The Chlamydomonas mating type plus fertilization tubule, a prototypic cell fusion organelle: isolation, characterization, and in vitro adhesion to mating type minus gametes.
In the biflagellated alga Chlamydomonas, adhesion and fusion of the plasma membranes of gametes during fertilization occurs via an actin-filled, microvillus-like cell protrusion. Formation of this approximately 3-microm-long fusion organelle, the Chlamydomonas fertilization tubule, is induced in mating type plus (mt+) gametes during flagellar adhesion with mating type minus (mt-) gametes. Subsequent adhesion between the tip of the mt+ fertilization tubule and the apex of a mating structure on mt- gametes is followed rapidly by fusion of the plasma membranes and zygote formation. In this report, we describe the isolation and characterization of fertilization tubules from mt+ gametes activated for cell fusion. Fertilization tubules were detached by homogenization of activated mt+ gametes in an EGTA-containing buffer and purified by differential centrifugation followed by fractionation on sucrose and Percoll gradients. As determined by fluorescence microscopy of samples stained with a fluorescent probe for filamentous actin, the method yielded 2-3 x 10(6) fertilization tubules/microg protein, representing up to a 360-fold enrichment of these organelles. Examination by negative stain electron microscopy demonstrated that the purified fertilization tubules were morphologically indistinguishable from fertilization tubules on intact, activated mt+ gametes, retaining both the extracellular fringe and the internal array of actin filaments. Several proteins, including actin as well as two surface proteins identified by biotinylation studies, copurified with the fertilization tubules. Most importantly, the isolated mt+ fertilization tubules bound to the apical ends of activated mt- gametes between the two flagella, the site of the mt- mating structure; a single fertilization tubule bound per cell, binding was specific for gametes, and fertilization tubules isolated from trypsin-treated, activated mt+ gametes did not bind to activated mt- gametes.


Baseline differences in the HF-ACTION trial by sex.
In patients with heart failure (HF), assessment of functional capacity plays an important prognostic role. Both 6-minute walk and cardiopulmonary exercise testing have been used to determine physical function and to determine prognosis and even listing for transplantation. However, as in HF trials, the number of women reported has been small, and the cutoffs for transplantation have been representative of male populations and extrapolated to women. It is also well known that peak VO(2) as a determinant of fitness is inherently lower in women than in men and potentially much lower in the presence of HF. Values for a female population from which to draw for this important determination are lacking. The HF-ACTION trial randomized 2,331 patients (28% women) with New York Heart Association class II-IV HF due to systolic dysfunction to either a formal exercise program in addition to optimal medical therapy or to optimal medical therapy alone without any formal exercise training. To characterize differences between men and women in the interpretation of final cardiopulmonary exercise testing models, the interaction of individual covariates with sex was investigated in the models of (1) VE/VCO(2), (2) VO(2) at ventilatory threshold (VT), (3) distance on the 6-minute walk, and (4) peak VO(2). The women were younger than the men and more likely to have a nonischemic etiology and a higher ejection fraction. Dose of angiotensin converting enzyme inhibitor (ACEI) was lower in the women, on average. The lower ACEI dose may reflect the higher use of angiotensin II receptor blocker (ARB) in women. Both the peak VO(2) and the 6-minute walk distance were significantly lower in the women than in the men. Perhaps the most significant finding in this dataset of baseline characteristics is that the peak VO(2) for women was significantly lower than that for men with similar ventricular function and health status. Therefore, in a well-medicated, stable, class II-IV HF cohort of patients who are able to exercise, women have statistically significantly lower peak VO(2) and 6-minute walk distance than men with similar health status and ventricular function. These data should prompt careful thought when considering prognostic markers for women and listing for cardiac transplant.


The Shepherds' Tale: A Genome-Wide Study across 9 Dog Breeds Implicates Two Loci in the Regulation of Fructosamine Serum Concentration in Belgian Shepherds.
Diabetes mellitus is a serious health problem in both dogs and humans. Certain dog breeds show high prevalence of the disease, whereas other breeds are at low risk. Fructosamine and glycated haemoglobin (HbA1c) are two major biomarkers of glycaemia, where serum concentrations reflect glucose turnover over the past few weeks to months. In this study, we searched for genetic factors influencing variation in serum fructosamine concentration in healthy dogs using data from nine dog breeds. Considering all breeds together, we did not find any genome-wide significant associations to fructosamine serum concentration. However, by performing breed-specific analyses we revealed an association on chromosome 3 (pcorrected ≈ 1:68 × 10-6) in Belgian shepherd dogs of the Malinois subtype. The associated region and its close neighbourhood harbours interesting candidate genes such as LETM1 and GAPDH that are important in glucose metabolism and have previously been implicated in the aetiology of diabetes mellitus. To further explore the genetics of this breed specificity, we screened the genome for reduced heterozygosity stretches private to the Belgian shepherd breed. This revealed a region with reduced heterozygosity that shows a statistically significant interaction (p = 0.025) with the association region on chromosome 3. This region also harbours some interesting candidate genes and regulatory regions but the exact mechanisms underlying the interaction are still unknown. Nevertheless, this finding provides a plausible explanation for breed-specific genetic effects for complex traits in dogs. Shepherd breeds are at low risk of developing diabetes mellitus. The findings in Belgian shepherds could be connected to a protective mechanism against the disease. Further insight into the regulation of glucose metabolism could improve diagnostic and therapeutic methods for diabetes mellitus.


High affinity esterification of eicosanoid precursor fatty acids by platelets.
We have examined the relative rates of uptake of several fatty acids into washed, human platelets by measuring incorporation into cellular phospholipids. In the presence of 15 microM fatty acid-free albumin and with radioactive fatty acid concentrations of 5-500 nM, esterification into phospholipid was linear with time and platelet concentration and saturable with respect to fatty acid concentration. Two distinct classes of uptake rate were observed. Arachidonate and 5,8,11,14,17-eicosapentaenoate exhibited high affinity, relatively rapid incorporation into platelet phospholipids at pH 6.5: apparent Michaelis constant (Km) = 30 nM, apparent maximum velocity (Vmax) = 28 pmol/min per 10(9) platelets. Two other eicosanoid precursors, 5,8,11-eicosatrienoate and 8,11,14-eicosatrienoate, exhibited the same Vmax, but Km of 85 and 60 nM, respectively. Under the same conditions, stearate, oleate, and linoleate were incorporated into phospholipids much less efficiently (Vmax approximately 8 pmol/10(9) cells per min, apparent Km greater than or equal to 170 nM). Qualitatively similar results were found at pH 7.4. Uptake of radiolabeled, rapid-uptake fatty acids was not diminished by the presence of excess, unlabeled, slow-uptake fatty acids. Thus, the specificity of this esterification system resembles that of the arachidonate-specific, long-chain acyl-CoA synthetase present in platelets. It may represent the expression in vivo of the synthetase, although the apparent affinity of the synthetase for fatty acid is much less. This esterification system probably represents the physiologic mechanism for platelet arachidonate uptake, whereby arachidonate is collected from plasma, despite the fact that its concentration is considerably lower than that of other plasma fatty acids.


A forensic perspective of the AFL investigation into peptides: an antidoping investigation case study.
The World Anti-Doping Agency (WADA) is introducing enhancements to doping investigations in its 2015 Code, which include improved sharing of information between antidoping organisations (including sporting bodies) and enhanced accountability of athlete support staff. These additions will improve the control of links between sports doping and organised crime. In February 2013 the Australian Crime Commission released a report that linked several professional sporting codes, professional athletes with links to organised crime, performance enhancing drugs and illicit substances. Following this report the Australian Football League (AFL) partnered the Australian national antidoping organisation to investigate peptide use in Australian football. This review compared the model proposed by Marclay, a hypothetical model for anti-doping investigations that proposed a forensic intelligence and analysis approach, to use the forensic capabilities of the AFL investigation to test the model's relevance to an actual case. The investigation uncovered the use of peptides used to enhance athlete performance. The AFL investigation found a high risk of doping where athlete support staff existed in teams with weak corporate governance controls. A further finding included the need for the investigation to provide a timely response in professional team sports that were sensitive to the competition timing. In the case of the AFL the team was sanctioned prior to the finals as an interim outcome for allowing the risk of use of performance-enhancing substances. Doping violation charges are still being considered. Antidoping strategies should include the investigation of corporate officers in team doping circumstances, the mandatory recording of all athlete substance use during competition and training phases, the wider sharing of forensic intelligence with non-sporting bodies particularly law enforcement and collaboration between antidoping and sporting organisations in doping investigations. The AFL investigation illustrated the importance of the 2015 WADA Code changes and highlighted the need for a systematic use of broad forensic intelligence activities in the investigation of doping violations.


Public exposure and attitudes about homelessness.
In this study, we conducted a survey among a large sample of U.S. adults to assess attitudes and beliefs about the causes of homelessness, policies to address homelessness, and programs for homeless individuals. In 2016, we surveyed a national sample of 541 adults from 47 different U.S. states using Amazon Mechanical Turk. Of the total sample, 78% reported that homelessness was a problem in their communities and 60% believed homelessness would increase in the next 5 years. The majority expressed compassion for homeless individuals and endorsed structural, intrinsic, and health factors as causes of homelessness. Most participants (73%-88%) believed the federal government should dedicate more funds and policies for homeless individuals. These attitudes were substantially more likely to be reported by participants who were female, lower income, Democrat, and personally exposed to homelessness. Most Americans care about homelessness as a major problem but there are divergent perspectives on solutions to address homelessness based on gender, income level, and political affiliation.


