Increased expression of costimulatory markers CD134 and CD80 on interleukin-17 producing T cells in patients with systemic lupus erythematosus.
There is growing evidence that interleukin 17 (IL-17) producing T cells are involved in the pathogenesis of systemic lupus erythematosus (SLE). Previous studies showed that increased percentages of T-cell subsets expressing the costimulatory molecules CD80 and CD134 are associated with disease activity and renal involvement in SLE. The aim of this study was to investigate the distribution and phenotypical characteristics of IL-17 producing T-cells in SLE, in particular in patients with lupus nephritis, with emphasis on the expression of CD80 and CD134. Thirty-four patients (3 male, 31 female, mean age 41 ± 15 years) fulfilling at least four of the American College of Rheumatology (ACR) revised criteria for the diagnosis of SLE and 24 healthy controls were enrolled. T-cells from the peripheral blood were analysed by fluorescence activated cell sorting (FACS) for their expression levels of CD80, CD134 and CCR6. In vitro stimulated CD3+IL17+ cells were also investigated for the expression of these costimulatory markers. Finally, renal biopsies from SLE patients were evaluated for the presence of CD134 expressing T-cells. Percentages of IL-17 expressing T-cells were significantly increased in patients with active disease as compared to healthy controls (1.46 ± 0.58% versus 0.93 ± 0.30%, P = 0.007). The percentage of IL-17 producing T-cells was correlated with disease activity as assessed by systemic lupus erythematosus disease activity index (SLEDAI) (r = 0.53, P = 0.003). In patients, most of the IL-17 producing T-cells were confined to the CCR6+ T-cell subset (80 ± 13%). Expression of CD80 and CD134 on the IL-17 producing T-cell subset was higher in SLE than in healthy controls (HC) (CD134: 71.78 ± 14.51% versus 51.45 ± 16.58%, P = 0.002; CD80: 25.5 ± 14.99% versus 14.99 ± 5.74%, P = 0.02). Also, patients with lupus nephritis expressed higher levels of CD134+ on CD3+IL-17+ cells as compared to HC (72.69 ± 11.54% versus 51.45 ± 16.58%, P = 0.006). Furthermore, renal biopsies of lupus nephritis patients showed infiltration of CD134+ T cells. Percentages of IL-17 expressing T-cells correlate with disease activity. Further, these cells show increased expression of costimulatory markers such as CD134 and CD80. The presence of CD134+ T-cells in renal biopsies of lupus nephritis patients suggest that these cells migrate to the kidney and might contribute to inflammatory processes through IL-17 secretion.


Perforated esophageal intervention focus (PERF) study: a multi-center examination of contemporary treatment.
The treatment of esophageal perforation (EP) remains a significant clinical challenge. While a number of investigators have previously documented efficient approaches, these were mostly single-center experiences reported prior to the introduction of newer technologies: specifically endoluminal stents. This study was designed to document contemporary practice in the diagnosis and management of EP at multiple institutions around the world and includes early clinical outcomes. A five-year (2009-2013) multicenter retrospective review of management and outcomes for patients with thoracic or abdominal esophageal perforation was conducted. Demographics, etiology, diagnostic modalities, treatments, subsequent early outcomes as well as morbidity and mortality were captured and analyzed. During the study period, 199 patients from 10 centers in the United States, Canada, and Europe were identified. Mechanisms of perforation included Boerhaave syndrome (60, 30.1%), iatrogenic injury (65, 32.6%), and penetrating trauma (25, 12.6%). Perforation was isolated to the thoracic segment alone in 124 (62.3%), with 62 (31.2%) involving the thoracoabdominal esophagus. Mean perforation length was 2.5 cm. Observation was selected as initial management in 65 (32.7%), with only two failures. Direct operative intervention was initial management in 65 patients (32.6%), while 29 (14.6%) underwent esophageal stent coverage. Compared to operative intervention, esophageal stent patients were significantly more likely to be older (61.3 vs. 48.3 years old, P < 0.001) and have sustained iatrogenic mechanisms of esophageal perforation (48.3% vs.15.4%). Secondary intervention requirement for patients with perforation was 33.7% overall (66). Complications included sepsis (56, 28.1%), pneumonia (34, 17.1%) and multi-organ failure (23, 11.6%). Overall mortality was 15.1% (30). In contemporary practice, diagnostic and management approaches to esophageal perforation vary widely. Despite the introduction of endoluminal strategies, it continues to carry a high risk of mortality, morbidity, and need for secondary intervention. A concerted multi-institutional, prospectively collected database is ideal for further investigation.


Access to health care for undocumented migrant children and pregnant women: the paradox between values and attitudes of health care professionals.
Access to health care for undocumented migrant children and pregnant women confronts human rights and professional values with political and institutional regulations that limit services. In order to understand how health care professionals deal with these diverging mandates, we assessed their attitudes toward providing care to this population. Clinicians, administrators, and support staff (n = 1,048) in hospitals and primary care centers of a large multiethnic city responded to an online survey about attitudes toward access to health care services. Analysis examined the role of personal and institutional correlates of these attitudes. Foreign-born respondents and those in primary care centers were more likely to assess the present access to care as a serious problem, and to endorse broad or full access to services, primarily based on human rights reasons. Clinicians were more likely than support staff to endorse full or broad access to health care services. Respondents who approved of restricted or no access also endorsed health as a basic human right (61.1%) and child development as a priority (68.6%). A wide gap separates attitudes toward entitlement to health care and the endorsement of principles stemming from human rights and the best interest of the child. Case-based discussions with professionals facing value dilemmas and training on children's rights are needed to promote equitable practices and advocacy against regulations limiting services.


