multiple correlated (intermediate) traits at the gene- or pathway-level by mining existing single GWAS or meta-analyzed GWAS data. For this purpose, we present an adaptive gene-based test and a pathway-based test for association analysis of multiple traits with GWAS summary statistics. The proposed tests are adaptive at both the SNP- and trait-levels; that is, they account for possibly varying association patterns (e.g. signal sparsity levels) across SNPs and traits, thus maintaining high power across a wide range of situations. Furthermore, the proposed methods are general: they can be applied to mixed types of traits, and to Z-statistics or P-values as summary statistics obtained from either a single GWAS or a meta-analysis of multiple GWAS. Our numerical studies with simulated and real data demonstrated the promising performance of the proposed methods. The methods are implemented in R package aSPU, freely and publicly available at: https://cran.r-project.org/web/packages/aSPU/ CONTACT: email@example.com information: Supplementary data are available at Bioinformatics online.


Benefits from Below: Silicon Supplementation Maintains Legume Productivity under Predicted Climate Change Scenarios.
Many studies demonstrate that elevated atmospheric carbon dioxide concentrations (eCO2) can promote root nodulation and biological nitrogen fixation (BNF) in legumes such as lucerne (Medicago sativa). But when elevated temperature (eT) conditions are applied in tandem with eCO2, a more realistic scenario for future climate change, the positive effects of eCO2 on nodulation and BNF in M. sativa are often much reduced. Silicon (Si) supplementation of M. sativa has also been reported to promote root nodulation and BNF, so could potentially restore the positive effects of eCO2 under eT. Increased nitrogen availability, however, could also increase host suitability for aphid pests, potentially negating any benefit. We applied eCO2 (+240 ppm) and eT (+4°C), separately and in combination, to M. sativa growing in Si supplemented (Si+) and un-supplemented soil (Si-) to determine whether Si moderated the effects of eCO2 and eT. Plants were either inoculated with the aphid Acyrthosiphon pisum or insect-free. In Si- soils, eCO2 stimulated plant growth by 67% and nodulation by 42%, respectively, whereas eT reduced these parameters by 26 and 48%, respectively. Aphids broadly mirrored these effects on Si- plants, increasing colonization rates under eCO2 and performing much worse (reduced abundance and colonization) under eT when compared to ambient conditions, confirming our hypothesized link between root nodulation, plant growth, and pest performance. Examined across all CO2 and temperature regimes, Si supplementation promoted plant growth (+93%), and root nodulation (+50%). A. pisum abundance declined sharply under eT conditions and was largely unaffected by Si supplementation. In conclusion, supplementing M. sativa with Si had consistent positive effects on plant growth and nodulation under different CO2 and temperature scenarios. These findings offer potential for using Si supplementation to maintain legume productivity under predicted climate change scenarios without making legumes more susceptible to insect pests.


Characterization of natural hybrids of Saccharomyces cerevisiae and Saccharomyces bayanus var. uvarum.
Nine yeast strains were isolated from spontaneous fermentations in the Alsace area of France, during the 1997, 1998 and 1999 grape harvests. Strains were characterized by pulsed-field gel electrophoresis, PCR-restriction fragment length polymorphism (RFLP) of the MET2 gene, delta-PCR, and microsatellite patterns. Karyotypes and MET2 fragments of the nine strains corresponded to mixed chromosomal bands and restriction patterns for both Saccharomyces cerevisiae and Saccharomyces bayanus var. uvarum. They also responded positively to amplification with microsatellite primers specific to both species and were demonstrated to be diploid. However, meiosis led to absolute nonviability of their spores on complete medium. All the results demonstrated that the nine yeast strains isolated were S. cerevisiaexS. bayanus var. uvarum diploid hybrids. Moreover, microsatellite DNA analysis identified strains isolated in the same cellar as potential parents belonging to S. bayanus var. uvarum and S. cerevisiae.