Evaluation of corneal topography and biomechanical parameters after use of systemic isotretinoin in acne vulgaris.
Purpose. We report the effect of isotretinoin on corneal topography, corneal thickness, and biomechanical parameters in patients with acne vulgaris. Method. Fifty-four eyes of 54 patients who received oral isotretinoin for treatment of acne vulgaris were evaluated. All patients underwent a corneal topographical evaluation with a Scheimpflug camera combined with Placido-disk (Sirius), ultrasonic pachymetry measurements, and corneal biomechanical evaluation with an ocular response analyzer at baseline, in the 1st, 3rd, and 6th months of treatment, and 6 months after isotretinoin discontinuation. Results. The thinnest corneal thickness measured with Sirius differed significantly in the 1st, 3rd, and 6th months compared with the baseline measurement; there was no significant change in ultrasonic central corneal thickness measurements and biomechanical parameters (corneal hysteresis and corneal resistance factor) throughout the study. Average simulated keratometry and surface asymmetry index increased significantly only in the first month of treatment according to the baseline. All changes disappeared 6 months after the end of treatment. Conclusion. Basal tear secretion and corneal morphologic properties were significantly influenced during the systemic isotretinoin treatment and the changes were reversible after discontinuation. No statistical important biomechanical differences were found to be induced by isotretinoin.


Interleukin 4 induces cultured monocytes/macrophages to form giant multinucleated cells.
Giant multinucleated cells (GMCs) are associated with granulomatous lesions that form in response to various infectious and noninfectious agents. The present study shows that mouse IL-4 induces the in vitro formation of GMCs by factor-dependent bone marrow and alveolar monocytes via cell fusion. GMCs appear 2 d after incubation of cell cultures with 20 U/ml or more of IL-4. Anti-IL-4 mAbs block the appearance of GMCs in these cultures, indicating that IL-4 acts directly on monocytes to promote fusion and does not secondarily induce the production of other soluble fusion factors. In soft agar cultures, IL-4 also causes the aggregation of macrophages and diminishes their migration. The role of IL-4 in a granulomatous inflammatory response is discussed.


Mining Arabidopsis thaliana RNA-seq data with Integrated Genome Browser reveals stress-induced alternative splicing of the putative splicing regulator SR45a.
High-throughput sequencing of cDNA libraries prepared from diverse samples (RNA-seq) can reveal genome-wide changes in alternative splicing. Using RNA-seq data to assess splicing at the level of individual genes requires the ability to visualize read alignments alongside genomic annotations. To meet this need, we added RNA-seq visualization capability to Integrated Genome Browser (IGB), a free desktop genome visualization tool. To illustrate this capability, we present an in-depth analysis of abiotic stresses and their effects on alternative splicing of SR45a (AT1G07350), a putative splicing regulator from Arabidopsis thaliana. cDNA libraries prepared from Arabidopsis plants that were subjected to heat and dehydration stresses were sequenced on an Illumina GAIIx sequencer, yielding more than 511 million high-quality 75-base, single-end sequence reads. Reads were aligned onto the reference genome and visualized in IGB. Using IGB, we confirmed exon-skipping alternative splicing in SR45a. Exon-skipped variant AT1G07350.1 encodes full-length SR45a protein with intact RS and RNA recognition motifs, while nonskipped variant AT1G07350.2 lacks the C-terminal RS region due to a frameshift in the alternative exon. Heat and drought stresses increased both transcript abundance and the proportion of exon-skipped transcripts encoding the full-length protein. We identified new splice sites and observed frequent intron retention flanking the alternative exon. This study underlines the importance of visual inspection of RNA-seq alignments when investigating alternatively spliced genes. We showed that heat and dehydration stresses increase overall abundance of SR45a mRNA while also increasing production of transcripts encoding the full-length SR45a protein relative to other splice variants.


Phylogenetic analysis of the vertebrate excitatory/neutral amino acid transporter (SLC1/EAAT) family reveals lineage specific subfamilies.
The composition and expression of vertebrate gene families is shaped by species specific gene loss in combination with a number of gene and genome duplication events (R1, R2 in all vertebrates, R3 in teleosts) and depends on the ecological and evolutionary context. In this study we analyzed the evolutionary history of the solute carrier 1 (SLC1) gene family. These genes are supposed to be under strong selective pressure (purifying selection) due to their important role in the timely removal of glutamate at the synapse. In a genomic survey where we manually annotated and analyzing sequences from more than 300 SLC1 genes (from more than 40 vertebrate species), we found evidence for an interesting evolutionary history of this gene family. While human and mouse genomes contain 7 SLC1 genes, in prototheria, sauropsida, and amphibia genomes up to 9 and in actinopterygii up to 13 SLC1 genes are present. While some of the additional slc1 genes in ray-finned fishes originated from R3, the increased number of SLC1 genes in prototheria, sauropsida, and amphibia genomes originates from specific genes retained in these lineages.Phylogenetic comparison and microsynteny analyses of the SLC1 genes indicate, that theria genomes evidently lost several SLC1 genes still present in the other lineage. The genes lost in theria group into two new subfamilies of the slc1 gene family which we named slc1a8/eaat6 and slc1a9/eaat7. The phylogeny of the SLC1/EAAT gene family demonstrates how multiple genome reorganization and duplication events can influence the number of active genes. Inactivation and preservation of specific SLC1 genes led to the complete loss of two subfamilies in extant theria, while other vertebrates have retained at least one member of two newly identified SLC1 subfamilies.


Biotin tagging deletion analysis of domain limits involved in protein-macromolecular interactions. Mapping the tau binding domain of the DNA polymerase III alpha subunit.
The tau subunit dimerizes DNA polymerase III via interaction with the alpha subunit, allowing DNA polymerase III holoenzyme to synthesize both leading and lagging strands simultaneously at the DNA replication fork. Here, we report a general method to map the limits of domains required for heterologous protein-protein interactions using surface plasmon resonance. The method employs fusion of a short biotinylation sequence at either the NH2 or COOH terminus of the protein to be immobilized on streptavidin-derivatized biosensor chips. Inclusion of a hexahistidine sequence permits rapid purification and separation of the fusion protein from the endogenous Escherichia coli biotin carboxyl carrier protein. Ten deletions of the alpha subunit were constructed and purified by Ni2+-nitrilotriacetic acid chromatography and, when required, monomeric avidin chromatography. Each alpha deletion protein was captured by streptavidin immobilized on a Pharmacia Biosensor BIAcore chip, and the tau binding activity of each alpha deletion was analyzed using surface plasmon resonance. The tau subunit bound very tightly to a full-length amino-terminal fusion of the biotinylation sequence with alpha (KD approximately 70 pm). Four additional NH2-terminal alpha deletion proteins (60, 240, 360, and 542 residues deleted) retained strong binding activity to the tau subunit (KD = 0.19-0.39 nM), whereas deletion of 705 residues or more from the NH2 terminus of the alpha subunit abolished tau binding activity. Full-length alpha that contained a carboxyl-terminal fusion with the biotinylation sequence bound tau strongly (KD = 0.37 nM). However, deletion of 48 amino acids from the COOH terminus totally eliminated tau binding. These results indicate that the COOH-terminal half of the alpha subunit is involved in tau interaction.


Investigations into resting-state connectivity using independent component analysis.
Inferring resting-state connectivity patterns from functional magnetic resonance imaging (fMRI) data is a challenging task for any analytical technique. In this paper, we review a probabilistic independent component analysis (PICA) approach, optimized for the analysis of fMRI data, and discuss the role which this exploratory technique can take in scientific investigations into the structure of these effects. We apply PICA to fMRI data acquired at rest, in order to characterize the spatio-temporal structure of such data, and demonstrate that this is an effective and robust tool for the identification of low-frequency resting-state patterns from data acquired at various different spatial and temporal resolutions. We show that these networks exhibit high spatial consistency across subjects and closely resemble discrete cortical functional networks such as visual cortical areas or sensory-motor cortex.


Who Benefits from Aggressive Rapid Response System Treatments Near the End of Life? A Retrospective Cohort Study.
Many patients near the end of life are subject to rapid response system (RRS) calls. A study was conducted in a large Sydney teaching hospital to identify a cutoff point that defines nonbeneficial treatment for older hospital patients receiving an RRS call, describe interventions administered, and measure the cost of hospitalization. This was a retrospective cohort of 733 adult inpatients with data for the period three months before and after their last placed RRS call. Subgroup analysis of patients aged ≥ 80 years was conducted. Log-rank, chi-square, and t-tests were used to compare survival, and logistic regression was used to examine predictors of death. Overall, 65 (8.9%) patients had a preexisting not-for-resuscitation (NFR) or not-for-RRS order; none of those patients survived to three months. By contrast, patients without an NFR or not-for-RRS order had three-month survival probability of 71% (log-rank χ2 145.63; p < 0.001). Compared with survivors, RRS recipients who died were more likely to be older, to be admitted to a medical ward, and to have a larger mean number of admissions before the RRS. The average cost of hospitalization for the very old transferred to the ICU was higher than for those not requiring treatment in the ICU (US$33,990 vs. US$14,774; p = 0.045). Identifiable risk factors clearly associated with poor clinical outcomes and death can be used as a guide to administer less aggressive treatments, including reconsideration of ICU transfers, adherence to NFR orders, and transition to end-of-life management instead of calls to the RRS team.


VIP and noradrenaline act synergistically to increase cyclic AMP in cerebral cortex.
There is growing evidence that two, or possibly more, neurotransmitters can coexist within the same neurone. In particular, the presence of a peptide and a biogenic amine has been demonstrated in the same terminals of central and peripheral neurones. These findings have led to the hypothesis that neurotransmitters, coexisting within the same neurones, can interact at pre- or postsynaptic sites in a functionally coordinated manner. However, interactions between neurotransmitters contained in distinct neuronal systems terminating within the same region of the central nervous system (CNS) can be envisaged. We have examined this last possibility in the cerebral cortex, an area of the CNS where the two neurotransmitters vasoactive intestinal polypeptide and noradrenaline are contained in separate neuronal systems and where they both stimulate the formation of cyclic AMP. We report here that vasoactive intestinal polypeptide and noradrenaline act synergistically to stimulate the formation of cyclic AMP and that this synergistic interaction is antagonized by the specific alpha-adrenergic antagonist phentolamine.


