Climate services for society: origins, institutional arrangements, and design elements for an evaluation framework.
Climate services involve the generation, provision, and contextualization of information and knowledge derived from climate research for decision making at all levels of society. These services are mainly targeted at informing adaptation to climate variability and change, widely recognized as an important challenge for sustainable development. This paper reviews the development of climate services, beginning with a historical overview, a short summary of improvements in climate information, and a description of the recent surge of interest in climate service development including, for example, the Global Framework for Climate Services, implemented by the World Meteorological Organization in October 2012. It also reviews institutional arrangements of selected emerging climate services across local, national, regional, and international scales. By synthesizing existing literature, the paper proposes four design elements of a climate services evaluation framework. These design elements include: problem identification and the decision-making context; the characteristics, tailoring, and dissemination of the climate information; the governance and structure of the service, including the process by which it is developed; and the socioeconomic value of the service. The design elements are intended to serve as a guide to organize future work regarding the evaluation of when and whether climate services are more or less successful. The paper concludes by identifying future research questions regarding the institutional arrangements that support climate services and nascent efforts to evaluate them.


Toward functional pi-conjugated organophosphorus materials: design of phosphole-based oligomers for electroluminescent devices.
The photophysical, electrochemical, and optoelectronic properties of conjugated systems incorporating dibenzophosphole or phosphole moieties are described. Dibenzophosphole derivatives are not suitable materials for OLEDs due to their weak photoluminescence (PL) in the solid state and the instability of the devices. Variation of the substitution pattern of phospholes and chemical modification of their P atoms afford thermally stable derivatives, which are photo- and electroluminescent. Comparison of the optical properties of solution and thin film of thioxophospholes shows that these compounds do not form aggregates in the solid state. This property, which is also supported by an X-ray diffraction study of three novel derivatives, results in an enhancement of the fluorescence quantum yields in the solid state. In contrast, (phosphole)gold(I) complexes exhibit a broad emission in thin film, which is due to the formation of aggregates. Single- and multilayer OLEDs using these P derivatives as the emissive layer have been fabricated. The emission color of these devices and their performances vary with the nature of the P material. Interestingly, di(2-thienyl)thiooxophosphole is an efficient host for the red dopant DCJTB, and devices using the gold complexes have broad emission spectra.


Sebaceous carcinoma of the face following irradiation.
Malignant neoplasms of sebaceous glands are uncommon but generally occur about the head and neck, where these glands are concentrated. The sites of most frequent occurrence are the ocular adnexae, where sebaceous carcinoma presents as an infiltrating malignancy with a poor prognosis. Cutaneous sebaceous carcinoma is rare at other sites but has a better prognosis. We have located eight cases in the literature of sebaceous carcinoma arising in a previously irradiated field: five in the ocular adnexae and three elsewhere on the face. We here report a fourth case of recurrent sebaceous carcinoma with metastases that occurred on the face of an 82-year-old woman who had received radiation to the area for cosmetic epilation 35 years previously.


Invited Review: The spectrum of neuropathology in COVID-19.
There is increasing evidence that patients with Coronavirus disease 19 (COVID-19) present with neurological and psychiatric symptoms. Anosmia, hypogeusia, headache, nausea and altered consciousness are commonly described, although there are emerging clinical reports of more serious and specific conditions such as acute cerebrovascular accident, encephalitis and demyelinating disease. Whether these presentations are directly due to viral invasion of the central nervous system (CNS) or caused by indirect mechanisms has yet to be established. Neuropathological examination of brain tissue at autopsy will be essential to establish the neuro-invasive potential of the SARS-CoV-2 virus but, to date, there have been few detailed studies. The pathological changes in the brain probably represent a combination of direct cytopathic effects mediated by SARS-CoV-2 replication or indirect effects due to respiratory failure, injurious cytokine reaction, reduced immune response and cerebrovascular accidents induced by viral infection. Further large-scale molecular and cellular investigations are warranted to clarify the neuropathological correlates of the neurological and psychiatric features seen clinically in COVID-19. In this review, we summarize the current reports of neuropathological examination in COVID-19 patients, in addition to our own experience, and discuss their contribution to the understanding of CNS involvement in this disease.


Effects of acute tryptophan depletion on three different types of behavioral impulsivity.
While central nervous system serotonin has been implicated in a variety of problematic impulsive behaviors, biological manipulation of brain serotonin using acute tryptophan depletion for studying changes in impulsive behavior has received little attention. Using identical treatment conditions, we examined the effects of reduced serotonin synthesis for each of three matched groups using acute tryptophan depletion. Thirty healthy men and women (ages 18-45) were assigned to perform one of three tasks assessing different types of behavioral impulsivity: response initiation, response inhibition, and consequence sensitivity (N = 90). Participants completed two experimental days during which each consumed either a tryptophan-depletion or balanced-placebo amino-acid formulation and completed 5 sessions of their respective tasks at 0.25 h before and 1.5, 4.0, 5.0, and 6.0 h after beverage consumption. During peak effectiveness (5.0 h to 6.0 h following amino-acid consumption), depletion produced selective differences dependent on the type of impulsivity being tested. Specifically, relative to baseline testing (pre-depletion), response initiation impulsivity was significantly increased during the peak effects of depletion. And, when compared to placebo control, both response initiation and consequence sensitivity impulsivity were increased during the peak effects of depletion. Though response initiation and consequence sensitivity impulsivity were affected by tryptophan depletion, response inhibition impulsivity was not, suggesting that other biological processes may underlie this specific component of impulsivity. Future research in other populations or using different pharmacological agents is warranted to further examine the biological processes underlying these components of impulsivity.


Dosing strategies for lithium monotherapy in children and adolescents with bipolar I disorder.
The primary goal of this exploratory study was to obtain data that could lead to evidence-based dosing strategies for lithium in children and adolescents suffering from bipolar I disorder. Outpatients aged 7-17 years meeting Diagnostic and Statistical Manual of Mental Disorders, 4th edition, diagnostic criteria for bipolar I disorder (manic or mixed) were eligible for 8 weeks of open label treatment with lithium in one of three dosing arms. In Arm I, participants began treatment at a dose of 300 mg of lithium twice daily. The starting dose of lithium in Arms II and III was 300 mg thrice daily. Patients in Arms I and II could have their dose increased by 300 mg/day, depending on clinical response, at weekly visits. Patients in Arm III also had mid-week telephone interviews after which they could also have their dose of lithium increased by 300 mg per day. Youths weighing <30 kg were automatically assigned to Arm I, whereas youths weighing ≥30 kg were randomly assigned to Arm I, II, or III. Randomization was balanced by age (7-11 years, 12-17 years) and sex in approximately equal numbers. A priori response criteria were defined as a Clinical Global Impressions-Improvement scale score of ≤ 2 and a 50% decrease from baseline on the Young Mania Rating Scale. Of the 61 youths [32 males (52.5%)] who received open-label lithium, 60 youths completed at least 1 week of treatment and returned for a postbaseline assessment. Most patients had a ≥ 50% improvement in Young Mania Rating Scale score, and more than half of the patients (58%) achieved response. Overall, lithium was well tolerated. All three treatment arms had similar effectiveness, side effect profiles, and tolerability of lithium. On the basis of these results, a dosing strategy in which pediatric patients begin lithium at a dose of 300 mg thrice daily (with an additional 300 mg increase during the first week), followed by 300 mg weekly increases until a priori stopping criteria are met, will be used in an upcoming randomized, placebo-controlled trial.


RANTES and SDF-1 Are Keys in Cell-based Therapy of TMJ Osteoarthritis.
The present study aimed to investigate the therapeutic effect of injections of local bone marrow mesenchymal stem cells (BMSCs) on osteoarthritis (OA) of the temporomandibular joint (TMJ) and to explore the role of stromal cell-derived factor 1 (SDF-1) and regulated on activation, normal T-cell expressed and secreted (RANTES) in this effect. Fundamentally, OA of the TMJ was induced by unilateral anterior crossbite in mice. Exogenous green fluorescent protein-labeled BMSCs (GFP-BMSCs) were weekly injected into the TMJ region for 4, 8, and 12 wk. The reparative effects of exogenous GFP-BMSCs were investigated by morphological observation and micro-computed tomography. The differentiation of GFP-BMSCs in the cartilage was examined by double immunofluorescence of GFPs with type II collagen, and the expression of related factors in the condylar cartilage was quantified by real-time polymerase chain reaction. The role of RANTES and SDF-1 in the therapeutic effect of exogenous BMSCs was examined by both in vitro and in vivo studies. The OA cartilage of the TMJ displays a synchronous increase in SDF-1 and RANTES expression and a higher capability of attracting the migration of GFP-BMSCs. The implanted GFP-BMSCs differentiated into type II collagen-positive cells and reversed cartilage degradation and subchondral bone loss in mice with OA of the TMJ. The migration of GFP-BMSCs towards OA cartilage and the rescuing effect of GFP-BMSC injections were impaired by the inhibitors of C-X-C chemokine receptor type 4 (CXCR4) and C-C chemokine receptor type 1 (CCR1), which are the receptors of SDF-1 and RANTES, respectively. Our data indicated that SDF-1/CXCR4 and RANTES/CCR1 signals are pivotal and function synergistically in the recruitment of GFP-BMSCs towards degraded cartilage in mice OA of the TMJ.


Vital signs: Repeat births among teens - United States, 2007-2010.
Teen childbearing has potential negative health, economic, and social consequences for mother and child. Repeat teen childbearing further constrains the mother's education and employment possibilities. Rates of preterm and low birth weight are higher in teens with a repeat birth, compared with first births. To assess patterns of repeat childbearing and postpartum contraceptive use among teens, CDC analyzed natality data from the National Vital Statistics System (NVSS) and the Pregnancy Risk Assessment Monitoring System (PRAMS) from 2007-2010. Based on 2010 NVSS data from all 50 states and the District of Columbia, of more than 367,000 births to teens aged 15-19 years, 18.3% were repeat births. The percentage of teen births that represented repeat births decreased by 6.2% between 2007 and 2010. Disparities in repeat teen births exist by race/ethnicity, with the highest percentages found among American Indian/Alaska Natives (21.6%), Hispanics (20.9%), and non-Hispanic blacks (20.4%) and lowest among non-Hispanic whites (14.8%). Wide geographic disparities in the percentage of teen births that were repeat births also exist, ranging from 22% in Texas to 10% in New Hampshire. PRAMS data from 16 reporting areas (15 states and New York City) indicate that 91.2% of teen mothers used a contraceptive method 2-6 months after giving birth, but only 22.4% of teen mothers used the most effective methods. Teens with a previous live birth were significantly more likely to use the most effective methods postpartum compared with those with no prior live birth (29.6% versus 20.9%, respectively). Non-Hispanic white and Hispanic teens were significantly more likely to use the most effective methods than non-Hispanic black teens (24.6% and 27.9% versus 14.3%, respectively). The percentage of teens reporting postpartum use of the most effective methods varied greatly geographically across the PRAMS reporting areas, ranging from 50.3% in Colorado to 7.2% in New York State. Although the prevalence of repeat teen birth has declined in recent years, nearly one in five teen births is a repeat birth. Large disparities exist in repeat teen births and use of the most effective contraceptive methods postpartum, which was reported by fewer than one out of four teen mothers. Evidence-based approaches are needed to reduce repeat teen childbearing. These include linking pregnant and parenting teens to home visiting and similar programs that address a broad range of needs, and offering postpartum contraception to teens, including long-acting methods of reversible contraception.


