Adult bacterial meningitis: earlier treatment and improved outcome following guideline revision promoting prompt lumbar puncture.
In suspected acute bacterial meningitis (ABM), cerebral computerized tomography (CT) is recommended before lumbar puncture (LP) if mental impairment. Despite guideline emphasis on early treatment, performing CT prior to LP implies a risk of delayed treatment and unfavorable outcome. Therefore, Swedish guidelines were revised in 2009, deleting impaired mental status as a contraindication for LP without prior CT scan. The aim of the present study was to evaluate the guideline revision. The Swedish quality registry for community-acquired ABM was analyzed retrospectively. Door-to-antibiotic time and outcome were compared among patients treated 2005-2009 (n=394) and 2010-2012 (n=318). The effect of different LP-CT sequences was analyzed during 2008-2012. Adequate treatment was started 1.2 hours earlier, and significantly more patients were treated <2 hours from admission 2010-2012 than 2005-2009. Compared with CT before LP, immediate LP resulted in 1.6 hours earlier treatment, significant increase in door-to-antibiotic times of <1 and <2 hours, and a favorable outcome. In 2010-2012, mortality was lower (6.9% vs 11.7%) and the risk of sequelae at follow-up decreased (38% vs 49%) in comparison with 2005-2009. Treatment delay resulted in a significantly increased risk for fatal outcome, with a relative increase in mortality of 12.6% per hour of delay. The deletion of impaired mental status as contraindication for prompt LP and LP without prior CT scan are associated with significantly earlier treatment and a favorable outcome. A revision of current international guidelines should be considered.


Cooling of Tropical Brazil (5{degrees}C) During the Last Glacial Maximum.
A 30,000-year paleotemperature record derived from noble gases dissolved in carbon-14-dated ground water indicates that the climate in lowland Brazil (Piaui Province, 7 degrees S, 41.5 degrees W; altitude, 400 meters) was 5.4 degrees +/- 0.6 degrees C cooler during the last glacial maximum than today. This result suggests a rather uniform cooling of the Americas between 40 degrees S and 40 degrees N. A 5.4 degrees C cooling of tropical South America is consistent with pollen records, snow line reconstructions, and strontium/calcium ratios and delta(18)O coral records but is inconsistent with the sea-surface temperature reconstruction of CLIMAP (Climate: Long-Range Investigation, Mapping and Prediction). On the basis of these results, it appears that the tropical Americas are characterized by a temperature sensitivity comparable to that found in higher latitudes.


Modulation of Ras signaling alters the toxicity of hydroquinone, a benzene metabolite and component of cigarette smoke.
Benzene is an established human leukemogen, with a ubiquitous environmental presence leading to significant population exposure. In a genome-wide functional screen in the yeast Saccharomyces cerevisiae, inactivation of IRA2, a yeast ortholog of the human tumor suppressor gene NF1 (Neurofibromin), enhanced sensitivity to hydroquinone, an important benzene metabolite. Increased Ras signaling is implicated as a causal factor in the increased pre-disposition to leukemia of individuals with mutations in NF1. Growth inhibition of yeast by hydroquinone was assessed in mutant strains exhibiting varying levels of Ras activity. Subsequently, effects of hydroquinone on both genotoxicity (measured by micronucleus formation) and proliferation of WT and Nf1 null murine hematopoietic precursors were assessed. Here we show that the Ras status of both yeast and mammalian cells modulates hydroquinone toxicity, indicating potential synergy between Ras signaling and benzene toxicity. Specifically, enhanced Ras signaling increases both hydroquinone-mediated growth inhibition in yeast and genotoxicity in mammalian hematopoetic precursors as measured by an in vitro erythroid micronucleus assay. Hydroquinone also increases proliferation of CFU-GM progenitor cells in mice with Nf1 null bone marrow relative to WT, the same cell type associated with benzene-associated leukemia. Together our findings show that hydroquinone toxicity is modulated by Ras signaling. Individuals with abnormal Ras signaling could be more vulnerable to developing myeloid diseases after exposure to benzene. We note that hydroquinone is used cosmetically as a skin-bleaching agent, including by individuals with cafe-au-lait spots (which may be present in individuals with neurofibromatosis who have a mutation in NF1), which could be unadvisable given our findings.


Proliferation of rhesus ovarian surface epithelial cells in culture: lack of mitogenic response to steroid or gonadotropic hormones.
Ovarian cancer is the most lethal gynecological cancer, and approximately 90% of ovarian cancers derive from the ovarian surface epithelium (OSE), yet the biology of the OSE is poorly understood. Factors associated with increased risk of non-hereditary ovarian cancer include the formation of inclusion cysts, effects of reproductive hormones and the number of ovulations experienced in a woman's lifetime. Distinguishing between these factors is difficult in vivo, but cultured OSE cells are viable tools for some avenues of research. Here we establish rhesus macaque OSE cultures and demonstrate that these cells express cytokeratin, vimentin, N-cadherin, ER-alpha, and PR but are negative for E-cadherin. We show that these cells activate MAPK and proliferate in response to extracellular calcium, as do human and rat OSE. In contrast, the gonadotropic hormones FSH (4-400 IU/liter), LH (8.5-850 IU/liter), and human CG (10-1000 IU/liter) fail to stimulate proliferation. We find that concentrations of progesterone and estrogen normally present in follicles just before ovulation ( approximately 1000 ng/ml) significantly decrease the number of mitotically active rhesus macaque OSE cells as determined by PCNA labeling, total cell count, and (3)H-thymidine uptake, whereas lower steroid concentrations have no effect.


Fetal cardiac screening: why bother?
Congenital heart disease (CHD) is a common form of congenital malformation associated with significant morbidity and mortality. Antenatal diagnosis of life-threatening forms of CHD may help to improve survival and morbidity as well as allowing parental choice and preparation. The diagnosis of CHD during fetal life can be made with a very high level of diagnostic accuracy in tertiary centres dealing with high-risk pregnancies. However, most cases of CHD will occur in low-risk groups and will only be detected by screening at the time of obstetric ultrasound scans. The concept of antenatal screening for CHD was introduced in the UK over 20 years ago, and current national guidelines recommend that the heart should be examined at the time of the obstetric anomaly scan. However, there remains a large regional variation in prenatal detection rates of CHD. Widespread teaching and training is required to achieve a more uniform standard but this will be associated with significant resource implications.


Programmable assembly of DNA-functionalized liposomes by DNA.
Bionanotechnology involves the use of biomolecules to control both the structure and property of nanomaterials. One of the most studied examples is DNA-directed assembly of inorganic nanoparticles such as gold nanoparticles (AuNPs). However, systematic studies on DNA-linked soft nanoparticles, such as liposomes, are still lacking. We herein report the programmable assembly and systematic characterization of DNA-linked liposomes as a function of liposome size, charge, fluidity, composition, DNA spacer, linker DNA sequence, and salt concentration for direct comparison to DNA-directed assembly of AuNPs. Similar to the assemblies of AuNPs, sharp melting transitions were observed for liposomes where the first derivative of the melting curve full width at half-maximum (fwhm) is equal to or less than 1 °C for all of the tested liposomes, allowing sequence specific DNA detection. We found that parameters such as liposome size, charge, and fluidity have little effect on the DNA melting temperature. Cryo-TEM studies showed that programmable assemblies can be obtained and that the majority of the liposomes maintained a spherical shape in the assembled state. While liposome and AuNP systems are similar in many aspects, there are also important differences that can be explained by their respective physical properties.


A theory of waveguide design for plasmonic nanolasers.
We propose a theory for the waveguide design and analysis for plasmonic nanolasers by reformulating the fundamental waveguide requirements. This theory does not rely on further optimizing previously used structures, but examines each possible design without prejudice. Our exploration of one-dimensional (i.e., layered) plasmonic nanowaveguide geometries and the subsequent extension to 2D structures not only provides a deep understanding of the characteristics of currently used designs, but also leads to superior structures with the potential to address long-standing challenges in plasmonic nanolasers. In addition, we discover analogies between the reformulated fundamental requirements for the waveguide for nanolasers and nanoscale four-wave mixing (FWM) devices. Therefore, after a slight modification, our theory can also be applied to the waveguide design for plasmonic FWM devices.


Use of anti-asthma drugs in New Zealand.
Increased sales of anti-asthma drugs, and a second "epidemic" of asthma mortality, raised concerns about the management of asthma in New Zealand. To study this, prescriptions were obtained from randomly selected pharmacies to identify 235 patients receiving one common anti-asthma drug, 175 of whom were willing to be interviewed. The authors considered that 80% had asthma, and only 20% suffered primarily from chronic bronchitis or emphysema. The increased sales of anti-asthma drugs could not therefore be explained by their increasing use in treatment of other respiratory disorders. One third of the identified asthmatic subjects experienced daily symptoms despite regular drug treatment. Inhaled corticosteroids were used by only 42% of this group with persistent symptoms. Regular or short course oral corticosteroids, with or without inhaled steroids, had been required by 49%. All patients with domiciliary nebulisers appeared to use these appropriately, and most had peak expiratory flow meters. Despite the increased sales of anti-asthma drugs, corticosteroids appear to be as much underused in patients with chronic asthma in the community as in those who die of their disease.


Establishment of an osteocyte-like cell line, MLO-Y4.
Although osteocytes are the most abundant cells in bone, their functional role remains unclear. In part, this is due to lack of availability of osteocyte cell lines which can be studied in vitro. Since others have shown that cell lines can be readily developed from transgenic mice in which the SV40 large T-antigen oncogene is expressed under the control of a promoter which targets the cells of interest, we used this approach to develop an osteocyte cell line. We chose as a promoter osteocalcin, whose expression is essentially limited to bone cells and which is expressed more abundantly in osteocytes than in osteoblasts. From these transgenic mice, we isolated cells from the long bones using sequential collagenase digestion and maintained these cells on collagen-coated surfaces which are optimal for osteocyte maintenance and growth. We describe here the properties of a cell line cloned from these cultures, called MLO-Y4 (for murine long bone osteocyte Y4). The properties of MLO-Y4 cells are very similar to primary osteocytes. Like primary osteocytes and unlike primary osteoblasts, the cell line produces large amounts of osteocalcin but low amounts of alkaline phosphatase. The cells produce extensive, complex dendritic processes and are positive for T-antigen, for osteopontin, for the neural antigen CD44, and for connexin 43, a protein found in gap junctions. This cell line also produces very small amounts of type I collagen mRNA compared with primary osteoblasts. MLO-Y4 cells lack detectable mRNA for osteoblast-specific factor 2, which appears to be a positive marker for osteoblasts but may be a negative marker for osteocytes. This newly established cell line should prove useful for studying the effects of mechanical stress on osteocyte function and for determining the means whereby osteocytes communicate with other bone cells such as osteoblasts and osteoclasts.


