Role of mutational reversions and fitness restoration in Zika virus spread to the Americas.
Zika virus (ZIKV) emerged from obscurity in 2013 to spread from Asia to the South Pacific and the Americas, where millions of people were infected, accompanied by severe disease including microcephaly following congenital infections. Phylogenetic studies have shown that ZIKV evolved in Africa and later spread to Asia, and that the Asian lineage is responsible for the recent epidemics in the South Pacific and Americas. However, the reasons for the sudden emergence of ZIKV remain enigmatic. Here we report evolutionary analyses that revealed four mutations, which occurred just before ZIKV introduction to the Americas, represent direct reversions of previous mutations that accompanied earlier spread from Africa to Asia and early circulation there. Our experimental infections of Aedes aegypti mosquitoes, human cells, and mice using ZIKV strains with and without these mutations demonstrate that the original mutations reduced fitness for urban, human-amplifed transmission, while the reversions restored fitness, increasing epidemic risk. These findings include characterization of three transmission-adaptive ZIKV mutations, and demonstration that these and one identified previously restored fitness for epidemic transmission soon before introduction into the Americas. The initial mutations may have followed founder effects and/or drift when the virus was introduced decades ago into Asia.


Comparative analysis of canine monocyte- and bone-marrow-derived dendritic cells.
Dendritic cells (DC) represent a heterogeneous cell family of major importance for innate immune responses against pathogens and antigen presentation during infection, cancer, allergy and autoimmunity. The aim of the present study was to characterize canine DC generated in vitro with respect to their phenotype, responsiveness to toll-like receptor (TLR) ligands and T-cell stimulatory capacity. DC were derived from monocytes (MoDC) and from bone marrow hematopoietic cells cultured with either Flt3-ligand (FL-BMDC) or with GM-CSF (GM-BMDC). All three methods generated cells with typical DC morphology that expressed CD1c, CD11c and CD14, similar to macrophages. However, CD40 was only found on DC, CD206 on MPhi and BMDC, but not on monocytes and MoDC. CD1c was not found on monocytes but on all in vitro differentiated cells. FL-BMDC and GM-BMDC were partially positive for CD4 and CD8. CD45RA was expressed on a subset of FL-BMDC but not on MoDC and GM-BMDC. MoDC and FL-DC responded well to TLR ligands including poly-IC (TLR2), Pam3Cys (TLR3), LPS (TLR4) and imiquimod (TLR7) by up-regulating MHC II and CD86. The generated DC and MPhi showed a stimulatory capacity for lymphocytes, which increased upon maturation with LPS. Taken together, our results are the basis for further characterization of canine DC subsets with respect to their role in inflammation and immune responses.


[Acute peritonitis as a complication of Crohn's disease of the colon (author's transl)].
Acute peritonitis is a rare complication of Crohn's disease of the colon. We present the case of a young woman with peritonitis complicating an acute exacerbation of Crohn's disease of the colon. Seventeen other cases from the literature show that perforation may be the first manifestation of the disease or may occur during an acute exacerbation of a chronic form. With the presently limited experience, the optimal surgical treatment has not been defined.


[Effect of music on children with cancer].
This study looked for a relationship between immunity and one's spirits while investigating the effect musical therapy produces on Immunoglobulina A (IgA) found in saliva and a Patient's Opinion on the Likert Scale (OPEL). There were 30 children as patients, 15 in a control group and 15 in an experimental group. They were 5 or 16 years of age and checked into the Sant Joan de Deu Hospital in Barcelona due to neoplastic illness.


The Effect of H2 Receptor Antagonist in Acid Inhibition and Its Clinical Efficacy.
The first histamine H2 receptor antagonists (H2RAs) were developed in the early 1970s. They played a dominant role in treating peptic ulcer disease and gastroesophageal reflux disease (GERD). H2RAs block the production of acid by H+, K+-ATPase at the parietal cells and produce gastric luminal anacidity for varying periods. H2RAs are highly selective, and they do not affect H1 receptors. Moreover, they are not anticholinergic agents. Sequential development of H2RAs, proton pump inhibitors (PPIs), and discovery of Helicobacter pylori infection changed the paradigm of peptic ulcer disease with marked decrease of morbidity and mortality. PPIs are known to be the most effective drugs that are currently available for suppressing gastric acid secretion. Many studies have shown its superiority over H2RAs as a treatment for acid-related disorders, such as peptic ulcer disease, GERD, and Zollinger-Ellison syndrome. However, other studies have reported that PPIs may not be able to render stomach achlorhydric and have identified a phenomenon of increasing gastric acidity at night in individuals receiving a PPI twice daily. These nocturnal acid breakthrough episodes can be eliminated with an addition of H2RAs at night. The effectiveness of nighttime dose of H2RA suggests a major role of histamine in nocturnal acid secretion. H2RAs reduce secretion of gastric acid, and each H2RA also has specific effects. For instance, nizitidine alleviates not only symptoms of GERD, but also provokes gastric emptying, resulting in clinical symptom improvement of functional dyspepsia. The aim of this paper was to review the characteristics and role of H2RAs and assess the future strategy and treatment of upper gastrointestinal disease, including acid related disorders.


Confidence intervals for proportion difference from two independent partially validated series.
Partially validated series are common when a gold-standard test is too expensive to be applied to all subjects, and hence a fallible device is used accordingly to measure the presence of a characteristic of interest. In this article, confidence interval construction for proportion difference between two independent partially validated series is studied. Ten confidence intervals based on the method of variance estimates recovery (MOVER) are proposed, with each using the confidence limits for the two independent binomial proportions obtained by the asymptotic, Logit-transformation, Agresti-Coull and Bayesian methods. The performances of the proposed confidence intervals and three likelihood-based intervals available in the literature are compared with respect to the empirical coverage probability, confidence width and ratio of mesial non-coverage to non-coverage probability. Our empirical results show that (1) all confidence intervals exhibit good performance in large samples; (2) confidence intervals based on MOVER combining the confidence limits for binomial proportions based on Wilson, Agresti-Coull, Logit-transformation, Bayesian (with three priors) methods perform satisfactorily from small to large samples, and hence can be recommended for practical applications. Two real data sets are analysed to illustrate the proposed methods.


Enamel matrix derivative and titanium implants.
The aim of present study was to evaluate if an enamel matrix derivative (Emdogain) may enhance bone formation and osseointegration of titanium implants, using a well-documented rabbit model. Thirty-six threaded commercially pure titanium (cp.ti.) implants were inserted in six New Zealand white rabbits. One implant was placed in each femur and two in each tibia. Prior to implant insertion approximately 0.5 mL of Emdogain (EMD) (test) or the vehicle gel (PGA: propylene glycol alginate) (control) was injected into the surgically prepared implant site. The follow-up time was 6 weeks. Biomechanical evaluations by resonance frequency analysis (RFA) and removal torque measurements (RTQ) were performed. Histomorphometrical quantifications were made on ground sections by measurements of the percentage of bone-to-metal contact, bone area inside the threads as well as outside the threads (mirror image). Bone lengths along the implant surface were also measured and used for shear strength calculations. The results demonstrated no beneficial effects from the EMD treatment on bone formation around titanium implants in any of the tested parameters. Significant differences were demonstrated with removal torque test and shear force calculations for the control implants. No other parameter demonstrated a statistically significant difference. The results of the present study may indicate that EMD does not contribute to bone formation around titanium implants. This observation may indicate that the bone formation that occurs after EMD treatment in periodontal defects is the result of functional adaptation. However, further research is required to evaluate the effect of EMD treatment on bone formation.


Anisomycin injection in area CA3 of the hippocampus impairs both short-term and long-term memories of contextual fear.
Protein synthesis is involved in the consolidation of short-term memory into long-term memory. Previous electrophysiological data concerning LTP in CA3 suggest that protein synthesis in that region might also be necessary for short-term memory. We tested this hypothesis by locally injecting the protein synthesis inhibitor anisomycin in hippocampal area CA1 or CA3 immediately after contextual fear conditioning. As previously shown, injections in CA1 impaired long-term memory but spared short-term memory. Conversely, injections in CA3 impaired both long-term and short-term memories. We conclude that early steps of experience-induced plasticity occurring in CA3 and underlying short-term memory require protein synthesis.


Prediction of off-target specificity and cell-specific fitness of CRISPR-Cas System using attention boosted deep learning and network-based gene feature.
CRISPR-Cas is a powerful genome editing technology and has a great potential for in vivo gene therapy. Successful translational application of CRISPR-Cas to biomedicine still faces many safety concerns, including off-target side effect, cell fitness problem after CRISPR-Cas treatment, and on-target genome editing side effect in undesired tissues. To solve these issues, it is needed to design sgRNA with high cell-specific efficacy and specificity. Existing single-guide RNA (sgRNA) design tools mainly depend on a sgRNA sequence and the local information of the targeted genome, thus are not sufficient to account for the difference in the cellular response of the same gene in different cell types. To incorporate cell-specific information into the sgRNA design, we develop novel interpretable machine learning models, which integrate features learned from advanced transformer-based deep neural network with cell-specific gene property derived from biological network and gene expression profile, for the prediction of CRISPR-Cas9 and CRISPR-Cas12a efficacy and specificity. In benchmark studies, our models significantly outperform state-of-the-art algorithms. Furthermore, we find that the network-based gene property is critical for the prediction of cell-specific post-treatment cellular response. Our results suggest that the design of efficient and safe CRISPR-Cas needs to consider cell-specific information of genes. Our findings may bolster developing more accurate predictive models of CRISPR-Cas across a broad spectrum of biological conditions as well as provide new insight into developing efficient and safe CRISPR-based gene therapy.


Stem Cell-Derived Endothelial Cell Model that Responds to Tobacco Smoke Like Primary Endothelial Cells.
To clarify how smoking leads to heart attack and stroke, we developed an endothelial cell model (iECs) generated from human induced Pluripotent Stem Cells (iPSC) and evaluated its responses to tobacco smoke. These iECs exhibited a uniform endothelial morphology, and expressed markers PECAM1/CD31, VWF/ von Willebrand Factor, and CDH5/VE-Cadherin. The iECs also exhibited tube formation and acetyl-LDL uptake comparable to primary endothelial cells (EC). RNA sequencing (RNA-Seq) revealed a robust correlation coefficient between iECs and EC (R = 0.76), whereas gene responses to smoke were qualitatively nearly identical between iECs and primary ECs (R = 0.86). Further analysis of transcriptional responses implicated 18 transcription factors in regulating responses to smoke treatment, and identified gene sets regulated by each transcription factor, including pathways for oxidative stress, DNA damage/repair, ER stress, apoptosis, and cell cycle arrest. Assays for 42 cytokines in HUVEC cells and iECs identified 23 cytokines that responded dynamically to cigarette smoke. These cytokines and cellular stress response pathways describe endothelial responses for lymphocyte attachment, activation of coagulation and complement, lymphocyte growth factors, and inflammation and fibrosis; EC-initiated events that collectively lead to atherosclerosis. Thus, these studies validate the iEC model and identify transcriptional response networks by which ECs respond to tobacco smoke. Our results systematically trace how ECs use these response networks to regulate genes and pathways, and finally cytokine signals to other cells, to initiate the diverse processes that lead to atherosclerosis and cardiovascular disease.