Evaluation of four single-locus markers for Leishmania species discrimination by sequencing.
Several genetic markers have been described for discriminating Leishmania species. In most reported cases, one or a few polymorphisms are the basis of species identification, and the methods were validated on a limited number of strains from a particular geographical region. Therefore, most techniques may underestimate the global intraspecies variability and are applicable only in certain areas. In addition, interlaboratory standardization is mostly absent, complicating comparisons among different studies. Here, we compared species typing results from all sequence polymorphisms found in four popular markers that can be applied directly on clinical samples: the miniexon or spliced leader, the internal transcribed spacer of the ribosomal DNA array, the 7SL RNA gene, and the heat shock protein 70 gene. Clustering was evaluated among 74 Leishmania strains, selected to represent a wide geographic distribution and genetic variability of the medically relevant species of the genus. Results were compared with a multilocus sequence typing (MLST) approach using 7 single-copy household genes and with multilocus enzyme electrophoresis (MLEE), still considered the gold standard by some. We show that strain groupings are highly congruent across the four different single-locus markers, MLST, and MLEE. Overall, the heat shock protein 70 gene and the miniexon presented the best resolutions for separating medically relevant species. As gene sequence analysis is validated here on a global scale, it is advocated as the method of choice for use in genetic, clinical, and epidemiological studies and for managing patients with unknown origins of infection, especially in Western infectious disease clinics dealing with imported leishmaniasis.


Link-centric analysis of variation by demographics in mobile phone communication patterns.
We present a link-centric approach to study variation in the mobile phone communication patterns of individuals. Unlike most previous research on call detail records that focused on the variation of phone usage across individual users, we examine how the calling and texting patterns obtained from call detail records vary among pairs of users and how these patterns are affected by the nature of relationships between users. To demonstrate this link-centric perspective, we extract factors that contribute to the variation in the mobile phone communication patterns and predict demographics-related quantities for pairs of users. The time of day and the channel of communication (calls or texts) are found to explain most of the variance among pairs that frequently call each other. Furthermore, we find that this variation can be used to predict the relationship between the pairs of users, as inferred from their age and gender, as well as the age of the younger user in a pair. From the classifier performance across different age and gender groups as well as the inherent class overlap suggested by the estimate of the bounds of the Bayes error, we gain insights into the similarity and differences of communication patterns across different relationships.


Adenylate cyclase activity during phenotypic variation of Bordetella pertussis.
During MgSO4-induced modulation of Bordetella pertussis, adenylate cyclase activity, histamine-sensitizing activity (HSA) and the major cell-envelope polypeptides with Mr 28000 and 30000 (X polypeptides) were lost synchronously at a rate which could be accounted for by a simple growth-dilution effect. MgSO4 and other compounds which induced the above phenotypic change caused little inhibition of adenylate cyclase activity. Nicotinic acid was the sole exception and at 4.1 mM-caused 60% inhibition of activity. Lysates of modulated cells, mixed with lysates of unmodulated cells, had no effect on either adenylate cyclase activity or HSA. Protein synthesis was a prerequisite for MgSO4-induced modulation and also for the reversal of this process. Exogenous cAMP and dibutyryl cAMP (5 mM) had no counteracting effect on MgSO4- or nicotinic acid-induced modulation. The concentration of MgSO4 required to induce loss of the X polypeptides (10 to 11 mM) was not altered by promoting adenylate cyclase activity by including an activator in the growth medium. In one culture containing 10 mM-MgSO4 and activator, partial loss of the X polypeptides occurred and yet the extracellular cAMP concentration was twice that of cultures without activator and where full expression of the X polypeptides occurred. [3H]cAMP-binding activity was detected in cell extracts of several strains of B. pertussis, but antiserum against purified Escherichia coli catabolite repressor protein gave no reaction with B. pertussis cell extracts. Respiration rates with amino acids were similar for modulated and unmodulated variants and an avirulent strain of B. pertussis. These results are discussed in relation to a possible causal role for adenylate cyclase in modulation of B. pertussis.


Recommended nomenclature for five mammalian carboxylesterase gene families: human, mouse, and rat genes and proteins.
Mammalian carboxylesterase (CES or Ces) genes encode enzymes that participate in xenobiotic, drug, and lipid metabolism in the body and are members of at least five gene families. Tandem duplications have added more genes for some families, particularly for mouse and rat genomes, which has caused confusion in naming rodent Ces genes. This article describes a new nomenclature system for human, mouse, and rat carboxylesterase genes that identifies homolog gene families and allocates a unique name for each gene. The guidelines of human, mouse, and rat gene nomenclature committees were followed and "CES" (human) and "Ces" (mouse and rat) root symbols were used followed by the family number (e.g., human CES1). Where multiple genes were identified for a family or where a clash occurred with an existing gene name, a letter was added (e.g., human CES4A; mouse and rat Ces1a) that reflected gene relatedness among rodent species (e.g., mouse and rat Ces1a). Pseudogenes were named by adding "P" and a number to the human gene name (e.g., human CES1P1) or by using a new letter followed by ps for mouse and rat Ces pseudogenes (e.g., Ces2d-ps). Gene transcript isoforms were named by adding the GenBank accession ID to the gene symbol (e.g., human CES1_AB119995 or mouse Ces1e_BC019208). This nomenclature improves our understanding of human, mouse, and rat CES/Ces gene families and facilitates research into the structure, function, and evolution of these gene families. It also serves as a model for naming CES genes from other mammalian species.


MC3-R as a novel target for antiinflammatory therapy.
To date five melanocortin receptors (MC-R) have been cloned, identified and shown to have a wide distribution throughout the body and likely many diverse functions. MC1-R, found on melanocytes, is involved in pigmentation, while MC2-R is the classic adrenocorticotropic (ACTH) receptor found on the adrenal cortex and adipocytes. MC3-R, MC4-R and MC5-R are in their infancy with regard to their characterization. MC4-R has generated wide interest for its involvement in obesity, whereas our own studies have indicated a role for MC3-R in experimental inflammation. An ACTH fragment unable to alter circulating corticosterone, ACTH-4-10, acts at murine MC3-R present on peritoneal macrophage to inhibit cytokine formation and subsequent neutrophil extravasation. These findings were confirmed using agonists with a higher degree of selectivity toward MC3-R, such as gamma-2-MSH and the synthetic mixed MC3/4-R agonist MTII. In vitro, all these agents were able to affect macrophage functions, including phagocytosis and production of the CXC chemokine KC. Besides using RT-PCR and cAMP formation assays, the involvement of MC3-R in the antiinflammatory actions of these melanocortins was validated with the antagonist SHU-9119. Together these experimental data support the notion that agonism at MC3-R can be used for the design of novel therapeutics for inflammatory conditions.


On the nature of Acket's characteristic parameter C in semiconductor lasers.
Quasi-static interferometric signals in lasers under feedback arise from slowly varying perturbations of the intracavity electric field resulting from the reinjection of a portion of the emitted field into the cavity. Such interferometric signals are well described by the steady-state solution to the Lang-Kobayashi rate equation model. We give an exact series expansion for this steady-state solution that shows precisely how Acket's characteristic parameter C and Henry's linewidth enhancement factor α influence such signals. We show how the series coefficients can be extracted easily and explain how to determine C and α directly from them. Moreover, we draw a precise analogy between self-mixing and FM signals, showing that C plays exactly the same role in self-mixing as the modulation index does in FM.


NMDA receptors in retrosplenial cortex are necessary for retrieval of recent and remote context fear memory.
Over time, memory retrieval is thought to transfer from the hippocampus to a distributed network of neocortical sites. Of these sites, the retrosplenial cortex (RSC) is robustly activated during retrieval of remotely acquired, emotionally valenced memories. It is unclear, however, whether RSC is specifically involved in memory storage or retrieval, and which neurotransmitter receptor mechanisms serve its function. We addressed these questions by inhibiting NMDARs in RSC via infusions of APV before tests for context fear in mice. Anterior cingulate cortex (ACC) and dorsal hippocampus (DH), which have been implicated in the retrieval of remote and recent memory, respectively, served as neuroanatomical controls. Surprisingly, infusion of APV only into RSC, but not ACC or DH, abolished retrieval of remote memory, as revealed by lack of freezing to the conditioning context. APV infused into RSC also impaired retrieval of recent memory, but had no effect on conditioning or memory storage. Within-subject experiments confirmed that the role of RSC in memory retrieval is not time limited. RSC-dependent context fear memory retrieval was mediated by NR2A, but not NR2B, subunit-containing NMDARs. Collectively, these data are the first demonstration that NMDARs in RSC are necessary for the retrieval of remote and recent memories of fear-evoking contexts. Dysfunction of RSC may thereby contribute significantly to the reexperiencing of traumatic memories in patients with posttraumatic stress disorder.


Neurochemical phenotype of cytoglobin-expressing neurons in the rat hippocampus.
Cytoglobin (Cygb), a novel oxygen-binding protein, is expressed in the majority of tissues and has been proposed to function in nitric oxide (NO) metabolism in the vasculature and to have cytoprotective properties. However, the overall functions of Cygb remain elusive. Cygb is also expressed in a subpopulation of brain neurons. Recently, it has been shown that stress upregulates Cygb expression in the brain and the majority of neuronal nitric oxide synthase (nNOS)-positive neurons, an enzyme that produces NO, co-express Cygb. However, there are more neurons expressing Cygb than nNOS, thus a large number of Cygb neurons remain uncharacterized by the neurochemical content. The aim of the present study was to provide an additional and more detailed neurochemical phenotype of Cygb-expressing neurons in the rat hippocampus. The rat hippocampus was chosen due to the abundance of Cygb, as well as this limbic structure being an important target in a number of neurodegenerative diseases. Using triple immunohistochemistry, it was demonstrated that nearly all the parvalbumin- and heme oxygenase 1-positive neurons co-express Cygb and to a large extent, these neuron populations are distinct from the population of Cygb neurons co-expressing nNOS. Furthermore, it was shown that the majority of neurons expressing somastostatin and vasoactive intestinal peptide also co-express Cygb and nNOS. Detailed information regarding the neurochemical phenotype of Cygb neurons in the hippocampus can be a valuable tool in determining the function of Cygb in the brain.