[Does ultrasonographic volume of the thyroid gland correlate with difficult intubation? An observational study].
Preoperative ultrasonographic evaluation of the thyroid gland done by surgeons could let us foresee airway management challenges. The aim of this observational study was to evaluate the effects of thyroid-related parameters assessed preoperatively by surgeons via ultrasonography and chest X-ray on intubation conditions. Fifty patients undergoing thyroid surgery were enrolled. Thyromental distance, Mallampati score, neck circumference and range of neck movement were evaluated before the operation. Thyroid volume, signs of invasion or compression and tracheal deviation on chest X-ray were also noted. The intubation conditions were assessed with Cormack and Lehane score and the intubation difficulty scale. Statistical analyses were done with SPSS 15.0 software. The mean thyroid volume of the patients was 26.38±14mL. The median intubation difficulty scale was 1 (0-2). Thyromental distance (p=0.011; r=0.36; 95% CI 0.582-0.088), Mallampati score (p=0.041; r=0.29; 95% CI 0.013-0.526), compression or invasion signs (p=0.041; r=0.28; 95% CI 0.006-0.521) and tracheal deviation on chest X-ray (p=0.041; r=0.52; 95% CI 0.268-0.702) were correlated with intubation difficulty scale. Also patients were classified into two groups related to their intubation difficulty scale (Group I, n=19: intubation difficulty scale=0; Group II, n=31: 1<intubation difficulty scale≤5) and difficult intubation predictors and thyroid-related parameters were compared. Only Mallampati score was significantly different between groups (p=0.025). The thyroid volume is not associated with difficult intubation. However clinical assessment parameters may predict difficult intubation.


Insulin-like growth factor (IGF)-I and -II and IGF-binding proteins-1, -2, and -3 in children and adolescents with diabetes mellitus: correlation with metabolic control and height attainment.
The putative effects of diabetes and metabolic control on circulating levels of insulin-like growth factors (IGFs) and their binding proteins (IGFBPs) remain controversial. In the present study, serum levels of IGF-I and IGF-II and IGFBP-1, -2, and -3 were measured in 58 patients (age, 0.8-17 yr) with treated (51 subjects) or untreated (7 subjects) insulin-dependent diabetes mellitus (IDDM) and were compared with the levels in normal subjects. In the untreated patients IGF-I and IGF-II were decreased as compared with the healthy controls. In the treated diabetics IGF-I and IGF-II were reduced; IGFBP-2 (only in prepubertal subjects) and IGFBP-3 were increased. Furthermore, age-adjusted values of IGF-I, IGF-II, and IGFBP-3 were lower in prepubertal than in pubertal patients. Regression analysis revealed a negative correlation between hemoglobin (Hb)A1c and standard deviation scores (SDS) of IGF-I and a positive association between HbA1c and IGFBP-1 SDS or IGFBP-2 SDS. In the treated patients HbA1c was positively related to IGFBP-1 SDS and IGFBP-2 SDS when applying simple regression analysis and to IGFBP-2 SDS when using a multiple regression model. Strong correlations were observed between height SDS and IGF-I SDS, IGF-II SDS, and IGFBP-3 SDS in prepubertal subjects who had had IDDM for at least 2 yr, but not in adolescents. Such correlations have also been found in healthy children and adolescents. In conclusion; 1) IDDM is associated with alterations of the IGF-IGFBP system, which are partially accounted for by differences in metabolic control and pubertal status; 2) the lower plasma concentrations of serum IGF-I may play a role in the pathogenesis of growth impairment of poorly controlled prepubertal, but not pubertal, children and adolescents with IDDM; and 3) in addition, a potential role of the altered IGF-IGFBP system for the development of diabetic late complications is hypothesized.


Providing comprehensive health services for young key populations: needs, barriers and gaps.
Adolescence is a time of physical, emotional and social transitions that have implications for health. In addition to being at high risk for HIV, young key populations (YKP) may experience other health problems attributable to high-risk behaviour or their developmental stage, or a combination of both. We reviewed the needs, barriers and gaps for other non-HIV health services for YKP. We searched PubMed and Google Scholar for articles that provided specific age-related data on sexual and reproductive health; mental health; violence; and substance use problems for adolescent, youth or young sex workers, men who have sex with men, transgender people, and people who inject drugs. YKP experience more unprotected sex, sexually transmitted infections including HIV, unintended pregnancy, violence, mental health disorders and substance use compared to older members of key populations and youth among the general population. YKP experience significant barriers to accessing care; coverage of services is low, largely because of stigma and discrimination experienced at both the health system and policy levels. YKP require comprehensive, integrated services that respond to their specific developmental needs, including health, educational and social services within the context of a human rights-based approach. The recent WHO Consolidated Guidelines on HIV Prevention, Diagnosis, Treatment and Care for Key Populations are an important first step for a more comprehensive approach to HIV programming for YKP, but there are limited data on the effective delivery of combined interventions for YKP. Significant investments in research and implementation will be required to ensure adequate provision and coverage of services for YKP. In addition, greater commitments to harm reduction and rights-based approaches are needed to address structural barriers to access to care.


Hormonal regulation of mammary gland development and lactation.
Lactation is critical to infant short-term and long-term health and protects mothers from breast cancer, ovarian cancer and type 2 diabetes mellitus. The mammary gland is a dynamic organ, regulated by the coordinated actions of reproductive and metabolic hormones. These hormones promote gland development from puberty onwards and induce the formation of a branched, epithelial, milk-secreting organ by the end of pregnancy. Progesterone withdrawal following placental delivery initiates lactation, which is maintained by increased pituitary secretion of prolactin and oxytocin, and stimulated by infant suckling. After weaning, local cytokine production and decreased prolactin secretion trigger large-scale mammary cell loss, leading to gland involution. Here, we review advances in the molecular endocrinology of mammary gland development and milk synthesis. We discuss the hormonal functions of the mammary gland, including parathyroid hormone-related peptide secretion that stimulates maternal calcium mobilization for milk synthesis. We also consider the hormonal composition of human milk and its associated effects on infant health and development. Finally, we highlight endocrine and metabolic diseases that cause lactation insufficiency, for example, monogenic disorders of prolactin and prolactin receptor mutations, maternal obesity and diabetes mellitus, interventions during labour and delivery, and exposure to endocrine-disrupting chemicals such as polyfluoroalkyl substances in consumer products and other oestrogenic compounds.