Induction of p53 contributes to apoptosis of HCT-116 human colon cancer cells induced by the dietary compound fisetin.
Fisetin, or 3,3',4',7-tetrahydroxyflavone, is present in fruits and vegetables and has been previously reported to inhibit the proliferation of a variety of cancer cells (Lu X, Jung J, Cho HJ, Lim do Y, Lee HS, Chun HS, Kwon DY, Park JH. J Nutr 135: 2884-2890, 2005). We have demonstrated in a previous work that 20-60 micromol/l fisetin inhibits cyclin-dependent kinase activities resulting in cell cycle arrest in HT-29 colon cancer cells. In the present study, we attempted to characterize the mechanisms by which fisetin induces apoptosis in HCT-116 cells. DNA condensations, cleavage of poly(ADP-ribose) polymerase (PARP), and cleavage of caspases 9, 7, and 3 were induced in HCT-116 cells treated with 5-20 micromol/l of fisetin. Fisetin induced a reduction in the protein levels of antiapoptotic Bcl-xL and Bcl-2 and an increase in the levels of proapoptotic Bak and Bim. Fisetin did not affect the Bax protein levels, but induced the mitochondrial translocation of this protein. Fisetin also enhanced the permeability of the mitochondrial membrane and induced the release of cytochrome c and Smac/Diablo. Additionally, fisetin caused an increase in the protein levels of cleaved caspase-8, Fas ligand, death receptor 5, and TNF-related apoptosis-inducing ligand, and the caspase-8 inhibitor Z-IETD-FMK suppressed fisetin-induced apoptosis and the activation of caspase-3. Furthermore, fisetin increases p53 protein levels, and the inhibition of p53 expression by small interference RNA resulted in a decrease in the fisetin-induced translocation of Bax to the mitochondria, release of mono- and oligonucleosome in the cytoplasm, and PARP cleavage. These results show that fisetin induces apoptosis in HCT-116 cells via the activation of the death receptor- and mitochondrial-dependent pathway and subsequent activation of the caspase cascade. The induction of p53 results in the translocation of Bax to the mitochondria, which contributes to fisetin-induced apoptosis in HCT-116 cells.


An altered tissue distribution of flaxseed lignans and their metabolites in Abcg2 knockout mice.
Lignans are dietary polyphenols, which are metabolized by gut microbiota into the phytoestrogenic metabolites enterolignans, mainly enterolactone and enterodiol. Breast Cancer Resistance Protein (BCRP/ABCG2) is an efflux transporter that affects the plasma and milk secretion of several drugs and natural compounds. We hypothesized here that Abcg2 could influence the levels of lignans and their derived metabolites in target tissues. Consequently, we aimed to evaluate the role of Abcg2 in the tissue distribution of these compounds. We used Abcg2-/- knockout and wild-type male mice fed with a lignan-enriched diet for one week and analysed their plasma, small intestine, colon, liver, kidneys and testicles. High levels of lignans as well as enterolignans and their glucuronide and sulfate conjugates in the small intestine and colon were detected, with higher concentrations of the conjugates in the wild-type compared with Abcg2-/- mice. Particularly relevant was the detection of 24-fold and 8-fold higher concentrations of enterolactone-sulfate and enterolactone-glucuronide, respectively, in the kidney of Abcg2-/- compared with wild-type mice. In conclusion, our study showed that lignans and their derived metabolites were in vivo substrates of Abcg2, which affected their plasma and tissue levels. These results highlight the role of Abcg2 in influencing the health-beneficial properties of dietary lignans.


Combined stimulant and antipsychotic treatment in adolescents with attention-deficit/hyperactivity disorder: a cross-sectional observational structural MRI study.
Meta-analyses suggest normalizing effects of methylphenidate on structural fronto-striatal abnormalities in patients with attention-deficit/hyperactivity disorder (ADHD). A subgroup of patients receives atypical antipsychotics concurrent with methylphenidate. Long-term safety and efficacy of combined treatment are unknown. The current study provides an initial investigation of structural brain correlates of combined methylphenidate and antipsychotic treatment in patients with ADHD. Structural magnetic resonance imaging was obtained in 31 patients who had received combined methylphenidate and antipsychotic treatment, 31 matched patients who had received methylphenidate but not antipsychotics, and 31 healthy controls (M age 16.7 years). We analyzed between-group effects in total cortical and subcortical volume, and in seven frontal cortical and eight subcortical-limbic volumes of interest, each involved in dopaminergic neurotransmission. Patients in the combined treatment group, but not those in the methylphenidate only group, showed a reduction in total cortical volume compared to healthy controls (Cohen's d = 0.69, p < 0.004), which was apparent in most frontal volumes of interest. Further, the combined treatment group, but not the methylphenidate group, showed volume reduction in bilateral ventral diencephalon (Left Cohen's d = 0.48, p < 0.04; Right Cohen's d = 0.46, p < 0.05) and the left thalamus (Cohen's d = 0.47, p < 0.04). These findings may indicate antipsychotic treatment counteracting the normalizing effects of methylphenidate on brain structure. However, it cannot be ruled out that pre-existing clinical differences between both patient groups may have resulted in anatomical differences at the time of scanning. The absence of an untreated ADHD group hinders unequivocal interpretation and implications of our findings.


A P2rx7 Passenger Mutation Affects the Vitality and Function of T cells in Congenic Mice.
Among laboratory mouse strains many genes are differentially expressed in the same cell population. As consequence, gene targeting in 129-derived embryonic stem cells (ESCs) and backcrossing the modified mice onto the C57BL/6 background can introduce passenger mutations in the close proximity of the targeted gene. Here, we demonstrate that several transgenic mice carry a P2rx7 passenger mutation that affects the function of T cells. By the example of P2rx4tm1Rass we demonstrate that P2X4ko T cells express higher levels of P2X7 and are more sensitive toward the P2X7 activators ATP and NAD+, rendering these cells more vulnerable toward NAD-induced cell death (NICD) compared with wild type (WT). The enhanced NICD sensitivity confounded functional assays e.g. cytokine production and cell migration. Our results need to be considered when working with P2rx4tm1Rass mice or other 129-based transgenic strains that target P2rx7 neighboring genes.


Energy transduction between membranes. TonB, a cytoplasmic membrane protein, can be chemically cross-linked in vivo to the outer membrane receptor FepA.
TonB, a cytoplasmic membrane protein, couples cytoplasmic membrane protonmotive force to active transport across the outer membrane of Escherichia coli. In vivo cross-linking studies were initiated to analyze TonB interactions with other cell envelope proteins. Four TonB-specific cross-linked complexes were detected with apparent molecular masses of 195, 77, 59, and 43.5 kDa. The 195-kDa complex was shown to contain both TonB and FepA, the outer membrane receptor for the siderophore enterochelin. The 195-kDa complex is absent in strains missing either TonB or FepA and can be detected by either TonB-specific or FepA-specific monoclonal antibodies. This is the first direct in vivo evidence that TonB can span the periplasmic space to interact physically with outer membrane receptors. Consistent with that observation, the outer membrane protease OmpT was shown to play a role in TonB turnover, both in the presence and absence of ExbB results in the rapid degradation of TonB. The absence of OmpT could be used to stabilize TonB in an exbB::Tn10 strain such that steady state levels of TonB protein are identical to a wild-type strain. Under those conditions, the absence of ExbB results in greatly reduced TonB activity, indicating that ExbB plays a direct role in energy transduction and probably secondarily protects TonB protein from proteolysis. The 59-kDa complex was absent in an exbB::Tn10 strain, suggesting either that ExbB is in the complex with TonB or that ExbB is required to form the 59-kDa complex. A tolQ nonsense mutation had no effect on the cross-linking profile observed, confirming that its participation in TonB-dependent phenomena is minor and most likely the result of evolutionary cross-talk.


Simplified quantitative methods for bacteriuria and pyuria.
Although pyelonephritis is a common disease, it escapes clinical detection in an undesirably high proportion of patients. The present unsatisfactory diagnostic position would be much improved by widespread screening of patients by simple yet reasonably accurate methods. Bacterial counts by the pour-plate technique and estimates of the white cell excretion per hour or day, while undoubtedly of diagnostic value, are probably unsuitable for use on a wide scale. In an attempt to find more convenient procedures a simplified stroke-plate method of bacterial counting and a simplified quantitative white cell count method were devised and applied to over 1,000 mid-stream urine samples from 398 patients. Good correlation was obtained between the simpler stroke-plate method of bacterial counting and the more time-consuming pour-plate method. The quantitative white cell procedure was a much more sensitive index of pyuria than wet-film microscopy, and comparison with the bacterial count results showed that it gave a useful indication of urinary infection. It is suggested that a quantitative bacterial count should replace non-quantitative culture methods when urinary infection is suspected and that the quantitative white cell count should be performed as a routine part of the initial clinical and laboratory assessment of all patients, followed by a bacterial count if pyuria is revealed. Experience has shown that routine urine microscopy by a precise method leads to the detection of many cases of occult urinary infection.


Are Unit Costs the Same? A Case Study Comparing Different Valuation Methods for Unit Cost Calculation of General Practitioner Consultations.
To inform allocation decisions in any healthcare system, robust cost data are indispensable. Nevertheless, recommendations on the most appropriate valuation approaches vary or are nonexistent, and no internationally accepted gold standard exists. This costing analysis exercise aims to assess the impact and implications of different calculation methods and sources based on the unit cost of general practitioner (GP) consultations in Austria. Six costing methods for unit cost calculation were explored, following 3 Austrian methodological approaches (AT-1, AT-2, AT-3) and 3 approaches applied in 3 other European countries (Germany, The Netherlands, United Kingdom). Drawing on Austrian data, mean unit costs per GP consultation were calculated in euros for 2015. Mean unit costs ranged from €15.6 to €42.6 based on the German top-down costing approach (DE) and the Austrian Physicians' Chamber's price recommendations (AT-3), respectively. The mean unit cost was estimated at €18.9 based on Austrian economic evaluations (AT-1) and €17.9 based on health insurance payment tariffs (AT-2). The Dutch top-down (NL) and the UK bottom-up approaches (UK) yielded higher estimates (NL: €25.3, UK: €29.8). Overall variation reached 173%. Our study is the first to systematically investigate the impact of differing calculation methods on unit cost estimates. It shows large variations with potential impact on the conclusions in an economic evaluation. Although different methodological choices may be justified by the adopted study perspective, different costing approaches introduce variation in cross-study/cross-country cost estimates, leading to decreased confidence in data quality in economic evaluations.