An exhaustive, non-euclidean, non-parametric data mining tool for unraveling the complexity of biological systems--novel insights into malaria.
Complex, high-dimensional data sets pose significant analytical challenges in the post-genomic era. Such data sets are not exclusive to genetic analyses and are also pertinent to epidemiology. There has been considerable effort to develop hypothesis-free data mining and machine learning methodologies. However, current methodologies lack exhaustivity and general applicability. Here we use a novel non-parametric, non-euclidean data mining tool, HyperCube®, to explore exhaustively a complex epidemiological malaria data set by searching for over density of events in m-dimensional space. Hotspots of over density correspond to strings of variables, rules, that determine, in this case, the occurrence of Plasmodium falciparum clinical malaria episodes. The data set contained 46,837 outcome events from 1,653 individuals and 34 explanatory variables. The best predictive rule contained 1,689 events from 148 individuals and was defined as: individuals present during 1992-2003, aged 1-5 years old, having hemoglobin AA, and having had previous Plasmodium malariae malaria parasite infection ≤10 times. These individuals had 3.71 times more P. falciparum clinical malaria episodes than the general population. We validated the rule in two different cohorts. We compared and contrasted the HyperCube® rule with the rules using variables identified by both traditional statistical methods and non-parametric regression tree methods. In addition, we tried all possible sub-stratified quantitative variables. No other model with equal or greater representativity gave a higher Relative Risk. Although three of the four variables in the rule were intuitive, the effect of number of P. malariae episodes was not. HyperCube® efficiently sub-stratified quantitative variables to optimize the rule and was able to identify interactions among the variables, tasks not easy to perform using standard data mining methods. Search of local over density in m-dimensional space, explained by easily interpretable rules, is thus seemingly ideal for generating hypotheses for large datasets to unravel the complexity inherent in biological systems.


Radiosensitivity of different aged human lymphocytes following electron irradiation in vitro.
Cytochalasin B-blocking micronucleus test and chromosomal aberration analysis were used in this study to estimate the yield of individual variability in radiation response of different aged human lymphocytes. Both analyses were performed in three groups of adults, aged 18-65 years, on two sampling times, following irradiation by therapeutical dose of 2 Gy e- in vitro. No statistically significant difference in the induced yield of exchange aberrations between individuals under consideration was found. The yield of total aberration data showed greater variability and was statistically significant in the oldest group against two other adult groups. Regarding to fixation times no statistically significant differences in the induced yield of chromosomal aberrations (exchanges as well as total aberrations) were observed. The study has shown a slight increase in spontaneously occurring micronuclei with age. Almost equal mean number of radiation induced micronuclei was observed in the groups of adults aged 18-20 and 45-55 years. The highest mean number was observed in the oldest group. Evident variation in number of radiation induced micronuclei among individuals from the same age group was observed. The results of micronuclei assay for all individuals under consideration show statistically significant difference in the yield of radiation-induced micronuclei regarding the second fixation time. This study has shown that cytochalasin-B blocking micronucleus test is more sensitive assay than chromosomal aberration analysis for the estimation of individual radiosensitivity.


Lymphatic endothelial cell sphingosine kinase activity is required for lymphocyte egress and lymphatic patterning.
Lymphocyte egress from lymph nodes (LNs) is dependent on sphingosine-1-phosphate (S1P), but the cellular source of this S1P is not defined. We generated mice that expressed Cre from the lymphatic vessel endothelial hyaluronan receptor 1 (Lyve-1) locus and that showed efficient recombination of loxP-flanked genes in lymphatic endothelium. We report that mice with Lyve-1 CRE-mediated ablation of sphingosine kinase (Sphk) 1 and lacking Sphk2 have a loss of S1P in lymph while maintaining normal plasma S1P. In Lyve-1 Cre+ Sphk-deficient mice, lymphocyte egress from LNs and Peyer's patches is blocked. Treatment with pertussis toxin to overcome Galphai-mediated retention signals restores lymphocyte egress. Furthermore, in the absence of lymphatic Sphks, the initial lymphatic vessels in nonlymphoid tissues show an irregular morphology and a less organized vascular endothelial cadherin distribution at cell-cell junctions. Our data provide evidence that lymphatic endothelial cells are an in vivo source of S1P required for lymphocyte egress from LNs and Peyer's patches, and suggest a role for S1P in lymphatic vessel maturation.


Human Rights as Political Determinants of Health: A Retrospective Study of North Korean Refugees.
The gravity, scale, and nature of human rights violations are severe in North Korea. Little is known about the mental health consequences of the lifelong exposures to these violations. In 2014-2015, a retrospective study was conducted among 383 North Korean refugees in South Korea using respondent-driven sampling to access this hidden population. This study collected information on the full range of political and economic rights violations and measured post-traumatic stress disorder, anxiety and depression symptoms, and social functioning by standard instruments. Multivariate regression analysis was performed with the adjustment of political, economic, and demographic variables in 2016-2017. The results indicate elevated symptoms of anxiety (60.1%, 95% CI=54.3%, 65.7%), depression (56.3%, 95% CI=50.8%, 61.9%), and post-traumatic stress disorder (22.8%, 95% CI=18.6%, 27.4%), which are significantly associated with exposures to political rights violations (ten to 19 items versus non-exposure: anxiety AOR=16.78, p<0.001, depression AOR=12.52, p<0.001, post-traumatic stress disorder AOR=16.71, p<0.05), and economic rights violations (seven to 13 items versus non-exposure: anxiety AOR=5.68, p<0.001, depression AOR=4.23, p<0.01, post-traumatic stress disorder AOR=5.85, p<0.05). The mean score of social functioning was also lower in those who were exposed to political (adjusted difference= -13.29, p<0.001) and economic rights violations (adjusted difference= -11.20, p<0.001). This study highlights mental health consequences of lifelong human rights violations in North Korea. Beyond the conventional approach, it suggests the need for a collaborative preventive response from global health and human rights activists to address human rights in regard to mental health determinants of the 20 million people in North Korea.


Pilot Study Evaluating the Feasibility of Comparing Computer Game Play with Close Work During Occlusion in Children Aged 2-7 Years with Amblyopia.
Computer games have been used to stimulate vision in amblyopia with varying degrees of success. The aim of this pilot study was to evaluate the feasibility of conducting a randomised controlled trial to test the effectiveness of computer game play compared to close work during occlusion treatment in children. Children aged 2-7 years with amblyopia and no prior amblyopia treatment were invited to participate. Participants were randomised to a computer game group or close work group and asked to complete two hours occlusion per day, incorporating one hour of their allocated activity. LogMAR visual acuity (VA) was assessed before treatment commenced and after 7(±1) weeks. The same examiner, who was unaware of the allocated treatment, assessed the participant using the same VA test. Eighteen participants (mean age of 4.2 ± 1.3 years) completed the study. After seven weeks the mean VA of the amblyopic eye in the computer game group improved by 0.147 ± 0.182 logMAR, and in the close work group improved by 0.181 ± 0.124 logMAR. The difference in VA improvement between the computer game and the close work groups was not statistically significant (F(1,32) = 3.71; p = 0.06). No significant difference was found in visual outcomes between the two groups, but a larger sample size would be needed to draw conclusions regarding the amblyopic population. Evaluation of the study design suggests it would be feasible to conduct a randomised controlled trial comparing computer games and close work during occlusion to determine if a significant difference in visual outcome exists.


Transient and sustained bacterial adaptation following repeated sublethal exposure to microbicides and a novel human antimicrobial peptide.
Microbicides (biocides) play an important role in the prevention and treatment of infections. While there is currently little evidence for in-use treatment failures attributable to acquired reductions in microbicide susceptibility, the susceptibility of some bacteria can be reduced by sublethal laboratory exposure to certain agents. In this investigation, a range of environmental bacterial isolates (11 genera, 18 species) were repeatedly exposed to four microbicides (cetrimide, chlorhexidine, polyhexamethylene biguanide [PHMB], and triclosan) and a cationic apolipoprotein E-derived antimicrobial peptide (apoEdpL-W) using a previously validated exposure system. Susceptibilities (MICs and minimum bactericidal concentrations [MBCs]) were determined before and after 10 passages (P10) in the presence of an antimicrobial and then after a further 10 passages without an antimicrobial to determine the stability of any adaptations. Bacteria exhibiting >4-fold increases in MBCs were further examined for alterations in biofilm-forming ability. Following microbicide exposure, ≥4-fold decreases in susceptibility (MIC or MBC) occurred for cetrimide (5/18 bacteria), apoEdpL-W (7/18), chlorhexidine (8/18), PHMB (8/18), and triclosan (11/18). Of the 34 ≥4-fold increases in the MICs, 15 were fully reversible, 13 were partially reversible, and 6 were nonreversible. Of the 26 ≥4-fold increases in the MBCs, 7 were fully reversible, 14 were partially reversible, and 5 were nonreversible. Significant decreases in biofilm formation in P10 strains occurred for apoEdpL-W (1/18 bacteria), chlorhexidine (1/18), and triclosan (2/18), while significant increases occurred for apoEdpL-W (1/18), triclosan (1/18), and chlorhexidine (2/18). These data indicate that the stability of induced changes in microbicide susceptibility varies but may be sustained for some combinations of a bacterium and a microbicide.