A microfabricated platform to form three-dimensional toroidal multicellular aggregate.
Techniques that allow cells to self-assemble into three-dimensional (3D) spheroid microtissues provide powerful in vitro models that are becoming increasingly popular in fields such as stem cell research, tissue engineering, and cancer biology. Appropriate simulation of the 3D environment in which tissues normally develop and function is crucial for the engineering of in vitro models that can be used for the formation of complex tissues. We have developed a unique multicellular aggregate formation platform that utilizes a maskless gray-scale photolithography. The cellular aggregate formed using this platform has a toroidal-like geometry and includes a micro lumen that facilitates the supply of oxygen and growth factors and the expulsion of waste products. As a result, this platform was capable of rapidly producing hundreds of multicellular aggregates at a time, and of regulating the diameter of aggregates with complex design. These toroidal multicellular aggregates can grow as long-term culture. In addition, the micro lumen can be used as a continuous channel and for the insertion of a vascular system or a nerve system into the assembled tissue. These platform characteristics highlight its potential to be used in a wide variety of applications, e.g. as a bioactuator, as a micro-machine component or in drug screening and tissue engineering.


Tissue expression of components of the renin-angiotensin system in experimental post-infarction heart failure in rats: effects of heart failure and angiotensin-converting enzyme inhibitor treatment.
1. It has been suggested that local tissue renin-angiotensin systems may be activated in heart failure and that effects on such systems may, at least partially, explain the beneficial effects of angiotensin-converting enzyme (ACE) inhibitors in this syndrome. To investigate these hypotheses, we examined expression of renin-angiotensin system components in several tissues in a rodent model of post-myocardial infarction (MI) heart failure, and analysed whether such expression is modified by ACE inhibitor treatment. 2. Four groups of rats (n = 8 - 12 per group) were studied 30 days after surgery: (A) sham-operated rats with no treatment, (B) rats with post-MI heart failure induced by ligation of the left coronary artery, (C) sham-operated rats treated with the ACE inhibitor perindopril (1.5 mg day-1 kg-1), and (D) rats as per B, but treated with perindopril. Expression of renin, angiotensinogen, ACE and angiotensin subtype 1 receptor was assessed by quantification of their respective mRNAs by Northern blotting. 3. Renal renin mRNA increased 2-fold in animals with MI (group B) compared with controls (group A) (P < 0.05) and between 50 and 100-fold after ACE inhibitor treatment (P < 0.001). No change in renin gene expression was found in any extra-renal site either following MI or after ACE inhibitor treatment. Hepatic angiotensinogen mRNA level was similar in all groups, but kidney angiotensinogen mRNA level was increased 1.6-fold (P < 0.01) in the groups receiving perindopril. ACE mRNA level in the lung was not affected by ACE inhibitor treatment but decreased by 50% following MI (groups B and D, P < 0.01). This was associated with a similar (50%, P < 0.01) fall in lung ACE activity and was correlated with the severity of heart failure. Angiotensin subtype 1 receptor mRNA level was not affected in any tissue by either MI or ACE inhibitor treatment. 4. We did not find a systematic activation of tissue renin-angiotensin systems, as assessed by steady-state mRNA levels of key components of the system in experimental post-MI heart failure, or a major effect of ACE inhibitor treatment on expression of these components. However, we observed tissue-specific changes in expression of selected components of the renin-angiotensin system in the kidney and the lung in post-MI heart failure and after ACE inhibitor treatment, which may be of relevance to the pathophysiology of the syndrome and the effects of ACE inhibition.


Mental health and HIV sexual risk behavior among patrons of alcohol serving venues in Cape Town, South Africa.
Alcohol-serving venues in South Africa provide a location for HIV prevention interventions due to risk factors of patrons in these establishments. Understanding the association between mental health and risk behaviors in these settings may inform interventions that address alcohol use and HIV prevention. Participants (n = 738) were surveyed in 6 alcohol-serving venues in Cape Town to assess post-traumatic stress disorder (PTSD) and depression symptoms, traumatic experiences, sexual behavior, and substance use. Logistic regression models examined whether traumatic experiences predicted PTSD and depression. Generalized linear models examined whether substance use, PTSD, and depressive symptoms predicted unprotected sexual intercourse. Men and women were analyzed separately. Participants exhibited high rates of traumatic experiences, PTSD, depression, alcohol consumption, and HIV risk behaviors. For men, PTSD was associated with being hit by a sex partner, physical child abuse, sexual child abuse and HIV diagnosis; depression was associated with being hit by a sex partner, forced sex and physical child abuse. For women, both PTSD and depression were associated with being hit by a sex partner, forced sex, and physical child abuse. Unprotected sexual intercourse was associated with age, frequency and quantity of alcohol use, drug use, and PTSD for men and frequency and quantity of alcohol use, depression, and PTSD for women. Mental health in this setting was poor and was associated with sexual risk behavior. Treating mental health and substance-use problems may aid in reducing HIV infection. Sexual assault prevention and treatment after sexual assault may strengthen HIV prevention efforts.


Pain assessment in the critically ill mechanically ventilated adult patients: Comparison between skin conductance algesimeter index and physiologic indicators.
Critically ill patients are unable to communicate effectively, so it is difficult to assess their pain and analgesic requirement. Skin conductance algesimeter (SCA) index is a device that primarily measures changes in skin conductance real time to assess pain. We planned this quantitative prospective observational study to assess pain in the critically ill mechanically ventilated patients in comparison to physiologic indicators such as blood pressure and heart rate. A repeated measures design was chosen, and a sample size of 180 was taken from 60 patients with sepsis, acute exacerbations of chronic obstructive pulmonary disease, community-acquired pneumonia, and postsurgical patients in the Intensive Care Unit (ICU). The two painful procedures chosen were tracheal suction and patient positioning. The data were collected at rest, at tracheal suctioning, 20 min later at positioning of the patient, and final reading 20 min later. Three testing periods, each including 4 assessments for a total of 12 pain assessments with sixty patients, were completed during each patient's ICU course. A total of six assessments were done with the patient at rest and three each with pain stimulus of tracheal suctioning and patient positioning. There was a significant increase in both hemodynamic variables during painful procedures except for the heart rate during positioning. The correlation between the SCA index and Ramsay scale was negative and significant. This instrument might prove useful to measure pain in uncommunicative critically ill patients and to evaluate the effectiveness of analgesic treatment and adapt it.