The Role of Attention Shifting in Orthographic Competencies: Cross-Sectional Findings from 1st, 3rd, and 8th Grade Students.
Attention shifting refers to one core component of executive functions, a set of higher-order cognitive processes that predict different aspects of academic achievement. To date, few studies have investigated the role of attention shifting in orthographic competencies during middle childhood and early adolescence. In the present study, 69 first-grade, 121 third-grade, and 85 eighth-grade students' attention shifting was tested with a computer version of the Dimensional Change Card Sort (DCCS; Zelazo, 2006). General spelling skills and specific writing and spelling strategies were assessed with the Hamburger Writing Test (May, 2002). Results suggested associations between attention shifting and various orthographic competencies that differ across age groups and by sex. Across all age groups, better attention shifting was associated with less errors in applying alphabetical strategies. In third graders, better attention shifting was furthermore related to better general spelling skills and less errors in using orthographical strategies. In this age group, associations did not differ by sex. Among first graders, attention shifting was negatively related to general spelling skills, but only for boys. In contrast, attention shifting was positively related to general spelling skills in eighth graders, but only for girls. Finally, better attention shifting was associated with less case-related errors in eighth graders, independent of students' sex. In sum, the data provide insight into both variability and consistency in the pattern of relations between attention shifting and various orthographic competencies among elementary and middle school students.


Preexisting melanoma and hematological malignancies, prognosis, and timing to solid organ transplantation: A consensus expert opinion statement.
Patients undergoing evaluation for solid organ transplantation (SOT) frequently have a history of malignancy. Only patients with treated cancer are considered for SOT but the benefits of transplantation need to be balanced against the risk of tumor recurrence, taking into consideration the potential effects of immunosuppression. Prior guidelines on timing to transplant in patients with a prior treated malignancy do not account for current staging, disease biology, or advances in cancer treatments. To update these recommendations, the American Society of Transplantation (AST) facilitated a consensus workshop to comprehensively review contemporary literature regarding cancer therapies, cancer stage specific prognosis, the kinetics of cancer recurrence, as well as the limited data on the effects of immunosuppression on cancer-specific outcomes. This document contains prognosis, treatment, and transplant recommendations for melanoma and hematological malignancies. Given the limited data regarding the risk of cancer recurrence in transplant recipients, the goal of the AST-sponsored conference and the consensus documents produced are to provide expert opinion recommendations that help in the evaluation of patients with a history of a pretransplant malignancy for transplant candidacy.


Consensus statement: Eriksholm workshop on wideband absorbance measures of the middle ear.
The participants in the Eriksholm Workshop on Wideband Absorbance Measures of the Middle Ear developed statements for this consensus article on the final morning of the Workshop. The presentations of the first 2 days of the Workshop motivated the discussion on that day. The article is divided into three general areas: terminology; research needs; and clinical application. The varied terminology in the area was seen as potentially confusing, and there was consensus on adopting an organizational structure that grouped the family of measures into the term wideband acoustic immittance (WAI), and dropped the term transmittance in favor of absorbance. There is clearly still a need to conduct research on WAI measurements. Several areas of research were emphasized, including the establishment of a greater WAI normative database, especially developmental norms, and more data on a variety of disorders; increased research on the temporal aspects of WAI; and methods to ensure the validity of test data. The area of clinical application will require training of clinicians in WAI technology. The clinical implementation of WAI would be facilitated by developing feature detectors for various pathologies that, for example, might combine data across ear-canal pressures or probe frequencies.


The effect of exercise-induced arousal on chosen tempi for familiar melodies.
Many previous studies have shown that arousal affects time perception, suggesting a direct influence of arousal on the speed of the pacemaker of the internal clock. However, it is unknown whether arousal influences the mental representation of tempo (speed) for highly familiar and complex stimuli, such as well-known melodies, that have long-term representations in memory. Previous research suggests that mental representations of the tempo of familiar melodies are stable over time; the aim of the present study was to investigate whether these representations can be systematically altered via an increase in physiological arousal. Participants adjusted the tempo of 14 familiar melodies in real time until they found a tempo that matched their internal representation of the appropriate tempo for that piece. The task was carried out before and after a physiologically arousing (exercise) or nonarousing (anagrams) manipulation. Participants completed this task both while hearing the melodies aloud and while imagining them. Chosen tempi increased significantly following exercise-induced arousal, regardless of whether a melody was heard aloud or imagined. These findings suggest that a change in internal clock speed affects temporal judgments even for highly familiar and complex stimuli such as music.


Prediction of Pathological Subjects Using Genetic Algorithms.
This paper aims at estimating pathological subjects from a population through various physical information using genetic algorithm (GA). For comparison purposes, K-Means (KM) clustering algorithm has also been used for the estimation. Dataset consisting of some physical factors (age, weight, and height) and tibial rotation values was provided from the literature. Tibial rotation types are four groups as RTER, RTIR, LTER, and LTIR. Each tibial rotation group is divided into three types. Narrow (Type 1) and wide (Type 3) angular values were called pathological and normal (Type 2) angular values were called nonpathological. Physical information was used to examine if the tibial rotations of the subjects were pathological. Since the GA starts randomly and walks all solution space, the GA is seen to produce far better results than the KM for clustering and optimizing the tibial rotation data assessments with large number of subjects even though the KM algorithm has similar effect with the GA in clustering with a small number of subjects. These findings are discovered to be very useful for all health workers such as physiotherapists and orthopedists, in which this consequence is expected to help clinicians in organizing proper treatment programs for patients.


Cell sources for nucleus pulposus regeneration.
There is increasing interest in the development of cell therapy as a possible approach for the treatment of degenerative disc disease. To regenerate nucleus pulposus tissue, the cells must produce an appropriate proteoglycan-rich matrix, as this is essential for the functioning of the intervertebral disc. The natural environment within the disc is very challenging to implanted cells, particularly if they have been subcultured in normal laboratory conditions. The purpose of this work is to discuss parameters relevant to translating different proposed cell therapies of IVD into clinical use. Several sources of cells have been proposed, including nucleus pulposus cells, chondrocytes and mesenchymal stem cells derived from bone marrow or adipose tissue. There are some clinical trials and reports of attempts to regenerate nucleus pulposus utilising either autologous or allogenic cells. While the published results of clinical applications of these cell therapies do not indicate any safety issues, additional evidence will be needed to prove their long-term efficacy. This article discusses parameters relevant for successful translation of research on different cell sources into clinically applicable cell therapies: the influence of the intervertebral disc microenvironment on the cell phenotype, issues associated with cell culture and technical preparation of cell products, as well as discussing current regulatory requirements. There are advantages and disadvantages of each proposed cell type, but no strong evidence to favour any one particular cell source at the moment.


Immune regulatory and neuroprotective properties of preimplantation factor: From newborn to adult.
Embryonic-maternal interaction from the earliest stages of gestation has a key, sustained role in neurologic development, persisting into adulthood. Early adverse events may be detrimental in adulthood. Protective factors present during gestation could significantly impact post-natal therapy. The role of PreImplantation Factor (PIF) within this context is herein examined. Secreted by viable early embryos, PIF establishes effective embryonic-maternal communication and exerts essential trophic and protective roles by reducing oxidative stress and protein misfolding and by blunting the nocive let-7 microRNA related pathway. PIF's effects on systemic immunity lead to comprehensive immune modulation, not immune suppression. We examine PIF's role in protecting embryos from adverse maternal environment, which can lead to neurological disorders that may only manifest post-nataly: Synthetic PIF successfully translates endogenous PIF features in both pregnant and non-pregnant clinically relevant models. Specifically PIF has neuroprotective effects in neonatal prematurity. In adult relapsing-remitting neuroinflammation, PIF reverses advanced paralysis while promoting neurogenesis. PIF reversed Mycobacterium smegmatis induced brain infection. In graft-vs.-host disease, PIF reduced skin ulceration, liver inflammation and colon ulceration while maintaining beneficial anti-cancer, graft-vs.-leukemia effect. Clinical-grade PIF has high-safety profile even at supraphysiological doses. The FDA awarded Fast-Track designation, and university-sponsored clinical trials for autoimmune disorder are ongoing. Altogether, PIF properties point to its determining regulatory role in immunity, inflammation and transplant acceptance. Specific plans for using PIF for the treatment of complex neurological disorders (ie. traumatic brain injury, progressive paralysis), including neuroprotection from newborn to adult, are presented.


Safety and tolerability of aripiprazole for irritability in pediatric patients with autistic disorder: a 52-week, open-label, multicenter study.
Evaluate the long-term safety and tolerability of aripiprazole in the treatment of irritability in pediatric subjects (6-17 years) with autistic disorder. A 52-week, open-label, flexibly dosed (2-15 mg/d) study of the safety and tolerability of aripiprazole in outpatients with a DSM-IV-TR diagnosis of autistic disorder who either had completed 1 of 2 antecedent, 8-week randomized trials or were enrolled de novo (ie, not treated in the randomized trials). Safety and tolerability measures included incidences of adverse events, extrapyramidal symptoms, weight, metabolic measures, vital signs, and other clinical assessments. Subjects were enrolled between September 2006 and June 2009. Three hundred thirty subjects entered the treatment phase: 86 de novo, 174 prior aripiprazole, and 70 prior placebo. A total of 199 (60.3%) subjects completed 52 weeks of treatment. Adverse events were experienced by 286/330 subjects (86.7%). Common adverse events included weight increase, vomiting, nasopharyngitis, increased appetite, pyrexia, upper respiratory tract infection, and insomnia. Discontinuations due to adverse events occurred in 35/330 randomized subjects (10.6%)-most commonly aggression and weight increase. One patient discontinued from the study due to a laboratory-related adverse event (moderately increased alanine transaminase and aspartate transaminase). Nine subjects experienced serious adverse events-most frequently aggression. Extrapyramidal symptoms-related adverse events occurred in 48/330 subjects (14.5%)-most commonly tremor (3.0%), psychomotor hyperactivity (2.7%), akathisia (2.4%), and dyskinesia (not tardive, 2.4%). At > 9 months' aripiprazole exposure (n = 220), mean change in body weight z score was 0.33 and body mass index z score was 0.31. The percentages of subjects with clinically significant fasting metabolic abnormalities at > 9 months were 2% for glucose, 5% for total cholesterol, 7% for low-density lipoprotein cholesterol, 30% for high-density lipoprotein cholesterol, and 5% for triglycerides. Aripiprazole was generally safe and well tolerated in the long-term treatment of irritability associated with autistic disorder in pediatric subjects. Weight should be proactively monitored during long-term treatment. clinical trials.gov Identifier: NCT00365859.