Antimicrobial susceptibility testing of Shiga toxin-producing Escherichia coli from various samples by using a spiral gradient endpoint technique.
Shiga toxin-producing Escherichia coli (STEC) remains a major public health concern. Microbial resistance may be due to use of antimicrobial agents (AAs) as a growth promoter in food animals or overuse of AAs in humans. The objective of the current study was to determine the antimicrobial susceptibility patterns of STEC strains isolated from food, veterinary, and clinical sources against 14 AAs by using the spiral gradient endpoint method. One hundred ten isolates from three sources were characterized. Results of the current study showed that all strains were resistant to the folate pathway inhibiting AAs including tylosin tartrate (gradient minimum inhibitory concentration [GMIC] ranges from ≥180.00 to 256.00 μg/mL; end concentration [EC] ranges from ≥130.00 to 151.22 μg/mL; and tail-end concentration [TEC] ≥145.00 μg/mL). All the strains isolated from three sources were susceptible to the fluoroquinolone class of AAs (GMIC ranges from ≤1.00 to 64.30 μg/mL; EC ranges from ≤3.33 to 72.00 μg/mL; and TEC ranges from ≤12.13 to 45.00 μg/mL). Among the food isolates, less resistance was found within the aminoglycoside and amphenicol group (GMIC ≥256.00 μg/mL; EC=161.00 μg/mL). Eight strains were resistant to one to three, 44 strains were resistant to four to six, and two strains were resistant to seven or more AAs. All the clinical isolates (100%) were susceptible to the fluoroquinolones and gentamycin. Results also showed that antimicrobial resistance was observed between four and six AAs among the isolates. Some veterinary isolates were resistant to five AAs. Least AAs resistance was shown by 3.7% of isolates to gentamycin and 7.45% to chloramphenicol. This study showed an increasing trend of antimicrobial resistant strains of STEC, and we suggest that periodic surveillance of the antimicrobial susceptibility may be a useful measure to detect the antimicrobial resistant pathogens.


Repurposing of phentolamine as a potential anticancer agent against human castration-resistant prostate cancer: A central role on microtubule stabilization and mitochondrial apoptosis pathway.
Drug repurposing of phentolamine, an α-adrenoceptor antagonist, as an anticancer agent has been studied in human castration-resistant prostate cancer (CRPC). Cell proliferation was examined by sulforhodamine B and CFSE staining assays. Cell cycle progression and mitochondrial membrane potential (ΔΨm) were detected by flow cytometric analysis. Protein expression was detected by Western blotting. Effect on tubulin/microtubule was determined using confocal immunofluorescence microscopic examination, microtubule assembly detection, tubulin turbidity assay, and binding assay. Several assessments were used to characterize apoptotic signaling pathways and combinatory effect. Phentolamine induced anti-proliferative effect in PC-3 and DU-145, two CRPC cell lines, and P-glycoprotein (P-gp) overexpressing cells. This effect was not significantly reduced in paclitaxel-resistant cells. Rhodamine 123 efflux assay showed that phentolamine was not a P-gp substrate. Phentolamine induced mitotic arrest of the cell cycle and formation of hyperdiploid cells, followed by an increase of apoptosis. Mitotic arrest was confirmed by cyclin B1 up-regulation, Cdk1 activation, and a dramatic increase of mitotic protein phosphorylation. Both in vitro and cellular identification demonstrated that phentolamine, similar to paclitaxel, induced tubulin polymerization and formation of multiple nuclei. Besides, it did not compete with paclitaxel binding on tubulin. Phentolamine induced the phosphorylation and degradation of Bcl-2 and Bcl-xL, two anti-apoptotic Bcl-2 family members, and the loss of ΔΨm indicating the induction of mitochondrial damage. It ultimately induced the activation of caspase-9, -8, and -3 and apoptotic cell death. Moreover, combination treatment with phentolamine and paclitaxel caused a synergistic apoptosis. The data suggest that phentolamine is a potential anticancer agent. In contrast to a wide variety of microtubule disrupting agents, phentolamine induces microtubule assembly, leading to mitotic arrest of the cell cycle which "in turn" induces subsequent mitochondrial damage and activation of related apoptotic signaling pathways in CRPC cells. Furthermore, combination between phentolamine and paclitaxel induces a synergistic apoptotic cell death. Phentolamine has a simple chemical structure and is not a P-gp substrate. Optimization of phentolamine structure may also be a potential approach for further development.


Comparative Effectiveness of Tumor Necrosis Factor Agents and Disease-modifying Antirheumatic Therapy in Children with Enthesitis-related Arthritis: The First Year after Diagnosis.
To characterize the effect of anti-tumor necrosis factor (TNF) therapy compared to conventional synthetic disease-modifying antirheumatic drugs (csDMARD) in children with enthesitis-related arthritis (ERA) over the first year after diagnosis. We conducted a multicenter retrospective comparative effectiveness study of children diagnosed with ERA. We estimated the effect of anti-TNF therapy on clinical variables (active joint count, tender entheses count) and patient-reported pain and global assessment of disease activity over the first year after diagnosis using state-of-the-art comparative effectiveness analytic methods. During the study period, 217 patients newly diagnosed with ERA had a total of 965 clinic visits the first year after disease diagnosis. Children [median age 11.6 yrs, interquartile range 10-14] were treated with anti-TNF monotherapy (n = 33, 15.2%), csDMARD monotherapy (n = 73, 33.6%), or both (n = 52, 23.9%) in the first year after disease diagnosis. There was a statistically significant improvement in the primary outcome, active joint count, over time in children who received an anti-TNF drug versus those who did not (p = 0.03). Additionally, use of anti-TNF therapy versus no anti-TNF therapy was associated with less patient-reported pain (p < 0.01) and improved disease activity over time as assessed by the clinical Juvenile Arthritis Disease Activity Score (p < 0.01). The magnitude of estimated effect on clinical outcomes was uniformly greater, with the exception of tender entheses count, in children treated with an anti-TNF drug versus a csDMARD. During the first year after diagnosis, anti-TNF exposure was associated with benefits for several clinically meaningful outcomes in children with enthesitis-related arthritis.


The effect of matrix characteristics on fibroblast proliferation in 3D gels.
Engineering synthetic hydrogels on a molecular basis to introduce natural features that are important in instructing cell behavior is becoming increasingly crucial in biomaterial-based approaches for regenerative medicine and in cell biology to study cell-matrix interactions in three-dimensions (3D). Here, we used collagen gels and exploited the design flexibility of the biological, biochemical and physical characteristics offered by a PEG-based hydrogel system to systematically study the effect of specific extracellular microenvironments on the behavior of primary human fibroblasts in 3D. We firstly found that the proliferation profiles of fibroblasts from different patients cultured within collagen gels (3D) differed significantly from their behavior observed on tissue culture plastic (2D). Furthermore, using the biomimetic PEG-based matrix we showed that cell proliferation in 3D could be selectively manipulated via alteration of the gel characteristics. In particular, this study revealed that, in spite of matrix sensitivity to proteases (e.g. MMP) and the presence of cell-integrin binding sites, at high stiffness (elastic modulus, G' >1200 Pa) the matrix acts as a barrier for cells cultured in 3D. Finally, a comparison between the biomimetic PEG-based and collagen gels indicated that differences in their viscoelastic behaviours, determined by the nature of network structures and cross-links, may influence the mechanism(s) cells employ to remodel their 3D extracellular microenvironment. In conclusion, these studies highlight that for proliferation in 3D, compared to 2D, cells require strategies to overcome the physical impediment posed by the matrix. We also demonstrate that by exploiting the design flexibility of the characteristics offered by these biomimetic hydrogels, it is possible to separately investigate complex aspects characterizing the cell-matrix interactions in 3D; this has the potential to have great impact in regenerative medicine, as well as in cell biology and cancer research.


Hyperglycemia-induced beta-cell apoptosis in pancreatic islets of Psammomys obesus during development of diabetes.
The gerbil Psammomys obesus develops nutrition-dependent diabetes associated with moderate obesity. The disease is characterized by initial hyperinsulinemia, progressing to hypoinsulinemia associated with depleted pancreatic insulin stores. The contribution of changes in beta-cell turnover to insulin deficiency was investigated in vivo during transition to overt diabetes. Normo glycemic diabetes-prone P. obesus animals who were given a high-calorie diet developed hyperglycemia within 4 days, which was found to be associated with a progressive decline in pancreatic insulin content. This was accompanied by a transient increase in beta-cell proliferative activity and by a prolonged increase in the rate of beta-cell death, culminating in disruption of islet architecture. The hypothesis that "glucotoxicity" was responsible for these in vivo changes was investigated in vitro in primary islet cultures. Exposure of islets from diabetes-prone P. obesus to high glucose levels resulted in a dose-dependent increase in beta-cell DNA fragmentation. In contrast, high glucose levels did not induce DNA fragmentation in rat islets, whereas islets from a diabetes-resistant P. obesus line exhibited a reduced and delayed response. Aminoguanidine did not prevent glucose-induced beta-cell DNA fragmentation in vitro, suggesting that formation of nitric oxide and/or advanced glycation end products plays no major role. Elevated glucose concentrations stimulated beta-cell proliferation in both rat and P. obesus islets. However, unlike the marked long-lasting effect in rat islets, only a transient and reduced proliferative response was observed in P. obesus islets; furthermore, beta-cell proliferation was inhibited after prolonged exposure to elevated glucose levels. These results suggest that hyperglycemia-induced beta-cell death coupled with reduced proliferative capacity may contribute to the insulin deficiency and deterioration of glucose homeostasis in P. obesus. Similar adverse effects of hyperglycemia could play a role in the evolution of type 2 diabetes in genetically susceptible individuals.


U.K. Prospective Diabetes Study 22. Effect of age at diagnosis on diabetic tissue damage during the first 6 years of NIDDM.
To assess the effect of age at diagnosis on the initial prevalence and subsequent risk of the progression of diabetic tissue damage in patients with NIDDM. The prevalence of Q-wave myocardial infarction, absent dorsalis pedis pulses, retinopathy, absent ankle jerks, hypertension, and microalbuminuria were determined at baseline and at 3 and 6 years of follow-up in five consecutive 6-year age-cohorts of 3,027 newly diagnosed white patients aged between 36 and 65 years recruited to the U.K. Prospective Diabetes Study. The effect of age at diagnosis on the initial prevalence and the risk of progression of these complications and associated conditions was analyzed using logistic regression and proportional odds methods, respectively. Q-wave myocardial infarction and hypertension were more prevalent in older patients at presentation, but age at diagnosis did not have a significant effect on the increased risk of either after 6 years of NIDDM. Absent dorsalis pedis pulses and ankle jerks were also more prevalent in the older age-groups at presentation, but age at diagnosis was a significant predictor of the increasing prevalence of both during follow-up. The baseline prevalence of retinopathy and microalbuminuria was not related to age. The subsequent risk of retinopathy, but not microalbuminuria, increased significantly with age at diagnosis. Age at diagnosis has a variable impact on different types of diabetic tissue damage and may thus be an important variable in epidemiological and intervention studies in NIDDM. Regular ophthalmologic surveillance and examination of the feet increase in importance with increasing age since the diagnosis of NIDDM.