Paroxysmal nocturnal hemoglobinuria and the risk of venous thrombosis: review and recommendations for management of the pregnant and nonpregnant patient.
Paroxysmal nocturnal hemoglobinuria is a rare, clonal primitive hematopoietic cell disorder, often affecting middle-aged adults, including women of reproductive age. Major morbidity and mortality with this disease are often ascribed to the development of venous thromboembolism. We reviewed the current literature on the risk of venous thrombosis among nonpregnant and pregnant patients, and generated recommendations for the prevention of venous thromboembolism, as well as duration of treatment for affected patients who develop thrombotic disease. We searched Medline for papers published between January 1966 and April 1999. We also requested relevant unpublished data from speakers who attended a recent international workshop of paroxysmal nocturnal hemoglobinuria. References from all primary data and review publications were also examined. Only English language publications were included. Event rates for venous thromboembolism and death were pooled using a random effect technique. Reports of paroxysmal nocturnal hemoglobinuria during pregnancy were summarized using descriptive statistics only. Thirteen retrospective studies of paroxysmal nocturnal hemoglobinuria in nonpregnant individuals were found. The rates of venous thrombosis varied considerably, but were reported to affect 14.4% of all individuals [95% confidence interval (CI) 7.6-25.5]. Among patients from western nations, venous thromboembolism seemed to develop at a higher rate (30.3%, 95% CI 26. 1-34.9). The majority of venous thromboembolic events were intra-abdominal, principally within the hepatic and mesenteric veins. The likely cause of death among patients with paroxysmal nocturnal hemoglobinuria was described in nine studies: 22.2% of fatalities were due to venous thrombosis (95% CI 11.8-38.0), more commonly in western countries (event rate 37.2%, 95% CI 21.6-56.0). Another 20 published reports described the outcome of 33 pregnant women with paroxysmal nocturnal hemoglobinuria. Two women developed venous thromboembolism during pregnancy and another 2 during the postpartum state for a combined event rate of 12.1% (95% CI 3.4-25.2), three of which resulted in death. The all-cause mortality rate was 20.8% (95% CI 7.3-39.0). Both anemia (event rate 72.7%, 95% CI 56.5-86.3), and thrombocytopenia (event rate 27.3%, 95% CI 13.7-43.5) were common, often necessitating red cell or platelet transfusions. Almost half of all infants (54.8%, 95% CI 36.1-72.7) were delivered preterm, and had a mean live birth weight of 2,800 g. Three of 34 reported births ended in death (perinatal mortality rate 8.8%, 95% C 1.9-23.7). In accordance with the apparently high rate of venous thrombosis among pregnant and nonpregnant individuals with paroxysmal nocturnal hemoglobinuria, especially for fatal thrombosis, we developed practical recommendations for the prevention and treatment of venous thromboembolic disease in these groups.


Acute effects of cigarette smoking on microcirculation of the thumb.
The acute effect of smoking on the microcirculation of the skin of the thumb was investigated in healthy volunteers. Twenty-two were smokers and 10 were non-smokers. The flow was assessed by means of laser Doppler flowmetry. The smokers inhaled 2 cigarettes. During smoking of their first and second cigarette respectively, a mean decrease in laser Doppler flow of 23.8% and 29.0% was seen (p = 0.03; p = 0.01). Ten minutes after smoking this decrease was recovered by half. This experiment confirms that one should prohibit smoking of cigarettes pre- and postoperatively for optimal wound healing conditions.


ST segment elevation pattern of acute myocardial infarction induced by acute pancreatitis.
The authors report the first emergent angiographic assessment of the coronaries with accompanying echocardiography in a 64-year-old man with dermatomyositis, who presented with ST segment elevation and cardiac specific enzyme derangements highly suggestive of myocardial infarction in the presence of acute pancreatitis. Both studies revealed no anatomical or functional evidence of obstructive coronary disease. Although the mechanism of electrocardiogram abnormalities found in acute pancreatitis remains to be elucidated fully, the authors propose a direct cardiac toxic effect by the pancreatic proteolytic enzymes to account for these changes and we recommend an angiographic approach as the first step to avoid the potentially lethal administration of thrombolytic therapy or potent anticoagulation.


Asymmetric T lymphocyte division in the initiation of adaptive immune responses.
A hallmark of mammalian immunity is the heterogeneity of cell fate that exists among pathogen-experienced lymphocytes. We show that a dividing T lymphocyte initially responding to a microbe exhibits unequal partitioning of proteins that mediate signaling, cell fate specification, and asymmetric cell division. Asymmetric segregation of determinants appears to be coordinated by prolonged interaction between the T cell and its antigen-presenting cell before division. Additionally, the first two daughter T cells displayed phenotypic and functional indicators of being differentially fated toward effector and memory lineages. These results suggest a mechanism by which a single lymphocyte can apportion diverse cell fates necessary for adaptive immunity.


Validation of Candidate Gene-Based Markers and Identification of Novel Loci for Thousand-Grain Weight in Spring Bread Wheat.
Increased thousand-grain weight (TGW) is an important breeding target for indirectly improving grain yield (GY). Fourteen reported candidate genes known to enhance TGW were evaluated in two independent and existing datasets of wheat at CIMMYT, the Elite Yield Trial (EYT) from 2015 to 2016 (EYT2015-16) and the Wheat Association Mapping Initiative (WAMI) panel, to study their allele effects on TGW and to verify their suitability for marker-assisted selection. Of these, significant associations were detected for only one gene (TaGs3-D1) in the EYT2015-16 and two genes (TaTGW6 and TaSus1) in WAMI. The reported favorable alleles of TaGs3-D1 and TaTGW6 genes decreased TGW in the datasets. A haplotype-based genome wide association study was implemented to identify the genetic determinants of TGW on a large set of CIMMYT germplasm (4,302 lines comprising five EYTs), which identified 15 haplotype blocks to be significantly associated with TGW. Four of them, identified on chromosomes 4A, 6A, and 7A, were associated with TGW in at least three EYTs. The locus on chromosome 6A (Hap-6A-13) had the largest effect on TGW and additionally GY with increases of up to 2.60 g and 258 kg/ha, respectively. Discovery of novel TGW loci described in our study expands the opportunities for developing diagnostic markers and for multi-gene pyramiding to derive new allele combinations for enhanced TGW and GY in CIMMYT wheat.


Sleep Disturbance and Disorders within Adult Inpatient Rehabilitation Settings: A Systematic Review to Identify Both the Prevalence of Disorders and the Efficacy of Existing Interventions.
Older people who are admitted to inpatient rehabilitation settings often experience sleep disturbance or disorders. Both intrinsic and environmental factors may contribute to reduced sleep quality. Poor sleep quality has been reported to be associated with poorer rehabilitation outcomes. The aim of this review was (1) to describe the prevalence of sleep disturbance or disorder among older people participating in inpatient rehabilitation, and (2) to describe interventions that have been trialed to increase sleep quality and/or quantity in older people in rehabilitation settings and report on their efficacy. Systematic review involving search of 3 electronic databases and gray literature. Two authors independently reviewed citations and reviewed full text and agreed on included studies. Data were extracted and synthesized and risk of bias was assessed. Studies were included if they provided quantitative data about the prevalence of sleep disturbance or disorder in older people in a rehabilitation setting or if they reported the results of a randomized trial evaluating an intervention to improve sleep quality in older people in a rehabilitation setting. Studies were included if they reported data from monitoring (such as polysomnography or actigraphy), clinical assessments, or questionnaires. 16 studies reporting prevalence data and 3 studies reporting evaluations of interventions were included in this review. Studies reported data from stroke and mixed population rehabilitation settings. The prevalence of obstructive sleep apnea was high and ranged from 12% to 92% in stroke rehabilitation settings. Other types of sleep disturbance, such as difficulty initiating sleep, were also common. Many older people participating in inpatient rehabilitation have sleep disturbance or sleep disorders. Poor sleep quality is associated with poorer health status and recovery; therefore, it is important that rehabilitation settings take steps to enhance sleep quality for inpatients drawing on the principles of good sleep hygiene.


Active breathing control (ABC) for Hodgkin's disease: reduction in normal tissue irradiation with deep inspiration and implications for treatment.
Active breathing control (ABC) temporarily immobilizes breathing. This may allow a reduction in treatment margins. This planning study assesses normal tissue irradiation and reproducibility using ABC for Hodgkin's disease. Five patients underwent CT scans using ABC obtained at the end of normal inspiration (NI), normal expiration (NE), and deep inspiration (DI). DI scans were repeated within the same session and 1-2 weeks later. To simulate mantle radiotherapy, a CTV1 was contoured encompassing the supraclavicular region, mediastinum, hila, and part of the heart. CTV2 was the same as CTV1 but included the whole heart. CTV3 encompassed the spleen and para-aortic lymph nodes. The planning target volume (PTV) was defined as CTV + 9 mm. PTVs were determined at NI, NE, and DI. A composite PTV (comp-PTV) based on the range of NI and NE PTVs was determined to represent the margin necessary for free breathing. Lung dose-mass histograms (DMH) for PTV1 and PTV2 and cardiac dose-volume histograms (DVH) for PTV3 were compared at the three different respiratory phases. ABC was well-tolerated by all patients. DI breath-holds ranged from 34 to 45 s. DMHs determined for PTV1 revealed a median reduction in lung mass irradiated at DI of 12% (range, 9-24%; n = 5) compared with simulated free-breathing. PTV2 comparisons also showed a median reduction of 12% lung mass irradiated (range, 8-28%; n = 5). PTV3 analyses revealed the mean volume of heart irradiated decreased from 26% to 5% with deep inspiration (n = 5). Lung volume comparisons between intrasession and intersession DI studies revealed mean variations of 4%. ABC is well tolerated and reproducible. Radiotherapy delivered at deep inspiration with ABC may decrease normal tissue irradiation in Hodgkin's disease patients.