Preclinical rationale for combining radiation therapy and immunotherapy beyond checkpoint inhibitors (i.e., CART).
An increasing appreciation for the role of the immune system in targeting cancer cells over the last decade has led to the development of several immunomodulatory agents aimed at enhancing the systemic antitumor immune response. One such method is the use of T cells that are genetically engineered to express chimeric antigen receptors (CARs). The remarkable success of this approach in advanced hematologic malignancies has garnered much enthusiasm for using this novel tool in treating other cancers. However, multiple challenges have hampered the application of this therapy to a broader set of solid tumors, most notably lung cancer. Immunotherapy has already shown great success in lung cancer, and is now the first-line treatment in PD-L1 expressing metastatic disease. Given the mounting evidence that radiation therapy plays a crucial role in amplifying the immune response elicited by immunomodulatory agents, there is potential for radiation to help in overcoming some of these challenges. In this review, we describe the basic principles of CAR T cell therapy and examine its successes and challenges to date. We then discuss the preclinical and clinical data supporting the use of radiation with immunomodulatory agents with a focus on preclinical rationale for combining CAR T cells and radiation therapy in future experiments with a focus on lung cancer.


Neuromuscular and microvascular changes associated with chronic administration of an extract of Teucrium stocksianum in mice.
This work examines the effect on the weights of vital body organs, on blood biochemical variables, on neuromuscular coordination and on cerebral microcirculation of aqueous extracts of Teucrium stocksianum, given to mice in drinking water at concentrations of 2 and 4% for 56 days. The treatment caused progressive impairment of neuromuscular coordination, as evidenced by the time spent on the rota-rod. After photochemical challenge, the time for first observable platelet aggregation in arterioles was shorter than for the control group by 22 and 45% in the 2 and 4% T. stocksianum-treated groups, respectively. Platelet aggregation on the venular side was not affected by the treatment nor were microvascular diameters. Treatment with the plant extract produced no statistically significant effect on the plasma biochemical variables that are considered indices of liver and kidney function. Histologically, brains obtained from mice treated with T. stocksianum showed loss of cerebellar Purkinje cells. Although it is likely that the accelerated platelet aggregation might have contributed to an ischaemic effect which could, at least in part, have caused the cytotoxicological changes, this does not exclude the possibility of a direct cytotoxicological effect of the plant extract. Further pharmacological and toxicological investigations on Teucrium species seem warranted.


PAFit: A Statistical Method for Measuring Preferential Attachment in Temporal Complex Networks.
Preferential attachment is a stochastic process that has been proposed to explain certain topological features characteristic of complex networks from diverse domains. The systematic investigation of preferential attachment is an important area of research in network science, not only for the theoretical matter of verifying whether this hypothesized process is operative in real-world networks, but also for the practical insights that follow from knowledge of its functional form. Here we describe a maximum likelihood based estimation method for the measurement of preferential attachment in temporal complex networks. We call the method PAFit, and implement it in an R package of the same name. PAFit constitutes an advance over previous methods primarily because we based it on a nonparametric statistical framework that enables attachment kernel estimation free of any assumptions about its functional form. We show this results in PAFit outperforming the popular methods of Jeong and Newman in Monte Carlo simulations. What is more, we found that the application of PAFit to a publically available Flickr social network dataset yielded clear evidence for a deviation of the attachment kernel from the popularly assumed log-linear form. Independent of our main work, we provide a correction to a consequential error in Newman's original method which had evidently gone unnoticed since its publication over a decade ago.


[Energy-dependent Ca2+-transport in intracellular smooth muscle structures].
In experiments performed on digitonin-treated myometrium cells the existence of intracellular energy-dependent Ca(2+)-transporting system has been identified. The mitochondria-linked energy-dependent accumulation of Ca2+ was inhibited by ruthenium red and sodium azide with Ki of 0.7 +/- 0.3 microM and 1.1 +/- 0.4 mM, respectively. The Ca2+ ionophore A23187 added to the incubation medium following the termination of the accumulation process by ruthenium red caused a discharge of Ca2+ accumulated from the mitochondria. In the endoplasmic reticulum there exists a Mg2+,ATP-dependent Ca(2+)-pump which is potentiated by oxalate, immune to ruthenium red and sodium azide and inhibited by thapsigargin with Ki < 20 nM. In the absence of oxalate, the component of mitochondrial accumulation of Ca2+ was 3-4 times higher that of the endoplasmic reticulum. These results indicate that under the conditions used the calcium capacity of mitochondria was significantly higher than that of the endoplasmic reticulum. It is suggested that mitochondria serve as a high capacity intracellular Ca(2+)-accumulating store protecting smooth muscle cells from Ca2+ overload under some abnormal states. Under normal physiological conditions the endoplasmic reticulum serves as the major intracellular store involved in the pharmacomechanical coupling in the smooth muscle of the myometrium.


Descriptions of five new species of Metriaclima (Teleostei: Cichlidae) from Lake Malaŵi, Africa.
Lake Malaŵi is known for its endemic haplochromine species flock, most notably the rock-dwelling cichlids known as mbuna. One of the larger genera of mbuna is Metriaclima, a group consisting of 31 described species (including the five described herein) and approximately 45 recognized unique populations. Metriaclima is diagnosed by its feeding behavior and several morphological characteristics including the angle of the vomer and the presence of bicuspid teeth in the outer row of both the upper and lower jaws. Metriaclima zebra, the type species for the genus, was described based on a single specimen. While the collection location of this holotype is not known, based on the travel records of its collector, it is likely that the specimen originated from Likoma Island. The holotype was therefore compared to specimens from several localities around this island and was found to be morphologically indistinguishable from some of these. This study includes the morphological analysis of 496 specimens of Metriaclima belonging to 31 collections from Lake Malaŵi. Morphometric differences were analyzed and the relationships among several distinguishable populations of Metriaclima zebra were investigated. Our study further resulted in the description of the following five new species belonging to the M. zebra species complex: M. pambazuko, M. lundoense, M. midomo, M. tarakiki, and M. nigrodorsalis. These species were distinguished and described based on color patterns, morphometric, meristic, and ecological differences. These new species were compared with and distinguished from nearby populations of Metriaclima having similar pigmentation patterns and/or similar ecological niches. An artificial dichotomous key to the described species of Metriaclima is presented.


Molecular signal suppression by in situ microextraction in nuclear magnetic resonance spectroscopy.
The detailed characterization of complex mixtures by NMR is often hampered by the presence of signals from uninformative compounds, the resonances of which overlap with those of the molecules of interest. We provide here a proof of principle for an approach to NMR signal suppression in complex samples using Molecularly Imprinted Polymers (MIPS). Addition of a few milligrams of polymer to a solution traps the target molecule in typical micromolar to millimolar concentration, thus achieving in situ signal suppression, without altering any other spectral features. This method minimized any manipulation or perturbation of the spectrum and was applied to a complex mixture of known compounds and to a plant extract, in both cases spiked with a compound (bisphenol A), which was subsequently removed by selective binding to a complementary MIP. What is described in this report is comparable with microextraction and may in due course be applied to a large number of analytical challenges.


Rethinking knowledge and pedagogy in dental education.
Dentistry as a profession has often been considered both art and science. Traditional dental education has attempted to address both; however, in many places only the science of dentistry is emphasized. The move toward competency-based curricula in dental education requires an expansion of what constitutes meaningful knowledge in the curriculum and what pedagogies best support that curriculum. The scientific and technical knowledge considered foundational to clinical practice is not sufficient to teach competencies associated with the art of dentistry. Habermas, a social scientist, offers a way of looking beyond technical knowledge to consider two other forms of knowledge: practical and emancipatory. Pedagogy that supports development of practical and emancipatory knowledge includes problem-based learning and case methods, heuristics, reflective practica, journals, storytelling, and performance-based assessment methods. These important teaching strategies are being integrated into various dental curricula including a new competency-based dental curriculum at Marquette University's School of Dentistry. It will be critical for dental educators to continue developing these methods to provide efficient and effective education for future practitioners in both the art and science of dentistry.


Prevalence of common phobias and their socio-demographic correlates in children and adolescents in a traditional developing society.
The aim of this study was to identify the most common phobias in children and adolescents and to determine the prevalence, age distribution, and socio-demographic correlates of phobias. This was a prospective cross-sectional study conducted at public and private schools from July 2009 to February 2009. The questionnaire included socio-demographic information, extra-curricular activities and hobbies, behaviour at home and various phobic fears and it was distributed among children aged 6 to 18 years. Psychiatrists determined the definitive diagnosis for various phobias by checking and screening their symptoms. Of the studied subjects, 44% were males and 56% were females. The overall prevalence of phobia in children and adolescents was 19.7%. Among children with phobia, females had higher rates of phobias (62.4% vs 37.6%) than males. Nearly half of total sufferers were in the 12-15 year age group (46.3%). Social phobia (12.7%) was the commonest phobia found followed by agoraphobia (8.6%). Secondary school children were highly afflicted with social phobia (14.9%), agoraphobia (11.7%) and specific phobia (9.6%), while preparatory students (8.3%) were more likely to have 'medical' phobia (fear of physical illness, medical tests and procedures). A significant difference was observed between the age groups in children with agoraphobia (p=0.002). The overall prevalence of phobia in children and adolescents in Qatar was higher than rates found in other epidemiologic studies, with the most common phobias observed being social phobia, agoraphobia and specific phobia.