Extracting the temperature of hot carriers in time- and angle-resolved photoemission.
The interaction of light with a material's electronic system creates an out-of-equilibrium (non-thermal) distribution of optically excited electrons. Non-equilibrium dynamics relaxes this distribution on an ultrafast timescale to a hot Fermi-Dirac distribution with a well-defined temperature. The advent of time- and angle-resolved photoemission spectroscopy (TR-ARPES) experiments has made it possible to track the decay of the temperature of the excited hot electrons in selected states in the Brillouin zone, and to reveal their cooling in unprecedented detail in a variety of emerging materials. It is, however, not a straightforward task to determine the temperature with high accuracy. This is mainly attributable to an a priori unknown position of the Fermi level and the fact that the shape of the Fermi edge can be severely perturbed when the state in question is crossing the Fermi energy. Here, we introduce a method that circumvents these difficulties and accurately extracts both the temperature and the position of the Fermi level for a hot carrier distribution by tracking the occupation statistics of the carriers measured in a TR-ARPES experiment.


Potential inherited causes of recurrent prosthetic mitral valve thrombosis in a pregnant patient suffering from recurrent miscarriage.
An effective anticoagulation is critical in pregnant patients with prosthetic heart valves. Inherited disorders may interfere with the coagulation cascade and may be associated with obstetrical complications as well as with prosthetic valve-derived complications. The patient in the present case had a history of recurrent prosthetic heart valve thrombosis (PHVT) despite an effective anticoagulation. She underwent a thrombolysis with low-dose prolonged infusion of tissue-type plasminogen activator for the management of her recurrrent prosthetic valve thrombosis. The genetic testing showed homozygous mutations of methylenetetrahydrofolate reductase (MTHFR) A 1298 C and heterozygous mutations of β-fibrinogen 455 G-A. Inherited disorders such as MTHFR A 1298 C and fibrinogen 455G/A polymorphisms may be involved in the pathogenesis of recurrent PHVT and/or pregnancy loss.


Challenges in the Assessment of Bycatch: Postmortem Findings in Harbor Porpoises (Phocoena phocoena) Retrieved From Gillnets.
Bycatch is considered one of the most significant threats affecting cetaceans worldwide. In the North Sea, bottom-set gillnets are a specific risk for harbor porpoises (Phocoena phocoena). Methods to estimate bycatch rates include on-board observers, remote electronic monitoring, and fishermen voluntarily reporting; none of these are systematically conducted. Additionally, necropsies of stranded animals can provide insights into bycatch occurrence and health status of individuals. There are, however, uncertainties when it comes to the assessment of bycatch in stranded animals, mainly due to the lack of diagnostic tools specific for underwater entrapment. We conducted a literature review to establish criteria that aid in the assessment of bycatch in small cetaceans, and we tested which of these criteria applied to harbor porpoises retrieved from gillnets in the Netherlands (n = 12). Twenty-five criteria were gathered from literature. Of these, "superficial incisions," "encircling imprints," and "recent ingestion of prey" were observed in the vast majority of our confirmed bycatch cases. Criteria like "pulmonary edema," "pulmonary emphysema," and "organ congestion" were also frequently observed, although considered unspecific as an indicator of bycatch. Notably, previously mentioned criteria as "favorable health status," "absence of disease," or "good nutritional condition" did not apply to the majority of our bycaught porpoises. This may reflect an overall reduced fitness of harbor porpoises inhabiting the southern North Sea or a higher chance of a debilitated porpoise being bycaught, and could result in an underestimation of bycatch rates when assessing stranded animals.


Comparing air dispersion model predictions with measured concentrations of VOCs in urban communities.
Air concentrations of nine volatile organic compounds were measured over 48-h periods at 23 locations in three communities in the Minneapolis-St. Paul metropolitan area. Concentrations at the same times and locations were modeled using a standard regulatory air dispersion model (ISCST3). The goal of the study was to evaluate model performance by comparing predictions with measurements using linear regression and estimates of bias. The modeling, done with mobile and area source emissions resolved to the census tract level and characterized as model area sources, represents an improvement over large-scale airtoxics modeling analyses done to date. Despite the resolved spatial scale, the model did not fully capture the spatial resolution in concentrations in an area with a sharp gradient in emissions. In a census tract with a major highway at one end of the tract (i.e., uneven distribution of emissions within the tract), model predictions atthe opposite end of the tract overestimated measured concentrations. This shortcoming was seen for pollutants emitted mainly by mobile sources (benzene, ethylbenzene, toluene, and xylenes). We suggest that major highways would be better characterized as line sources. The model also failed to fully capture the temporal variability in concentrations, which was expected since the emissions inventory comprised annual average values. Based on our evaluation metrics, model performance was best for pollutants emitted mainly from mobile sources and poorest for pollutants emitted mainlyfrom area sources. Important sources of error appeared to be the source characterization (especially location) and emissions quantification. We expect that enhancements in the emissions inventory would give the greatest improvement in results. As anticipated for a Gaussian plume model, performance was dramatically better when compared to measurements that were not matched in space or time. Despite the limitations of our analysis, we found thatthe regulatory air dispersion model was generally able to predict space and time matched 48-h average ambient concentrations of VOC species within a factor of 2 on average, results that meet regulatory model acceptance criteria.


Design and development of functionalized cyclometalated ruthenium chromophores for light-harvesting applications.
The syntheses and the electrochemical spectroscopic properties of a suite of asymmetrical bistridentate cyclometalated Ru(II) complexes bearing terminal triphenylamine (TPA) substituents are reported. These complexes, which contain structural design elements common to both inorganic and organic dyes that exhibit superior power conversion efficiencies in the dye-sensitized solar cell (DSSC), are broadly formulated as [Ru(II)(L-2,5'-thiophene-TPA-R(1))(L-R(2))](+) [L = tridentate chelating ligand (e.g., 2,2':6',2''-terpyridine (tpy); deprotonated forms of 1,3-di(pyridin-2-yl)benzene (Hdpb) or 6-phenyl-2,2'-bipyridine (Hpbpy)); R(1) = -H, -Me, -OMe; R(2) = -H, -CO(2)Me, -CO(2)H]. The following structural attributes were systematically modified for the series: (i) electron-donating character of the terminal substituents (e.g., R(1) = -H, -Me, -OMe) placed para to the amine of the "L-2,5'-thiophene-TPA-R(1)" ligand framework; (ii) electron-withdrawing character of the tridentate chelate distal to the TPA-substituted ligand (e.g., R(2) = -H, -CO(2)Me, -CO(2)H); and (iii) position of the organometallic bond about the Ru(II) center. UV-vis spectra reveal intense and broad absorption bands arising from a collection of metal-to-ligand charge-transfer (MLCT) and TPA-based intraligand charge-transfer (ILCT) transitions that, in certain cases, extend beyond 800 nm. Electrochemical data indicate that the oxidative behavior of the TPA and metal chelate units can be independently modulated except in cases where the anionic phenyl ring is in direct conjugation with the TPA unit. In most cases, the anionic character of the cyclometalating ligands renders a metal-based oxidation event prior to the oxidation of the TPA unit. This situation can, however, be reversed with an appropriately positioned Ru-C bond and electron-rich R(1) group. This finding is important in that this arrangement confines the highest occupied molecular orbital (HOMO) to the TPA unit rather than the metal, which is optimal for sensitizing TiO(2); indeed, a remarkably high power conversion efficiency (η) in the DSSC (i.e., 8.02%) is measured for the TPA-substituted pbpy(-) chelate where R(1) = -OMe. These results provide a comprehensive strategy for improving the performance of bistridentate Ru sensitizers devoid of NCS(-) groups for the DSSC.


A Comparative Study Between Traditional Microscopic Surgeries and Endoscopic Endonasal Surgery for Skull Base Chordomas.
Skull base chordomas (SBCs) are rare clinically aggressive neoplasms, developing local recurrences after surgical resection. Although SBCs have traditionally been resected by craniotomy or microscopic transsphenoidal surgery (TSS), the recent development of the endoscopic endonasal approach (EEA) has revolutionized treatment strategies through minimally invasive techniques. This study aimed to evaluate clinical outcomes after traditional microsurgeries or EEAs for SBCs. The present retrospective study investigated 66 patients with primary SBCs who underwent surgery between 1977 and 2019. Resection was performed via EEA in 17 cases, craniotomy in 23, transoral approach in 8, TSS in 12, staged surgery in 4, and others in 2. The median follow-up period for progression-free survival (PFS) was 19.5 months. There were no significant differences in preoperative tumor volume or resection rate among these approaches. The incidence of postoperative cranial nerve palsy was significantly lower in EEA than that in craniotomy (P < 0.05). Although total resection was observed in 4 cases of EEA expanding into the superior and inferior part of the clivus, no cases of transoral approach or TSS achieved total resection for both parts. No significant difference in PFS was found among these approaches. Multivariate analysis showed that being female and the absence of radiotherapy were significantly associated with shorter PFS (P < 0.05 and P < 0.001, respectively). The resection rate was not associated with PFS. EEA is a less invasive surgical approach for SBCs. The development of surgical instruments and postoperative radiotherapy will further improve patients' outcomes.


Intraarterial autologous implantation of adult stem cells for patients with Parkinson disease.
To evaluate the feasibility, safety, and effectiveness of intraarterial autologous implantation of adult stem cells for Parkinson disease (PD). From June 2006 to August 2008, 36 men and 14 women (mean age, 62.5 years +/- 10.4; range, 38-81 y) with PD (mean duration, 9.3 years; range, 1-28 y) underwent autologous implantation of stem cells with superselective arterial catheterization. Patients were evaluated with clinical and neurologic examinations; internationally recognized scales for the evaluation of PD, disability, activities of daily living, depression, and quality of life (QOL); as well as videos, magnetic resonance (MR) imaging, and MR spectroscopy. Stem cells were implanted in the posterior region of the circle of Willis. Patients were evaluated according to clinical measures. Comparison was made versus data collected from all scales before treatment, as well as videos and spectroscopy in eight patients. In a mean follow-up of 7.4 months +/- 4.5 (range, 1-18 months), patients showed a median improvement of 51.1% and quartile deviation (QD) of 24.8% on the Unified PD Rating Scale. They showed significant improvement in disability, activities of daily living, depression, and QOL (P < .5). No complications were observed. In eight patients, follow-up MR spectroscopy revealed mean improvements in n-acetylaspartate/creatine ratio from 1.805 to 2.07 (12.8%) and from 1.25 to 1.88 (43.56%) in right and left basal ganglia, respectively, versus preprocedural values (P < .05). Treatment of PD with intraarterial autologous implantation of adult stem cells is feasible and safe and results in improved severity of disease and QOL.