Ultrasonic Courtship Vocalizations of Adult Male Mice: A Laboratory Exercise Illustrating Comparable Activation by either Estradiol or Testosterone.
The importance of testosterone (T) in maintaining full expression of male mammal reproductive behavior is well documented and widely acknowledged. However, the role of estradiol (E2) in the regulation of male reproductive behaviors is much less widely recognized or appreciated. Indeed, T can be enzymatically converted to E2 via aromatase. Many regions within the brain of males and females are rich in aromatase, thereby offering the possibility that many of T's actions in the brain require conversion to E2. According to this aromatization hypothesis, full expression of male reproductive behaviors is mediated via the action of E2 in the brain. This paper describes a laboratory exercise that illustrates this phenomenon as it relates to 70 kHz ultrasonic courtship vocalizations by male mice in response to either a female conspecific or the odor of female urine. The results indicate that while castration reduces male vocalizations to either female stimulus, subcutaneous implants of estradiol benzoate (EB) or testosterone propionate (TP) are equivalent in maintaining high levels of vocalization. I find this exercise extremely useful in illustrating to students why the characterization of hormones as either masculine or feminine is erroneous. Background information is presented that will help guide class discussions. Modifications to the laboratory exercise are described that do not require surgical castration or the use of controlled substances.


Topographies of retinal cone photoreceptors in two Australian marsupials.
Microspectrophotometry indicates the presence of at least three cone visual pigments in two Australian marsupials, the fat-tailed dunnart (Sminthopsis crassicaudata) and honey possum (Tarsipes rostratus). Here we have examined the distribution of cone types using antisera, JH455 and JH492, that recognize short-wavelength-sensitive (SWS) and medium-to-long-wavelength-sensitive (M/LWS) cone opsins, respectively. SWS cones were concentrated in dorso-temporal retina in the dunnart with a shallow decreasing gradient extending to the periphery (2300-1500/mm2). In the honey possum, SWS cones showed a uniform distribution (2700/mm2), except for a slight increase in a narrow peripheral band (3100/mm2). In both species, M/LWS cones dominated and their distributions were similar to those of retinal ganglion cells: a horizontal streak in the dunnart (31,000-21,000/mm2) and a shallow mid-ventral to peripheral gradient in the honey possum (37,000-26,000/mm2). A low number of cones remained unlabeled when the antisera were combined revealing further minority cone population(s). We discuss cone distributions in relation to visual capabilities and requirements of the species.


Tuberculosis screening in internationally adopted children: the need for initial and repeat testing.
Because most internationally adopted children come from areas of high tuberculosis prevalence, an initial tuberculin skin test is recommended after arrival to the United States. We evaluated whether repeat testing of children >or=3 months after arrival to the United States would identify additional children with latent tuberculosis infection. Internationally adopted children who were seen at our International Adoption Center and had a tuberculin skin test within 2 months of arrival to the United States were eligible for the study. Children not diagnosed with tuberculosis with initial testing were retested at least 3 months later. The prevalence of tuberculosis on arrival and after repeat testing was determined, and potential risk factors for infection were examined. Of the 527 internationally adopted children with an initial tuberculin skin test completed, 111 (21%) had evidence of latent tuberculosis infection. Repeat tuberculosis testing was complete for 191 internationally adopted children (46.9% of those who had an initially negative tuberculin skin test). Latent tuberculosis infection was found in 20% of those who were retested. No children were found to have active tuberculosis disease. Children with an initially positive tuberculin skin test result had slightly higher weight-for-age z scores at their initial clinic visit, whereas those whose tuberculin skin test result was positive after repeat testing had slightly lower weight-for-age z scores. A strong correlation between BCG immunization and tuberculin skin test result was observed. Latent tuberculosis infection is common in internationally adopted children. A high proportion of internationally adopted children had an initially false-negative tuberculin skin test. Repeat tuberculosis testing of all internationally adopted children with an initially negative tuberculin skin test should be the standard of care for identifying tuberculosis infection and preventing tuberculosis disease in this high-risk population.


Cognitive reserve moderates decline in information processing speed in multiple sclerosis patients.
Cognitive reserve is widely recognized as a moderator of cognitive decline in patients with senile dementias such as Alzheimer's disease. The same effect may occur in multiple sclerosis (MS), an immunologic disorder affecting the central nervous system. While MS is traditionally considered an inflammatory, white matter disease, degeneration of gray matter is increasingly recognized as the primary contributor to progressive cognitive decline. Our aim was to determine if individual differences in estimated cognitive reserve protect against the progression of cognitive dysfunction in MS. Ninety-one patients assessed twice roughly 5 years apart were identified retrospectively. Cognitive testing emphasized mental processing speed. Cognitive reserve was estimated by years of education and by performance on the North American Adult Reading Test (NAART). After controlling for baseline characteristics, both years of education (p = .013) and NAART scores (p = .049) significantly improved regression models predicting cognitive decline. Symbol Digit Modalities Test (SDMT) performance showed no significant change in patients with > 14 years of education, whereas it declined significantly in patients with ≤ 14 years of education. We conclude that greater cognitive reserve as indexed by either higher premorbid intelligence or more years of education protects against the progression of cognitive dysfunction in MS.


Treatment burden experienced by patients with lung cancer.
Patients' burden from lung cancer treatment is not well researched, but this understanding can facilitate a patient-centred treatment approach. Current models of treatment burden suggest it is influenced by a patient's perception of their disease and treatment and their capacity to do the work required to treat their disease. Sixteen patients and 1 carer who were undergoing or had completed conventional or stereotactic ablative radiotherapy, chemotherapy or immunotherapy for lung cancer in the last 6 months participated in a semi-structured interview. A treatment burden framework was used with three main themes: a) treatment work, b) consequences of treatment and c) psychosocial factors affecting treatment burden. The majority of patients did not feel unduly burdened by treatment tasks, despite having a large treatment-associated workload. Many saw treatment as a priority, causing them to restructure their life to accommodate for it. Patients wished that they would have been better informed about the lifestyle changes that they would have to make before treatment for lung cancer commenced and that the health service would provide services to assist them with this task. While there was a large burden associated with lung cancer treatment, patients felt motivated and equipped to manage the workload because the disease was considered severe and life-threatening, and the treatment was seen as beneficial. Before initiating treatment for lung cancer, patients should be informed about lifestyle changes they likely have to make and should be offered assistance.


A population-based approach for implementing change from opt-out to opt-in research permissions.
Due to recently proposed changes in the Common Rule regarding the collection of research preferences, there is an increased need for efficient methods to document opt-in research preferences at a population level. Previously, our institution developed an opt-out paper-based workflow that could not be utilized for research in a scalable fashion. This project was designed to demonstrate the feasibility of implementing an electronic health record (EHR)-based active opt-in research preferences program. The first phase of implementation required creating and disseminating a patient questionnaire through the EHR portal to populate discreet fields within the EHR indicating patients' preferences for future research study contact (contact) and their willingness to allow anonymised use of excess tissue and fluid specimens (biobank). In the second phase, the questionnaire was presented within a clinic nurse intake workflow in an obstetrical clinic. These permissions were tabulated in registries for use by investigators for feasibility studies and recruitment. The registry was also used for research patient contact management using a new EHR encounter type to differentiate research from clinical encounters. The research permissions questionnaire was sent to 59,670 patients via the EHR portal. Within four months, 21,814 responses (75% willing to participate in biobanking, and 72% willing to be contacted for future research) were received. Each response was recorded within a patient portal encounter to enable longitudinal analysis of responses. We obtained a significantly lower positive response from the 264 females who completed the questionnaire in the obstetrical clinic (55% volunteers for biobank and 52% for contact). We demonstrate that it is possible to establish a research permissions registry using the EHR portal and clinic-based workflows. This patient-centric, population-based, opt-in approach documents preferences in the EHR, allowing linkage of these preferences to health record information.


Sources of heavy metal pollution in agricultural soils of a rapidly industrializing area in the Yangtze Delta of China.
The rapid industrialization and urbanization in developing countries have increased pollution by heavy metals, which is a concern for human health and the environment. In this study, 230 surface soil samples (0-20cm) were collected from agricultural areas of Jiaxing, a rapidly industrializing area in the Yangtze Delta of China. Sequential Gaussian simulation (SGS) and multivariate factorial kriging analysis (FKA) were used to identify and explore the sources of heavy metal pollution for eight metals (Cu, Zn, Pb, Cr, Ni, Cd, Hg and As). Localized hot-spots of pollution were identified for Cu, Zn, Pb, Cr, Ni and Cd with area percentages of 0.48 percent, 0.58 percent, 2.84 percent, 2.41 percent, 0.74 percent, and 0.68 percent, respectively. The areas with Hg pollution covered approximately 38 percent whereas no potential pollution risk was found for As. The soil parent material and point sources of pollution had significant influences on Cr, Ni, Cu, Zn and Cd levels, except for the influence of agricultural management practices also accounted for micro-scale variations (nugget effect) for Cu and Zn pollution. Short-range (4km) diffusion processes had a significant influence on Cu levels, although they did not appear to be the dominant sources of Zn and Cd variation. The short-range diffusion pollution arising from current and historic industrial emissions and urbanization, and long-range (33km) variations in soil parent materials and/or diffusion jointly determined the current concentrations of soil Pb. The sources of Hg pollution risk may be attributed to the atmosphere deposition of industrial emission and historical use of Hg-containing pesticides.


Training modalities: impact on endurance capacity.
Endurance athletes demonstrate an exceptional resistance to fatigue when exercising at high intensity. Much research has been devoted to the contribution of aerobic capacity for the economy of endurance performance. Important aspects of the fine-tuning of metabolic processes and power output in the endurance athlete have been overlooked. This review addresses how training paradigms exploit bioenergetic pathways in recruited muscle groups to promote the endurance phenotype. A special focus is laid on the genome-mediated mechanisms that underlie the conditioning of fatigue resistance and aerobic performance by training macrocycles and complements. The available data on work-induced muscle plasticity implies that different biologic strategies are exploited in athletic and untrained populations to boost endurance capacity. Olympic champions are probably endowed with a unique constitution that renders the conditioning of endurance capacity for competition particularly efficient.