Enhanced rate of nerve regeneration and directional errors after sciatic nerve injury in receptor protein tyrosine phosphatase sigma knock-out mice.
The receptor protein tyrosine phosphatase sigma (PTPsigma) is a member of the mammalian leukocyte common antigen-related (LAR) family. Its expression is developmentally regulated in neuronal tissues. The Drosophila homolog of the mammalian LAR family of phosphatases (DLAR) controls axon guidance during Drosophila embryogenesis. We have demonstrated previously that mice deficient in PTPsigma have CNS and peripheral nervous system abnormalities. The sciatic nerve in the PTPsigma(-/-) mice demonstrates an increased number of small diameter fibers and slower nerve conduction velocities compared with PTPsigma(+/+) or PTPsigma(+/-) controls. To study whether peripheral nerve regeneration is affected by PTPsigma activity, we assessed nerve regeneration in the PTPsigma(-/-) mouse after three standard models of sciatic nerve injury. We report that after sciatic nerve crush injury, nerve regeneration was significantly faster in the PTPsigma(-/-) animals, as determined by histologic, electrophysiologic, and neuromuscular testing. After sciatic nerve transection with immediate microsurgical repair or allografting, PTPsigma(-/-) nerve fibers demonstrated errors in directional growth compared with controls. We propose that PTPsigma regulates the axonal regeneration rate and guidance of regenerating fibers.


[Azygos vein aneurysm occurring simultaneously with hemoptysis].
A 64-year-old man was admitted with hemoptysis. A chest X-ray showed a well-defined round nodule at the right tracheobronchial angle. Enhanced computed tomography revealed the superior vena cava to be completely occluded while the azygos vein was also observed to have formed an aneurysm. Bronchoscopy showed the submucosal bronchial vessels at the right second carina to be markedly dilated, and thus considered them to be the likely cause of hemoptysis. Based on the above findings, we considered the following events as the most likely to have taken place: The superior vena cava was completely occluded due to the long-term placement of a pacemaker. The blood flow of head and upper limbs could not sufficiently return to the heart, but instead flowed into the azygos vein via the collateral circulation. This resulted in both the formation of the azygos vein aneurysm and an elevated venous pressure of the bronchial vein. Due to this pressure elevation, the bronchial vein and small vessels had both ruptured, thereby inducing the onset of hemoptysis.


A unique antigen, RT11, in the placenta of the rat.
In the course of exploring the antibody response in the unsensitized WF (u) female pregnant by a DA (a) male, we prepared a hybridoma that secreted an antibody (mAb 213) that was specific to the a haplotype but identified an antigen different from Pa. This antigen was designated RT11. It is present from the twelfth day of gestation on the collagen fibers of the placenta and of all organs in fetal and adult rats. It is particularly prominent on red blood cells; in the yolk sac epithelium; in the walls of the endodermal sinus, blood vessels and bronchioles; and in capsules and trabeculae. A very small amount is present on DA lymphocytes, since 17-20% of them react with mAb 213 by cytofluorimetry. The RT11 antigen is absent from the basal and labyrinthine trophoblast cells, from the parenchymal cells of all organs, and from T and B cells. This distribution pattern is completely different from that of the Aa and Pa antigens. Inhibition and absorption studies showed that RT11 is not an integral part of the collagen molecule. The SDS-PAGE analysis of the immunoprecipitates of RT11 from radioiodinated whole-membrane extracts of red blood cells and from the glycoprotein fraction thereof showed that it is an unglycosylated protein of molecular weight 29,000. The evidence to date suggests that RT11 is a blood group antigen. Studies on the genetic control of the expression of RT11 were undertaken to determine whether a gene linked to the MHC was involved and whether the control mechanism was unigenic or polygenic. Backcrosses generated using inbred strains--(DA x BN)F1 x DA-- and using complementary congenic strains--(DA.1N x BN.1A) F1 x BN.1A--showed that the expression of RT11 was under polygenic control, and that both an MHC-linked gene (1.2 cM from RT1.Aa) and genes not linked to the MHC are involved. By contrast, the expression of the Pa antigen is under the control of an MHC gene only.