Modulation of spermatozoon acrosome reaction.
Spermatozoon acrosome reaction is an exocytotic event of the utmost importance for the development of mammalian fertilisation. Current evidence shows that the triggering of the acrosome reaction (AR) could be regulated by the action of diverse compounds, namely, metabolites, neurotransmitters and hormones. The aim of the present review is to describe the modulating effects of several compounds that have been classified as inductors or inhibitors of acrosome reaction. Among AR inductors, it is necessary to mention progesterone, angiotensin II, atrial natriuretic peptide, cathecolamines, insulin, leptin, relaxin and other hormones. Regarding the inhibitors, oestradiol and epidermal growth factor are among the substances that retard AR. It is worth mentioning that gamma-aminobutyric acid, a neurotransmitter known to be an inhibitor in the central nervous system, has been shown to induce AR. The multiple hormones located in the fluids of the female reproductive tract are also likely to act as subtle regulators of AR, constituting a fundamental aspect for the development of successful fertilisation. Finally, it is necessary to emphasise that the study of regulation exerted by hormones and other compounds on AR is essential for further understanding of mammalian reproductive biology, especially spermatozoon physiology.


Multiple docking adapter window for the s-190 experiment.
Martin Marietta/NASA subcontracted the design, fabrication, testing, and installation of the multiple docking adapter S-190 experiment window assembly to Actron. In this paper we primarily treat the thermal-optical analysis aspect of the design phase. The window is a pane of BK 7 glass 59.18 cm x 44.91 cm x 4.06 cm. Though it must meet many mechanical specifications, the optical requirements, dictated by the high acuity, distortion-free S-190 multiband camera system, are the most severe. Under operating conditions over any 7.6-cm circular area, the maximum rms deviation from the best-fitting plane must be less than 12.0 nm, and from the reference plane through the entire window, 60.0 nm.


Successes and Misses of Global Health Development: Detecting Temporal Concept Drift of Under-5 Mortality Prediction Models with Bias Scan.
Under-5 Mortality rates have been decreasing across Africa for the past two decades. Contributing factors include policy changes, technology, and health investments. This study identifies sub-populations that have experienced more-than-expected change in mortality rates (either increasing or decreasing) during this time period. We train under-5 mortality predictive models on Demographic and Health Survey (DHS) datasets from the early 2000s and apply those models to data collected in more recent versions of the survey. This provides an estimate of the risk current families would have faced in the past. We then apply techniques from anomalous pattern detection to identify sub-populations that have the most divergence between their predicted and observed mortality rates; higher and lower. These detected groups are examples of successes and possible misses of the health progress observed in Africa over the course of decades. Identifying these groups through data-driven discovery may lead to a better understanding of health policies in developing countries.


Osteogenesis and angiogenesis induced by porous β-CaSiO(3)/PDLGA composite scaffold via activation of AMPK/ERK1/2 and PI3K/Akt pathways.
As a potential bioactive material, β-calcium silicate (β-CS) has attracted particular attention in the field of bone regeneration. In this study, porous β-CS/Poly-D,L-Lactide-Glycolide (PDLGA) composite scaffolds were developed with the goals of controlling the degradation rate and improving the mechanical and biological properties. The compressive strength and toughness were significantly enhanced by PDLGA modification of porous β-CS ceramic scaffolds. The effects of the ionic extract from β-CS/PDLGA composite scaffolds on osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells (rBMSCs), proliferation of human umbilical vein endothelial cells (HUVECs) and the related mechanisms were investigated. It was shown that bioactive ions from β-CS/PDLGA scaffolds could enhance cell viability, alkaline phosphatase (ALP) activity, calcium mineral deposition, and mRNA expression levels of osteoblast-related genes of rBMSCs without addition of extra osteogenic reagents. The activation in AMP-activated protein kinase (AMPK), extracellular signal-related kinases (ERK) 1/2 and RUNX-2 were observed in rBMSCs cultured in the extract of β-CS/PDLGA, and these effects could be blocked by AMPK inhibitor Compound C. The extracts of β-CS/PDLGA composites stimulated HUVECs proliferation that was associated with phosphorylation of protein kinase B (Akt) and endothelial nitric oxide synthase (eNOS) as well as an increase in nitric oxide (NO) production and secretion of vascular endothelial growth factor (VEGF). The inductions were abolished by the addition of phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002. The composite scaffolds were implanted in critical sized rabbit femur defects (6 × 10 mm) for 4, 12 and 20 weeks with β-tricalcium phosphate (β-TCP) as controls. Sequential histological evaluations and radiographs revealed that β-CS/PDLGA dramatically stimulated new bone formation and angiogenesis. The biodegradation rate of the β-CS/PDLGA scaffolds was lower than that of β-TCP at each time point examined, and matched the new bone formation rates. These data suggest that β-CS/PDLGA could promote bone regeneration in vivo, which might be ascribed to the enhanced osteogenic differentiation of mesenchymal stem cells (MSCs) and increased angiogenic activity of endothelial cells (ECs).


Mouse HORMAD1 and HORMAD2, two conserved meiotic chromosomal proteins, are depleted from synapsed chromosome axes with the help of TRIP13 AAA-ATPase.
Meiotic crossovers are produced when programmed double-strand breaks (DSBs) are repaired by recombination from homologous chromosomes (homologues). In a wide variety of organisms, meiotic HORMA-domain proteins are required to direct DSB repair towards homologues. This inter-homologue bias is required for efficient homology search, homologue alignment, and crossover formation. HORMA-domain proteins are also implicated in other processes related to crossover formation, including DSB formation, inhibition of promiscuous formation of the synaptonemal complex (SC), and the meiotic prophase checkpoint that monitors both DSB processing and SCs. We examined the behavior of two previously uncharacterized meiosis-specific mouse HORMA-domain proteins--HORMAD1 and HORMAD2--in wild-type mice and in mutants defective in DSB processing or SC formation. HORMADs are preferentially associated with unsynapsed chromosome axes throughout meiotic prophase. We observe a strong negative correlation between SC formation and presence of HORMADs on axes, and a positive correlation between the presumptive sites of high checkpoint-kinase ATR activity and hyper-accumulation of HORMADs on axes. HORMADs are not depleted from chromosomes in mutants that lack SCs. In contrast, DSB formation and DSB repair are not absolutely required for depletion of HORMADs from synapsed axes. A simple interpretation of these findings is that SC formation directly or indirectly promotes depletion of HORMADs from chromosome axes. We also find that TRIP13 protein is required for reciprocal distribution of HORMADs and the SYCP1/SC-component along chromosome axes. Similarities in mouse and budding yeast meiosis suggest that TRIP13/Pch2 proteins have a conserved role in establishing mutually exclusive HORMAD-rich and synapsed chromatin domains in both mouse and yeast. Taken together, our observations raise the possibility that involvement of meiotic HORMA-domain proteins in the regulation of homologue interactions is conserved in mammals.


LSD-associated "Alice in Wonderland Syndrome"(AIWS): A Hallucinogen Persisting Perception Disorder (HPPD) Case Report.
A side effect associated with the use of LSD is the return of perceptual disturbances which anteriorly emerged during intoxication, despite absence of present use. Here we present the case of a patient with a previous history of sporadic and recreational cannabis, alcohol and LSD consumption who reported LSD associated "Alice in Wonderland Syndrome" (AIWS) or Todd's syndrome. AIWS is basically characterized by four frequent visual illusions: macropsia, micropsia, pelopsia and teleopsia. AIWS only appeared during LSD consumption and continued after LSD suspension, namely, Hallucinogen Persisting Perception Disorder (HPPD). This phenomenon did not cause a major functional impairment but provoked sufficient worry and concern due to its persistent continuation. The patient refused medical treatment and continued psychiatric follow-up. At the one year follow-up he reported complete remission. To the best of our knowledge this is the first reported case of AIWS which persist after LSD interruption (HPPD) in the professional literature. Reasons for this intriguing, benign, reversible and apparently harmless side effect are proposed.


Variations in Morning Serum Cortisol Levels Based on Sex and Pubertal Status.
Patients with suspected adrenal insufficiency undergo screening with a serum morning cortisol level and confirmatory testing with an adrenocorticotropic hormone (ACTH) stimulation test. However, much of the data collected to determine appropriate values for morning cortisol levels are derived from adult populations and may not accurately represent pediatric physiology. The purpose of this study was to evaluate the mean morning cortisol level in the pediatric population based on pubertal status and sex in order to better understand such influences on laboratory evaluation of adrenal insufficiency. A retrospective chart review was conducted using electronic medical records of patients seen at Children's Mercy Kansas City from 11/01/2007 to 11/01/2017. Patients between 2 and 18 years of age who had pubertal staging assessed by a pediatric endocrinologist and confirmed adrenal sufficiency by high-dose ACTH stimulation testing were included. Two-sample Wilcoxon rank sum (Mann-Whitney) tests or t tests were used to compare morning cortisol levels between females and males - both independent of Tanner stage and by Tanner stage. Multivariable regression models were used to evaluate associations among covariates on two outcomes: morning cortisol levels and peak cortisol values with ACTH stimulation. Morning cortisol levels were greater in females than males independent of Tanner staging (p = 0.0054) and also in Tanner stage 1 (p = 0.0042). No differences in mean morning cortisol levels between Tanner stage 2-5 females and males were found (p = 0.4652). Morning cortisol levels were not significantly different between Tanner 1 patients and Tanner 2-5 patients independent of sex (p = 0.0575). Sex was predictive of serum morning cortisol levels (p = 0.015), and morning cortisol levels were predictive of peak cortisol levels during ACTH stimulation testing (p < 0.001). These data suggest that different normative cortisol values may need to be established for pediatric females and males, and by pubertal status. Larger prospective studies are needed to evaluate the role of sex and pubertal status in identifying adrenal insufficiency in the -pediatric population.