Beliefs and attitudes of male and female adolescents and the risk of smoking behavior.
Adolescent smoking relates to numerous risk factors, of which beliefs and attitudes toward smoking may play a role. The study aimed to investigate the association between beliefs and attitudes and the risk of adolescent smoking. In a school-based cross-sectional study, 3,400 students were recruited from 34 intermediate and secondary schools in Madinah City, Al Madinah Region, Saudi Arabia. Data about sociodemographics, smoking-related factors, and beliefs and attitudes toward smoking were collected using a valid and reliable self-administered questionnaire. Prevalence of smoking was estimated and the studied beliefs and attitudes were compared by smoking status and sex using appropriate statistical analyses including multivariate logistic regression. Of the 3,322 respondents, 33.02% (38.9% males and 26.4% females) were current smokers. Beliefs and attitudes toward smoking significantly differed between smokers and nonsmokers in the studied male and female students. The adjusted risk of smoking was significantly increased among female adolescents who believed that male smokers were more attractive [odds ratio (OR) = 2.2; 95% confidence interval (CI) = 1.6-2.9] and among male smokers who believed that female smokers are more attractive (OR = 1.7; 95% CI = 1.2-2.2). The risk was also increased among all adolescents who believed that smoking lent comfort in social gatherings. Belief that smoking is harmful, however, was negatively associated with the risk of smoking, particularly among females (OR = 0.55; 95% CI = 0.35-0.91). The study revealed a considerable high prevalence of smoking among male and female adolescents. Addressing the beliefs and knowledge about smoking during childhood is crucial in any antismoking program.


Hematopoietic cells expressing the peripheral cannabinoid receptor migrate in response to the endocannabinoid 2-arachidonoylglycerol.
Cb2 is a novel protooncogene encoding the peripheral cannabinoid receptor. Previous studies demonstrated that 2 distinct noncoding first exons exist: exon-1A and exon-1B, which both splice to protein-coding exon-2. We demonstrate that in retrovirally induced murine myeloid leukemia cells with proviral insertion in Cb2, exon-1B/exon-2 Cb2 messenger RNA levels have been increased, resulting in high receptor numbers. In myeloid leukemia cells without virus insertion in this locus, low levels of only exon-1A/exon-2 Cb2 transcripts were present and receptors could not be detected. To elucidate the function of Cb2 in myeloid leukemia cells, a set of in vitro experiments was carried out using 32D/G-CSF-R (granulocyte colony-stimulating factor receptor) cells transfected with exon-1B/exon-2 Cb2 complementary DNA and a myeloid cell line carrying a virus insertion in Cb2 (ie, NFS 78). We demonstrate that a major function of the Cb2 receptor is stimulation of migration as determined in a transwell assay. Exposure of Cb2-expressing cells to different cannabinoids showed that the true ligand for Cb2 is 2-arachidonoylglycerol (2-AG), which may act as chemoattractant and as a chemokinetic agent. Furthermore, we observed a significant synergistic activity between 2-AG and interleukin-3 or G-CSF, suggesting cross-talk between the different receptor systems. Radioactive-ligand binding studies revealed significant numbers of Cb2 receptors in normal spleen. Transwell experiments carried out with normal mouse spleen cells showed 2-AG-induced migration of B220-, CD19-, immunoglobulin M-, and immunoglobulin D-expressing B lymphocytes. Our study demonstrates that a major function of Cb2 receptor expressed on myeloid leukemia cells or normal splenocytes is stimulation of migration.


Injectate temperature: effects on thermodilution CO measurements.
The present study has confirmed the findings of other research and indicates that current technology permits accurate CO determination using room temperature injectate. Some patients, especially those with high output, temperature, or blood pressure, may continue to require iced injectate measures to ensure accuracy; more research is needed here. Room temperature measures may be less reproducible than iced. Several ways of compensating for this have been suggested, depending on the technology available. At present, these methods are preferable to using a correction formula based on regression analyses.


Non-spindle microtubule organizing centers in metaphase II-arrested mouse oocytes.
A human autoantiserum (5051) directed against pericentriolar material (PCM) was used to study the distribution of microtubule-organizing centers (MTOCs) in the oocyte and during the first cell cycle of mouse development. In oocytes, the PCM was found not only at the poles of the barrel-shaped metaphase II spindle but also at many discrete loci around the cytoplasm near the cell cortex. The spindle poles were also composed of several PCM foci. In metaphase-arrested eggs only the PCM foci located near the chromosomes acted as MTOCs. However, after reduction of the critical concentration for tubulin polymerization by taxol, the cytoplasmic PCM foci were also found to be associated with nucleation of microtubules. After fertilization the cortical PCM foci remained in a peripheral position until the end of the S phase, when they appeared to migrate centrally towards the pronuclei. At prometaphase of the first mitotic division, numerous MTOCs were found around the two sets of chromosomes; these MTOCs then aligned to form two bands on either side of the metaphase plate of the first mitosis.


The catalytic site of serine proteinases as a specific binding cavity for xenon.
Under moderate pressure, xenon can bind to proteins and form weak but specific interactions. Such protein-xenon complexes can be used as isomorphous derivatives for phase determination in X-ray crystallography. Investigation of the serine proteinase class of enzymes shows that the catalytic triad, the common hydrolytic motif of these enzymes, is a specific binding site for one xenon atom and shows high occupancy at pressures below 12 bar. Complexes of xenon with two different serine proteinases, elastase and collagenase, were analyzed and refined to 2.2 A and 2.5 A resolution, respectively. In both cases, a single xenon atom with a low temperature factor is located in the active site at identical positions. Weak interactions exist with several side chains of conserved amino acids at the active site. Xenon binding does not induce any major changes in the protein structure and, as a consequence, crystals of the xenon complexes are highly isomorphous with the native protein structures. Xenon is also found to bind to the active site of subtilisin Carlsberg, a bacterial serine proteinase, that also has a catalytic triad motif. As the region around the active site shows conserved structural homology in all serine proteinases, it is anticipated that xenon binding will prove to be a general feature of this class of proteins.


Shapes of early change in psychotherapy under routine outpatient conditions.
Although improvement of clients' state is a central concern for psychotherapy, relatively little is known about how change in outcome variables unfolds during psychotherapy. Client progress may follow highly variable temporal courses, and this variation in treatment courses may have important clinical implications. By analyzing treatment progress using growth mixture modeling up to the 6th session in a sample of 192 outpatients treated under routine clinic conditions, the authors identified 5 client groups based on similar progress on the short form versions of the Clinical Outcomes in Routine Evaluation-Outcome Measure. The shapes of early change typical for these client groups were characterized by (a) high initial impairment, (b) low initial impairment, (c) early improvement, (d) medium impairment with continuous treatment progress, or (e) medium impairment with discontinuous treatment progress. Moreover, the shapes of early change were associated with different treatment outcomes and durations, and several intake variables (depression, anxiety, and age) enabled prediction of the shape of early change and/or prediction of individual treatment progress within client groups with similar shapes of change.


Ataxia associated with Hashimoto's disease: progressive non-familial adult onset cerebellar degeneration with autoimmune thyroiditis.
Acquired cerebellar ataxia has been described with hypothyroidism, and is typically reversible by thyroid hormone replacement therapy. The cerebellar dysfunction has been attributed to metabolic and physiological effects of the endocrine disorder. In a few patients, however, ataxia has persisted despite thyroid replacement therapy. Other mechanisms may be involved in ataxia associated with thyroid disorders. To document progressive non-familial adult onset cerebellar degeneration (PNACD) occurring in six patients with raised antithyroid antibodies (Hashimoto's/autoimmune thyroiditis), and other autoimmune manifestations, in the absence of hypothyroidism; and to document the independence of the cerebellar disorder from the endocrine dysfunction. A case study of six patients with PNACD reviewing the clinical course and relation to endocrine and autoimmune status. All six patients were euthyroid when they developed their symptoms; had raised antithyroid antibodies consistent with Hashimoto's autoimmune thyroiditis; and had strong personal or family histories of organ specific autoimmune diatheses. Brain MRI disclosed atrophy of the cerebellar vermis in four patients and olivopontocerebellar atrophy in two. Other possible causes of cerebellar degeneration were excluded. De novo treatment (two patients) or continued treatment (three patients) with L-thyroxine did not modify the progression of the ataxia. Cerebellar degeneration in these patients with raised antithyroid antibodies may be immune mediated. The presence of antithyroid antibodies may signal or cause the autoimmune process producing cerebellar degeneration. "Hashimoto's associated ataxia" seems to represent a recognisable and not uncommon condition; a trial of immunomodulating therapy should be considered in these patients.


Prevention of progressive back-specific dysfunction during pregnancy: an assessment of osteopathic manual treatment based on Cochrane Back Review Group criteria.
Back pain during pregnancy may be associated with deficits in physical functioning and disability. Research indicates that osteopathic manual treatment (OMT) slows the deterioration of back-specific functioning during pregnancy. To measure the treatment effects of OMT in preventing progressive back-specific dysfunction during the third trimester of pregnancy using criteria established by the Cochrane Back Review Group. A randomized sham-controlled trial including 3 parallel treatment arms: usual obstetric care and OMT (UOBC+OMT), usual obstetric care and sham ultrasound therapy (UOBC+SUT), and usual obstetric care (UOBC). The Osteopathic Research Center within the University of North Texas Health Science Center in Fort Worth. A total of 144 patients were randomly assigned and included in intention-to-treat analyses. Progressive back-specific dysfunction was defined as a 2-point or greater increase in the Roland-Morris Disability Questionnaire (RMDQ) score during the third trimester of pregnancy. Risk ratios (RRs) and 95% confidence intervals (CIs) were used to compare progressive back-specific dysfunction in patients assigned to UOBC+OMT relative to patients assigned to UOBC+SUT or UOBC. Numbers needed to treat (NNTs) and 95% CIs were also used to assess UOBC+OMT vs each comparator. Subgroup analyses were performed using median splits of baseline scores on a numerical rating scale for back pain and the RMDQ. Overall, 68 patients (47%) experienced progressive back-specific dysfunction during the third trimester of pregnancy. Patients who received UOBC+OMT were significantly less likely to experience progressive back-specific dysfunction (RR, 0.6; 95% CI, 0.3-1.0; P=.046 vs UOBC+SUT; and RR, 0.4; 95% CI, 0.2-0.7; P<.0001 vs UOBC). The effect sizes for UOBC+OMT vs UOBC+SUT and for UOBC+OMT vs UOBC were classified as medium and large, respectively. The corresponding NNTs for UOBC+OMT were 5.1 (95% CI, 2.7-282.2) vs UOBC+SUT; and 2.5 (95% CI, 1.8-4.9) vs UOBC. There was no statistically significant interaction between subgroups in response to OMT. Osteopathic manual treatment has medium to large treatment effects in preventing progressive back-specific dysfunction during the third trimester of pregnancy. The findings are potentially important with respect to direct health care expenditures and indirect costs of work disability during pregnancy.