The role of rearrests in juvenile offenders' and their mothers' attitudes toward police.
Both personal experience and parental attitudes shape youths' attitudes toward the justice system. The present study tested the influence of (a) youth rearrests and (b) parents' attitudes toward police on trajectories of youthful offenders' attitudes toward police over 3 years. Among a sample of 317 first-time male juvenile offenders and their mothers, group-based trajectory modeling identified 4 trajectories of youths' attitudes toward police over the 3 years since the youths' first arrests. Mothers with more positive initial attitudes toward the police were more likely to have sons who were part of 1 of the 2 positive attitude trajectory groups than the 2 negative attitude trajectory groups. In contrast, youth rearrests were rarely associated with trajectory group membership, suggesting that a mother's attitude toward the police may outweigh a youth's own experience (via arrests) with the justice system in determining trajectories of his attitudes toward the police. Additionally, the present study tests whether parents' own attitudes toward police are predicted by their children's rearrests. We found that mothers whose sons were rearrested during the study period had more negative attitudes toward the police. The results add a novel component to our understanding of family attitude dynamics as related to perceptions of law enforcement and juvenile arrests. Overall, a family level perspective is necessary when considering the effects of rearrests on youths'-and their parents'-attitudes toward the police. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


A photobleaching resistant polymer supported hexanuclear molybdenum iodide cluster for photocatalytic oxygenations and photodynamic inactivation of Staphylococcus aureus.
The ability of a hexanuclear molybdenum cluster, [Mo6I8(CH3COO)6]2-, to carry out, upon irradiation, singlet oxygen mediated photocatalytic oxygenation reactions has been tested. This complex has been also supported on a solid polymeric matrix, yielding an immobilized photosensitizer with remarkable photostability and recyclability. Preliminary studies of antibacterial photoinactivation of Staphylococcus aureus have been conducted, with positive results.


Clinical Nurse Specialist Practice Strategies for Children With Medical Complexity.
Clinical nurse specialists are advanced practice nurses that influence patient care and nursing clinical practice through expert consultation and ongoing support. They play a vital role in caring for children with medical complexity. The purpose of this project is to highlight the practice of the clinical nurse specialist in identifying complex patient care needs and describe the resources needed to successfully transition a patient through the continuum of care by using a case example of a neonate with epidermolysis bullosa. Caring for children with medical complexity can be challenging for healthcare providers and overwhelming for families once they are ready for discharge. Patient outcomes rely heavily on a specific and detailed plan of care, multidisciplinary collaboration, provider education, and safe transitions. The clinical nurse specialist can greatly influence these 4 components and positively impact outcomes for this patient population. A patient with complex medical needs was successfully transferred from the intensive care unit to a medical floor and eventually discharged home. This framework can be used for any child with medical complexity as they transition across care settings to ensure safe and effective care is consistently provided.


[On the improvement of the legal support of the food safety in the conditions of trade and economic integration of states-members of the Customs union and the Russian Federation's accession to the WTO].
In the article priority activities of The Federal Service for the Oversight of Consumer Protection and Welfare on improvement of standard legal support of safety of foodstuff and control of compliance of foodstuff to legislation requirements are reported. The main documents directed on harmonization of the international requirements with national ones and requirements of the Customs union on safety of foodstuff are submitted. Work within a framework of Russian Federation's accession to the WTO is described. And data on control of quality and safety of foodstuff are provided also.


Cerebrospinal fluid viral load is related to cortical atrophy and not to intracerebral calcifications in children with symptomatic HIV disease.
The relationships between viral load in plasma and cerebrospinal fluid (CSF) and computed tomography (CT) brain scan abnormalities were studied in 39 children between 0.5 and 13 years of age with symptomatic HIV-1 disease. Quantitative RNA PCR was used to determine HIV-1 RNA levels and a semiquantitative analog rating technique was used to evaluate non-contrast CT brain scans. CSF HIV-1 RNA copy number correlated significantly with CT brain scan ratings for severity of cortical atrophy (r = 0.36; P < 0.05) but not with ratings of intracerebral calcifications (r = -12; NS). The difference between these two correlations was significant (P < 0.05). Plasma HIV-1 RNA copy number did not correlate significantly with any CT brain scan ratings or with CSF viral load (r = 0.05; NS). Severity of cortical atrophy appeared to reflect the level of viral load in the CSF, supporting the notion that active HIV-1 replication in the CNS is at least in part responsible for such brain abnormalities in children. The lack of correlation of intracerebral calcifications with other CT brain scan abnormalities as well as with CSF viral load suggests that this lesion is relatively independent and may reflect a different neuropathologic process.


Differentiation between solid-ankle cushioned heel and energy storage and return prosthetic foot based on step-to-step transition cost.
Decreased push-off power by the prosthetic foot and inadequate roll-over shape of the foot have been shown to increase the energy dissipated during the step-to-step transition in human walking. The aim of this study was to determine whether energy storage and return (ESAR) feet are able to reduce the mechanical energy dissipated during the step-to-step transition. Fifteen males with a unilateral lower-limb amputation walked with their prescribed ESAR foot (Vari-Flex, Ossur; Reykjavik, Iceland) and with a solid-ankle cushioned heel foot (SACH) (1D10, Ottobock; Duderstadt, Germany), while ground reaction forces and kinematics were recorded. The positive mechanical work on the center of mass performed by the trailing prosthetic limb was larger (33%, p = 0.01) and the negative work performed by the leading intact limb was lower (13%, p = 0.04) when walking with the ESAR foot compared with the SACH foot. The reduced step-to-step transition cost coincided with a higher mechanical push-off power generated by the ESAR foot and an extended forward progression of the center of pressure under the prosthetic ESAR foot. Results can explain the proposed improvement in walking economy with this kind of energy storing and return prosthetic foot.


The role of stereotactic body radiation therapy (SBRT) in the treatment of oligometastatic disease in the elderly.
To report about clinical outcome of stereotactic body radiation therapy (SBRT) in the treatment of oligometastatic disease in elderly patients. Patients with 1-4 inoperable metastases were treated with SBRT. Dose prescription ranged from 40 to 75 Gy in 3-8 fractions. SBRT was delivered using the volumetric modulated arc therapy technique with flattening filter-free photon beams. The primary end points were in-field local control (LC) and toxicity. Secondary end points were overall survival (OS) and disease-specific survival (DSS). 82 patients with 111 total metastases were treated. Median age was 79 years. 64 patients (78%) had a single lesion; the remaining patients had 2-4 lesions. 16 (14.4%) lesions were localized in the abdomen, 50 (45.0%) in the liver and 45 (40.5%) in the lungs. Local response was observed for 87 lesions (78.4%) while local progression was observed in 24 lesions (21.6%). Actuarial 1-year LC was 86.8% ± 3.3%. Actuarial 1-year OS was 93.6% ± 2.7%. 2-year findings were 76.3% ± 4.4% and 72.0% ± 5.6%, respectively. Actuarial 1- and 2-year DSS results were 97.5% ± 2.0% and 81.6% ± 4.9%, respectively. Treatment-related Grade 2-3 toxicity was observed in five patients (4.2%); Grade 1 toxicity in seven patients (5.9%) and no toxicity was observed in 85.4% of the cases. SBRT is a safe and effective therapeutic option for the treatment of oligometastatic disease in the elderly with acceptable rates of LC and low treatment-related toxicity. The use of SBRT for oligometastatic disease in the elderly can be considered as a valuable approach, particularly for patients with fragile status or refusing other approaches.


A sex difference in the specificity of sexual arousal.
Sexual arousal is category-specific in men; heterosexual men are more aroused by female than by male sexual stimuli, whereas homosexual men show the opposite pattern. There is reason to believe that female sexual arousal is organized differently. We assessed genital and subjective sexual arousal to male and female sexual stimuli in women, men, and postoperative male-to-female transsexuals. In contrast to men, women showed little category specificity on either the genital or the subjective measure. Both heterosexual and homosexual women experienced strong genital arousal to both male and female sexual stimuli. Transsexuals showed a category-specific pattern, demonstrating that category specificity can be detected in the neovagina using a photoplethysmographic measure of female genital sexual arousal. In a second study, we showed that our results for females are unlikely to be explained by ascertainment biases. These findings suggest that sexual arousal patterns play fundamentally different roles in male and female sexuality.


The stability of myocardial area at risk estimated electrocardiographically in patients with ST elevation myocardial infarction.
In patients with ST-elevation myocardial infarction (STEMI) the amount of myocardial area at risk (MaR) indicates the maximal potential loss of myocardium if the coronary artery remains occluded. During the time course of infarct evolution ischemic MaR is replaced by necrosis, which results in a decrease in ST segment elevation and QRS complex distortion. Recently it has been shown that combining the electrocardiographic (ECG) Aldrich ST and Selvester QRS scores result in a more accurate estimate of MaR than using either method alone. Therefore, we hypothesized that the combined Aldrich and Selvester score, indicating MaR, is stable until myocardial reperfusion therapy. In a retrospective analysis of a study population of 114 patients, 33 patients were included. The combined Aldrich and Selvester score was determined in ECGs recorded in the ambulance (ECG1) and in the hospital before reperfusion (ECG2). The combined Aldrich and Selvester score was considered stable if the difference between ECG1 and ECG2 was <4.5-percentage point. Stability of the combined Aldrich and Selvester score was observed in 12/33 patients (36.4%), and in regards to anterior and inferior ST elevation in 4/14 patients (28.6%) and 8/19 patients (42.1%), respectively. The median time between the recording of ECG1 and ECG2 was 75 minutes, however the changes in ECG scores were independent of the time between ECG recordings. Patients not meeting the stability criterion either had a decrease (9 patients) or increase (12 patients) of the combined Aldrich and Selvester score. In conclusion, the ECG estimated MaR was stable between the earliest recording time and initiation of reperfusion treatment only in a subgroup of the patients with STEMI. The findings of this study may suggest heterogeneity in regards to the development of the MaR and could indicate a potential need for differentiation in the acute treatment.