Comprehensive hormone profiling of the developing seeds of four grain legumes.
Developmental context and species-specific hormone requirements are of key importance in the advancement of in vitro protocols and manipulation of seed development. Improvement of in vitro tissue and cell culture protocols in grain legumes such as embryo rescue, interspecific hybridization, and androgenesis requires an understanding of the types, activity, and balance of hormones within developing seeds. Towards this goal, the concentration of auxin, cytokinin, gibberellin, and abscisic acid (ABA) and their precursors and derivatives were measured in the developing seeds of field pea (Pisum sativum L.), chickpea (Cicer arietinum L.), lentil (Lens culinaris Medik.), and faba bean (Vicia faba L.) from 4 days after anthesis until 8 days after reaching maximum fresh weight. The importance of developmental context (developmental time and space) is demonstrated in both the differences and similarities between species for hormone profiles, especially with regard to cytokinin and ABA biosynthesis during the embryo formation. Auxin and its conjugates are significant during the pattern formation stage of all legumes; however, IAA-Asparagine appears important in the Vicieae species and its concentrations are greater than IAA from the globular stage of embryo development on in multi-seed fruits. Finally, the significance of non-polar gibberellins during lentil seed development is highlighted.


Obesity prevalence in Nepal: public health challenges in a low-income nation during an alarming worldwide trend.
The future toll of the obesity epidemic will likely hit hardest in low- and middle-income countries. Ongoing urbanization promotes risk factors including sedentary lifestyle and fat- and sugar-laden diets. Low-income countries like Nepal experience a double disease burden: infectious diseases as well as rising incidence of noncommunicable diseases (e.g., cardiovascular disease and diabetes mellitus) frequently characterized by obesity. Nepal currently directs efforts towards curing disease but pays little attention to preventive actions. This article highlights obesity prevalence in Nepal, delineates the challenges identified by our pilot study (including low health literacy rates), and suggests strategies to overcome this trend.


Principle considerations for the risk assessment of sprayed consumer products.
In recent years, the official regulation of chemicals and chemical products has been intensified. Explicitly for spray products enhanced requirements to assess the consumers'/professionals' exposure to such product type have been introduced. In this regard the Aerosol-Dispensers-Directive (75/324/EEC) with obligation for marketing aerosol dispensers, and the Cosmetic-Products-Regulation (1223/2009/EC) which obliges the insurance of a safety assessment, have to be mentioned. Both enactments, similar to the REACH regulation (1907/2006/EC), require a robust chemical safety assessment. From such assessment, appropriate risk management measures may be identified to adequately control the risk of these chemicals/products to human health and the environment when used. Currently, the above-mentioned regulations lack the guidance on which data are needed for preparing a proper hazard analysis and safety assessment of spray products. Mandatory in the process of inhalation risk and safety assessment is the determination and quantification of the actual exposure to the spray product and more specifically, its ingredients. In this respect the current article, prepared by the European Aerosol Federation (FEA, Brussels) task force "Inhalation Toxicology", intends to introduce toxicological principles and the state of the art in currently available exposure models adapted for typical application scenarios. This review on current methodologies is intended to guide safety assessors to better estimate inhalation exposure by using the most relevant data.


[Assessment of the Family Health Program in municipalities in Santa Catarina State, Brazil].
This evaluative study aimed to classify the municipalities (counties) in the State of Santa Catarina, Brazil, as to implementation of the Family Health Program, using three categories (satisfactory, intermediate, and unsatisfactory) and two different periods (2001 e 2004). The indicators were coverage, evidence of change in the healthcare model, and impact. According to the results, coverage increased significantly from 2001 to 2004 (p<0.0001). Evidence of change in the healthcare model and impact did not change significantly. In 2001, 48% of the municipalities showed an unsatisfactory situation, as compared to 45% with intermediate and 7% satisfactory performance, while in 2004 the figures were 29%, 58%, and 13%, respectively. Improvement in classification from 2001 to 2004 was statistically significant (p=0.0061). Analysis of the data as a whole showed that changes in the final classification resulted basically from the coverage indicator, demonstrating effective expansion in access. However, these indicators did not show significant changes in the healthcare model during the period analyzed.


Self-Perception of HIV Risk and Candidacy for Pre-Exposure Prophylaxis Among Men Who Have Sex with Men Testing for HIV at Commercial Sex Venues in New York City.
The purpose of this study was to evaluate the ability of men who have sex with men (MSM) testing for HIV at commercial sex venues to assess the following: their candidacy for pre-exposure chemoprophylaxis (PrEP) as defined by meeting entry criteria for the iPrEx (Iniciativa Profilaxis Pre-Exposición) phase III clinical trial of PrEP, and their perception of their own HIV risk and candidacy for PrEP. Interviewers surveyed 629 MSM at three NYC commercial sex venues from June 2011 through June 2012. Questions focused on demographics, sexual activity, and drug use in the three months prior to testing, as well as perceived risk of HIV acquisition and perceived candidacy for PrEP use. Data were analyzed by Chi square and Fisher's exact test. Results show that a majority of clients (80.3%) met entry criteria for the iPrEX. Most of these men (78.0%), however, did not perceive their risk to be significant enough to warrant PrEP use (P=.000). Factors were identified which associated with a risk perception that correlated with eligibility for iPrEX.


Views of older people in the Netherlands on wellbeing: A Q-methodology study.
Population ageing and restricted budgets result in the need for an efficient allocation of scarce resources in care services for older people. As these services tend to address more than only health, diverse wellbeing measures have been developed to assess their benefits in economic evaluations. These measures are grounded in research on wellbeing of older people and its determinants. Little is known about possible heterogeneity in this context and the extent to which wellbeing measures cover the aspects of wellbeing that are most important to older people with different views on wellbeing. We conducted a Q-methodology study between December 2016 and October 2017 to investigate the variety in views among people aged 65 and older in the Netherlands on what is important to their wellbeing. A purposive sample of 53 respondents ranked 34 opinion statements according to importance to their wellbeing and explained their ranking during a follow-up interview. Data were analysed using by-person factor analysis to identify common patterns in the rankings of the statements. Five distinct views were extracted in which different aspects were considered important: (I) health, financial security and a life partner; (II) family, support and physical functioning (III); autonomy, mental health and helping others; (IV) social contacts, support, mental health and religion; and (V) a life partner, social contacts, living environment and adaptation. This heterogeneity in views of older people on what constitutes wellbeing supports the use of person-centered approaches in care services for older people. Arguably, (evaluations of) policies and services for older people should take this plurality into consideration.


Cognitive Safety is Largely Ignored in Clinical Drug Trials: A Study of Registered Study Protocols.
The number of reports on suspected drug-induced memory impairment submitted to the US Food and Drug Administration increased 30-fold from 2000 to 2022. Drugs are the most common cause of reversible dementia. However, there is very little research on drug-induced cognitive impairment. The aim of this study was to investigate if and how an assessment of cognitive safety was included in recent, registered, controlled, clinical drug trials. The clinical trials registry ( www. gov ) was searched for randomized controlled clinical trials with available study protocols. After excluding irrelevant trials such as surgical procedures, local or short-term treatment, and dietary supplements, 803 trials were included in this study. The protocols were manually reviewed for information on if, and how, cognitive safety had been assessed. Trial drugs were categorized into those targeting the central nervous system or not, as well as older and newer drugs. Methods used for the assessment of cognitive function were categorized into questionnaires, screening instruments, and neuropsychological tests. If the trial results were published, we examined whether the publication contained any data on cognitive safety that had emerged from the trial. The start dates of the screened trials ranged from 31 July, 2009, to 4 April, 2021. Out of the 803 trials, 52 (6.5%) actively assessed cognitive safety. The remaining trials relied solely on spontaneous reporting. Of 429 trials studying a new drug, 32 (7.5%) actively assessed cognitive safety. One hundred and fifty-eight trials examined drugs intended to, or known to have, pharmacological effects on the central nervous system. Of these, 21 (13.5%) assessed cognitive safety. Most of the trials that assessed cognitive safety used either crude screening tools or questionnaires. Cognitive safety is largely ignored by recent controlled clinical trials. This applies even to trials assessing new drugs and trials assessing central nervous system drugs. There is an urgent need for drug manufacturers, regulatory authorities, and the medical profession to address the cognitive safety of drugs.


Serenoa repens (Permixon®) reduces intra- and postoperative complications of surgical treatments of benign prostatic hyperplasia.
The aim of this study was to evaluate the efficacy of pre-treatment with Serenoa repens (Permixon®) in reducing intra- and postoperative complications of surgical treatments of benign prostatic hyperplasia (BPH). A total of 144 patients with BPH, who were candidates for transurethral resection of the prostate (TURP) and open prostatectomy (OP), were admitted to the study and randomized to receive either a daily pre-treatment with 320 mg of Serenoa repens, Permixon® for two months prior to surgery (G1) or to undergo surgery without any pre-treatment (G2). Preoperative and postoperative courses were carefully monitored for each patient. Of the 144 patients, 24 dropped out of the trial: 10 in G1 and 14 in G2. In G1, the duration of surgery was significantly shorter than in G2 (59.8 min vs. 77.6 min, respectively; P<0.001), no intraoperative complications were observed (0% vs. 15% in the control group; P=0.001) and transfusion needs were remarkably lower (0% in G1vs. 38.33% in G2; P<0.001). The postoperative course was significantly more favorable in G1 (with a shorter duration of catheterization, 64.95 h vs. 91.7 h in G2; P<0.001). There was also a significant change in hemodynamic parameters and the length of hospitalization was significantly shorter in the pretreated group (5.92 d in G1 vs. 7.92 d in G2; P<0.001). These data suggest that pre-treatment with Serenoa repens before surgery (TURP or OP) for BPH is effective in reducing intra- and postoperative complications.