Electronic health record-based triggers to detect potential delays in cancer diagnosis.
Delayed diagnosis of cancer can lead to patient harm, and strategies are needed to proactively and efficiently detect such delays in care. We aimed to develop and evaluate 'trigger' algorithms to electronically flag medical records of patients with potential delays in prostate and colorectal cancer (CRC) diagnosis. We mined retrospective data from two large integrated health systems with comprehensive electronic health records (EHR) to iteratively develop triggers. Data mining algorithms identified all patient records with specific demographics and a lack of appropriate and timely follow-up actions on four diagnostic clues that were newly documented in the EHR: abnormal prostate-specific antigen (PSA), positive faecal occult blood test (FOBT), iron-deficiency anaemia (IDA), and haematochezia. Triggers subsequently excluded patients not needing follow-up (eg, terminal illness) or who had already received appropriate and timely care. Each of the four final triggers was applied to a test cohort, and chart reviews of randomly selected records identified by the triggers were used to calculate positive predictive values (PPV). The PSA trigger was applied to records of 292 587 patients seen between 1 January 2009 and 31 December 2009, and the CRC triggers were applied to 291 773 patients seen between 1 March 2009 and 28 February 2010. Overall, 1564 trigger positive patients were identified (426 PSA, 355 FOBT, 610 IDA and 173 haematochezia). Record reviews revealed PPVs of 70.2%, 66.7%, 67.5%, and 58.3% for the PSA, FOBT, IDA and haematochezia triggers, respectively. Use of all four triggers at the study sites could detect an estimated 1048 instances of delayed or missed follow-up of abnormal findings annually and 47 high-grade cancers. EHR-based triggers can be used successfully to flag patient records lacking follow-up of abnormal clinical findings suspicious for cancer.


Defects in G protein-coupled signal transduction in human disease.
G proteins couple receptors for many hormones and neurotransmitters to effectors that regulate second messenger metabolism. G protein-coupled receptors comprise a superfamily with the common structural feature of a single polypeptide with seven membrane-spanning domains. G proteins themselves are heterotrimers with an alpha subunit that binds guanine nucleotides. In the basal state, G proteins tightly bind GDP; receptor activation allows exchange of bound GDP for GTP that activates the G protein and causes it to modulate effector activity. An intrinsic GTPase activity hydrolyzes bound GTP to GDP thereby deactivating the G protein. The effects (cholera, whooping cough) of bacterial toxins that target G proteins for covalent modification signal the potential importance of G protein dysfunction as a cause of human disease. Conceptually, G protein dysfunction could involve gain or loss of function. For Gs, examples of both types have already been defined. Mutations in G protein-coupled receptors have also been identified in several human diseases. Germline loss of function mutations in rhodopsin, cone opsins, the V2 vasopressin receptor, ACTH receptor, and calcium-sensing receptor are responsible for retinitis pigmentosa, color blindness, nephrogenic diabetes insipidus, familial ACTH resistance, and familial hypocalciuric hypercalcemia, respectively. Missense mutations that cause constitutive receptor activation have been identified in the TSH and LH receptors.


Perspectives of primary health care staff on the implementation of a sexual health quality improvement program: a qualitative study in remote aboriginal communities in Australia.
Young people living in remote Australian Aboriginal communities experience high rates of sexually transmissible infections (STIs). STRIVE (STIs in Remote communities, ImproVed and Enhanced primary care) was a cluster randomised control trial of a sexual health continuous quality improvement (CQI) program. As part of the trial, qualitative research was conducted to explore staff perceptions of the CQI components, their normalisation and integration into routine practice, and the factors which influenced these processes. In-depth semi-structured interviews were conducted with 41 clinical staff at 22 remote community clinics during 2011-2013. Normalisation process theory was used to frame the analysis of interview data and to provide insights into enablers and barriers to the integration and normalisation of the CQI program and its six specific components. Of the CQI components, participants reported that the clinical data reports had the highest degree of integration and normalisation. Action plan setting, the Systems Assessment Tool, and the STRIVE coordinator role, were perceived as adding value to the program, but were less readily integrated or normalised. The remaining two components (dedicated funding for health promotion and service incentive payments) were seen as least relevant. Our analysis also highlighted factors which enabled greater integration of the CQI components. These included familiarity with CQI tools, increased accountability of health centre staff and the translation of the CQI program into guideline-driven care. The analysis also identified barriers, including high staff turnover, limited time involved in the program and competing clinical demands and programs. Across all of the CQI components, the clinical data reports had the highest degree of integration and normalisation. The action plans, systems assessment tool and the STRIVE coordinator role all complemented the data reports and allowed these components to be translated directly into clinical activity. To ensure their uptake, CQI programs must acknowledge local clinical guidelines, be compatible with translation into clinical activity and have managerial support. Sexual health CQI needs to align with other CQI activities, engage staff and promote accountability through the provision of clinic specific data and regular face-to-face meetings. Australian and New Zealand Clinical Trials Registry ACTRN12610000358044 . Registered 6/05/2010. Prospectively Registered.


Impact of combining indoor residual spraying and long-lasting insecticidal nets on Anopheles arabiensis in Ethiopia: results from a cluster randomized controlled trial.
Indoor residual house spraying (IRS) and long-lasting insecticidal nets (LLINs) are the key front-line malaria vector interventions against Anopheles arabiensis, the sole primary malaria vector in Ethiopia. Universal coverage of both interventions has been promoted and there is a growing demand in combinations of interventions for malaria control and elimination. This study compared the impact on entomological outcomes of combining IRS and LLINs with either intervention alone in Adami Tullu district, south-central Ethiopia. The epidemiological outcomes were recently published on a separate paper. This factorial, cluster-randomized, controlled trial randomized villages to four study arms: IRS + LLIN, IRS, LLIN, and control. LLINs (PermaNet 2.0) were provided free of charge. IRS with propoxur was applied before the main malaria transmission season in 2014, 2015 and 2016. Adult mosquitoes were collected in randomly selected villages in each arm using CDC light trap catch (LTC) set close to a sleeping person, pyrethrum spray catch (PSC), and artificial pit shelter (PIT), for measuring mosquito host-seeking density (HSD), indoor resting density (IRD), and outdoor resting density (ORD), respectively. Human landing catch (HLC) was performed in a sub-set of villages to monitor An. arabiensis human biting rates (HBR). Mean vector densities and HBR were compared among study arms using incidence rate ratio (IRR) calculated by negative binomial regression. There were no significant differences in mean densities (HSD, IRD, ORD) and HBR of An. arabiensis between the IRS + LLIN arm and the IRS arm (p > 0.05). However, mean HSD, IRD, ORD, and HBR were significantly lower in the IRS + LLIN arm than in the LLIN alone arm (p < 0.05). All An. arabiensis tested for malaria infection were negative for Plasmodium species. For this reason, the entomological inoculation rate could not be determined. The IRS + LLIN were as effective as IRS alone in reducing densities and HBR of An. arabiensis. However, the effectiveness of the two interventions combined was higher than LLINs alone in reducing densities and HBR of the vector. Added impact of the combination intervention against malaria infectivity rates of An. arabiensis compared to either intervention alone remains unknown and warrants further research. Trial registration PACTR201411000882128. Registered 8 September 2014, https://trialsjournal.biomedcentral.com/articles/10.1186/s13063-016-1154-2.


Relationships between nurse- and physician-to-population ratios and state health rankings.
To evaluate the relationship between nurse-to-population ratios and population health, as indicated by state health ranking, and to compare the findings with physician-to-population ratios. Secondary analysis correlational design. The sample consisted of all 50 states in the United States. Data sources included the United Health Foundation's 2006 state health rankings, the 2004 National Sample Survey for Registered Nurses, and the U.S. Health Workforce Profile from the New York Center for Health Workforce Studies. Significant relationships between nurse-to-population ratio and overall state health ranking (rho=-.446, p tf?>=.001) and 11 of the 18 components of that ranking were found. Significant components included motor vehicle death rate, high school graduation rate, violent crime rate, infectious disease rate, percentage of children in poverty, percentage of uninsured residents, immunization rate, adequacy of prenatal care, number of poor mental health days, number of poor physical health days, and premature death rate, with higher nurse-to-population ratios associated with higher health rankings. Specialty (public health and school) nurse-to-population ratios were not as strongly related to state health ranking. Physician-to-population ratios were also significantly related to state health ranking, but were associated with different components than nurses. These findings suggest that greater nurses per capita may be uniquely associated with healthier communities; however, further multivariate research is needed.


Incidental findings on brain magnetic resonance imaging in the elderly:the PROOF study.
To prospectively evaluate the prevalence of incidental findings on standardized brain MRI scans in a cohort of elderly subjects. All participants provided written informed consent to participate in this prospective study, which was approved by the institutional review board and was HIPAA compliant. There were 503 subjects (75.3 ± 0.9 years of age, 58 % women) who received brain MRIs on a 1.5-T scanner using a standard acquisition protocol. All scans were reviewed by an experienced neuroradiologist. Incidental findings were stratified as follows: 1, no incidental findings; 2, incidental findings without clinical significance; 3, incidental findings with clinical significance or requiring clinical follow-up. Incidental findings were identified in 77.9 % of subjects. Among 392 scans that exhibited incidental findings, 494 abnormalities were identified. The most common findings in our study were cysts (45.9 % of subjects) and ear, nose, and throat (ENT) lesions (24.8 %) followed by stroke lesions (6.6 %) and neoplasms (3.8 %). There were 472 incidental findings that lacked clinical significance (group 2), and 22 incidental findings that required follow-up evaluation (group 3). Incidental findings on brain MRIs were commonly observed in this cohort of elderly subjects, but clinical follow-up was rarely indicated.


Organic solvent-tolerant bacterium which secretes an organic solvent-stable proteolytic enzyme.
A bacterial strain which can be grown in a medium containing organic solvents and can secrete a proteolytic enzyme was isolated and identified as Pseudomonas aeruginosa. The strain was derived by the following two-step procedures: high proteolytic enzyme producers were first isolated by the usual method, and then the organic solvent-tolerant microorganism was selected from these high-rate proteolytic enzyme producers. The proteolytic activity of the supernatant of the culture was stable in the presence of various organic solvents. The stability of the enzyme in the presence of organic solvents, of which the values of the logarithm of the partition coefficient (log P) were equal to or more than 3.2, was almost the same as that in the absence of organic solvents. It is expected that both the solvent-tolerant microorganism and the solvent-stable enzyme produced by this strain can be used as catalysts for reactions in the presence of organic solvents.