Analysis of the immune landscape of small bowel neuroendocrine tumors.
Immune checkpoint inhibitors have shown promising results in different cancers, and correlation between immune infiltration, expression of programmed death-ligand 1 (PD-L1) by tumor cells and response to immunotherapy has been reported. There is limited knowledge regarding the immune microenvironment of small bowel (SB) neuroendocrine tumors (NETs). This work was aimed at characterizing the immune landscape of SB NETs. Expression of PD-L1 and programmed death-1 (PD-1) was evaluated by immunohistochemistry in 102 surgically resected, primary NETs of the duodenum, jejunum and ileum. Extent and characteristics of the tumor-associated immune infiltrate were also assessed and investigated in their prognostic potential. We detected the expression of PD-L1 in ≥1 and ≥50% of tumor cells in 40/102 (39%; 95% CI, 30-49%) and 14/102 (14%; 95% CI, 8-22%) cases respectively. Intratumor host immune response was apparently absent in 35/102 cases (34%; 95% CI, 25-44%), mild to moderate in 46/102 samples (45%, 95% CI, 35-55%), intense in 21/102 tumors (21%, 95% CI, 13-30%). Expression of PD-L1 and extent of immune infiltration were significantly higher in duodenal NETs as compared with jejunal/ileal NETs. A marked peritumoral host response was organized as ectopic lymph node-like structures in 18/102 cases (18%; 95% CI, 11-26%). Neither PD-L1 expression nor the degree of immune infiltration showed any prognostic significance. Overall, the immune landscape of SB NETs is heterogeneous, with adaptive immune resistance mechanisms prevailing in duodenal NETs. Clinical trials of immune checkpoint inhibitors should take into account the immune heterogeneity of SB NETs.


Small-animal 360-deg fluorescence diffuse optical tomography using structural prior information from ultrasound imaging.
We demonstrate dual modality of free-space fluorescence diffuse optical tomography (FDOT) and handheld ultrasound (US) imaging to reveal both functional and structural information in small animals. FDOT is a noninvasive method for examining the fluorophore inside an object from the light distribution of the surface. In FDOT, a 660-nm continuous wave diode laser was used as an excitation source and an electron-multiplying charge-coupled device (EMCCD) was used for fluorescence data acquisition. Both the laser and EMCCD were mounted on a 360-deg rotation gantry for the transmission optical data collection. The structural information is obtained from a 6- to 17-MHz handheld US linear transducer by single-side access and conducts in the reconstruction as soft priors. The rotation ranges from 0 deg to 360 deg; different rotation degrees, object positions, and parameters were determined for comparison. Both phantom and tissue phantom results demonstrate that fluorophore distribution can be recovered accurately and quantitatively using this imaging system. Finally, an animal study confirms that the system can extract a dual-modality image, validating its feasibility for further in vivo experiments. In all experiments, the error and standard deviation decrease as the rotation degree is increased and the error was reduced to 10% when the rotation degree was increased over 135 deg.


A description of classroom help networks, individual network position, and their associations with academic achievement.
This study examined how classroom peer relations can be described in terms of the network of help relations among students, and the positions students take up in this help network, and whether the structure of adolescent classroom help networks and individual network positions were associated with academic achievement. Help networks were based on the peer nomination question "Who helps you with problems?" Building on previous studies on classroom climate and individual network position, higher academic achievement was expected in classrooms with: a dense help network; no or a few network isolates (referring to students that did not give or receive help at all); less segmentation in help relations; equally distributed help nominations. In addition, higher achievement was expected for individuals with more helpers and a more central position in the help network. Using the Dutch SNARE data (54 classrooms; 1,144 students), the multilevel models suggested that lower achievement was related to an unequal distribution of help relations in a classroom. Moreover, the centrality of individuals in the help network was linked to higher achievement. Classrooms varied strongly on network dimensions, and networks that would theoretically be expected to be most beneficial for achievement (with high density, a few isolates, low segmentation, and high equality) turned out to be highly uncommon. The findings demonstrated that subtle network processes were relevant for academic success, and that classroom network characteristics are associated with classroom-level variation in academic achievement. Descriptive results underlined the complexity of the social context of classrooms, and the absence of 'beneficial' classrooms suggests that researchers should adjust their notion of what is a beneficial or detrimental classroom environment for adolescents.


Non-alcoholic fatty liver disease and periodontal disease: A systematic review and meta-analysis of cross-sectional studies.
Non-alcoholic fatty liver disease (NAFLD) is a chronic liver disease, comprising hepatic steatosis, and non-alcoholic steatohepatitis. Periodontal disease (PD) may be a risk factor for the evolution of liver cirrhosis. This study aimed to evaluate the association between NAFLD and PD.We searched in Pubmed, Scopus, Cochrane, and Lilacs databases with descriptors (Non-alcoholic Fatty Liver Disease OR (non-alcoholic AND Fatty Liver AND disease) OR Nonalcoholic Steatohepatitis) AND (Periodontal Disease OR Gingivitis OR Periodontitis) from January 2021 to September 2021. We selected, by the abstract, cross-sectional, cohort (prospective and retrospective), and case-control studies that address periodontal disease in patients with Non-alcoholic Fatty Liver Disease, and aged ≥ 18 years. The search was without the restriction of language and publication time.The search resulted in 954 articles. After applying the selection criteria, five cross-sectional studies remained. A metanalysis combined the study estimates of periodontal disease in NAFLD, by using the random effects. The Odds Ratio (1.91; 95% CI 1.21-3.02; P = 0.006) indicates that the chance of presenting Periodontal disease is 91% higher in individuals with NAFLD when compared with individuals without NAFLD. There are few studies with appropriate methodology to produce sound evidence about the causal relationship between the use of NAFLD and PD, however, studies support the association. So, dental staff must be aware of this association for better management of periodontal disease in patients with NAFLD.


Quantitative proteomics and metabolomics analysis reveals the response mechanism of alfalfa (Medicago sativa L.) to o-coumaric acid stress.
O-coumaric acid (OCA), as a significant phenolic allelochemical found in hairy vetch (Vicia villosa Roth.), that can hinder the growth of alfalfa (Medicago sativa L.), particularly the growth of alfalfa roots. Nonetheless, the mechanism by which OCA inhibits alfalfa root growth remains unclear. In this study, a liquid chromatography tandem mass spectrometry (LC-MS/MS)-based quantitative proteomics analysis was carried out to identify differentially accumulated proteins (DAPs) under OCA treatment. The findings indicated that 680 proteins were DAPs in comparison to the control group. Of those, 333 proteins were up-regulated while 347 proteins were down-regulated. The enrichment analysis unveiled the significance of these DAPs in multiple biological and molecular processes, particularly in ribosome, phenylpropanoid biosynthesis, glutathione metabolism, glycolysis/gluconeogenesis and flavonoid biosynthesis. The majority of DAPs reside in the cytoplasm (36.62%), nucleus (20.59%) and extracellular space (14.12%). In addition, phenylalanine deaminase was identified as a potential chemical-induced regulation target associated with plant lignin formation. DAPs were mainly enriched in flavonoid biosynthesis pathways, which were related to plant root size. Using the UPLC-ESI-MS/MS technique and database, a total of 87 flavonoid metabolites were discovered. The metabolites were predominantly enriched for biosynthesizing naringenin chalcone, which was linked to plant lignin formation, aligning with the enrichment outcomes of DAPs. Consequently, it was deduced that OCA impacted the structure of cell walls by mediating the synthesis of lignin in alfalfa roots, subsequently inducing wilt. Furthermore, a range of proteins have been identified as potential candidates for the breeding of alfalfa strains with enhanced stress tolerance.


Cell surface features associated with differentiation-induction of methylcholanthrene-transformed AKR-2B fibroblasts by N,N-dimethylformamide.
Methylcholanthrene-transformed AKR-2B mouse embryonal fibroblasts (AKR-MCA cells) were examined for cell surface alterations after growth in culture medium containing N,N-dimethylformamide (DMF) using the lactoperoxidase-glucose oxidase radioiodination procedure with subsequent electrophoresis. DMF has been shown to induce maturational changes in a variety of transformed cells in vitro and has been reported to produce a more normal phenotype when applied to cultured AKR-MCA cells. The electrophoretic profile of 125I-labeled surface proteins from AKR-MCA cells exhibited a prominent peak of labeled material with a molecular weight of approximately 85,000. After growth of AKR-MCA cells in medium containing DMF, the Mr 85,000 peak was substantially reduced, while there was a large increase in Mr 200,000 to 250,000 radioiodinated surface material. This cell surface labeling pattern was virtually identical to that of the nontransformed AKR-2B fibroblasts from which AKR-MCA cells were derived. The cell surface alterations observed upon exposure of AKR-MCA cells to DMF occurred as a function of time of growth in DMF and DMF concentration. Growth of AKR-MCA cells in DMF resulted in a steady increase in cell surface 125I incorporation up to the fourth day of exposure to DMF. At this time, the incorporation level was 22.9-fold greater than that for untreated AKR-MCA cells. Incorporation of radiolabel was decreased after the fifth and sixth days of AKR-MCA exposure to DMF. This trend was also manifested by AKR-2B fibroblasts grown in the presence of DMF. The data suggest that there was increased expression of the Mr 200,000 to 250,000 surface protein on both AKR-2B and AKR-MCA cells when grown in DMF. DMF also inhibited morphological transformation and the cell surface changes associated with transformation of AKR-2B cells by exogenous transforming growth factors.


A population-based analysis of the discrepancy between potential and realized access to trauma center care.
To explore whether a discrepancy between the availability of trauma services (potential access) and trauma center utilization rates (realized access) exists, with the aim of informing strategies to improve access. Lack of access to trauma center care has frequently been attributed to the geographic distribution of trauma centers. Alternatively, impeded access to trauma center care might be due to suboptimal triage practices in the setting of appropriate resources. Population-based retrospective cohort study of severely injured adult patients (2002-2010). Potential access to trauma center care was evaluated using network-based spatial analysis of census data and was defined as residing within 1 hour of a trauma center. Realized access to trauma center care was evaluated using population-based data sources and was defined as direct transport from the scene of injury to a trauma center. Concordance between potential and realized access (high, moderate, or low) was evaluated at the county level. Of the population in the study region, 7,340,711 persons (60%) had potential access to trauma center care; persons in 11 counties (22%) had high potential access. Of 26,861 severely injured patients, 10,237 (38%) had realized access to trauma center care; patients in only 4 counties (8%) had high realized access. The concordance between potential and realized access was moderate (weighted κ = 0.49); 63% of counties (n = 7) with high potential access performed worse than expected and had moderate or low realized access. There is limited concordance between potential and realized access. Regions with high potential access had low realized access, and vice versa. This evaluation suggests that strategies to improve access must be based on understanding the distribution of centers and the triage practices used to access trauma care.