Embryonic stem cells facilitate the isolation of persistent clonal cardiovascular progenitor cell lines and leukemia inhibitor factor maintains their self-renewal and myocardial differentiation potential in vitro.
Compelling evidence for the existence of somatic stem cells in the heart of different mammalian species has been provided by numerous groups; however, so far it has not been possible to maintain these cells as self-renewing and phenotypically stable clonal cell lines in vitro. Thus, we sought to identify a surrogate stem cell niche for the isolation and persistent maintenance of stable clonal cardiovascular progenitor cell lines, enabling us to study the mechanism of self-renewal and differentiation in these cells. Using postnatal murine hearts with a selectable marker as the stem cell source and embryonic stem cells and leukemia inhibitory factor (LIF)-secreting fibroblasts as a surrogate niche, we succeeded in the isolation of stable clonal cardiovascular progenitor cell lines. These cell lines self-renew in an LIF-dependent manner. They express both stemness transcription factors Oct4, Sox2, and Nanog and early myocardial transcription factors Nkx2.5, GATA4, and Isl-1 at the same time. Upon LIF deprivation, they exclusively differentiate to functional cardiomyocytes and endothelial and smooth muscle cells, suggesting that these cells are mesodermal intermediates already committed to the cardiogenic lineage. Cardiovascular progenitor cell lines can be maintained for at least 149 passages over 7 years without phenotypic changes, in the presence of LIF-secreting fibroblasts. Isolation of wild-type cardiovascular progenitor cell lines from adolescent and old mice has finally demonstrated the general feasibility of this strategy for the isolation of phenotypically stable somatic stem cell lines.


Evaluation of a 12-color flow cytometry panel to study lymphocyte, monocyte, and dendritic cell subsets in humans.
Monitoring changes in human immune cell populations such as lymphocytes, monocytes, and dendritic cells (DCs) during infectious diseases like human immunodeficiency virus (HIV) is crucial. However, difficulties to identify rare or heterogeneous cell populations can be limiting. For example, to accurately measure DC subsets, eight flow cytometry parameters are ideal. The aim of this work was to analyze the phenotype of human lymphocyte, monocyte, and DC subsets using a single 12-color flow cytometry panel. After erythrocyte lysis, blood from healthy human volunteers was washed and labeled with a cocktail of 12 antibodies. Samples were analyzed on a Becton-Dickinson FACSAria equipped with three lasers. Data were compared with lineage-specific panels using 5-8 Ab combinations per lineage. Acquired data were analyzed using FlowJo software. Our 12-color panel allows for the identification of the following major subsets of circulating cells in a single tube: CD4+ and CD8+ T lymphocytes, B lymphocytes, NK cells, NKT cells, monocyte subsets (CD14 and/or CD16), and five nonoverlapping HLA-DR+Lin- subsets: CD34+ hematopoietic stem cells, CD123+ plasmacytoid DC, and three subsets of CD11c+ myeloid DC expressing either CD16, CD1c (BDCA-1), or CD141 (BDCA-3). We have developed a single flow cytometry panel that allows for simultaneous detection of the lymphocyte and monocyte cell populations and all known DC subsets. Studying these major players of the immune system in one single panel may give us a broader view of the immune response during HIV infection and the ability to better define the role of individual cell types in Acquired Immune Deficiency Syndrome (AIDS) pathogenesis. (c) 2010 International Society for Advancement of Cytometry.


Engineering microbes for tolerance to next-generation biofuels.
A major challenge when using microorganisms to produce bulk chemicals such as biofuels is that the production targets are often toxic to cells. Many biofuels are known to reduce cell viability through damage to the cell membrane and interference with essential physiological processes. Therefore, cells must trade off biofuel production and survival, reducing potential yields. Recently, there have been several efforts towards engineering strains for biofuel tolerance. Promising methods include engineering biofuel export systems, heat shock proteins, membrane modifications, more general stress responses, and approaches that integrate multiple tolerance strategies. In addition, in situ recovery methods and media supplements can help to ease the burden of end-product toxicity and may be used in combination with genetic approaches. Recent advances in systems and synthetic biology provide a framework for tolerance engineering. This review highlights recent targeted approaches towards improving microbial tolerance to next-generation biofuels with a particular emphasis on strategies that will improve production.


Dysregulation of CRMP2 Post-Translational Modifications Drive Its Pathological Functions.
Collapsin response mediator proteins (CRMPs) are a family of ubiquitously expressed, homologous phosphoproteins best known for coordinating cytoskeletal formation and regulating cellular division, migration, polarity, and synaptic connection. CRMP2, the most studied of the five family members, is best known for its affinity for tubulin heterodimers and function in regulating the microtubule network. These functions are tightly regulated by post-translational modifications including phosphorylation, SUMOylation, oxidation, and O-GlcNAcylation. While CRMP2's physiological functions rely mostly on its non-phosphorylated state, dysregulation of CRMP2 phosphorylation and SUMOylation has been reported to be involved in the pathophysiology of multiple diseases including cancer, chronic pain, spinal cord injury, neurofibromatosis type 1, and others. Here, we provide a consolidated update on what is known about CRMP2 signaling and function, first focusing on axonal growth and neuronal polarity, then illustrating the link between dysregulated CRMP2 post-translational modifications and diseases. We additionally discuss the roles of CRMP2 in non-neuronal cells, both in the CNS and regions of the periphery. Finally, we offer thoughts on the therapeutic implications of modulating CRMP2 function in a variety of diseases.


Taxonomic and Biological Characterization of Steinernema rarum Found in the Southeastern United States.
Two Steinernema isolates found in Louisiana and Mississippi were later identified as isolates of S. rarum. DNA sequences of ITS regions of the United States isolates are identical with sequences of Argentinean S. rarum strains Samiento and Noetinger and differ by two bases from the Arroyo Cabral isolate from Córdoba, Argentina. SEM observations revealed several new structures in the isolates from the US: female face views have a hexagonal-star perioral disc and eye-shaped lips; some females do not have cephalic papillae; lateral fields of infective juveniles are variable; there are two openings observed close to the posterior edge of the cloaca. Virulence of the US isolates to Anthonomus grandis, Diaprepes abbreviatus, Solenopsis invicta, Coptotermes formosanus, Agrotis ipsilon, Spodoptera frugiperda, and Trichoplusia ni and reproductive potential were evaluated in comparison with other heterorhabditid and steinernematid nematodes. Results such as particularly high virulence to S. frugiperda indicate that the biocontrol potential of the new S. rarum strains merits further study.


"Did I Say Cherry?" Error Patterns on a Blocked Cyclic Naming Task for Bilingual Children With and Without Developmental Language Disorder.
Purpose Using a blocked cyclic picture-naming task, we compared accuracy and error patterns across languages for Spanish-English bilingual children with and without developmental language disorder (DLD). Method Pictured stimuli were manipulated for semantic similarity across two (Same and Mixed) category contexts. Children's productions were scored off-line for accuracy, error frequency, and error type. Results Typically developing children were more accurate and produced fewer errors than their peers with DLD; however, this was moderated by task language and semantic context. Children were generally more accurate when naming pictures in English compared to Spanish and in the Mixed-category context compared to the Same-category context. Analyses of error types further showed that children with less English language exposure specifically produced more nonresponses in English than in Spanish. Children with DLD produced more of each error type than their typically developing peers, particularly in Spanish. Conclusions Regardless of language ability, bilingual children demonstrated greater difficulty with lexical retrieval for pictured items in the semantically related context than in the unrelated context. However, bilingual children with DLD produced more errors of all types than is typical for children developing more than one language. Their greater error rates are not secondary to limited second language exposure but instead reflect deficits inherent to the nature of language impairment. Results from this study are discussed using a framework of semantic constraint, where we propose that because bilingual children with DLD have impoverished semantic networks, and this knowledge insufficiently constrains activation for lexical selection, thereby increasing error production.


Humidity and pressure sensor based on internal reflection.
A low-cost humidity and pressure optical sensor, based on the internal reflection phenomenon, is presented. It takes advantage of the phase difference acquired by s- and p-polarized light undergoing internal reflection to generate an easily detectable minimum in the reflected profile, in a position corresponding to the critical angle. The apparatus presents good sensitivity to relative humidity changes above 70% and a response time below one second. The same device is also capable of measuring changes in pressure and can be used as a vacuum gauge between 1 and 1000 mbar.


AP-2alpha knockout mice exhibit optic cup patterning defects and failure of optic stalk morphogenesis.
Appropriate development of the retina and optic nerve requires that the forebrain-derived optic neuroepithelium undergoes a precisely coordinated sequence of patterning and morphogenetic events, processes which are highly influenced by signals from adjacent tissues. Our previous work has suggested that transcription factor activating protein-2 alpha (AP-2alpha; Tcfap2a) has a non-cell autonomous role in optic cup (OC) development; however, it remained unclear how OC abnormalities in AP-2alpha knockout (KO) mice arise at the morphological and molecular level. In this study, we show that patterning and morphogenetic defects in the AP-2alpha KO optic neuroepithelium begin at the optic vesicle stage. During subsequent OC formation, ectopic neural retina and optic stalk-like tissue replaced regions of retinal pigment epithelium. AP-2alpha KO eyes also displayed coloboma in the ventral retina, and a rare phenotype in which the optic stalk completely failed to extend, causing the OCs to be drawn inward to the midline. We detected evidence of increased sonic hedgehog signaling in the AP-2alpha KO forebrain neuroepithelium, which likely contributed to multiple aspects of the ocular phenotype, including expansion of PAX2-positive optic stalk-like tissue into the OC. Our data suggest that loss of AP-2alpha in multiple tissues in the craniofacial region leads to severe OC and optic stalk abnormalities by disturbing the tissue-tissue interactions required for ocular development. In view of recent data showing that mutations in human TFAP2A result in similar eye defects, the current findings demonstrate that AP-2alpha KO mice provide a valuable model for human ocular disease.


The Evolution of Mass Cell Suicide in Bacterial Warfare.
Behaviors that cause the death of an actor are typically strongly disfavored by natural selection, and yet many bacteria undergo cell lysis to release anti-competitor toxins [1-5]. This behavior is most easily explained if only a small proportion of cells die to release toxins and help their clonemates, but the frequency of cells that actually lyse during bacterial warfare is unknown. The challenge is finding a way to distinguish cells that have undergone programmed suicide from those that were simply killed by a competitor's toxin. We developed a two-color fluorescence reporter assay in Escherichia coli to overcome this problem. This revealed conditions where nearly all cells undergo programmed lysis. Specifically, adding a DNA-damaging toxin (DNase colicin) from another strain induced mass cell suicide where ∼85% of cells lysed to release their own toxins. Time-lapse 3D confocal microscopy showed that self-lysis occurs locally at even higher frequencies (∼94%) at the interface between toxin-producing colonies. By exposing E. coli that do not perform lysis to the DNase colicin, we found that mass lysis occurs when cells are going to die anyway from toxin exposure. From an evolutionary perspective, this renders the behavior cost-free as these cells have zero reproductive potential. This helps to explain how mass cell suicide can evolve, as any small benefit to surviving clonemates can lead to this retaliatory strategy being favored by natural selection. Our findings have parallels to the suicidal attacks of social insects [6-9], which are also performed by individuals with low reproductive potential.