Variations in Amazon forest productivity correlated with foliar nutrients and modelled rates of photosynthetic carbon supply.
The rate of above-ground woody biomass production, W(P), in some western Amazon forests exceeds those in the east by a factor of 2 or more. Underlying causes may include climate, soil nutrient limitations and species composition. In this modelling paper, we explore the implications of allowing key nutrients such as N and P to constrain the photosynthesis of Amazon forests, and also we examine the relationship between modelled rates of photosynthesis and the observed gradients in W(P). We use a model with current understanding of the underpinning biochemical processes as affected by nutrient availability to assess: (i) the degree to which observed spatial variations in foliar [N] and [P] across Amazonia affect stand-level photosynthesis; and (ii) how these variations in forest photosynthetic carbon acquisition relate to the observed geographical patterns of stem growth across the Amazon Basin. We find nutrient availability to exert a strong effect on photosynthetic carbon gain across the Basin and to be a likely important contributor to the observed gradient in W(P). Phosphorus emerges as more important than nitrogen in accounting for the observed variations in productivity. Implications of these findings are discussed in the context of future tropical forests under a changing climate.


Do most practices have an age-sex register? Results of the West Midlands age-sex register study.
A cross sectional study based on all (1260) general practices in the West Midlands was conducted in late 1984 and early 1985. Responses to a postal questionnaire were received from 762 (60.5%) practices, and of these, 387 (50.8%) stated that they had an age-sex register; 104 (13.6%) practices replied that they had definite plans to establish an age-sex register within six months, and a random sample of these was later telephoned to see if the plans had come to fruition. A random telephone sample was also made from the 498 (39.5%) non-responders. It was concluded that the current prevalence of age-sex registers in the West Midlands is 52% (+/- 5% for 95% confidence) of all practices. A register has now been established of 422 practices (35.5% of the total number in the West Midlands) that have an age-sex register. This is being used as a database for other research. The most common reason stated for not having an age-sex register was that it would be too time consuming. Most of the practices that had one stated that it was used for research, disease recording, screening, surveillance, or health education, which implies that the registers are being used.


Fecal shedding of, antimicrobial resistance in, and serologic response to Salmonella Typhimurium in dairy calves.
To determine the duration of fecal shedding of and serologic response to Salmonella spp after natural infection in dairy calves and characterize Salmonella organisms recovered from these herds. Longitudinal study. Calves from 2 dairy herds (A and B) in the northeast United States that were identified at the beginning of a Salmonella outbreak. Fecal samples were collected twice per week (herd A) or once per week (herd B); blood samples were collected for serologic testing once per week in both herds. Bacteriologic culture of fecal samples was performed, and Salmonella isolates were characterized by serotype, pulsed-field gel electrophoresis (PFGE) pattern, and antimicrobial resistance profile. All Salmonella isolates from herd A were serovar Typhimurium var Copenhagen, had the same PFGE pattern, and were resistant to at least 9 antimicrobials. All isolates from herd B were Salmonella Typhimurium, represented 2 PFGE patterns, and were susceptible to all antimicrobials evaluated. The estimated duration of fecal shedding was 14 days in herd A and 9 days in herd B. Few calves were seropositive for antibody against Salmonella lipopolysaccharide within the first week after birth (0 of 20 in herd A and 13 of 79 in herd B) or seroconverted (6 in herd A and 4 in herd B). Fecal shedding was more common in calves that seroconverted, but overall, there was not a strong association between seropositivity and fecal shedding of Salmonella organisms. Although the herds differed in serologic response and Salmonella subtype, the duration of fecal shedding among calves was similar between herds.


Van der Waals interactions dominate ligand-protein association in a protein binding site occluded from solvent water.
In the present study we examine the enthalpy of binding of 2-methoxy-3-isobutylpyrazine (IBMP) to the mouse major urinary protein (MUP), using a combination of isothermal titration calorimetry (ITC), NMR, X-ray crystallography, all-atom molecular dynamics simulations, and site-directed mutagenesis. Global thermodynamics data derived from ITC indicate that binding is driven by favorable enthalpic contributions, rather than a classical entropy-driven signature that might be expected given that the binding pocket of MUP-1 is very hydrophobic. The only ligand-protein hydrogen bond is formed between the side-chain hydroxyl of Tyr120 and the ring nitrogen of the ligand in the wild-type protein. ITC measurements on the binding of IBMP to the Y120F mutant demonstrate a reduced enthalpy of binding, but nonetheless binding is still enthalpy dominated. A combination of solvent isotopic substitution ITC measurements and all-atom molecular dynamics simulations with explicit inclusion of solvent water suggests that solvation is not a major contributor to the overall binding enthalpy. Moreover, hydrogen/deuterium exchange measurements suggest that there is no significant contribution to the enthalpy of binding derived from "tightening" of the protein structure. Data are consistent with binding thermodynamics dominated by favorable dispersion interactions, arising from the inequality of solvent-solute dispersion interactions before complexation versus solute-solute dispersion interactions after complexation, by virtue of poor solvation of the binding pocket.


The role of effort in moderating the anxiety-performance relationship: Testing the prediction of processing efficiency theory in simulated rally driving.
We tested some of the key predictions of processing efficiency theory using a simulated rally driving task. Two groups of participants were classified as either dispositionally high or low anxious based on trait anxiety scores and trained on a simulated driving task. Participants then raced individually on two similar courses under counterbalanced experimental conditions designed to manipulate the level of anxiety experienced. The effort exerted on the driving tasks was assessed though self-report (RSME), psychophysiological measures (pupil dilation) and visual gaze data. Efficiency was measured in terms of efficiency of visual processing (search rate) and driving control (variability of wheel and accelerator pedal) indices. Driving performance was measured as the time taken to complete the course. As predicted, increased anxiety had a negative effect on processing efficiency as indexed by the self-report, pupillary response and variability of gaze data. Predicted differences due to dispositional levels of anxiety were also found in the driving control and effort data. Although both groups of drivers performed worse under the threatening condition, the performance of the high trait anxious individuals was affected to a greater extent by the anxiety manipulation than the performance of the low trait anxious drivers. The findings suggest that processing efficiency theory holds promise as a theoretical framework for examining the relationship between anxiety and performance in sport.


Intraindividual Reaction Time Variability, Falls, and Gait in Old Age: A Systematic Review.
Intraindividual variability (IIV) refers to the variation in reaction time (RT) performance across a given cognitive task. As greater IIV may reflect compromise of the frontal circuitry implicated in falls and gait impairment in older adults, we conducted a systematic review of the literature relating to this issue. Searches were conducted of electronic databases that identified empirical investigations of IIV, falls, and gait in older adult samples with a mean age of 65 years or older. Data were extracted relating to IIV measures, study population, and outcomes. Of 433 studies initially identified, 9 met inclusion criteria for IIV and falls (n = 5), and gait (n = 4). Representing a total of 2,810 older participants, all of the studies of IIV and falls showed that elevated variability was associated with increased risk of falling, and half of the studies of gait indicated greater IIV was related to gait impairment. Across studies, IIV measures were consistently associated with falls in older persons and demonstrated some potential in relation to gait. IIV metrics may, therefore, have considerable potential in clinical contexts and supplement existing test batteries in the assessment of falls risk and gait impairment in older populations.


Design, synthesis and pharmacological evaluation of 5-hydroxytryptamine(1a) receptor ligands to explore the three-dimensional structure of the receptor.
In this work, we evaluate the structural differences of transmembrane helix 3 in rhodopsin and the 5-hydroxytryptamine 1A (5-HT1A) receptor caused by their different amino acid sequence. Molecular dynamics simulations of helix 3 in the 5-HT1A receptor tends to bend toward helix 5, in sharp contrast to helix 3 in rhodopsin, which is properly located within the position observed in the crystal structure. The relocation of the central helix 3 in the helical bundle facilitates the experimentally derived interactions between the neurotransmitters and the Asp residue in helix 3 and the Ser/Thr residues in helix 5. The different amino acid sequence that forms helix 3 in rhodopsin (basically the conserved Gly(3.36)Glu(3.37) motif in the opsin family) and the 5-HT1A receptor (the conserved Cys(3.36)Thr(3.37) motif in the neurotransmitter family) produces these structural divergences. These structural differences were experimentally checked by designing and testing ligands that contain comparable functional groups but at different interatomic distance. We have estimated the position of helix 3 relative to the other helices by systematically changing the distance between the functional groups of the ligands (1 and 2) that interact with the residues in the receptor. Thus, ligand 1 optimally interacts with a model of the 5-HT1A receptor that matches rhodopsin template, whereas ligand 2 optimally interacts with a model that possesses the proposed conformation of helix 3. The lack of affinity of 1 (K(i) > 10,000 nM) and the high affinity of 2 (K(i) = 24 nM) for the 5-HT1A receptor binding sites, provide experimental support to the proposed structural divergences of helix 3 between the 5-HT1A receptor and rhodopsin.


Influence of viraemia and genotype upon serological reactivity in screening assays for antibody to hepatitis C virus.
Detection of antibody to recombinant proteins derived from hepatitis C virus (HCV) genotype 1 represents the principal method for diagnosis of HCV infection. A method was developed for quantifying antibody reactivity in two third-generation enzyme immunoassays (Ortho EIA 3.0 and Murex VK48), and the influence of viraemia, HCV genotype, and host factors such as age, gender, and risk group upon antibody levels were investigated in a consecutive series of 117 anti-HCV-positive volunteer blood donors. Viraemic donors (as assessed by the polymerase chain reaction; PCR) showed significantly higher levels of anti-HCV by the Ortho EIA than those who were nonviraemic (adjusted mean difference of 10.1 fold after multiple regression analysis). The only other factor to influence significantly antibody level was genotype, where it was found that donors infected with type 1 showed 4 to 4.5 times greater serological reactivity by the Ortho assay than those infected with type 2 or 3. Antibody levels by the Ortho assay correlated closely to those detected by the Murex VK48 assay, and similar differences between PCR-positive and negative donors and between those infected with different genotypes were found. Differences in serological reactivity between genotypes indicate that a large proportion of epitopes of the type 1a or 1b recombinant proteins used in current assays are genotype specific. Variation in sensitivity of screening assays for different genotypes is of potential concern when used in countries where non-type 1 genotypes predominate in the blood donor or patient population.