Delayed onset of experimental autoimmune encephalomyelitis in Olig1 deficient mice.
Olig1 is a basic helix-loop-helix (bHLH) transcription factor that is essential for oligodendrogenesis and efficient remyelination. However, its role in neurodegenerative disorders has not been well-elucidated. Here we investigated the effects of Olig1 deficiency on experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS). We show that the mean disease onset of myelin oligodendrocyte glycoprotein (MOG)-induced EAE in Olig1(-/-) mice is significantly slower than wide-type (WT) mice (19.8 ± 2.2 in Olig1(-/-) mice and 9.5 ± 0.3 days in WT mice). In addition, 10% of Olig1(-/-) mice did not develop EAE by the end of the observation periods (60 days). The severity of EAE, the extent of demyelination, and the activation of microglial cells and astrocytes in spinal cords, were significantly milder in Olig1(-/-) mice compared with WT mice in the early stage. Moreover, the visual function, as assessed by the second-kernel of multifocal electroretinograms, was better preserved, and the number of degenerating axons in the optic nerve was significantly reduced in Olig1(-/-) mice. Interestingly, Olig1 deficiency had no effect on T cell response capability, however, it reduced the expression of myelin proteins such as MOG, myelin basic protein (MBP) and myelin-associated glycoprotein (MAG). The expression of Olig2 remained unchanged in the optic nerve and brain, and it was reduced in the spinal cord of Olig1(-/-) mice. Our results suggest that the Olig1 signaling pathways may be involved in the incidence rate and the severity of neurological symptoms in MS.


First clinical implementation of real-time, real anatomy tracking and radiation beam control.
We describe the acceptance testing, commissioning, periodic quality assurance, and workflow procedures developed for the first clinically implemented magnetic resonance imaging-guided radiation therapy (MR-IGRT) system for real-time tracking and beam control. The system utilizes real-time cine imaging capabilities at 4 frames per second for real-time tracking and beam control. Testing of the system was performed using an in-house developed motion platform and a commercially available motion phantom. Anatomical tracking is performed by first identifying a target (a region of interest that is either tissue to be treated or a critical structure) and generating a contour around it. A boundary contour is also created to identify tracking margins. The tracking algorithm deforms the anatomical contour (target or a normal organ) on every subsequent cine frame and compares it to the static boundary contour. If the anatomy of interest moves outside the boundary, the radiation delivery is halted until the tracked anatomy returns to treatment portal. The following were performed to validate and clinically implement the system: (a) spatial integrity evaluation; (b) tracking accuracy; (c) latency; (d) relative point dose and spatial dosimetry; (e) development of clinical workflow for gating; and (f) independent verification by an outside credentialing service. The spatial integrity of the MR system was found to be within 2 mm over a 45-cm diameter field-of-view. The tracking accuracy for geometric targets was within 1.2 mm. The average system latency was measured to be within 394 ms. The dosimetric accuracy using ionization chambers was within 1.3% ± 1.7%, and the dosimetric spatial accuracy was within 2 mm. The phantom irradiation for the outside credentialing service had satisfactory results, as well. The first clinical MR-IGRT system was validated for real-time tracking and gating capabilities and shown to be reliable and accurate. Patient workflow methods were developed for efficient treatment. Periodic quality assurance tests can be efficiently performed with commercially available equipment to ensure accurate system performance.


A family of cyclin D homologs from plants differentially controlled by growth regulators and containing the conserved retinoblastoma protein interaction motif.
A new family of three related cyclins has been identified in Arabidopsis by complementation of a yeast strain deficient in G1 cyclins. Individual members show tissue-specific expression and are conserved in other plant species. They form a distinctive group of plant cyclins, which we named delta-type cyclins to indicate their similarities with mammalian D-type cyclins. The sequence relationships between delta and D cyclins include the N-terminal sequence LXCXE. This motif was originally identified in certain viral oncoproteins and is strongly implicated in binding to the retinoblastoma protein pRb. By analogy to mammalian cyclin D, these plant homologs may mediate growth and phytohormonal signals into the plant cell cycle. In support of this hypothesis, we show that, on restimulation of suspension-cultured cells, cyclin delta 3 is rapidly induced by the plant growth regulator cytokinin and cyclin delta 2 is induced by carbon source.


A stratified random sampling design in space and time for regional to global scale burned area product validation.
The potential research, policy and management applications of global burned area products place a high priority on rigorous, quantitative assessment of their accuracy. Such an assessment can be achieved by implementing validation methods employing design-based inference in which the independent reference data are selected via a probability sampling design. The majority of global burned area validation exercises use Landsat data to derive the independent reference data. This paper presents a three-dimensional sampling grid that allows for probability sampling of Landsat data in both space and time. To sample the globe in the spatial domain with non-overlapping sampling units, the Thiessen Scene Area (TSA) tessellation of the Landsat path/row geometry is used. The TSA grid is combined in time with the 16-day Landsat acquisition calendar to provide three-dimensional elements (voxels).This allows for implementation of stratified random sampling designs, where not only the location but also the time interval of the independent reference data is explicitly drawn by probability sampling. To illustrate this, we use a stratification methodology based on the Olson global ecoregion map and on the MODIS global active fire product. Using the global MODIS burned area product to establish a hypothetical population of reference data, we show that a sampling scheme based on the proposed stratification with equal sample allocation among strata is effective in reducing the standard errors of accuracy and area estimators compared to simple random sampling. Globally, the standard errors were reduced by 63%, 54%, 22% and 53% for overall accuracy, omission error, commission error and total burned area estimates respectively. By incorporating probability sampling in both the spatial and temporal domains, the present study establishes the foundation for rigorous design-based validation of global burned area products and, more generally, of terrestrial thematic products that have high temporal variability.