Randomized Comparison of Helmet CPAP Versus High-Flow Nasal Cannula Oxygen in Pediatric Respiratory Distress.
The current study aimed to compare the efficacy and safety of 2 noninvasive respiratory support methods, which included helmet CPAP and high-flow nasal cannula (HFNC) in children with respiratory distress admitted to a pediatric intermediate care unit. This study was a prospective observational study conducted on children with respiratory distress (age 1-24 months) who were admitted to our acute and emergency operative unit. All included subjects were randomly treated with helmet CPAP or HFNC in a 1:1 fashion until their clinical picture, oxygen saturation, and arterial blood gas (ABG) parameters resolved. The efficiencies of helmet CPAP and HFNC were evaluated by breathing frequency, SpO2 , ABG pH, ABG PaCO2 , ABG PaO2 , and PaO2 /FIO2 , recorded once at baseline and then after 1 and 6 h of treatment. Both noninvasive respiratory support modalities were compared with a control group of subjects with respiratory distress under standard therapeutic pharmaceutical protocols. We found that both helmet CPAP and HFNC were efficient in improving the clinical conditions of subjects with mild-to-moderate respiratory distress, although clinical response to helmet CPAP was more efficient and rapid compared with HFNC. Children who received respiratory support had a better clinical course in terms of hospitalization, days of intravenous rehydration therapy, and days of drug administration compared with the control group (P < .001). Based on our knowledge, the present study is the first research comparing the effects of CPAP and HFNC in respiratory distress resolution in a pediatric intermediate care setting. It aims to identify the most efficient treatment to avoid pediatric ICU admissions and endotracheal intubation and reduce the administration of drugs and days of hospitalization.


Carpatizine, a novel bridged oxazine derivative generated by non-enzymatic reactions.
Carpatizine (1), a new bridged oxazine derivative, was isolated from a marine-derived Streptomyces strain SNE-011. The structure was fully determined by spectroscopic analysis, ECD calculations and chemical methods. A plausible non-enzymatic reaction mechanism from daryamide D leading to carpatizine was presented, which was confirmed by chemical transformation.


Neighbourhood disadvantage and individual adversities in adolescence and total alcohol consumption up to mid-life-Results from the Northern Swedish Cohort.
This study tests if neighbourhood socioeconomic disadvantage and family social and material adversities during adolescence are independently related to total alcohol consumption from adolescence through to mid-life. Self-reports from the Northern Swedish Cohort (effective sample=950) at ages 16, 18, 21, 30 and 42 was combined with register data on the socioeconomic composition of neighbourhoods at age 16. Total volume of alcohol consumed between age 16-42 was estimated based on the five survey waves, and self-reported social and material adversities were computed as composite variables. Neighbourhood socioeconomic disadvantage at age 16 was associated with alcohol consumption age 16-42 for men but not for women. Social adversities at age 16 were associated with alcohol consumption age 16-42 for both women and men, but material adversity or parental class was not. In conclusion, neighbourhood socioeconomic disadvantage in adolescence has a significant relationship with later alcohol consumption among men, even independently from individual factors. On family level, social factors but not socioeconomic factors in adolescence independently predict later alcohol consumption.


Attention-deficit hyperactivity disorder.
Attention-deficit hyperactivity disorder (ADHD), like other psychiatric disorders, represents an evolving construct that has been refined and developed over the past several decades in response to research into its clinical nature and structure. The clinical presentation and course of the disorder have been extensively characterised. Efficacious medication-based treatments are available and widely used, often alongside complementary psychosocial approaches. However, their effectiveness has been questioned because they might not address the broader clinical needs of many individuals with ADHD, especially over the longer term. Non-pharmacological approaches to treatment have proven less effective than previously thought, whereas scientific and clinical studies are starting to fundamentally challenge current conceptions of the causes of ADHD in ways that might have the potential to alter clinical approaches in the future. In view of this, we first provide an account of the diagnosis, epidemiology, and treatment of ADHD from the perspective of both the fifth edition of the Diagnostic and Statistical Manual of Mental Disorders and the eleventh edition of the International Classification of Diseases. Second, we review the progress in our understanding of the causes and pathophysiology of ADHD on the basis of science over the past decade or so. Finally, using these discoveries, we explore some of the key challenges to both the current models and the treatment of ADHD, and the ways in which these findings can promote new perspectives.


Implementation of a mHealth solution to remotely monitor patients on a cardiac surgical waiting list: service evaluation.
The emergence of COVID-19 resulted in postponement of nonemergent surgical procedures for cardiac patients in London. mHealth represented a potentially viable mechanism for highlighting deteriorating patients on the lengthened cardiac surgical waiting lists. To evaluate the deployment of a digital health solution to support continuous triaging of patients on a cardiac surgical waiting list. An NHS trust utilized an app-based mHealth solution (Huma Therapeutics) to help gather vital information on patients awaiting cardiac surgery (valvular and coronary surgery). Patients at a tertiary cardiac center on a waiting list for elective surgery were given the option to be monitored remotely via a mobile app until their date of surgery. Patients were asked to enter their symptoms once a week. The clinical team monitored this information remotely, prompting intervention for those patients who needed it. Five hundred and twenty-five patients were on boarded onto the app. Of the 525 patients using the solution, 51 (9.71%) were identified as at risk of deteriorating based on data captured via the remote patient monitoring platform and subsequently escalated to their respective consultant. 81.7% of patients input at least one symptom after they were on boarded on the platform. Although not a generalizable study, this change in practice clearly demonstrates the feasibility and potential benefit digital remote patient monitoring can have in triaging large surgical wait lists, ensuring those that need care urgently receive it. We recommend further study into the potential beneficial outcomes from preoperative cardiac mHealth solutions.


A licorice ethanolic extract with peroxisome proliferator-activated receptor-gamma ligand-binding activity affects diabetes in KK-Ay mice, abdominal obesity in diet-induced obese C57BL mice and hypertension in spontaneously hypertensive rats.
The metabolic syndrome, including type 2 diabetes, insulin resistance, obesity/abdominal obesity, hypertension and dyslipidemia, is a major public health problem. Peroxisome proliferator-activated receptor-gamma (PPAR-gamma) ligands such as thiazolidinediones are effective against this syndrome. In this study, we showed that nonaqueous fractions of licorice (Glycyrrhiza uralensis Fisher) extracted with ethanol, ethyl acetate and acetone, but not an aqueous extract, had PPAR-gamma ligand-binding activity with a GAL4-PPAR-gamma chimera assay. Some prenylflavonoids including glycycoumarin, glycyrin, dehydroglyasperin C and dehydroglyasperin D, a newly found compound, were identified as active compounds with PPAR-gamma ligand-binding activity in the nonaqueous fraction of licorice. A licorice ethanolic extract contained these four active compounds at a total concentration of 16.7 g/100 g extract. Feeding the licorice ethanolic extract at 0.1-0.3 g/100 g diet [approximately 100 to 300 mg/(kg body x d)] for 4 wk decreased (P < 0.05) blood glucose level in younger (6 wk old) and older (13 wk old) diabetic KK-Ay mice and reduced (P < 0.05) weights of intra-abdominal adipose tissues in high fat diet-induced obese C57BL mice. An increase in blood pressure in spontaneously hypertensive rats was suppressed (P < 0.01) by 3 wk of oral administration of the licorice ethanolic extract at 300 mg/(kg body x d). These findings indicate that licorice ethanolic extract is effective in preventing and ameliorating diabetes, ameliorating abdominal obesity and preventing hypertension, and suggest that licorice ethanolic extract would be effective in preventing and/or ameliorating the metabolic syndrome.


The nuclear DNA base 5-hydroxymethylcytosine is present in Purkinje neurons and the brain.
Despite the importance of epigenetic regulation in neurological disorders, little is known about neuronal chromatin. Cerebellar Purkinje neurons have large and euchromatic nuclei, whereas granule cell nuclei are small and have a more typical heterochromatin distribution. While comparing the abundance of 5-methylcytosine in Purkinje and granule cell nuclei, we detected the presence of an unusual DNA nucleotide. Using thin-layer chromatography, high-pressure liquid chromatography, and mass spectrometry, we identified the nucleotide as 5-hydroxymethyl-2'-deoxycytidine (hmdC). hmdC constitutes 0.6% of total nucleotides in Purkinje cells, 0.2% in granule cells, and is not present in cancer cell lines. hmdC is a constituent of nuclear DNA that is highly abundant in the brain, suggesting a role in epigenetic control of neuronal function.


Recent initiatives and strategies to developing new drugs for tropical parasitic diseases.
Despite the toll of tropical parasitic diseases on human life in the developing world, present therapies still rely on drugs developed decades ago. In many cases, the clinical usefulness of these compounds is limited due to poor efficacy, toxicity and the constant attrition of drug resistance. The absence of a profit incentive regarding diseases afflicting the very poor has resulted in a lack of investment by the pharmaceutical industry in new chemotherapies. Given this background, this review addresses what alternative economic and scientific strategies have been implemented to procure novel drugs. The latest chemical, genetic and screening technologies to discover and develop drugs for tropical parasitic diseases are reviewed. In many cases these strategies are being implemented within the framework of public-private partnerships established to sustain dynamic drug development portfolios. Examples of public-private partnerships and their portfolios are discussed. Further, the contribution of dedicated academic screening centres to target discovery and preclinical prosecution of new small molecules is also highlighted. In every case, the latest scientific literature is cited, but also relevant press releases and website information to indicate the present vitality in the field. The tools, institutions and consortia are now in place and evolving to deliver new pharmaceuticals. Short-term results have already been realised in the clinic, mainly through the provision of new formulations of existing drugs. Long-term and consistent investment will be required, however, to identify, develop and clinically validate new chemical entities.