Plasma metabolomic profiles of breast cancer patients after short-term limonene intervention.
Limonene is a lipophilic monoterpene found in high levels in citrus peel. Limonene demonstrates anticancer properties in preclinical models with effects on multiple cellular targets at varying potency. While of interest as a cancer chemopreventive, the biologic activity of limonene in humans is poorly understood. We conducted metabolite profiling in 39 paired (pre/postintervention) plasma samples from early-stage breast cancer patients receiving limonene treatment (2 g QD) before surgical resection of their tumor. Metabolite profiling was conducted using ultra-performance liquid chromatography coupled to a linear trap quadrupole system and gas chromatography-mass spectrometry. Metabolites were identified by comparison of ion features in samples to a standard reference library. Pathway-based interpretation was conducted using the human metabolome database and the MetaCyc database. Of the 397 named metabolites identified, 72 changed significantly with limonene intervention. Class-based changes included significant decreases in adrenal steroids (P < 0.01), and significant increases in bile acids (P ≤ 0.05) and multiple collagen breakdown products (P < 0.001). The pattern of changes also suggested alterations in glucose metabolism. There were 47 metabolites whose change with intervention was significantly correlated to a decrease in cyclin D1, a cell-cycle regulatory protein, in patient tumor tissues (P ≤ 0.05). Here, oral administration of limonene resulted in significant changes in several metabolic pathways. Furthermore, pathway-based changes were related to the change in tissue level cyclin D1 expression. Future controlled clinical trials with limonene are necessary to determine the potential role and mechanisms of limonene in the breast cancer prevention setting.


Differential coral bleaching-Contrasting the activity and response of enzymatic antioxidants in symbiotic partners under thermal stress.
Mass coral bleaching due to thermal stress represents a major threat to the integrity and functioning of coral reefs. Thermal thresholds vary, however, between corals, partly as a result of the specific type of endosymbiotic dinoflagellate (Symbiodinium sp.) they harbour. The production of reactive oxygen species (ROS) in corals under thermal and light stress has been recognised as one mechanism that can lead to cellular damage and the loss of their symbiont population (Oxidative Theory of Coral Bleaching). Here, we compared the response of symbiont and host enzymatic antioxidants in the coral species Acropora millepora and Montipora digitata at 28°C and 33°C. A. millepora at 33°C showed a decrease in photochemical efficiency of photosystem II (PSII) and increase in maximum midday excitation pressure on PSII, with subsequent bleaching (declining photosynthetic pigment and symbiont density). M. digitata exhibited no bleaching response and photochemical changes in its symbionts were minor. The symbiont antioxidant enzymes superoxide dismutase, ascorbate peroxidase, and catalase peroxidase showed no significant upregulation to elevated temperatures in either coral, while only catalase was significantly elevated in both coral hosts at 33°C. Increased host catalase activity in the susceptible coral after 5days at 33°C was independent of antioxidant responses in the symbiont and preceded significant declines in PSII photochemical efficiencies. This finding suggests a potential decoupling of host redox mechanisms from symbiont photophysiology and raises questions about the importance of symbiont-derived ROS in initiating coral bleaching.


Creatine supplementation increases glycogen storage but not GLUT-4 expression in human skeletal muscle.
It has been speculated that creatine supplementation affects muscle glucose metabolism in humans by increasing muscle glycogen storage and up-regulating GLUT-4 protein expression. In the present study, we assessed the effects of creatine loading and prolonged supplementation on muscle glycogen storage and GLUT-4 mRNA and protein content in humans. A total of 20 subjects participated in a 6-week supplementation period during which creatine or a placebo was ingested. Muscle biopsies were taken before and after 5 days of creatine loading (20 g.day(-1)) and after 6 weeks of continued supplementation (2 g.day(-1)). Fasting plasma insulin concentrations, muscle creatine, glycogen and GLUT-4 protein content as well as GLUT-4, glycogen synthase-1 (GS-1) and glycogenin-1 (Gln-1) mRNA expression were determined. Creatine loading significantly increased total creatine, free creatine and creatine phosphate content with a concomitant 18 +/- 5% increase in muscle glycogen content (P<0.05). The subsequent use of a 2 g.day(-1) maintenance dose for 37 days did not maintain total creatine, creatine phosphate and glycogen content at the elevated levels. The initial increase in muscle glycogen accumulation could not be explained by an increase in fasting plasma insulin concentration, muscle GLUT-4 mRNA and/or protein content. In addition, neither muscle GS-1 nor Gln-1 mRNA expression was affected. We conclude that creatine ingestion itself stimulates muscle glycogen storage, but does not affect muscle GLUT-4 expression.


Race, presenting signs and symptoms, use of carotid artery imaging, and appropriateness of carotid endarterectomy.
We sought to determine whether there are racial differences in use of carotid artery imaging after controlling for clinical factors and to ascertain racial differences in presenting signs and symptoms and overall appropriateness for carotid endarterectomy (CE). We performed a retrospective cohort study of 803 patients older than 45 years, hospitalized between 1991 and 1994 at any of 4 Veterans Affairs Medical Centers, with a discharge diagnosis of transient ischemic attack or ischemic stroke. Clinical data were abstracted from the medical record, including presenting symptoms, diagnostic test results, and use of surgical procedures. Appropriateness for CE was determined according to RAND criteria. Black patients were more likely than white patients to present with stroke (78% versus 55%) but less likely to present with transient ischemic attack (22% versus 45%; P=0.001). There was no racial difference in medical comorbidity or preoperative risk. Black patients were less likely to have an imaging study of their carotid arteries (67% versus 79%; P=0.001). Race remained an independent predictor of imaging after adjustment for clinical factors (odds ratio=1.50; 95% CI, 1.06 to 2.13). Because of higher prevalence of significant carotid artery stenosis, whites were significantly more likely than blacks to be assessed as appropriate candidates for surgery with the use of RAND criteria (18% versus 4%; P=0.001). Use of carotid artery imaging, a critical step in determining eligibility for CE, is influenced by the patient's race after controlling for clinical presentation. Adjustment for appropriateness of CE reduces but does not eliminate the importance of race.


A Versatile Solid-Phase Approach to the Synthesis of Oligonucleotide Conjugates with Biodegradable Hydrazone Linker.
One of the ways to efficiently deliver various drugs, including therapeutic nucleic acids, into the cells is conjugating them with different transport ligands via labile or stable bonds. A convenient solid-phase approach for the synthesis of 5'-conjugates of oligonucleotides with biodegradable pH-sensitive hydrazone covalent bonds is proposed in this article. The approach relies on introducing a hydrazide of the ligand under aqueous/organic media to a fully protected support-bound oligonucleotide containing aldehyde function at the 5'-end. We demonstrated the proof-of-principle of this approach by synthesizing 5'-lipophilic (e.g., cholesterol and α-tocopherol) conjugates of modified siRNA and non-coding RNAs imported into mitochondria (antireplicative RNAs and guide RNAs for Mito-CRISPR/system). The developed method has the potential to be extended for the synthesis of pH-sensitive conjugates of oligonucleotides of different types (ribo-, deoxyribo-, 2'-O-methylribo-, and others) with ligands of different nature.


Novel flow quantification of the carotid bulb and the common carotid artery with vector flow ultrasound.
Abnormal blood flow is usually assessed using spectral Doppler estimation of the peak systolic velocity. The technique, however, only estimates the axial velocity component, and therefore the complexity of blood flow remains hidden in conventional ultrasound examinations. With the vector ultrasound technique transverse oscillation the blood velocities of both the axial and the transverse directions are obtained and the complexity of blood flow can be visualized. The aim of the study was to determine the technical performance and interpretation of vector concentration as a tool for estimation of flow complexity. A secondary aim was to establish accuracy parameters to detect flow changes/patterns in the common carotid artery (CCA) and the carotid bulb (CB). The right carotid bifurcation including the CCA and CB of eight healthy volunteers were scanned in a longitudinal plane with vector flow ultrasound (US) using a commercial vector flow ultrasound scanner (ProFocus, BK Medical, Denmark) with a linear 5 MHz transducer transverse oscillation vector flow software. CCA and CB areas were marked in one cardiac cycle from each volunteer. The complex flow was assessed by medical expert evaluation and by vector concentration calculation. A vortex with complex flow was found in all carotid bulbs, whereas the CCA had mainly laminar flow. The medical experts evaluated the flow to be mainly laminar in the CCA (0.82 ± 0.14) and mainly complex (0.23 ± 0.22) in the CB. Likewise, the estimated vector concentrations in CCA (0.96 ± 0.16) indicated mainly laminar flow and in CB (0.83 ± 0.07) indicated mainly turbulence. Both methods were thus able to clearly distinguish the flow patterns of CCA and CB in systole. Vector concentration from angle-independent vector velocity estimates is a quantitative index, which is simple to calculate and can differentiate between laminar and complex flow.


National gambling experiences in the United States: will history repeat itself?
The aim of this paper is to offer an account of the history and current status of gambling research in the United States. A review of the literature. Gambling has been a part of society in the United States since its early history. However, it was not until 1980 that the medical profession in the United States first recognized pathological gambling as a psychiatric disorder. Today, it is still rarely diagnosed or treated, and relatively little federal funding is available to support research in this area. With the upcoming fifth revision of the Diagnostic and Statistical Manual of Mental Disorders, pathological gambling is likely to be included alongside substance use disorders as the first non-substance-related addictive disorder. This change may represent an opportunity to expand research on gambling and treatment of pathological gambling. We provide 10 suggestions for reducing societal and personal harm associated with this disorder.