Implementing COVID-19 (SARS-CoV-2) Rapid Diagnostic Tests in Sub-Saharan Africa: A Review.
Introduction: For the COVID-19 (SARS-CoV-2) response, COVID-19 antigen (Ag), and antibody (Ab) rapid diagnostic tests (RDTs) are expected to complement central molecular testing particularly in low-resource settings. The present review assesses requirements for implementation of COVID-19 RDTs in sub-Saharan Africa. Methods: Review of PubMed-published articles assessing COVID-19 RDTs complemented with Instructions for Use (IFU) of products. Results: In total 47 articles on two COVID-19 Ag RDTs and 54 COVID-19 Ab RDTs and IFUs of 20 COVID-19 Ab RDTs were retrieved. Only five COVID-19 Ab RDTs (9.3%) were assessed with capillary blood sampling at the point-of-care; none of the studies were conducted in sub-Saharan Africa. Sampling: Challenges for COVID-19 Ag RDTs include nasopharyngeal sampling (technique, biosafety) and sample stability; for COVID-19 Ab RDTs equivalence of whole blood vs. plasma/serum needs further validation (assessed for only eight (14.8%) products). Sensitivity-Specificity: sensitivity of COVID-19 Ag and Ab RDTs depend on viral load (antigen) and timeframe (antibody), respectively; COVID-19 Ab tests have lower sensitivity compared to laboratory test platforms and the kinetics of IgM and IgG are very similar. Reported specificity was high but has not yet been assessed against tropical pathogens. Kit configuration: For COVID-19 Ag RDTs, flocked swabs should be added to the kit; for COVID-19 Ab RDTs, finger prick sampling materials, transfer devices, and controls should be added (currently only supplied in 15, 5, and 1/20 products). Usability and Robustness: some COVID-19 Ab RDTs showed high proportions of faint lines (>40%) or invalid results (>20%). Shortcomings were reported for buffer vials (spills, air bubbles) and their instructions for use. Stability: storage temperature was ≤ 30°C for all but one RDT, in-use and result stability were maximal at 1 h and 30 min, respectively. Integration in the healthcare setting requires a target product profile, landscape overview of technologies, certified manufacturing capacity, a sustainable market, and a stringent but timely regulation. In-country deployment depends on integration in the national laboratory network. Discussion/Conclusion: Despite these limitations, successful implementation models in triage, contact tracing, and surveillance have been proposed, in particular for COVID-19 Ab RDTs. Valuable experience is available from implementation of other disease-specific RDTs in sub-Saharan Africa.


Rhizosphere properties and heavy metal accumulation of plants growing in the fly ash dumpsite, Morupule power plant, Botswana.
Discarding fly ash from a coal power plant into a dumpsite does not only contribute to deforestation and loss of productive land but also leads to contamination of air, soil, and groundwater. Therefore, fly ash should be managed properly to avoid the migration of contaminants. One management option is phytoremediation using adapted plants and as a prerequisite, there is a need to identify suitable plants that can be used for revegetation of fly ash dumpsites. To identify prospective plants, a survey was carried out by assessing the plants growing in Morupule B fly ash dumpsite based on their ability to accumulate heavy metals and their bioconcentration (BAF) and translocation factors (TF). Of the twenty-two-plant species growing in the fly ash dumpsite of Morupule B power plant station, N. glauca is a potential phytoextraction agent for Cu (TFCu = 1.02; BAFCu = 2.16) and Pb (TFPb = 1.38; BAFPb = 1.65); P. burchellii for Pb (TFPb = 1.61, BAFPb = 0.9) and Zn (TFZn = 1.35; BAFZn = 5.74); I. pes-tigridis for Pb (TFPb = 1.35; BAFPb = 1.56) and Zn (TFZn = 1.62; BAFZn = 7.43); A. pungens for Cr (TFCr = 1.22; BAFCr = 0.11), Cu (TFCu = 2.18; BAFCu = 1.14), and Zn (TFZn = 1.04; BAFZn = 1.44); E. hirta for Zn (TFZn = 1.54, BAFZn = 2.44); A. spinosus for Pb (TFPb = 1.29; BAFPb = 1.55); C. dactylon for Cu (TFCu = 1.86; BAFCu = 1.07) and Zn (TFZn = 1.00; BAFZn = 2.46); and D. aegyptium for Pb (TFPb = 1.19; BAFPb = 2.57). Other plants growing in the fly ash dumpsite are potential candidates for phytostabilization as they can tolerate a high concentration of metals and low essential nutrients. Also, different plant groups variably modified the pH, EC, OM, and exchangeable fractions of metals in the rhizosphere wherein grasses can increase the OM at higher rates, and it has a higher capacity to acidify and solubilize heavy metals in the rhizosphere leading to higher EC and available metals compared to other plant groups. Overall, the information presented is useful in identifying plants or their combinations for the phytoremediation of fly ash and other heavy metal-polluted environments.


Clinical, Morphological, and Molecular Evaluations of Bone Regeneration With an Additive Manufactured Osteosynthesis Plate.
There is limited information on the biological status of bone regenerated with microvascular fibula flap combined with biomaterials. This paper describes the clinical, histological, ultrastructural, and molecular picture of bone regenerated with patient-customized plate, used for mandibular reconstruction in combination with microvascular osteomyocutaneous fibula flap. The plate was virtually planned and additively manufactured using electron beam melting. This plate was retrieved from the patient after 33 months. Microcomputed tomography, backscattered-scanning electron microscopy, histology, and quantitative-polymerase chain reaction were employed to evaluate the regenerated bone and the flap bone associated with the retrieved plate. At retrieval, the posterior two-thirds of the plate were in close adaptation with the underlying flap, whereas soft tissue was observed between the native mandible and the anterior one-third. The histological and structural analyses showed new bone regeneration, ingrowth, and osseointegration of the posterior two-thirds. The histological observations were supported by the gene expression analysis showing higher expression of bone formation and remodeling genes under the posterior two-thirds compared with the anterior one-third of the plate. The observation of osteocytes in the flap indicated its viability. The present data endorse the suitability of the customized, additively manufactured plate for the vascularized fibula mandibular reconstruction. Furthermore, the combination of the analytical techniques provides possibilities to deduce the structural and molecular characteristics of bone regenerated using this procedure.