The effect of the environment on participation of children and youth with disabilities: a scoping review.
The study's purpose was to identify and synthesize research evidence regarding the effect of the environment on community participation of children with disabilities. A scoping review of peer-reviewed studies published from 1990 to 2011 was performed. Two independent reviewers selected studies based on a systematic procedure. Inclusion criteria for studies were: participants with a disability, aged 5–21 years, whose environment was examined in relation to participation in out-of-school activities. Data were organized and synthesized based on environmental domains within the International Classification of Functioning, Disability and Health (ICF): Natural Environment/Products and Technology; Support and Relationships; Attitudes; and Services, Systems and Policies. Searching identified 1232 articles and 31 met the inclusion criteria. Each domain of the environment within the ICF influenced participation as a facilitator and/or barrier. The most common facilitators involved social support of family and friends and geographic location. The most common barriers included attitudes, physical environment, transportation, policies and the lack of support from staff and service providers. Knowledge derived from this review can assist practitioners in addressing the specific environmental domains that influence children's participation. Such awareness can also foster new research questions and assist policy makers in identifying the factors influencing participation. All domains of the environment, suggested by the ICF, have an influence on children's participation.Evidence regarding the effect of the environment on participation is focused primarily on children with physical disabilities; more studies are needed involving various health conditions and age groups. Practitioners and decision-makers can focus attention on specific aspects of the environment, e.g. attitudinal challenges and social support, in order to foster inclusion and participation-based communities.


A novel treatment of childhood lymphoblastic non-Hodgkin's lymphoma: early and intermittent use of teniposide plus cytarabine.
We treated 24 children and adolescents with stage III or IV lymphoblastic non-Hodgkin's lymphoma, using a protocol designed for patients with poor-prognosis acute lymphoblastic leukemia (ALL). Early therapy consisted of teniposide plus cytarabine administered before and immediately after prednisone, vincristine, and asparaginase. The two-drug combination was also given intermittently with continuous 6-mercaptopurine and methotrexate during the first year of continuation chemotherapy. Periodic intrathecal methotrexate and delayed cranial irradiation were used to prevent central nervous system involvement. Anthracycline compounds, alkylating agents, high-dose methotrexate, and involved-field irradiation were not used in any phase of treatment. Twenty-two (96%) of the 23 evaluable patients achieved complete remission. With a median follow-up of 2 1/2 years, only four patients have relapsed; the remainder have been disease-free for eight months to more than five years. The projected four-year continuous complete remission rate is 73% for all patients and 79% for the 19 with mediastinal involvement at diagnosis. These results demonstrate that use of teniposide plus cytarabine with an otherwise conventional plan of ALL therapy is an effective approach to the treatment of childhood lymphoblastic lymphoma.


Cortical adaptation staging system: a new and simple staging for result evaluation of functioning free-muscle transplantation for facial reanimation.
Movement-associated cortical reorganization occurs in patients after functioning free-muscle transplantation (FFMT), which is reinnervated by different neurotizers. Aiming to evaluate the process of recovery of the reinnervated muscle, we defined the cortical reorganization into 5 stages. This staging system has been applied during the past 25 years at our center with great convenience and accessibility. A standardized evaluation method for assessing the recovery after FFMT to reanimate the paralyzed face with at least a 1-year follow-up was applied. The evaluation included the following 5 stages: no movement, dependent movement, independent movement, and spontaneous movement with and without involuntary movement. Reliability of this technique was assessed by 3 examiners, who each evaluated the smiles of 30 unilateral facial paralysis patients 4 times, creating 360 sets of measurements. The intraclass correlation coefficients for interrater and intrarater reliability exceeded 0.929, which is considered excellent and reliable. Chuang's Cortical Adaptation Staging System is simple, quick,and accurate in evaluating patients after FFMT reanimation of the paralyzed face with no additional tools.


MECP2 mutations in Rett syndrome adversely affect lymphocyte growth, but do not affect imprinted gene expression in blood or brain.
Rett syndrome (RTT) is an X-linked dominant neurodevelopmental disorder caused by mutations in MECP2, encoding methyl-CpG-binding protein 2 (MeCP2). As female somatic cells are mosaic for expression of mutant MECP2, we performed single cell cloning of T lymphocytes from four RTT patients with MECP2 mutations to isolate cells expressing mutant MECP2. Mutant-expressing clones were present at a significantly lower frequency (P<0.0001) than wild-type clones. These results demonstrate that although MECP2 is not essential for lymphocyte growth, expression of the MECP2 mutation causes a growth disadvantage in cultured clonal T cells by reducing the response to mitogenic stimulation. Mutant MECP2 was expressed at normal transcript and protein levels, and exhibited no significant effect on acetylated histones or methyl-binding protein 3 (MBD3) levels. Since MeCP2 was predicted to silence transcription of methylated genes, we hypothesized that MeCP2 may be required for silencing imprinted or methylated gene expression. The allelic expression of three different imprinted genes (SNRPN, IPW and IGF2) was examined by RT-PCR and RFLP analysis, and demonstrated normal monoallelic expression of all RTT clones. We also examined the expression of five imprinted genes (SNRPN, IPW, NECDIN, H19 and IGF2) in RTT brain samples and observed exclusive monoallelic expression. Expression levels were also normal in MECP2 mutant-expressing T cells for IFNG, a non-imprinted, but methylated gene differentially expressed in T cells, and LINE-1 retrotransposons hypothesized to be silenced by MeCP2. The histone deacetylase inhibitor Trichostatin A did not alter SNRPN expression, but did reverse silencing of IFNG in a MECP2-mutant-expressing clone. In conclusion, our results do not support an essential role for either MeCP2 or HDAC in the silencing of several imprinted genes.


Response of periradicular tissues to growth factors introduced into the surgical site in the root-end filling material.
The objective of this study was to evaluate the healing of the periradicular tissues when exogenous growth factors were delivered to the respected root-end. The healing response was compared with that when Diaket was used as a control. Non-surgical root canal treatment was performed on mandibular teeth in mongrel dogs. Surgical treatment followed and included root-end resection and root-end cavity preparation. Insulin-like growth factor in combination with platelet-derived growth factor, or fibroblast growth factor alone, were then placed in the root-end preparations on a polylactic acid carrier (Atrisorb) with or without the incorporation of the carrier tetracalcium phosphate. The healing was evaluated at 60 days with regard to presence of inflammatory response, bone regeneration, periodontal ligament formation and cementum formation. Osseous regeneration in the excisional would and periodontal formation were significantly greater when Diaket was used as the root-end filling material. Likewise, cementum deposition occurred significantly more frequently in the Diaket group (P < 0.05). The polylactic carrier Atrisorb remained in the surgical sites for the duration of the study. The use of specific growth factors, FGF and a combination of IGF/PDGF, delivered to the prepared root end in a collagen carrier did not initiate the desired periradicular tissue response of regeneration. Diaket, as used in this study, did stimulate a periradicular tissue response compatible with regeneration.


Delivery of liposome-sequestered hydrophobic proteins to lysosomes of normal and Batten disease cells.
We have developed a method to deliver hydrophobic proteins such as ATP synthase subunit c and ubiquitin to lysosomes of PMN (polymorphonucleocytes) and fibroblasts. ATP synthase subunit c is stored in the lysosomes of various tissues in late infantile and juvenile forms of neuronal ceriod lipofuscinosis, also called Batten disease (BD). Whether this protein storage is due to an abbreviation in protein or in the lysosomal hydrolases of BD is still not clear. We have sequestered this protein and ubiquitin in the lipid membrane of liposomes. The liposomes coated with autologous heat-aggregated IgG or apolipoprotein E when presented to the PMN and fibroblasts, respectively, accumulated in the lysosomes. Both normal and BD PMN degraded 125I-ubiquitin; the rate of degradation was, however, slower by Batten PMN. These studies indicate that a hydrophobic molecule such as subunit c can be delivered to PMN and fibroblasts, and the sequestered proteins are accessible to lysosomal hydrolases. Therefore, this technique can be used to study the metabolism of highly hydrophobic proteins by lysosomes, especially the biochemical mechanism(s) of subunit c storage in BD.


Influence of Referral to a Combined Diabetology and Nephrology Clinic on Renal Functional Trends and Metabolic Parameters in Adults With Diabetic Kidney Disease.
To examine the impact of a diabetes renal clinic (DRC) on renal functional and metabolic indices in adults who have diabetes mellitus (DM) and chronic kidney disease (CKD). All patients evaluated at a DRC in a single tertiary referral center from January 1, 2008, to December 31, 2012, were identified. Serial renal and metabolic indices from January 1, 2004, to December 31, 2014, were recorded, and trends over time were analyzed by linear mixed-effects models. A total of 200 patients who had DM and CKD were identified and subdivided into 3 categories based on presumptive CKD etiology: 43 (21.5%) with type 1 DM (T1D) only, 127 (63.5%) with type 2 DM (T2D) only, and 30 (15.0%) with DM and an additional CKD etiology. Average annual absolute (mL/min per body surface area per year) and percentage (%/year) changes, respectively, in Chronic Kidney Disease Epidemiology Collaboration estimated glomerular filtration rate before vs after first DRC attendance were: -1.59 vs -3.10 (P=.31) and -1.22 vs -9.39 (P=.06) for T1D; -5.64 vs -3.07 (P=.004) and -10.88 vs -9.94 (P=.70) for T2D; and -6.50 vs +0.91 (P<.001) and -13.28 vs -2.29 (P=.001) for DM with an additional CKD etiology. Glycemic control worsened in those who had T2D, whereas trends in total cholesterol levels improved in those who had T1D. After first DRC attendance, the absolute rate of estimated glomerular filtration rate decline remained similar for those who had T1D, but it slowed for those who had T2D or DM with additional CKD etiology. Thus, benefits of combined diabetology and nephrology consultation may vary for different diabetic subpopulations.


Size dependence of efficiency of PbS quantum dots in NiO-based dye sensitised solar cells and mechanistic charge transfer investigation.
Quantum dots (QDs) are very attractive materials for solar cells due to their high absorption coefficients, size dependence and easy tunability of their optical and electronic properties due to quantum confinement. Particularly interesting are PbS QDs owing to their broad spectral absorption until long wavelengths, their easy processability and low cost. Here, we used control of the PbS QD size to understand charge transfer processes at the interfaces of a NiO semiconductor and explain the optimal QD size in photovoltaic devices. Towards this goal, we have synthesized a series of PbS QDs with different diameters (2.8 nm to 4 nm) and investigated charge transfer dynamics by time resolved spectroscopy and their ability to act as sensitizers in nanocrystalline NiO based solar cells using the cobalt tris(4,4'-ditert-butyl-2,2'-bipyridine) complex as a redox mediator. We found that PbS QDs with an average diameter of 3.0 nm show the highest performance in terms of efficient charge transfer and light harvesting efficiency. Our study showed that hole injection from the PbS QDs to the NiO valence band (VB) is an efficient process even with low injection driving force (-0.3 eV) and occurs in 6-10 ns. Furthermore we found that direct electrolyte reduction (photoinduced electron transfer to the cobalt redox mediator) also occurs in parallel to the hole injection with a rate constant of similar magnitude (10-20 ns). In spite of its large driving force, the rate constant of the oxidative quenching of PbS by Co(iii) diminishes more steeply than hole injection on NiO when the diameter of PbS increases. This is understood as the consequence of increasing the trap states that limit electron shift. We believe that our detailed findings will advance the future design of QD sensitized photocathodes.