Experiences With Medical Exemptions After a Change in Vaccine Exemption Policy in California.
: media-1vid110.1542/5804909532001PEDS-VA_2018-1051Video Abstract OBJECTIVES: In 2015, California passed Senate Bill 227 (SB277), eliminating nonmedical vaccine exemptions for school entry. Our objective for this study was to describe the experiences of health officers and immunization staff addressing medical exemption requests under SB277. We conducted semistructured telephone interviews between August 2017 and September 2017 with health officers and immunization staff from local health jurisdictions in California. Interviews were recorded, transcribed, and analyzed for key themes. We conducted 34 interviews with 40 health officers and immunization staff representing 35 of the 61 local health jurisdictions in California. Four main themes emerged related to experiences with medical exemptions: (1) the role of stakeholders, (2) reviewing medical exemptions received by schools, (3) medical exemptions that were perceived as problematic, and (4) frustration and concern over medical exemptions. Generally, local health jurisdictions described a narrow role in providing support and technical assistance to schools. Only 5 jurisdictions actively tracked medical exemptions received by schools, with 1 jurisdiction facing a lawsuit as a result. Examples were provided of medical exemptions that listed family history of allergies and autoimmune diseases as contraindications for immunization and of physicians charging steep fees for medical exemptions. Participants also reported concerns about the increase in medical exemptions after the implementation of SB277. Participants reported many challenges and concerns with medical exemptions under SB277. Without additional legal changes, including a standardized review of medical exemptions, some physicians may continue to write medical exemptions for vaccine-hesitant parents, potentially limiting the long-term impact of SB277.


Alkene Oxyamination Using Malonoyl Peroxides: Preparation of Pyrrolidines and Isoxazolidines.
Treatment of homoallylic N-tosyl amines or allylic N-tosyl hydroxylamines with 1.5 equiv of a malonoyl peroxide provides a stereoselective method to access functionalized pyrrolidines and isoxazolidines. This metal free alkene oxyamination proceeds in 50-85% yield and up to 13:1 trans-selectivity. In addition, the relative stereochemistry of the oxygen and nitrogen substituents can be inverted through an oxidation/reduction sequence or inverting the stereochemistry of the starting alkene. Mechanistic investigations show a higher reactivity for hydroxyl nucleophiles over sulfonamide nucleophiles revealing a preference for dioxygenation over oxyamination.


Ontogenetic shifts in brain scaling reflect behavioral changes in the life cycle of the pouched lamprey Geotria australis.
Very few studies have described brain scaling in vertebrates throughout ontogeny and none in lampreys, one of the two surviving groups of the early agnathan (jawless) stage in vertebrate evolution. The life cycle of anadromous parasitic lampreys comprises two divergent trophic phases, firstly filter-feeding as larvae in freshwater and secondly parasitism as adults in the sea, with the transition marked by a radical metamorphosis. We characterized the growth of the brain during the life cycle of the pouched lamprey Geotria australis, an anadromous parasitic lamprey, focusing on the scaling between brain and body during ontogeny and testing the hypothesis that the vast transitions in behavior and environment are reflected in differences in the scaling and relative size of the major brain subdivisions throughout life. The body and brain mass and the volume of six brain structures of G. australis, representing six points of the life cycle, were recorded, ranging from the early larval stage to the final stage of spawning and death. Brain mass does not increase linearly with body mass during the ontogeny of G. australis. During metamorphosis, brain mass increases markedly, even though the body mass does not increase, reflecting an overall growth of the brain, with particularly large increases in the volume of the optic tectum and other visual areas of the brain and, to a lesser extent, the olfactory bulbs. These results are consistent with the conclusions that ammocoetes rely predominantly on non-visual and chemosensory signals, while adults rely on both visual and olfactory cues.


Regulation of iro3 expression in the zebrafish spinal cord.
Specification of spinal cord neurons is regulated by several different transcription factors. In this study, we analyze expression and regulation of the transcription factor iro3 in zebrafish spinal cord. In addition to its broad expression in the progenitor domain of intermediate spinal cord, iro3 is also expressed in postmitotic ventral neurons, starting at early somitogenesis stages. Initially, this expression is only in two primary motoneurons, CaP and VaP, but by 24 hr postfertilization, iro3 is expressed by all classes of zebrafish spinal motoneurons as well as by a ventral interneuron called VeLD. iro3 expression in the progenitor domain of intermediate spinal cord is regulated independently from its expression in ventral neurons. Hedgehog (Hh) signaling is unnecessary for iro3 expression in intermediate spinal cord, but it is required to repress iro3 expression in the progenitor domain of ventral spinal cord. We also show that the basic helix-loop-helix transcription factor Olig2 is required for repression of iro3 expression in the progenitor domain of ventral spinal cord. We discuss our findings in the context of previous studies, suggesting that iro3 represses formation of motoneurons and promotes formation of interneurons.


Renal expression of osmotically responsive cation channel TRPV4 is restricted to water-impermeant nephron segments.
TRPV4, a nonselective cation channel of the transient receptor potential (TRP) family, is gated by hypotonicity. Expression of TRPV4 mRNA has been detected in the circumventricular organs of the brain responsible for sensing systemic tonicity and in the kidney distal convoluted tubule (DCT), among other sites. No analysis of TRPV4 expression at the protein level has been undertaken and no systematic analysis of expression of this channel has been reported in the kidney. Via RNAse protection assay and immunoblotting, abundant expression of TRPV4 was detected in the cortex, medulla, and papilla. The expression pattern of TRPV4 was characterized in both rat and mouse kidney, which revealed similar patterns of immunoreactivity. TRPV4 expression was absent from the proximal tubule (PT) and descending thin limb (DTL), whereas the strongest expression was observed in the ascending thin limb (ATL). The thick ascending limb (TAL) was strongly positive as was the DCT and connecting tubule. Importantly, the water-permeant cells of the macula densa were unstained. Moderate TRPV4 expression was noted in all collecting duct portions and in papillary epithelium; intercalated cells (type A) exhibited a particularly strong signal. In all positive segments, TRPV4 expression was concentrated at the basolateral membrane. Therefore, TRPV4 is expressed in only those nephron segments that are constitutively (i.e., ATL, TAL, and DCT) or conditionally (i.e., collecting duct) water impermeant and where generation of a substantial transcellular osmotic gradient could be expected. TRPV4 expression is absent from nephron segments exhibiting constitutive water permeability and unregulated apical aquaporin expression (i.e., PT and DTL). These data, although circumstantial, are consistent with a role for TRPV4 in the response to anisotonicity in the mammalian kidney.