Painful varicoceles: Role of varicocelectomy.
The incidence of varicocele in the general population is up to 15%. It is estimated that the prevalence of pain with varicoceles is around 2-10%. Till the year 2000, only two studies evaluated efficacy of varicocelectomy in painful varicoceles with conflicting results. Over the past decade many other studies have addressed this issue and reported on the treatment outcome and predictors of success. We critically appraised studies published from March 2000 to May 2013 evaluating surgical management in painful varicoceles to provide an evidence based review of effectiveness of varicocelectomy in relieving pain in patients with symptomatic painful varicoceles. The association between varicoceles and pain is not clearly established. Conservative treatment is warranted as the first line of treatment in men with painful clinical varicoceles. In carefully selected men with clinically palpable varicoceles and associated characteristic chronic dull ache, dragging or throbbing pain who do not respond to conservative therapy, varicocelectomy is warranted and is associated with approximately 80% success. However, surgical success does not always translate into resolution of pain and pain might persist even when no varicoceles are detected postoperatively.


Allosteric interactions at the m1, m2 and m3 muscarinic receptor subtypes.
The purpose of our study was to investigate the interactions of allosteric antagonists at the individual m1, m2 and m3 muscarinic receptor subtypes. This was achieved through the use of transformed Chinese hamster ovary cells stably expressing the rat m1 or m3 receptor genes. A homogeneous population of the m2 subtype was obtained from rat heart tissue. Our data indicate that the cardioselective antagonists (gallamine, methoctramine, AF-DX 116 and himbacine) display the following rank order of potency for both displacing ligand binding to the primary site on the receptor and allosterically decelerating ligand dissociation: m2 greater than m1 greater than m3. Schild analysis showed the following rank order of the magnitude of gallamine's cooperative interactions with the three receptor subtypes: m3 greater than m1 greater than m2. By comparison, the ion-channel blockers (verapamil, phencyclidine and quinidine) exhibited a rank order of potency for cooperative effects similar to that of cardioselective antagonists; however, these blockers did not show appreciable specificity in their interaction with the receptor primary binding site. There was a lack of correlation between the displacement of ligand binding and the allosteric potencies of the allosteric antagonists at each of the three muscarinic receptor subtypes, thus revealing the complex nature of interaction (both competitive and allosteric) between many of these compounds with the muscarinic receptor. Despite the fact that the majority of allosteric muscarinic antagonists are also K+ channel blockers, the use of pertussis toxin did not support the notion that this channel represents the allosteric site coupled to the receptor.


Disrupting Foxh1-Groucho interaction reveals robustness of nodal-based embryonic patterning.
The winged-helix transcription factor Foxh1 is an essential regulator of Nodal signaling during the key developmental processes of gastrulation, anterior-posterior (A-P) patterning, and the derivation of left-right (L-R) asymmetry. Current models have Foxh1 bound to phospho-Smad2/3 (pSmad2/3) as a central transcriptional activator for genes targeted by Nodal signaling including Nodal itself, the feedback inhibitor Lefty2, and the positive transcriptional effector Pitx2. However, the conserved Engrailed homology-1 (EH1) motif present in Foxh1 suggests that modulated interaction with Groucho (Grg) co-repressors would allow Foxh1 to function as a transcriptional switch, toggling between transcriptional on and off states via pSmad2-Grg protein-switching, to ensure the properly timed initiation and suppression, and/or amplitude, of expression of Nodal and its target genes. We minimally mutated the Foxh1 EH1 motif, creating a novel Foxh1(mEH1) allele to test directly the contribution of Foxh1-Grg-mediated repression on the transient, dynamic pattern of Nodal signaling in mice. All aspects of Nodal and its target gene expression in Foxh1(mEH1/mEH1) embryos were equivalent to wild type. A-P patterning and organ situs in homozygous embryos and adult mice were also unaffected. The finding that Foxh1-Grg-mediated repression is not essential for Nodal expression during mouse embryogenesis suggests that other regulators compensate for the loss of repressive regulatory input that is mediated by Grg interactions. We suggest that the pervasive inductive properties of Nodal signaling exist within the context of a strongly buffered regulatory system that contributes to resilience and accuracy of its dynamic expression pattern.


Towards accurate structural characterization of metal centres in protein crystals: the structures of Ni and Cu T(6) bovine insulin derivatives.
Using synchrotron radiation (SR), the crystal structures of T6 bovine insulin complexed with Ni(2+) and Cu(2+) were solved to 1.50 and 1.45 Å resolution, respectively. The level of detail around the metal centres in these structures was highly limited, and the coordination of water in Cu site II of the copper insulin derivative was deteriorated as a consequence of radiation damage. To provide more detail, X-ray absorption spectroscopy (XAS) was used to improve the information level about metal coordination in each derivative. The nickel derivative contains hexacoordinated Ni(2+) with trigonal symmetry, whereas the copper derivative contains tetragonally distorted hexacoordinated Cu(2+) as a result of the Jahn-Teller effect, with a significantly longer coordination distance for one of the three water molecules in the coordination sphere. That the copper centre is of type II was further confirmed by electron paramagnetic resonance (EPR). The coordination distances were refined from EXAFS with standard deviations within 0.01 Å. The insulin derivative containing Cu(2+) is sensitive towards photoreduction when exposed to SR. During the reduction of Cu(2+) to Cu(+), the coordination geometry of copper changes towards lower coordination numbers. Primary damage, i.e. photoreduction, was followed directly by XANES as a function of radiation dose, while secondary damage in the form of structural changes around the Cu atoms after exposure to different radiation doses was studied by crystallography using a laboratory diffractometer. Protection against photoreduction and subsequent radiation damage was carried out by solid embedment of Cu insulin in a saccharose matrix. At 100 K the photoreduction was suppressed by ∼15%, and it was suppressed by a further ∼30% on cooling the samples to 20 K.


Changes in Intraocular Pressure During Orbital Floor Fracture Repair.
To evaluate changes in intraocular pressure (IOP) during orbital floor fracture repair. Retrospective chart review of patients undergoing isolated orbital floor fracture repair with implant placement at a level 1 trauma center and tertiary care center. Patients with combined zygomaticomaxillary or medial wall fracture repair, or other significant ocular injuries were excluded. IOP measurements using a handheld applanation tonometer were recorded immediately after induction of anesthesia, immediately after orbital floor implant placement, and on postoperative day 1. Nine patients who underwent isolated orbital floor fracture repair were examined. There was a statistically significant mean decrease of 4.66 mmHg (p < 0.05) in IOP from prior to surgical to immediately after implant placement. There was also a significant mean increase in IOP of 7.44 mmHg (p < 0.05) at postoperative day 1 compared with immediately after implant placement. There was no statistically significant difference in IOP between prior to surgical incision and postoperative day 1. All patients who underwent orbital floor fracture repair in this study had a significant decrease in IOP during orbital floor fracture repair that returned to immediately preoperative levels by postoperative day 1. This initial decrease in IOP may be because of intraoperative manipulation of the globe.


Effect of Stress on Neuroimmune Processes.
Psychological stress worsens many diseases, especially those with inflammatory components, such as atopic dermatitis (AD) and autism spectrum disorder (ASD), conditions that are significantly correlated in large epidemiologic studies. However, how stress contributes to these conditions is still poorly understood. This narrative review of the relevant literature advances the premise that stress affects inflammatory processes in AD and ASD via stimulation of mast cells (MCs). MEDLINE was searched between 1980 and 2019 using the terms allergies, atopic dermatitis, autism spectrum disorder, brain, corticotropin-releasing hormone, inflammation, hypothalamic-pituitary-adrenal axis, mast cells, neuropeptides, stress, neurotensin, and substance P. Exposure to psychological stress is associated with onset and/or exacerbation of AD and ASD. This association could be attributable to activation of MCs, which are ubiquitous in the body, including the brain, and could contribute to inflammation. Understanding and addressing the connection between stress and MCs is important in clarifying the pathogenesis and developing effective treatments for diseases that worsen with stress and involve inflammation, such as AD and ASD.


Incidental gallbladder cancer at cholecystectomy: when should the surgeon be suspicious?
Preoperative predictors of incidental gallbladder cancer (iGBC) have been poorly defined despite the frequency with which cholecystectomy is performed. The objective of this study was to define the incidence of and consider risk factors for iGBC at cholecystectomy. The American College of Surgeons-National Surgical Quality Improvement Program (ACS-NSQIP) database (2005-2009) was used to identify all patients who underwent cholecystectomy (N = 91,260). Patients with an International Classification of Diseases, Ninth Revision, diagnosis of gallbladder malignancy who underwent a laparoscopic cholecystectomy (LC; n = 80,924) or open cholecystectomy (OC; n = 10,336) alone were included. The incidence of iGBC was 0.19% (n = 170) for all cholecystectomy cases, but 0.05% at LC, 0.60% at LC converted to OC (P < 0.001 vs LC), and 1.13% at OC (P < 0.001 vs others). Patients undergoing OC were 17.3 times more likely to have iGBC than LC patients. Age 65 years or older, Asian or African American race, ASA (American Society of Anesthesiologists) class 3 or more, diabetes mellitus, hypertension, weight loss more than 10%, alkaline phosphatase levels 120 units/L or more, and albumin levels 3.6 g/dL or less were associated with iGBC. Multiple logistic regression identified having an OC, age 65 years or older, Asian or African American race, an elevated alkaline phosphatase level, and female sex as independent risk factors. Patients with 1, 2, 3, and 4 of these factors had a 6.3-, 16.7-, 30.0-, and 47.4-fold risk of iGBC, respectively, from a zero-risk factor baseline of 0.03%. Surgeons' suspicion for GBC should be heightened when they are performing or converting from LC to OC and when patients are older, Asian or African American, female, and have an elevated alkaline phosphatase level.