A prognostic model and treatment strategy for intrahepatic recurrence of hepatocellular carcinoma after curative resection.
The aim of this study was to evaluate the prognostic factors for intrahepatic recurrence of hepatocellular carcinoma (HCC) after curative resection. Of 297 patients with HCC who underwent curative resection between 1998 and 2007, 145 had intrahepatic recurrence, and 125 of these were enrolled in this study. We analyzed the relationships between overall survival after HCC recurrence and 20 variables at initial hepatectomy and recurrence. Recurrent HCC was treated by repeat hepatectomy (Re-Hr, n = 29), radiofrequency ablation (RFA, n = 58), or transarterial chemoembolization (TAE, n = 38). Complete tumor control (CTC) by Re-He and RFA was selected for 70% of patients. RFA-treated patients had more tumors, smaller tumors, and poorer liver function at recurrence than the Re-Hr group. The overall 1-, 3-, and 5-year post-recurrence survival rates (SR) were 93.1, 66.8, 58.1%; 94.7, 75.1, 48.3%; and 80.1, 22.5, 0%, respectively, in the Re-Hr, RFA, and TAE groups. The SR was better for Re-Hr and RFA than for TAE (p < 0.0001). Outcomes were similar in Re-Hr and RFA, regardless of recurrent tumor size. Multivariate analysis identified Child-Pugh grade B, AFP ≥100 ng/ml at recurrence, recurrent tumor size ≥3 cm, tumor number ≥3, and CTC as significant prognostic factors for overall post-recurrence survival. A scoring system using 1 point for each patient-background factor provided a well-categorized predictive model. The overall 3-/5-year post-recurrence SRs were 83.1/59.3%, 64.1/41.9%, 42.0/18.0%, and 13.6/0% at risk number (R) R0, R1, R2, and R3/4, respectively (p < 0.05). Significant prognostic factors for intrahepatic recurrent HCC are poor hepatic reserve, AFP, recurrent tumor size and number, and CTC. Selection of treatment modality for intrahepatic recurrence requires the clinician to be mindful of the predictive factors and to control tumors aggressively by adequate treatment, selected by balancing various conditions.


Assessment of testing methods for drug-induced repolarization delay and arrhythmias in an iPS cell-derived cardiomyocyte sheet: multi-site validation study.
A prospective comparison study across 3 independent research laboratories of a pure IKr blocker E-4031 was conducted by using the same batch of human iPS cell-derived cardiomyocytes in order to verify the utility and reliability of our original standard protocol. Field potential waveforms were recorded with a multi-electrode array system to measure the inter-spike interval and field potential duration. The effects of E-4031 at concentrations of 1 to 100 nM were sequentially examined every 10 min. In each facility, E-4031 significantly prolonged the field potential duration corrected by Fridericia's formula and caused early afterdepolarizations occasionally resulting in triggered activities, whereas it tended to decrease the rate of spontaneous contraction. These results were qualitatively and quantitatively consistent with previous non-clinical in vitro and in vivo studies as well as clinical reports. There were inter-facility differences in some absolute values of the results, which were not observed when the values were normalized as percentage change. Information described in this paper may serve as a guide when predicting the drug-induced repolarization delay and arrhythmias with this new technology of stem cells.


Health behavior interests of adolescents with unhealthy diet and exercise: implications for weight management.
This study sought to determine individual factors that may influence adolescents' interests in various health behaviors and, by extension, their potential interest in programs that promote healthy lifestyles and reduce obesity. The sample consisted of 737 rural Pennsylvania (United States) middle and high school students not involved in either healthy exercise or dietary behaviors (a target group for health-promoting interventions). Participants completed a self-report measure of their general health functioning, including their interests in sports programs, outdoor recreation programs, weightlifting, weight loss and healthy eating/cooking. Nurses measured body mass indices (BMIs). The vast majority of the sample endorsed self-efficacy in healthy eating and physical activity, and this self-efficacy was associated with interest in a healthy diet and outdoor recreation. Interest in healthy activities was consistently higher among 7th graders (age mean = 12.6 years) than 11th graders (age mean = 16.3 years). Females were more interested in weight loss and healthy eating/cooking, whereas males were more interested in weightlifting. Higher BMI only predicted interest in weight loss. These results indicate that adolescent health interests vary on the basis of their gender, grade level, BMI and self-efficacy. These trends are potentially important to consider when seeking to match intervention programs to adolescent interests.


[Acute kidney injury].
'Acute kidney injury' is modern terminology for a sudden decline in kidney function, and is defined by the RIFLE classification (RIFLE is an acronym for Risk, Injury, Failure, Loss and End-stage kidney disease). Acute kidney injury occurs as a result of the combination of reduced perfusion in the kidney, systemic inflammation, and the administration of nephrotoxic drugs. Patients with chronic renal damage, advanced age, diabetes mellitus or heart failure are at an increased risk of acute kidney injury. In the short term, acute kidney injury leads to a markedly increased risk of death; the long-term effect of acute kidney injury is a permanent loss of renal microcirculation which could result in chronic renal disease. Certain biomarkers in the urine offer new possibilities for detecting acute kidney injury in its early stage. Treatment of patients with acute kidney injury is currently supportive in nature. The optimisation of a patient's haemodynamics results in a reduction of the occurrence of acute kidney injury during extensive surgical procedures. A promising treatment aimed at preventing acute kidney injury is called 'remote ischaemic pre-conditioning'.


Nelumbo nucifera leaves extracts inhibit mouse airway smooth muscle contraction.
Alkaloids extracted from lotus leaves (AELL) can relax vascular smooth muscle. However, whether AELL has a similar relaxant role on airway smooth muscle (ASM) remains unknown. This study aimed to explore the relaxant property of AELL on ASM and the underlying mechanism. Alkaloids were extracted from dried lotus leaves using the high temperature rotary evaporation extraction method. The effects of AELL on mouse ASM tension were studied using force measuring and patch-clamp techniques. It was found that AELL inhibited the high K+ or acetylcholine chloride (ACh)-induced precontraction of mouse tracheal rings by 64.8 ± 2.9%, or 48.8 ± 4.7%, respectively. The inhibition was statistically significant and performed in a dose-dependent manner. Furthermore, AELL-induced smooth muscle relaxation was partially mediated by blocking voltage-dependent Ca2+ channels (VDCC) and non-selective cation channels (NSCC). AELL, which plays a relaxant role in ASM, might be a new complementary treatment to treat abnormal contractions of the trachea and asthma.


The role of stromal cells in the persistence of chronic inflammation.
Inflammation is an unstable state; it either resolves or persists. Inflammatory reactions often have a propensity for specific anatomical sites. Why inflammation persists with specific tissue tropism remains obscure. Increasing evidence suggests that stromal cells which define tissue architecture are the key cells involved, and therefore make attractive therapeutic targets. Research on stromal cells in general and fibroblasts in particular has so far been hampered by a lack of fibroblast-specific cell markers. This review highlights our increasing understanding of the role of fibroblasts in inflammation, and suggests that these cells provide the cellular basis for site specific chronic inflammation.


The Efficacy of Neurofeedback in Patients with Major Depressive Disorder: An Open Labeled Prospective Study.
The purpose of this study was to evaluate the effect of neurofeedback on depressive symptoms and electrophysiological disturbances in patients with major depressive disorder. We recruited participants suffering from depression to evaluate efficacy of left prefrontal beta with alpha/theta training. An 8-week, prospective, open-label study was undertaken. Twenty participants were recruited. The treatment protocol was twice or three times a week training of beta at F3 with alpha/theta at Pz for 8 weeks. When every visit, patients were received beta training for 30 min, and then alpha/theta training for 30 min. Baseline, 4 and 8 week scores of; the Hamilton rating scale for Depression (HAM-D), the Hamilton rating scale for Anxiety (HAM-A), the Beck Depression Inventory (BDI)-II, the Beck Anxiety Inventory (BAI), Clinical global impression-severity (CGI-S), and pre- and post-treatment resting state EEGs were compared. Interhemispheric alpha power asymmetry (A score) was computed for homologous sites F3-F4. Pre- and post-training clinical assessments revealed significant improvements in HAM-D, HAM-A, BDI, and CGI-S scores. Cumulative response rates by HAM-D were 35.0 and 75.0 % at 4 and 8 weeks, respectively, corresponding cumulative remission rates by HAM-D were 15.0 and 55.0 %, respectively. No significant differences were found between pre- and post-treatment A score. Neurofeedback treatment could improve depressive symptoms significantly. In addition, anxiety symptoms and clinical illness severity decreased significantly after neurofeedback treatment. Despite its several limitations, such as, small sample size and lack of a control group, this study suggested neurofeedback has significant effects in patients with major depressive disorder.


An exposure-based framework for grouping pollutants for a cumulative risk assessment approach: case study of indoor semi-volatile organic compounds.
Humans are exposed to a large number of contaminants, many of which may have similar health effects. This paper presents a framework for identifying pollutants to be included in a cumulative risk assessment approach. To account for the possibility of simultaneous exposure to chemicals with common toxic modes of action, the first step of the traditional risk assessment process, i.e. hazard identification, is structured in three sub-steps: (1a) Identification of pollutants people are exposed to, (1b) identification of effects and mechanisms of action of these pollutants, (1c) grouping of pollutants according to similarity of their mechanism of action and health effects. Based on this exposure-based grouping we can derive "multi-pollutant" toxicity reference values, in the "dose-response assessment" step. The approach proposed in this work is original in that it is based on real exposures instead of a limited number of pollutants from a unique chemical family, as traditionally performed. This framework is illustrated by the case study of semi-volatile organic compounds in French dwellings, providing insights into practical considerations regarding the accuracy of the available toxicological information. This case study illustrates the value of the exposure-based approach as opposed to the traditional cumulative framework, in which chemicals with similar health effects were not always included in the same chemical class.


The stages in calcification of the rat kidney after the administration of uranium nitrate.
The anatomical and histochemical alterations in rat kidneys after the parenteral administration of uranium nitrate [UO(2)(NO(3))(2).6H(2)O] have been studied. The histological effects produced by this agent over a wide range of dose levels were nearly identical in character and differed principally in their speed of evolution. The deposition of calcium always began at foci in the cytoplasm of the cells of the proximal convoluted tubules of the inner cortex. It remained intracellular until the cell boundaries were destroyed. Deposits of calcium could be found before any other cellular damage could be demonstrated by histological examination. Later, when degeneration and necrosis were present, the foci of calcification were imperfectly related to them in location or degree. In contrast, the amount of calcification was correlated with the dose of uranium nitrate, being greatest in the kidneys of rats that received 20 mg./kg., next greatest in the 30 mg./kg. rats, less in the 10 mg./kg. rats, and slight in those that received 2 mg./kg. Histochemical stains for ferric and ferrous iron, chondroitinsulfate, and polysaccharides gave results that were negative or unrelated to the deposits of calcium, thus making it unlikely that these substances held any appreciable amount of calcium in the tissue. Yet it is clear that some anion other than phosphate must be combined with part of the calcium; the results with the alizarin and von Kóssa stains confirmed an earlier result (1) in showing that the first deposits of calcium are formed without comparable accumulations of phosphate.