Characterization of freshwater natural aquatic colloids by atomic force microscopy (AFM).
Atomic force microscopy (AFM) has been used to image and quantify riverine colloids in a quantitative and relatively nonperturbing manner. Three main classes of material have been imaged including fibrils (about 10 nm in diameter and 100 nm or more in length), discrete, near-spherical, small colloids primarily below 30-50 nm in diameter, and a surface film, of at least several nanometers thickness, which coatsthe entire mica surface within 30 min of exposure to river water. Colloid structure was found to vary as a function of pH, particularly at high pH. Substantially different structures were observed at high pH values, with the loss of the near-spherical colloids possibly due to rearrangement and aggregation. In addition,film thicknesses of up to 100 nm were estimated on the silicon nitride AFM cantilever after 30 h of deposition in the same water (unperturbed and size fractionated). The observation of these surface films has important implications for understanding the mechanisms by which colloids might bind trace elements. In particular, development of surface coatings implies that binding of pollutants (at least initial surface binding) may be dominated by adsorbed surface layers.


Questionnaire survey on pediatric hypertension in Japan and Korea.
Hypertension (HTN) is no longer viewed as an adult disease. The purpose of the present study was to understand how hypertensive children are evaluated and managed, by surveying pediatricians in Japan and South Korea. A questionnaire was mailed to 109 Japanese (JA) and 159 Korean (KO) pediatric cardiologists, pediatric nephrologists, and other pediatricians. A total of 127 replies were received (response rate 47%). Most of respondents did not check blood pressure (BP) routinely in outpatient clinics (JA 77%, KO 88%). A mercury sphygmomanometer was the most commonly used method for BP measurements (JA 72%, KO 62%). BP treatment goals were usually set at the 95th percentile for age, gender, and height (JA 47%, KO 54%). More KO used a lower goal in children with primary HTN than JA. KO respondents preferred angiotensin-converting enzyme inhibitors (ACEI) as initial agents regardless of underlying diseases whereas JA respondents chose various medications, that is, calcium channel blockers, diuretics, and ACEI. For BP monitoring, self-monitoring was found to be most frequent in both countries (JA 80%, KO 57%). Ambulatory BP monitoring was not frequently utilized in both countries (JA 33% KO 34%). The current assessment, management and differing trends in pediatric HTN in Japan and Korea have been identified in the present study. Pediatricians should be aware of the growing implications of HTN in children.


Effects of exposure to microwaves on cellular immunity and placental steroids in pregnant rats.
Microwaves produce various detrimental changes based on actions of heat or non-specific stress, although the effects of microwaves on pregnant organisms has not been uniform. This study was designed to clarify the effect of exposure to microwaves during pregnancy on endocrine and immune functions. Natural killer cell activity and natural killer cell subsets in the spleen were measured, as well as some endocrine indicators in blood--corticosterone and adrenocorticotrophic hormone (ACTH) as indices of the hypothalamic-pituitary-adrenal axis--beta-endorphin, oestradiol, and progesterone in six female virgin rats and six pregnant rats (nine to 11 days gestation) exposed to microwaves at 10 mW/cm2 incident power density at 2450 MHz for 90 minutes. The same measurements were performed in control rats (six virgin and six pregnant rats). Skin temperature in virgin and pregnant rats increased immediately after exposure to microwaves. Although splenic activity of natural killer cells and any of the subset populations identified by the monoclonal antibodies CD16 and CD57 did not differ in virgin rats with or without exposure to microwaves, pregnant rats exposed to microwaves showed a significant reduction of splenic activity of natural killer cells and CD16+CD57-. Although corticosterone and ACTH increased, and oestradiol decreased in exposed virgin and pregnant rats, microwaves produced significant increases in beta-endorphin and progesterone only in pregnant rats. Microwaves at the power of 10 mW/cm2 produced activation of the hypothalamic-pituitary-adrenal axis and increased oestradiol in both virgin and pregnant rats, suggesting that microwaves greatly stress pregnant organisms. These findings in pregnant rats suggest that--with exposure to microwaves--pregnancy induces immunosuppression, which could result in successful maintainance of pregnancy. This enhancement of adaptability to heat stress with pregnancy may be mediated by activation of placental progesterone and placental or pituitary beta-endorphin.


Quantitative cardiac magnetic resonance T2 imaging offers ability to non-invasively predict acute allograft rejection in children.
Monitoring for acute allograft rejection improves outcomes after cardiac transplantation. Endomyocardial biopsy is the gold standard test defining rejection, but carries risk and has limitations. Cardiac magnetic resonance T2 mapping may be able to predict rejection in adults, but has not been studied in children. Our aim was to evaluate T2 mapping in identifying paediatric cardiac transplant patients with acute rejection. Eleven paediatric transplant patients presenting 18 times were prospectively enrolled for non-contrast cardiac magnetic resonance at 1.5 T followed by endomyocardial biopsy. Imaging included volumetry, flow, and T2 mapping. Regions of interest were manually selected on the T2 maps using the middle-third technique in the left ventricular septal and lateral wall in a short-axis and four-chamber slice. Mean and maximum T2 values were compared with Student's t-tests analysis. Five cases of acute rejection were identified in three patients, including two cases of grade 2R on biopsy and three cases of negative biopsy treated for clinical symptoms attributed to rejection (new arrhythmia, decreased exercise capacity). A monotonic trend between increasing T2 values and higher biopsy grades was observed: grade 0R T2 53.4 ± 3 ms, grade 1R T2 54.5 ms ± 3 ms, grade 2R T2 61.3 ± 1 ms. The five rejection cases had significantly higher mean T2 values compared to cases without rejection (58.3 ± 4 ms versus 53 ± 2 ms, p = 0.001). Cardiac magnetic resonance with quantitative T2 mapping may offer a non-invasive method for screening paediatric cardiac transplant patients for acute allograft rejection. More data are needed to understand the relationship between T2 and rejection in children.