Culturability and toxicity of sick building syndrome-related fungi over time.
Two experiments were conducted regarding the culturability and toxicity of fungi located on building materials over time and the efficacy of seven laboratory techniques in recovering culturable fungi from sample swabs. In the first experiment, eight sections of drywall were inoculated with Stachybotrys chartarum and stored at 25 +/- 5 degrees Celsius and 20-60% relative humidity (RH) for up to two years. Another eight sections of ceiling tile were stored at 100% RH for 1 year. Six sections of ceiling tile and 15 swabs were also inoculated with Penicillium chrysogenum and S. chartarum respectively and stored under the same conditions for 8 months and 3.3 years. All materials were tested for culturability at the end of the storage period. S. chartarum-inoculated samples were also tested for toxicity. In the second experiment (replicated twice), S. chartarum and Chaetomium globosum were inoculated onto 84 swabs each. Storage was up to 266 days at 25 +/- 5 degrees Celsius and 20-60% RH. Seven techniques were compared regarding the recovery of culturable fungi from the swabs over different time points. Results for Experiment 1 showed that all samples were culturable after the storage period and that the S. chartarum-inoculated drywall samples were toxic. In Experiment 2, all techniques showed high rates of recovery. These data show that despite being without a water source, these organisms can be culturable and toxic after long periods of time under conditions similar to human-occupied dwellings and that a number of preparation techniques are suitable for the recovery of these fungi from inoculated swabs.


The influence of antisense oligonucleotide length on dystrophin exon skipping.
Antisense oligonucleotides (AOs) can be used to redirect dystrophin pre-messenger RNA (mRNA) processing, to remove selected exons from the mature dystrophin mRNA, to overcome nonsense mutations, and/or restore the reading frame. Redundancy within the dystrophin protein allows some domains to be removed without seriously compromising function. One of the challenges for splicing blockade is to design AOs that efficiently remove targeted exons across the dystrophin pre-mRNA. AOs are initially designed to anneal to the more obvious motifs implicated in the splicing process, such as acceptor or donor splice sites and in silico predicted exonic splicing enhancers. The AOs are evaluated for their ability to induce targeted exon skipping after transfection into cultured myoblasts. Although no single motif has been implicated in the consistent induction of exon skipping, the length of the AO has emerged as an important parameter in designing compounds that redirect dystrophin pre-mRNA processing. We present data from in vitro studies in murine and human cells showing that appropriately designed AOs of 25-31 nucleotides are generally more effective at inducing exon skipping than shorter counterparts. However, there appears to be an upper limit in optimal length, which may have to be established on a case-by-case basis.


Sole-parent work schedules and adolescent wellbeing:Evidence from Australia.
Extensive evidence has shown that working nonstandard hours, such as evening or night shifts, impacts negatively on workers' own health, and a growing literature suggests such impacts extend to the health of workers' children. Using matched parent and child data from a large Australian panel survey this paper explores the effects of parental work schedules on the mental and physical health of adolescents aged 15-20 in sole-parent families. Random-effects models indicate adolescents have marginally worse emotional and physical health when their parent works nonstandard hours, based on SF-36 component summary scores, associated primarily with emotional or physical role limitations. Parental weekend schedules are particularly detrimental to adolescent physical health. Evidence is found that the effects of nonstandard work schedules on adolescent wellbeing are transmitted through increased work-family conflict and exacerbated where parents have low job control.


Silent loss and the clinical encounter: Parents' and physicians' experiences of stillbirth-a qualitative analysis.
In the United States, an estimated 70 stillbirths occur each day, on average 25,000 each year. Research into the prevalence and causes of stillbirth is ongoing, but meanwhile, many parents suffer this devastating loss, largely in silence, due to persistent stigma and taboo; and many health providers report feeling ill equipped to support grieving parents. Interventions to address bereavement after neonatal death are increasingly common in U.S. hospitals, and there is growing data on the nature of parent bereavement after a stillbirth. However, further research is needed to evaluate supportive interventions and to investigate the parent-clinician encounter during hospitalization following a stillbirth. Qualitative inquiry offers opportunities to better understand the lived experience of parents against the backdrop of clinicians' beliefs, intentions, and well-meaning efforts to support grieving parents. We present a secondary qualitative analysis of transcript data from 3 semi-structured focus groups conducted with parents who had experienced a stillbirth and delivered in a hospital, and 2 focus groups with obstetrician-gynecologists. Participants were drawn from the greater Seattle region in Washington State. We examine parents' and physicians' experiences and beliefs surrounding stillbirth during the clinical encounter using iterative discourse analysis. Women reported that the cheery, bustling environment of the labor and delivery setting was a painful place for parents who had had a stillbirth, and that the well-meaning attempts of physicians to offer comfort often had the opposite effect. Parents also reported that their grief is deeply felt but not socially recognized. While physicians recognized patients' grief, they did not grasp its depth or duration. Physicians viewed stillbirth as an unexpected clinical tragedy, though several considered stillbirth less traumatic than the death of a neonate. In the months and years following a stillbirth, these parents continue to memorialize their children as part of their family. Hospitals need to examine the physical environment for deliveries and, wherever possible, offer designated private areas with staff trained in stillbirth care. Training programs in obstetrics need to better address the bereavement needs of parents following a stillbirth, and research is needed to evaluate effective bereavement interventions, accounting for cultural variation. Critical improvements are also needed for mental health support beyond hospitalization. Finally, medical professionals and parents can play an important role in reversing the stigma that surrounds stillbirth.


Lactoferrin conjugated iron oxide nanoparticles for targeting brain glioma cells in magnetic particle imaging.
Magnetic Particle Imaging (MPI) is a new real-time imaging modality, which promises high tracer mass sensitivity and spatial resolution directly generated from iron oxide nanoparticles. In this study, monodisperse iron oxide nanoparticles with median core diameters ranging from 14 to 26 nm were synthesized and their surface was conjugated with lactoferrin to convert them into brain glioma targeting agents. The conjugation was confirmed with the increase of the hydrodynamic diameters, change of zeta potential, and Bradford assay. Magnetic particle spectrometry (MPS), performed to evaluate the MPI performance of these nanoparticles, showed no change in signal after lactoferrin conjugation to nanoparticles for all core diameters, suggesting that the MPI signal is dominated by Néel relaxation and thus independent of hydrodynamic size difference or presence of coating molecules before and after conjugations. For this range of core sizes (14-26 nm), both MPS signal intensity and spatial resolution improved with increasing core diameter of nanoparticles. The lactoferrin conjugated iron oxide nanoparticles (Lf-IONPs) showed specific cellular internalization into C6 cells with a 5-fold increase in MPS signal compared to IONPs without lactoferrin, both after 24 h incubation. These results suggest that Lf-IONPs can be used as tracers for targeted brain glioma imaging using MPI.


Whole Genome Profiling provides a robust framework for physical mapping and sequencing in the highly complex and repetitive wheat genome.
Sequencing projects using a clone-by-clone approach require the availability of a robust physical map. The SNaPshot technology, based on pair-wise comparisons of restriction fragments sizes, has been used recently to build the first physical map of a wheat chromosome and to complete the maize physical map. However, restriction fragments sizes shared randomly between two non-overlapping BACs often lead to chimerical contigs and mis-assembled BACs in such large and repetitive genomes. Whole Genome Profiling (WGP™) was developed recently as a new sequence-based physical mapping technology and has the potential to limit this problem. A subset of the wheat 3B chromosome BAC library covering 230 Mb was used to establish a WGP physical map and to compare it to a map obtained with the SNaPshot technology. We first adapted the WGP-based assembly methodology to cope with the complexity of the wheat genome. Then, the results showed that the WGP map covers the same length than the SNaPshot map but with 30% less contigs and, more importantly with 3.5 times less mis-assembled BACs. Finally, we evaluated the benefit of integrating WGP tags in different sequence assemblies obtained after Roche/454 sequencing of BAC pools. We showed that while WGP tag integration improves assemblies performed with unpaired reads and with paired-end reads at low coverage, it does not significantly improve sequence assemblies performed at high coverage (25x) with paired-end reads. Our results demonstrate that, with a suitable assembly methodology, WGP builds more robust physical maps than the SNaPshot technology in wheat and that WGP can be adapted to any genome. Moreover, WGP tag integration in sequence assemblies improves low quality assembly. However, to achieve a high quality draft sequence assembly, a sequencing depth of 25x paired-end reads is required, at which point WGP tag integration does not provide additional scaffolding value. Finally, we suggest that WGP tags can support the efficient sequencing of BAC pools by enabling reliable assignment of sequence scaffolds to their BAC of origin, a feature that is of great interest when using BAC pooling strategies to reduce the cost of sequencing large genomes.


Association of Low-Carbohydrate and Low-Fat Diets With Mortality Among US Adults.
It is crucial to incorporate quality and types of carbohydrate and fat when investigating the associations of low-fat and low-carbohydrate diets with mortality. To investigate the associations of low-carbohydrate and low-fat diets with total and cause-specific mortality among US adults. This prospective cohort study used data from the US National Health and Nutrition Examination Survey from 1999 to 2014 from 37 233 adults 20 years or older with 24-hour dietary recall data. Data were analyzed from July 5 to August 27, 2019. Overall, unhealthy, and healthy low-carbohydrate-diet and low-fat-diet scores based on the percentage of energy as total and subtypes of carbohydrate, fat, and protein. All-cause mortality from baseline until December 31, 2015, linked to National Death Index mortality data. A total of 37 233 US adults (mean [SD] age, 49.7 [18.3] years; 19 598 [52.6%] female) were included in the present analysis. During 297 768 person-years of follow-up, 4866 total deaths occurred. Overall low-carbohydrate-diet and low-fat-diet scores were not associated with total mortality. The multivariable-adjusted hazard ratios for total mortality per 20-percentile increase in dietary scores were 1.07 (95% CI, 1.02-1.11; P = .01 for trend) for unhealthy low-carbohydrate-diet score, 0.91 (95% CI, 0.87-0.95; P < .001 for trend) for healthy low-carbohydrate-diet score, 1.06 (95% CI, 1.01-1.12; P = .04 for trend) for unhealthy low-fat-diet score, and 0.89 (95% CI, 0.85-0.93; P < .001 for trend) for healthy low-fat-diet score. The associations remained similar in the stratification and sensitivity analyses. In this study, overall low-carbohydrate-diet and low-fat-diet scores were not associated with total mortality. Unhealthy low-carbohydrate-diet and low-fat-diet scores were associated with higher total mortality, whereas healthy low-carbohydrate-diet and low-fat-diet scores were associated with lower total mortality. These findings suggest that the associations of low-carbohydrate and low-fat diets with mortality may depend on the quality and food sources of macronutrients.