Prognosis of patients with peritoneal metastatic colorectal cancer given systemic therapy: an analysis of individual patient data from prospective randomised trials from the Analysis and Research in Cancers of the Digestive System (ARCAD) database.
Patients with peritoneal metastatic colorectal cancer have reduced overall survival compared with patients with metastatic colorectal cancer without peritoneal involvement. Here we further investigated the effect of the number and location of metastases in patients receiving first-line systemic chemotherapy. We analysed individual patient data for previously untreated patients enrolled in 14 phase 3 randomised trials done between 1997 and 2008. Trials were included if protocols explicitly pre-specified and solicited for patients with peritoneal involvement in the trial data collection process or had done a formal peritoneum-focused review of individual pre-treatment scans. We used stratified multivariable Cox models to assess the prognostic associations of peritoneal metastatic colorectal cancer with overall survival and progression-free survival, adjusting for other key clinical-pathological factors (age, sex, Eastern Cooperative Oncology Group (ECOG) performance score, primary tumour location [colon vs rectum], previous treatment, and baseline BMI). The primary endpoint was difference in overall survival between populations with and without peritoneal metastases. Individual patient data were available for 10 553 patients. 9178 (87%) of 10 553 patients had non-peritoneal metastatic colorectal cancer (4385 with one site of metastasis, 4793 with two or more sites of metastasis), 194 (2%) patients had isolated peritoneal metastatic colorectal cancer, and 1181 (11%) had peritoneal metastatic colorectal cancer and other organ involvement. These groups were similar in age, ethnic origin, and use of targeted treatment. Patients with peritoneal metastatic colorectal cancer were more likely than those with non-peritoneal metastatic colorectal cancer to be women (565 [41%] of 1371 vs 3312 [36%] of 9169 patients; p=0·0003), have colon primary tumours (1116 [84%] of 1334 patients vs 5603 [66%]; p<0·0001), and have performance status of 2 (136 [10%] vs 521 [6%]; p<0·0001). We recorded a higher proportion of patients with mutated BRAF in patients with peritoneal-only (eight [18%] of 44 patients with available data) and peritoneal metastatic colorectal cancer with other sites of metastasis (34 [12%] of 289), compared with patients with non-peritoneal metastatic colorectal cancer (194 [9%] of 2230; p=0·028 comparing the three groups). Overall survival (adjusted HR 0·75, 95% CI 0·63-0·91; p=0·003) was better in patients with isolated non-peritoneal sites than in those with isolated peritoneal metastatic colorectal cancer. Overall survival of patients with two of more non-peritoneal sites of metastasis (adjusted HR 1·04, 95% CI 0·86-1·25, p=0.69) and those with peritoneal metastatic colorectal cancer plus one other site of metastasis (adjusted HR 1·10, 95% CI 0·89-1·37, p=0·37) was similar to those with isolated peritoneal metastases. Compared with patients with isolated peritoneal metastases, those with peritoneal metastases and two or more additional sites of metastasis had the shortest survival (adjusted HR 1·40; CI 1·14-1·71; p=0·0011). Patients with peritoneal metastatic colorectal cancer have significantly shorter overall survival than those with other isolated sites of metastases. In patients with several sites of metastasis, poor survival is a function of both increased number of metastatic sites and peritoneal involvement. The pattern of metastasis and in particular, peritoneal involvement, results in prognostic heterogeneity of metastatic colorectal cancer. None.


Multimodal particle size distributions emitted from HFA-134a solution pressurized metered-dose inhalers.
The purpose of this research was to investigate the measurement and in vitro delivery implications of multimodal distributions, occurring near or in the respirable range, emitted from pressurized metered-dose inhalers (pMDIs). Particle size distributions of solution pMDIs containing hydrofluoroalkane-134a (HFA-134a) and ethanol were evaluated using 2 complementary particle-sizing methods: laser diffraction (LD) and cascade impaction (CI). Solution pMDIs were formulated from mixtures of HFA-134a (50%-97.5% wt/wt) and ethanol. A range of propellant concentrations was selected for a range of vapor pressures. The fluorescent probe, Rhodamine B, was included for chemical analysis. The complementary nature of LD and CI allowed identification of 2 dominant particle size modes at 1 and 10 micro m or greater. Increasing propellant concentrations resulted in increases in the proportion of the size distributions at the 1- micro m mode and also reduced the particle size of the larger droplet population. Despite significant spatial differences and time scales of measurement between the particle-sizing techniques, the fine particle fractions obtained from LD and CI were practically identical. This was consistent with LD experiments, which showed that particle sizes did not decrease with increasing measurement distance, and may be explained by the absence of significant evaporation/disintegration of larger droplets. The fine particle fractions (FPFs) emitted from HFA-134a/ethanol solution pMDI can be predicted on the basis of formulation parameters and is independent of measurement technique. These results highlight the importance of presenting particle size distribution data from complementary particle size techniques.


CD40-mediated HIF-1α expression underlying microangiopathy in diabetic nerve pathology.
To understand the pathology and molecular signatures of microangiopathy in diabetic neuropathy, we systemically and quantitatively examined the morphometry of microvascular and nerve pathologies of sural nerves. In the endoneurium of diabetic nerves, prominent microangiopathy was observed, as evidenced by reduced capillary luminal area, increased capillary basement membrane thickness and increased proportion of fibrin(+) blood vessels. Furthermore, capillary basement membrane thickness and the proportion of fibrin(+) blood vessels were correlated with small myelinated fiber density in diabetic nerves. In diabetic nerves, there was also significant macrophage and T cell infiltration, and cluster of differentiation 40 (CD40) expression was increased. The molecular alterations observed were upregulation of hypoxia-inducible factor-1α (HIF-1α), mitogen-activated protein kinase-activated protein kinase 2 (MK2; MAPKAPK2) and phosphatase and tensin homolog (PTEN). In addition, HIF-1α was correlated with small myelinated fiber density and capillary luminal area, while both MK2 and PTEN were correlated with capillary basement membrane thickness. The molecular cascades were further demonstrated and replicated in a cell model of microangiopathy on human umbilical vein endothelial cells (HUVECs) exposed to high-glucose medium by silencing of CD40, PTEN and HIF-1α in HUVECs using shRNA. These data clarified the hierarchy of the molecular cascades, i.e. upregulation of CD40 leading to HIF-1α expression in endothelium and nerve fibers. In conclusion, this study revealed the association of microangiopathy, thrombosis and inflammatory infiltrates with nerve degeneration in diabetic nerves, demonstrating that CD40 is a key molecule for the upregulation of HIF-1α and PTEN underlying the severity of microangiopathy.


Limited detectability of linezolid-resistant Staphylococcus aureus by the Etest method and its improvement using enriched media.
The aim of this study was to evaluate Etest for detectability of linezolid-resistant meticillin-resistant Staphylococcus aureus (MRSA). The MIC of linezolid obtained by the Etest method in 18 linezolid-resistant strains of MRSA was compared with that obtained using standard agar and broth dilution methods according to Clinical and Laboratory Standards Institute guidelines. The mean linezolid MIC obtained by Etest in 18 linezolid-resistant strains of MRSA using Mueller-Hinton (MH) agar was 12.6-fold lower than that obtained by the agar dilution method, with the result that 78 % of the linezolid-resistant strains were incorrectly classified as linezolid-susceptible. The MIC of linezolid by Etest on brain-heart infusion (BHI) agar had a mean value 2.5-fold lower than that obtained by the agar dilution method, suggesting that replacing MH agar with BHI agar considerably improved the detectability of linezolid-resistant MRSA. Use of blood agar (MH agar supplemented with 5 % sheep blood) and 48 h of incubation resulted in 100 % agreement with the agar and broth dilution methods. Thus, this study revealed that the Etest on MH agar and BHI agar yielded false-negative results in a significant fraction of the linezolid-resistant MRSA. Hence, the use of blood agar and prolonged incubation is highly recommended for the accurate detection of linezolid-resistant MRSA using Etest.


Re-examining the effects of noncontingent success on self-handicapping behaviour.
Self-handicapping refers to the practice on the part of certain individuals to handicap their performance when poor performance is likely to reveal low ability. Noncontingent success (feedback that is inflated relative to performance) is more likely to promote self-handicapping behaviour than noncontingent failure (failure feedback based on false or misleading information). However, the reasons for the differing effects of these forms of performance feedback on self-handicapping behaviour remain obscure. The present study sought an explanation for the differing effects of these forms of performance feedback, testing the assumption that students high in self-handicapping behaviour would react more negatively following noncontingent success, reporting more unstable and external attributions, higher anxiety, and a greater propensity to claim handicaps than those low in self-handicapping behaviour. No differences were expected on any of these measures for high relative to low self-handicappers following either noncontingent failure or success. Participants were 72 undergraduate students, divided equally between high and low self-handicapping groups. High and low self-handicappers were assigned to one of three performance feedback conditions: noncontingent failure, success and noncontingent success. High and low self-handicappers were then given an opportunity to claim handicaps prior to completing measures of attributions and state anxiety. Subsequently, they completed 12 remote associate tasks, serving as an assessment of performance, and 16 unicursal tasks, assessing practice effort. Following noncontingent success, high self-handicappers reported greater anxiety, more unproductive attributions and claimed more handicaps than low self-handicappers. However no differences were evident for high and low self-handicappers following either noncontingent failure or success. High self-handicappers also performed poorly on the remote associates tasks and reduced practice effort on the unicursal tasks. These findings confirm the adverse effects of noncontingent success for high self-handicappers, while failing to provide evidence that noncontingent failure has any more adverse effects on high relative to low self-handicappers.


Old and New NICE Guidelines for the Evaluation of New Onset Stable Chest Pain: A Real World Perspective.
Stable chest pain is a common clinical presentation that often requires further investigation using noninvasive or invasive testing, resulting in a resource-consuming problem worldwide. At onset of 2016, the National Institute for Health and Care Excellence (NICE) published an update on its guideline on chest pain. Three key changes to the 2010 version were provided by the new NICE guideline. First, the new guideline recommends that the previously proposed pretest probability risk score should no longer be used. Second, they also recommend that a calcium score of zero should no longer be used to rule out coronary artery disease (CAD) in patients with low pretest probability. Third, the new guideline recommends that all patients with new onset chest pain should be investigated with a coronary computed tomographic angiography (CTA) as a first-line investigation. However, in real world the impact of implementation of CTA for the evaluation of new onset chest pain remains to be evaluated, especially regarding its cost effectiveness. The aim of the present report was to discuss the results of the studies supporting new NICE guideline and its comparison with European and US guidelines.