First isolation and cultivation of Borrelia burgdorferi sensu lato from Missouri.
Five Borrelia burgdorferi sensu lato isolates from Missouri are described. This represents the first report and characterization of such isolates from that state. The isolates were obtained from either Ixodes dentatus or Amblyomma americanum ticks that had been feeding on cottontail rabbits (Sylvilagus floridanus) from a farm in Bollinger County, Mo., where a human case of Lyme disease had been reported. All isolates were screened immunologically by indirect immunofluorescence by using monoclonal antibodies to B. burgdorferi-specific outer surface protein A (OspA) (antibodies H3TS and H5332), B. burgdorferi-specific OspB (antibody H6831), Borrelia (genus)-specific antiflagellin (antibody H9724), and Borrelia hermsii-specific antibody (antibody H9826). Analysis of the isolates also involved a comparison of their protein profiles by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Finally, the isolates were analyzed by PCR with six pairs of primers known to amplify selected DNA target sequences specifically found in the reference strain B. burgdorferi B-31. Although some genetic variability was detected among the five isolates as well as between them and the B-31 strain, enough similarities were found to classify them as B. burgdorferi sensu lato.


Physical and sexual abuse of wives in urban Bangladesh: husbands' reports.
Using data from 8,320 husbands'self reports for the 2006 Urban Health Survey, this article examines the prevalence of physical and sexual intimate partner violence (IPV) perpetrated by husbands against their wives in Bangladesh and identifies risk markers associated with such violence. Of the men included in the sample for this study, 55 percent reported perpetrating physical IPV against their wives at some point in their married lives, 23 percent reported perpetrating physical IPV in the past year, 20 percent reported ever perpetrating sexual IPV, and 60 percent reported ever perpetrating physical or sexual IPV. Bivariate analyses revealed that men residing in slums had a greater likelihood than those residing in nonslum areas and in district municipalities of perpetrating lifetime and past-year physical IPV, and any lifetime (physical or sexual) IPV. Lifetime sexual IPV prevalence, by contrast, was highest in district municipalities (26 percent), followed by slum (20 percent) and nonslum (17 percent) areas. Net of other factors, low socioeconomic levels were associated with men's increased likelihood of perpetrating IPV. Alcohol and drug use, sexually transmitted disease infection, poor mental health, and holding attitudes supportive of wife beating were predictive of IPV perpetration. These results suggest that IPV-prevention programs targeting men should consider spousal abuse, substance use, and sexual risk behaviors as social and public health problems and should also consider the sociocultural context within which men who abuse their partners are embedded.


Importance of auditory brainstem responses in the CT diagnosis of traumatic brainstem lesions.
Changes in brainstem auditory-evoked potential response were serially investigated in 20 head-injury patients with neurologic and computed tomographic signs of brainstem lesion. The reliability of computed tomography in the evaluation of indirect signs of brainstem lesion from the image was investigated with elaboration of the brainstem auditory-evoked potential response. The measurement of auditory brainstem response is thought to be useful in detecting the severity and predicting possible recovery in posttraumatic brainstem injury. Brainstem deformity on computed tomography was a bad prognostic sign, indicating irreversible structural change.


Manganese neurotoxicity: new perspectives from behavioral, neuroimaging, and neuropathological studies in humans and non-human primates.
Manganese (Mn) is an essential metal and has important physiological functions for human health. However, exposure to excess levels of Mn in occupational settings or from environmental sources has been associated with a neurological syndrome comprising cognitive deficits, neuropsychological abnormalities and parkinsonism. Historically, studies on the effects of Mn in humans and experimental animals have been concerned with effects on the basal ganglia and the dopaminergic system as it relates to movement abnormalities. However, emerging studies are beginning to provide significant evidence of Mn effects on cortical structures and cognitive function at lower levels than previously recognized. This review advances new knowledge of putative mechanisms by which exposure to excess levels of Mn alters neurobiological systems and produces neurological deficits not only in the basal ganglia but also in the cerebral cortex. The emerging evidence suggests that working memory is significantly affected by chronic Mn exposure and this may be mediated by alterations in brain structures associated with the working memory network including the caudate nucleus in the striatum, frontal cortex and parietal cortex. Dysregulation of the dopaminergic system may play an important role in both the movement abnormalities as well as the neuropsychiatric and cognitive function deficits that have been described in humans and non-human primates exposed to Mn.


IL-2 produced by CD8+ immune T cells can augment their IFN-γ production independently from their proliferation in the secondary response to an intracellular pathogen.
Chronic infection with Toxoplasma gondii induces a potent resistance against reinfection, and IFN-γ production by CD8(+) T cells is crucial for the protective immunity. However, the molecular mechanisms that regulate the secondary response remain to be elucidated. In the current study, we examined the role of IL-2 in IFN-γ production by CD8(+) immune T cells in their secondary responses using T. gondii-specific CD8(+) T cell hybridomas and splenic CD8(+) immune T cells from chronically infected mice. The majority (92%) of CD8(+) T cell hybridomas produced large amounts of IFN-γ only when a low amount (0.5 ng/ml) of exogenous IL-2 was provided in combination with T. gondii Ags. Inhibition of cell proliferation by mitomycin C did not affect the enhancing effect of IL-2 on the IFN-γ production, and significant increases in transcription factor T-bet expression were associated with the IL-2-mediated IFN-γ amplification. Splenic CD8(+) immune T cells produced similar low levels of IL-2 in the secondary response to T. gondii, and a blocking of IL-2 signaling by anti-IL-2Rα Ab or inhibitors of JAK1 and JAK3 significantly reduced IFN-γ production of the T cells. This IL-2-mediated upregulation of IFN-γ production was observed in mitomycin C-treated CD8(+) immune T cells, thus independent from their cell division. Therefore, endogenous IL-2 produced by CD8(+) immune T cells can play an important autocrine-enhancing role on their IFN-γ production in the secondary responses to T. gondii, suggesting an importance of induction of CD8(+) immune T cells with an appropriate IL-2 production for vaccine development.


Direct observation of mitotic spindle elongation in vitro.
Successful reactivation in vitro of anaphase B has recently been achieved with mitotic spindles isolated from the diatom Stephanopyxis turris. When a population of isolated spindles was studied indirectly by using immunofluorescence, nearly all of them were found to have elongated; however, when studied directly by using video microscopy, only a small proportion of spindles elongated. We report here conditions that allow nearly all of the spindles to elongate when observed directly with video microscopy. These direct observations validate previous ones made using indirect immunofluorescence. In addition, we find that the isolated spindles elongate with a linear rate, that the elongation is unchanged after the chromatin surrounding the spindles is digested with DNase I, and that during elongation a phase-dense matrix may accumulate in the spindle midzone.


HER2 CAR-T Cells Eradicate Uveal Melanoma and T-cell Therapy-Resistant Human Melanoma in IL2 Transgenic NOD/SCID IL2 Receptor Knockout Mice.
Chimeric antigen receptors (CAR) can transmit signals akin to those from activated T-cell receptors when bound to a cell surface target. CAR-expressing T cells against CD19 can cause curative effects in leukemia and lymphoma and is approved for clinical use. However, no CAR-T therapy is currently approved for use in solid tumors. We hypothesize that the resistance of solid tumors to CAR-T can be overcome by similar means as those used to reactivate tumor-infiltrating T lymphocytes (TIL), for example, by cytokines or immune checkpoint blockade. Here we demonstrate that CAR-T cells directed against HER2 can kill uveal and cutaneous melanoma cells in vitro and in vivo. Curative effects in vivo were only observed in xenografts grown in a NOD/SCID IL2 receptor gamma (NOG) knockout mouse strain transgenic for human IL2. The effect was target-specific, as CRISPR/Cas9-mediated disruption of HER2 in the melanoma cells abrogated the killing effect of the CAR-T cells. The CAR-T cells were also able to kill melanoma cells from patients resistant to adoptive T-cell transfer (ACT) of autologous TILs. Thus, CAR-T therapy represents an option for patients that do not respond to immunotherapy with ACT of TIL or immune checkpoint blockade. In addition, our data highlight the use of IL2 transgenic NOG mice as models to prove efficacy of CAR-T-cell products, possibly even in a personalized manner. SIGNIFICANCE: These findings demonstrate that a novel humanized mouse model can help clinical translation of CAR-T cells against uveal and cutaneous melanoma that do not respond to TIL therapy or immune checkpoint blockade.


Glucagon-like peptide-1 stimulates luteinizing hormone-releasing hormone secretion in a rodent hypothalamic neuronal cell line.
To examine the influence of the putative satiety factor (GLP-1) on the hypothalamo-pituitary-gonadal axis, we used GT1-7 cells as a model of neuronal luteinizing hormone- releasing hormone (LHRH) release. GLP-1 caused a concentration-dependent increase in LHRH release from GT1-7 cells. Specific, saturable GLP-1 binding sites were demonstrated on these cells. The binding of [125I]GLP-1 was time-dependent and consistent with a single binding site (Kd = 0.07+/-0.016 nM; binding capacity = 160+/-11 fmol/mg protein). The specific GLP-1 receptor agonists, exendin-3 and exendin-4, also showed high affinity (Ki = 0.3+/-0.05 and 0.32+/-0.06 nM, respectively) as did the antagonist exendin-(9-39) (Ki = 0.98+/-0.24 nM). At concentrations that increased LHRH release, GLP-1 (0.5-10 nM) also caused an increase in intracellular cAMP in GT1-7 cells (10 nM GLP-1: 7.66+/-0.4 vs. control: 0.23+/-0.02 nmol/mg protein; P < 0.001). Intracerebroventricular injection of GLP-1 at a single concentration (10 microg) produced a prompt increase in the plasma luteinizing hormone concentration in male rats (GLP-1: 1.09+/-0.11 vs. saline: 0.69+/-0.06 ng/ml; P < 0.005). GLP-1 levels in the hypothalami of 48-h-fasted male rats showed a decrease, indicating a possible association of the satiety factor with the low luteinizing hormone levels in animals with a negative energy balance.