Alzheimer's disease genes and cognition in the nondemented general population.
Genome-wide association studies have established 11 genes for late-onset Alzheimer's disease (AD). We investigated whether these genes jointly affect cognition in a nondemented population and improve prediction of AD. In 5171 nondemented people (age 45-99 years) from the population-based Rotterdam Study, we genotyped APOE-ε4 and single nucleotide polymorphisms from the genes CLU, PICALM, BIN1, CR1, ABCA7, MS4A6A, MS4A4E, CD2AP, EPHA1, and CD33. We constructed a genetic risk score by adding the risk alleles per individual weighted by the reported effect sizes. All people underwent cognitive testing. With linear regression we investigated the relationship between the genetic risk score and cognition, with and without APOE. In a subcohort, with more than 10 years of follow-up, we assessed whether the risk score predicted AD. The genetic risk score was primarily associated with memory (standardized difference [95% confidence interval] per SD increase in genetic risk score: -.05[-.07; -.02], p = 1.5 * 10(-4)). This association attenuated when APOE was excluded from the genetic risk score (-.03[-.05; .00], p = .047) and would not survive a multiple-testing correction. Similarly, we found that although the genetic risk score without APOE was associated with the development of AD (p = .010), it only marginally improved prediction of AD beyond age, sex, APOE (area under the curve: .8159 vs. .8148). In nondemented people, there is only a marginal joint effect of AD genes on memory independent from APOE. Moreover, although associated with AD, these genes jointly hardly improve prediction of AD.


Sustaining the rural workforce: nursing perspectives on worklife challenges.
Concerns have been raised about the sustainability of health care workforces in rural settings. According to the literature, rural nurses' work satisfaction varies with the resources and supports available to respond to specific challenges. Given the probable effects of stressors on retention, it is essential to understand the unique requirements of nurses in rural practice environments. To investigate whether nurses receive the resources and supports necessary to meet the challenges of rural practice. Semi-structured interviews were conducted with 21 managers and 44 staff nurses in 19 selected rural hospitals in Ontario, Canada. The interviews were taped and transcripts interpreted through a thematic analysis. Major worklife themes were identified and analyzed within a healthy work environment model based on the work of Kristensen. Three interrelated dimensions of the model were relevant to workforce sustainability: the balance between demands and the resources of the person, the level of social support, and the degree of influence. The availability of resources and supports affected whether the nurses perceived challenges as stimulating or overwhelming. Deficits interfered with practice and the well-being of the nurses and patients. The nurses felt frustrated and powerless when they lacked resources, support, and influence to manage negative situations. Strategies to achieve workforce sustainability include resources to reduce stress in the workplace, education to meet the needs of new and experienced nurses, and offering of employment preferences to the workforce. Addressing resources, support, and influence of rural nurses is essential to alleviate workplace challenges and sustain the rural nursing workforce.


Eosinophils in COPD: just another biomarker?
Eosinophils are innate immune cells that, under certain conditions, can be recruited to the lungs, where they have an incompletely understood role in health and disease. Eosinophils have been found in the airways, tissues, and circulation of patients with COPD, during both stable disease and exacerbations. Epidemiological studies and post-hoc analyses of clinical trials of corticosteroid treatment for COPD have shown that the blood eosinophil count is associated with the risk of COPD exacerbations, mortality, decline in FEV1, and response to both inhaled and systemic corticosteroids. Further studies are urgently needed to explore the contribution of eosinophils to the mechanism of disease in COPD and to identify their association with levels of clinical risk. In this review, we explore the role of the eosinophil as a biomarker and mediator of disease in COPD.


Three-Legged Tree Tensor Networks with SU(2) and Molecular Point Group Symmetry.
We extend the three-legged tree tensor network state (T3NS) [ J. Chem. Theory Comput. 2018 , 14 , 2026 - 2033 ] by including spin and the real abelian point group symmetries. T3NS intersperses physical tensors with branching tensors. Physical tensors have one physical index and at most two virtual indices. Branching tensors have up to three virtual indices and no physical index. In this way, T3NS combines the low computational cost of matrix product states and their simplicity for implementing symmetries, with the better entanglement representation of tree tensor networks. By including spin and point group symmetries, more accurate calculations can be obtained with lower computational effort. We illustrate this by presenting calculations on the bis(μ-oxo) and μ-η2:η2 peroxo isomers of [Cu2O2]2+. The used implementation is available on github .


Enzyme inhibitors for renin assay in rat plasma.
1. Degradation of 125I-labelled [Ile5]-angiotensin I and unlabelled angiotensin I was studied during incubation of rat plasma treated with disodium ethylenediaminetetra-acetate (disodium EDTA) at 37 degrees C, pH 6.5, for 1 h with 8-hydroxyquinoline, phenylmethylsulphonyl fluoride or di-isopropyl fluorophosphate. 2. Isoelectric focusing showed approximately 100, 59--66 and 10--31% preservation of 125I-labelled angiotensin I during incubation with 8-hydroxyquinoline (5.0 mmol/l), phenylmethylsulphonyl fluoride (15 mmol/l) and di-isopropyl fluorophosphate (1.09 mmol/l) respectively. Mean recovery of unlabelled angiotensin I was 102, 77 and 32% with 8-hydroxyquinoline, phenylmethylsulphonyl fluoride and di-isopropyl fluorophosphate respectively. 3. 8-Hydroxyquinoline appears the preferable enzyme inhibitor for renin assay in rat plasma.


Preschool selection considerations and experiences of school mistreatment among lesbian, gay, and heterosexual adoptive parents.
The current study is the first to investigate the school selection considerations and school-related experiences of sexual-minority parents with young children. The sample consisted of 210 parents in 105 couples, including 35 lesbian couples, 30 gay male couples, and 40 heterosexual couples, all of whom had adopted a child three years earlier. We found that parents with less income were more likely to consider cost in choosing a preschool, and parents with less education were more likely to consider location. More educated parents tended to emphasize racial diversity and the presence of adoptive families, and, among sexual-minority parents, the presence of other lesbian/gay parents. Sexual-minority parents were more likely to consider racial diversity than heterosexual parents. In reporting on their experiences with schools, heterosexual parents were more likely to perceive mistreatment due to their adoptive status than sexual-minority parents, and sexual-minority parents living in less gay-friendly communities were more likely to perceive mistreatment due to their sexual orientation than sexual-minority parents living in more gay-friendly communities. Our findings have implications for early childhood educators and administrators seeking to create an inclusive learning community for all types of families.