Calcium signaling and cell fate: how can Ca2+ signals contribute to wrong decisions for Chronic Lymphocytic Leukemic B lymphocyte outcome?
Ca(2+) signaling is a key regulator of B lymphocyte cell fate and defects in this signaling pathway have been reported in numerous diseases such as Chronic lymphocytic leukemia (CLL). CLL is a B cell clonal disorder characterized by the accumulation of mature monoclonal CD5(+) B cells. Although CLL could be considered to be a proliferative disease, most circulating CLL B cells are arrested in the G0 phase of the cell cycle and present both defects in calcium (Ca(2+)) homeostasis and signaling. The Ca(2+) response to antigen ligation is heterogeneous and related, in part, to defects arising from the incapacity to respond to B cell receptor (BCR) engagement (anergy), to the expression of T cell kinases (e.g. Zap70), and to the presence of negative feedback regulation by phosphatases (e.g. SHP-1). Anergic CD5(+) CLL B cells are characterized by an elevated basal Ca(2+) level, IgM/CD79 downregulation, a constitutive activation of BCR pathway kinases, and an activation of the nuclear factor of activated T cells (NF-AT). Based on the Ca(2+) response, patients are classified into three groups: unresponders, responders with apoptosis, and responders with entry in the cell cycle. Moreover, internal and direct interaction between leukemic BCR-HCDR3 epitopes at the plasma membrane and interaction between Bcl-2 and the IP3-receptor at the endoplasmic reticulum are also suspected to interfere with the intracellular Ca(2+) homeostasis in CLL-B cells. As a whole, the Ca(2+) pathway is emerging to play a key role in malignant CLL-B survival, disease progression, and last but not least, in the therapeutic response.


Prognostic role of dobutamine stress echocardiography in myocardial viability.
The aim of this review is to provide a perspective on the role of dobutamine stress echocardiography to evaluate myocardial viability and assess prognosis in patients with ischemic left ventricular dysfunction. Pooled analysis of currently available data demonstrates that dobutamine stress echocardiography provides useful information on recovery of dysfunctional but viable myocardium and prognosis. In fact, improvement of long-term prognosis may be the ultimate clinical challenge in the management of patients with ischemic left ventricular dysfunction. Dobutamine stress echocardiography can be used to assess residual ischemia and contractile reserve, which are significant determinants of prognosis. Additionally, other echocardiographic variables, such as end-diastolic wall thickness, end-systolic volume, and extent of viability, affect outcome after revascularization in patients with ischemic cardiomyopathy. Dobutamine stress echocardiography provides information on myocardial ischemia, contractile reserve, and other prognostically relevant parameters in patients with ischemic left ventricular dysfunction. This information can be used to select the optimal treatment strategy for these patients and to estimate and improve clinical outcome.


Medical marijuana: medical necessity versus political agenda.
Marijuana is classified by the Drug Enforcement Agency (DEA) as an illegal Schedule I drug which has no accepted medical use. However, recent studies have shown that medical marijuana is effective in controlling chronic non-cancer pain, alleviating nausea and vomiting associated with chemotherapy, treating wasting syndrome associated with AIDS, and controlling muscle spasms due to multiple sclerosis. These studies state that the alleviating benefits of marijuana outweigh the negative effects of the drug, and recommend that marijuana be administered to patients who have failed to respond to other therapies. Despite supporting evidence, the DEA refuses to reclassify marijuana as a Schedule II drug, which would allow physicians to prescribe marijuana to suffering patients. The use of medical marijuana has continued to gain support among states, and is currently legal in 16 states and the District of Columbia. This is in stark contrast to the federal government's stance of zero-tolerance, which has led to a heated legal debate in the United States. After reviewing relevant scientific data and grounding the issue in ethical principles like beneficence and nonmaleficence, there is a strong argument for allowing physicians to prescribe marijuana. Patients have a right to all beneficial treatments and to deny them this right violates their basic human rights.


The effect of phosphorylation on the electron capture dissociation of peptide ions.
The effect of site and frequency of phosphorylation on the electron capture dissociation of peptide ions has been investigated. The ECD of a suite of synthetic peptides (APLSFRGSLPKSYVK; one unmodified, three singly-phosphorylated, three-doubly phosphorylated, and one triply-phosphorylated); two tryptic phosphopeptides (YKVPQLEIVPN(p)SAEER, alpha-casein and FQ(p)SEEQQQTEDELQDK, beta-casein) and their unmodified counterparts, were determined over a range of ECD cathode potentials. The results show that, for doubly-charged precursor ions, the presence of phosphorylation has a deleterious effect on ECD sequence coverage. The fragmentation patterns observed suggest that for peptides with multiple basic residues, the phospho-groups exist in their deprotonated form and form salt-bridges with protonated amino acid side chains. The fragmentation observed for the acidic tryptic peptides suggested the presence of noncovalent interactions, which were perturbed on phosphorylation. Increasing the ECD electron energy significantly improves sequence coverage. Alternatively, improved sequence coverage can be achieved by performing ECD on triply-charged precursor ions. The findings are important for the understanding of gas-phase fragmentation of phosphopeptides.


The structure of liquid water beyond the first hydration shell.
To date there is a general consensus on the structure of the first coordination shells of liquid water, namely tetrahedral short range order of molecules. In contrast, little is known about the structure at longer distances and the influence of the tetrahedral molecular arrangement of the first shells on the order at these length scales. An expansion of the distance dependent excess entropy is used in this contribution to find out which molecular arrangements are important at each distance range. This was done by splitting the excess entropy into two parts: one connected to the relative position of two molecules and the other one related to their relative orientation. A transition between two previously unknown regimes in liquid water is identified at a distance of about ∼6 Å: from a predominantly orientational order at shorter distances to a regime at larger distances of up to ∼9 Å where the order is predominantly positional and molecules are distributed with the same tetrahedral symmetry as the very first molecules.


Disparities in the Context of Opportunities for Cancer Prevention in Early Life.
Persistent health disparities are a major contributor to disproportionate burden of cancer for some populations. Health disparities in cancer incidence and mortality may reflect differences in exposures to risk factors early in life. Understanding the distribution of exposures to early life risk and protective factors for cancer across different populations can shed light on opportunities to promote health equity at earlier developmental stages. Disparities may differentially influence risk for cancer during early life and create opportunities to promote health equity. Potential risk and protective factors for cancer in early life reveal patterns of disparities in their exposure. These disparities in exposures can manifest in downstream disparities in risk for cancer. These risk and protective factors include adverse childhood experiences; maternal alcohol consumption in pregnancy; childhood obesity; high or low birth weight; benzene exposure; use of assisted reproductive technologies; pesticide and insecticide exposure; isolated cryptorchidism; early pubertal timing; exposure to radiation; exposure to tobacco in utero and in early life; allergies, asthma, and atopy; and early exposure to infection. Disparities on the basis of racial and ethnic minority status, economic disadvantage, disability status, sex, geography, and nation of origin can occur in these risk and protective factors. Vulnerable populations experience disproportionally greater exposure to risk factors in early life. Addressing disparities in risk factors in early life can advance opportunities for prevention, promote health equity, and possibly reduce risk for subsequent development of cancer.


Organics captured from comet 81P/Wild 2 by the Stardust spacecraft.
Organics found in comet 81P/Wild 2 samples show a heterogeneous and unequilibrated distribution in abundance and composition. Some organics are similar, but not identical, to those in interplanetary dust particles and carbonaceous meteorites. A class of aromatic-poor organic material is also present. The organics are rich in oxygen and nitrogen compared with meteoritic organics. Aromatic compounds are present, but the samples tend to be relatively poorer in aromatics than are meteorites and interplanetary dust particles. The presence of deuterium and nitrogen-15 excesses suggest that some organics have an interstellar/protostellar heritage. Although the variable extent of modification of these materials by impact capture is not yet fully constrained, a diverse suite of organic compounds is present and identifiable within the returned samples.


Neuromotor development and the physiologic effects of positioning in very low birth weight infants.
To provide a comprehensive literature review of neuromotor development and related physiologic effects of positioning in very low birth weight infants. MEDLINE, CINHAL, Health Star, Current Contents, and the Australian Medical Index (1966-2000) databases were searched. Unpublished studies (e.g., dissertations, conference proceedings) and all relevant references listed in articles also were examined. One hundred and eighty theoretical writings, research studies, and clinical papers related to neuromotor development, the physiologic effects of positioning, and interventions to minimize or prevent short- and long-term effects of positioning in very low birth weight infants were reviewed. Studies were assessed for scientific rigor, evidence of theoretical foundation, and clinical relevance. Comparisons were made across data sources to determine the most reliable, valid, and consistent findings. Three compelling results emerged: (a) The development of posture and mobility in newborn infants requires an optimal balance between active and passive muscle tone, (b) the prone position is physiologically more beneficial for the preterm infant than supine and lateral positions, and (c) the prone position can lead to short- and long-term postural and associated developmental problems. Use of empirically tested postural interventions appropriate for an infant's gestational age, health status, and overall organizational capacity is recommended.


Effect of growth factors on cell proliferation and epithelialization in human skin.
The failure of chronic wounds to heal remains a major medical problem. Recent studies have suggested an important role for growth factors in promoting wound healing. We investigated the mitogenic effect of basic fibroblast growth factor (FGF), insulin-like growth factor-1 (IGF-1), and epidermal growth factor (EGF), comparing their effects with those of media alone (MEM) in a human skin explant model. A stable organ culture system for maintaining the histologic structure of human epidermis for 10 days in vitro was developed. DNA synthesis was measured on Days 1, 3, and 7 of organ culture using [3H]thymidine ([3H]thy) uptake and expressed as cpm/mg dry weight (mean +/- SEM). FGF, IGF-1, and EGF were each capable of stimulating [3H]thy uptake on Day 1 of culture (2372 +/- 335 FGF, 2226 +/- 193 IGF-1, 4037 +/- 679 EGF vs 1108 +/- 70 MEM, P < 0.05). IGF-1 and EGF also stimulated [3H]thy uptake on Days 3 and 7 of culture. The organ culture system was further employed to observe epidermal outgrowth. Longest keratinocyte outgrowth from the explant periphery (simulating epithelial regeneration from the wound edge) was observed on Day 7. EGF resulted in maximum stimulation of epithelial outgrowth (440 +/- 80 microns), followed by FGF (330 +/- 56 microns), IGF-1 (294 +/- 48 microns), and MEM (189 +/- 50 microns). We postulate, therefore, that FGF, IGF-1, and EGF are important mitogens for wound healing and that EGF in particular is capable of stimulating epithelialization.(ABSTRACT TRUNCATED AT 250 WORDS)