Extracortical origin of some murine subplate cell populations.
The subplate layer, the deepest cortical layer in mammals, has important roles in cerebral cortical development. The subplate contains heterogeneous cell populations that are morphologically diverse, with several projection targets. It is currently assumed that these cells are generated in the germinative zone of the earliest cortical neuroepithelium. Here we identify a pallial but extracortical area located in the rostromedial telencephalic wall (RMTW) that gives rise to several cell populations. Postmitotic neurons migrate tangentially from the RMTW toward the cerebral cortex. Most RMTW-derived cells are incorporated into the subplate layer throughout its rostrocaudal extension, with others contributing to the GABAergic interneuron pool of cortical layers V and VI.


Differential expression of VEGF isoforms in mouse during development and in the adult.
Vascular endothelial growth factor (VEGF), a factor that is critical for development of the vascular system in mouse embryos, exists as at least three isoforms, VEGF120, VEGF164, and VEGF188. The isoforms have different affinities for heparan sulfate as well as for the three known VEGF receptors, VEGFR-1 (Flt-1), VEGFR-2 (Flk-1), and neuropilin-1, suggesting that different VEGF isoforms may play distinct roles in vascular development. To determine whether there are differences in the organ-specific expression patterns that would support this concept, we used a quantitative RNase protection assay (RPA) to determine the distribution of different VEGF isoform mRNA in developing and adult mouse organs. Results revealed that the ratios of the three VEGF isoforms changed during organ development and that adult organs expressed different levels of the three VEGF isoforms. Because the lung expressed the highest levels of VEGF188 isoform, we used VEGF isoform-specific in situ hybridization in the developing lung and determined that type II alveolar epithelial cells were expressing high levels of VEGF188 mRNA. Finally, targeted exon deletion of the VEGF gene revealed that mice that developed in the absence of the heparan sulfate binding isoforms VEGF164 and VEGF188, displayed a variety of vascular defects, including abnormal pulmonary vascular development. Our results support the concept that different VEGF isoforms have distinct functions in vascular development.


Depression, anxiety, and smartphone addiction in university students- A cross sectional study.
The study aims to assess prevalence of smartphone addiction symptoms, and to ascertain whether depression or anxiety, independently, contributes to smartphone addiction level among a sample of Lebanese university students, while adjusting simultaneously for important sociodemographic, academic, lifestyle, personality trait, and smartphone-related variables. A random sample of 688 undergraduate university students (mean age = 20.64 ±1.88 years; 53% men) completed a survey composed of a) questions about socio-demographics, academics, lifestyle behaviors, personality type, and smartphone use-related variables; b) 26-item Smartphone Addiction Inventory (SPAI) Scale; and c) brief screeners of depression and anxiety (PHQ-2 and GAD-2), which constitute the two core DSM-IV items for major depressive disorder and generalized anxiety disorder, respectively. Prevalence rates of smartphone-related compulsive behavior, functional impairment, tolerance and withdrawal symptoms were substantial. 35.9% felt tired during daytime due to late-night smartphone use, 38.1% acknowledged decreased sleep quality, and 35.8% slept less than four hours due to smartphone use more than once. Whereas gender, residence, work hours per week, faculty, academic performance (GPA), lifestyle habits (smoking and alcohol drinking), and religious practice did not associate with smartphone addiction score; personality type A, class (year 2 vs. year 3), younger age at first smartphone use, excessive use during a weekday, using it for entertainment and not using it to call family members, and having depression or anxiety, showed statistically significant associations with smartphone addiction. Depression and anxiety scores emerged as independent positive predictors of smartphone addiction, after adjustment for confounders. Several independent positive predictors of smartphone addiction emerged including depression and anxiety. It could be that young adults with personality type A experiencing high stress level and low mood may lack positive stress coping mechanisms and mood management techniques and are thus highly susceptible to smartphone addiction.