Role of Ketogenic Diets in Neurodegenerative Diseases (Alzheimer's Disease and Parkinson's Disease).
The goal of this review was to assess the effectiveness of ketogenic diets on the therapy of neurodegenerative diseases. The ketogenic diet is a low-carbohydrate and fat-rich diet. Its implementation has a fasting-like effect, which brings the body into a state of ketosis. The ketogenic diet has, for almost 100 years, been used in the therapy of drug-resistant epilepsy, but current studies indicate possible neuroprotective effects. Thus far, only a few studies have evaluated the role of the ketogenic diet in the prevention of Parkinson's disease (PD) and Alzheimer's disease (AD). Single studies with human participants have demonstrated a reduction of disease symptoms after application. The application of the ketogenic diet to elderly people, however, raises certain concerns. Persons with neurodegenerative diseases are at risk of malnutrition, while food intake reduction is associated with disease symptoms. In turn, the ketogenic diet leads to a reduced appetite; it is not attractive from an organoleptic point of view, and may be accompanied by side effects of the gastrointestinal system. All this may lead to further lowering of consumed food portions by elderly persons with neurodegenerative diseases and, in consequence, to further reduction in the supply of nutrients provided by the diet. Neither data on the long-term application of the ketogenic diet in patients with neurodegenerative disease or data on its effects on disease symptoms are available. Further research is needed to evaluate the suitability of the ketogenic diet in the therapy of AD- or PD-affected persons.


Differential roles of JNK in ConA/GalN and ConA-induced liver injury in mice.
Tumor necrosis factor-alpha-mediated liver injury can be induced by several different means; however, the signaling events and mechanisms of cell death are likely different. We investigated the mechanism of both apoptotic and necrotic hepatocyte cell death as well as the role of c-Jun NH2-terminal kinase (JNK) in the ConA and ConA/D-galactosamine (GalN) models of murine liver injury. ConA alone induced primarily necrotic cell death with no caspase activation, whereas ConA/GalN induced apoptosis in addition to necrotic cell death. The bi-modal death pattern in the ConA/GalN model was confirmed by the use of transgenic mice expressing a dominant-negative form of Fas-associated death domain in which the mice were resistant to apoptotic but not necrotic cell death. JNK1 and, more significantly, JNK2 participated in the induction of hepatocyte apoptosis in response to ConA/GalN. Deletion of JNK led to the stabilization of FLIP L, reduced caspase-8 activation, decreased Bid cleavage, and inhibition of the mitochondrial apoptosis pathway. In contrast, JNK did not participate in necrotic death induced by ConA either alone or in combination with GalN. As such, JNK-deficient mice remained susceptible to necrotic liver injury in both model systems. Thus, ConA and ConA/GalN mouse models induce liver injury with different mechanisms of cell death, and JNK contributes to apoptotic but not necrotic cell death. These findings further elucidate the specific pathways involved in tumor necrosis factor-alpha-mediated liver injury.


The use of PBPK models to inform human health risk assessment: case study on perchlorate and radioiodide human lifestage models.
Physiologically-based pharmacokinetic (PBPK) models are often submitted to or selected by agencies, such as the U.S. Environmental Protection Agency (U.S. EPA) and Agency for Toxic Substances and Disease Registry, for consideration for application in human health risk assessment (HHRA). Recently, U.S. EPA evaluated the human PBPK models for perchlorate and radioiodide for their ability to estimate the relative sensitivity of perchlorate inhibition on thyroidal radioiodide uptake for various population groups and lifestages. The most well-defined mode of action of the environmental contaminant, perchlorate, is competitive inhibition of thyroidal iodide uptake by the sodium-iodide symporter (NIS). In this analysis, a six-step framework for PBPK model evaluation was followed, and with a few modifications, the models were determined to be suitable for use in HHRA to evaluate relative sensitivity among human lifestages. Relative sensitivity to perchlorate was determined by comparing the PBPK model predicted percent inhibition of thyroidal radioactive iodide uptake (RAIU) by perchlorate for different lifestages. A limited sensitivity analysis indicated that model parameters describing urinary excretion of perchlorate and iodide were particularly important in prediction of RAIU inhibition; therefore, a range of biologically plausible values available in the peer-reviewed literature was evaluated. Using the updated PBPK models, the greatest sensitivity to RAIU inhibition was predicted to be the near-term fetus (gestation week 40) compared to the average adult and other lifestages; however, when exposure factors were taken into account, newborns were found to be populations that need further evaluation and consideration in a risk assessment for perchlorate.


Adrenergic Receptor Signaling Regulates the Response of Tumors to Ionizing Radiation.
While ionizing radiation is a major form of cancer therapy, radioresistance remains a therapeutic obstacle. We have previously shown that the mandated housing temperature for laboratory mice (∼22°C) induces mild, but chronic, cold stress resulting in increased circulating norepinephrine, which binds to, and triggers activation of, beta-adrenergic receptors (β-AR) on tumor and immune cells. This adrenergic signaling increases tumor cell intrinsic resistance to chemotherapy and suppression of the anti-tumor immune response. These findings led us to hypothesize that adrenergic stress signaling increases radioresistance in tumor cells in addition to suppressing T-cell-mediated anti-tumor immunity, thus suppressing the overall sensitivity of tumors to radiation. We used three strategies to test the effect of adrenergic signaling on responsiveness to radiation. For one strategy, mice implanted with CT26 murine colon adenocarcinoma were housed at either 22°C or at thermoneutrality (30°C), which reduces physiological adrenergic stress. For a second strategy, we used a β-AR antagonist ("beta blocker") to block adrenergic signaling in mice housed at 22°C. In either case, tumors were then irradiated with a single 6 Gy dose and the response was compared to mice whose adrenergic stress signaling was not reduced. For the third strategy, we used an in vitro approach in which several different tumor cell lines were treated with a β-AR agonist and irradiated, and cell survival was then assessed by clonogenic assay. Overall, we found that adrenergic stress significantly impaired the anti-tumor efficacy of radiation by inducing tumor cell resistance to radiation-induced cell killing and by suppression of anti-tumor immunity. Treatment using beta blockers is a promising strategy for increasing the anti-tumor efficacy of radiotherapy.


Diminished A-type potassium current and altered firing properties in presympathetic PVN neurones in renovascular hypertensive rats.
Accumulating evidence supports a contribution of the hypothalamic paraventricular nucleus (PVN) to sympathoexcitation and elevated blood pressure in renovascular hypertension. However, the underlying mechanisms resulting in altered neuronal function in hypertensive rats remain largely unknown. Here, we aimed to address whether the transient outward potassium current (I(A)) in identified rostral ventrolateral medulla (RVLM)-projecting PVN neurones is altered in hypertensive rats, and whether such changes affected single and repetitive action potential properties and associated changes in intracellular Ca(2+) levels. Patch-clamp recordings obtained from PVN-RVLM neurons showed a reduction in I(A) current magnitude and single channel conductance, and an enhanced steady-state current inactivation in hypertensive rats. Morphometric reconstructions of intracellularly labelled PVN-RVLM neurons showed a diminished dendritic surface area in hypertensive rats. Consistent with a diminished I(A) availability, action potentials in PVN-RVLM neurons in hypertensive rats were broader, decayed more slowly, and were less sensitive to the K(+) channel blocker 4-aminopyridine. Simultaneous patch clamp recordings and confocal Ca(2+) imaging demonstrated enhanced action potential-evoked intracellular Ca(2+) transients in hypertensive rats. Finally, spike broadening during repetitive firing discharge was enhanced in PVN-RVLM neurons from hypertensive rats. Altogether, our results indicate that diminished I(A) availability constitutes a contributing mechanism underlying aberrant central neuronal function in renovascular hypertension.


Efficacy of a new image-enhancement technique for achieving hemostasis in endoscopic submucosal dissection.
Hemostasis during endoscopic submucosal dissection (ESD) can sometimes be challenging and stressful for the endoscopist. Therefore, we aimed to assess the usefulness of dual red imaging (DRI), a new image enhancement technique that uses 3 wavelengths (540, 600, and 630 nm) to visualize bleeding points and to examine the efficacy of DRI in shortening the time required to achieve hemostasis (hemostasis time) during ESD. DRI and white-light imaging (WLI) were used alternately for managing 378 bleeding events in 97 patients undergoing ESD. Hemostasis time for each hemostasis event was measured. Using portable eye-tracking glasses, 4 experienced endoscopists were shown random videos of intraoperative bleeding during ESD (20 cases each on WLI and DRI) and identified the bleeding point in each video. The mean distances of eye movement per unit of time until the bleeding point were identified in each video and compared between the WLI and DRI groups. Average hemostasis time was significantly shorter in the DRI group. The mean distance of eye movement was significantly shorter in the DRI group than in the WLI group for all endoscopists. DRI can offer useful images to help in clearly detecting bleeding points and in facilitating hemostasis during ESD. It is feasible and may help in successfully performing ESD that is safer and faster than WLI. (Clinical trial registration number: UMIN000018309.).


A polymorphism in the AMH type II receptor gene is associated with age at menopause in interaction with parity.
Anti-Müllerian hormone (AMH) inhibits primordial follicle recruitment in the mouse ovary. We hypothesize that in women AMH signaling also regulates the usage of the primordial follicle pool and hence influences the onset of menopause. Since age at menopause has a strong genetic component, we investigated the role of AMH signaling using a candidate gene approach. In two large population-based cohorts of Dutch post-menopausal women (n = 2381 and n = 248), we examined the association between two polymorphisms, one in the AMH gene and one in the AMH type II receptor (AMHR2) gene, and natural age at menopause. The AMH Ile(49)Ser polymorphism (rs10407022) was not associated with age at menopause in either cohort. In the Rotterdam cohort, the AMHR2 -482 A > G polymorphism (rs2002555) was associated with age at menopause in interaction with the number of offspring (P = 0.001). Nulliparous women homozygous for the G-allele entered menopause 2.6 years earlier compared with nulliparous women homozygous for the A-allele (P = 0.005). In the LASA cohort, women with the G/G genotype tended to enter menopause 2.8 years earlier compared with the A/A genotype (P = 0.063). The observed association of the AMHR2 -482 A > G polymorphism with natural age at menopause suggests a role for AMH signaling in the usage of the primordial follicle pool in women.