Reduction and transforaminal lumbar interbody fusion with posterior fixation versus transsacral cage fusion in situ with posterior fixation in the treatment of Grade 2 adult isthmic spondylolisthesis in the lumbosacral spine.
In situ transsacral fusion in the treatment of low-grade isthmic spondylolisthesis has rarely been reported. The authors treated 13 cases of L-5 Grade 2 isthmic spondylolisthesis associated with collapsed disc space and osteoporosis by using transsacral fusion and fixation, and compared its clinical and radiological outcomes with the results of transforaminal lumbar interbody fusion (TLIF) and instrumental reduction in 21 patients. The authors retrospectively analyzed 21 patients in Group A who were treated with reduction and TLIF, and 13 patients in Group B who were treated with transsacral cage fusion. Oswestry Disability Index and visual analog scale scores of back and leg pain were used to evaluate clinical outcomes. Radiological parameters for assessment included the percentage of slippage, whole lumbar lordosis, and lumbosacral angle. Operative data, fusion rate, and perioperative complications were recorded as well. The mean operation time and blood loss in Group B was less than that in Group A. Both groups realized good recovery from previous symptoms. The decrease in back and leg pain after surgery was significant within each group, without much difference between the 2 groups. No significant differences were found in lumbosacral angle, whole lumbar lordosis, visual analog scale score, and Oswestry Disability Index score between the 2 groups after surgery. The solid fusion rate was 95.2% in Group A and 92.3% in Group B. In Group A, 2 patients suffered from graft site pain, 1 had a superficial infection, and 1 had screw loosening; in Group B, dural tears were found in 2 patients, transient S-1 paresthesia in 2, and extensor hallucis longus muscle weakness in 1. For patients with a collapsed disc space and poor bone quality, posterior in situ transsacral cage fusion may be used as an alternative to the TLIF procedure. The short-term clinical and radiological outcomes in the transsacral cage group were comparable with those in the TLIF group, although with a relatively higher neurological complication rate.


Complete genome sequence of endophyte Bacillus flexus KLBMP 4941 reveals its plant growth promotion mechanism and genetic basis for salt tolerance.
Bacillus flexus KLBMP 4941 is a halotolerant endophyte isolated from the halophyte Limonium sinense. This strain can improve host seedling growth under salt stress conditions. We here report the complete genome information of endophyte KLBMP 4941. It has a circular chromosome and two plasmids for a total genome 4,104,242 bp in size with a G+C content of 38.09%. Genes related to plant growth promotion (PGP), such as those associated with nitrogen fixation, siderophore, spermidine, and acetoin synthesis were found in the KLBMP 4941 genome. Some genes responsible for high salinity tolerance, like genes associated with the Na+/H+ antiporter, glycine betaine transporter, and betaine-aldehyde dehydrogenase were also found in the KLBMP 4941 genome. The genome analysis will provide better understanding of the mechanisms underlying the promotion of plant growth in strain KLBMP 4941 under salt stress conditions and its ability to adapt to coastal salt marsh habitats, and provide a basis for its further biotechnological applications in agriculture.


Mechanisms of hepatocellular toxicity associated with dronedarone--a comparison to amiodarone.
Dronedarone is a new antiarrhythmic drug with an amiodarone-like benzofuran structure. Shortly after its introduction, dronedarone became implicated in causing severe liver injury. Amiodarone is a well-known mitochondrial toxicant. The aim of our study was to investigate mechanisms of hepatotoxicity of dronedarone in vitro and to compare them with amiodarone. We used isolated rat liver mitochondria, primary human hepatocytes, and the human hepatoma cell line HepG2, which were exposed acutely or up to 24h. After exposure of primary hepatocytes or HepG2 cells for 24h, dronedarone and amiodarone caused cytotoxicity and apoptosis starting at 20 and 50 µM, respectively. The cellular ATP content started to decrease at 20 µM for both drugs, suggesting mitochondrial toxicity. Inhibition of the respiratory chain required concentrations of ~10 µM and was caused by an impairment of complexes I and II for both drugs. In parallel, mitochondrial accumulation of reactive oxygen species (ROS) was observed. In isolated rat liver mitochondria, acute treatment with dronedarone decreased the mitochondrial membrane potential, inhibited complex I, and uncoupled the respiratory chain. Furthermore, in acutely treated rat liver mitochondria and in HepG2 cells exposed for 24h, dronedarone started to inhibit mitochondrial β-oxidation at 10 µM and amiodarone at 20 µM. Similar to amiodarone, dronedarone is an uncoupler and an inhibitor of the mitochondrial respiratory chain and of β-oxidation both acutely and after exposure for 24h. Inhibition of mitochondrial function leads to accumulation of ROS and fatty acids, eventually leading to apoptosis and/or necrosis of hepatocytes. Mitochondrial toxicity may be an explanation for hepatotoxicity of dronedarone in vivo.


Prospective comparison of the tuberculin skin test and 2 whole-blood interferon-gamma release assays in persons with suspected tuberculosis.
Interferon-gamma release assays (IGRAs) are attractive alternatives to the tuberculin skin test (TST) for detecting Mycobacterium tuberculosis infection. However, the inability to definitively confirm the presence of most M. tuberculosis infections hampers assessment of IGRA accuracy. Although IGRAs are primarily indicated for the detection of latent tuberculosis infection, we sought to determine the sensitivity of the TST and 2 whole-blood IGRAs (QuantiFERON-TB assay [QFT] and QuantiFERON-TB Gold assay [QFT-G]) in situations in which infection is confirmed by recovery of M. tuberculosis by culture. We conducted a prospective, multicenter, cross-sectional comparison study in which 148 persons suspected to have tuberculosis were tested simultaneously with the TST, QFT, and QFT-G. M. tuberculosis was cultured from samples from 69 (47%) of 148 persons suspected to have tuberculosis; the TST induration was > or = 5 mm for 51 (73.9%) of the 69 subjects (95% confidence interval [CI], 62.5%-82.8%). The QFT indicated tuberculosis infection for 48 (69.6%) of the 69 subjects (95% CI, 57.9%-79.2%) and was indeterminate for 7 (10.1%). The QFT-G yielded positive results for 46 (66.7%) of the 69 subjects (95% CI, 54.9%-76.7%) and indeterminate results for 9 subjects (13.0%). If subjects with indeterminate QFT-G results were excluded, 46 (76.7%) of 60 subjects (95% CI, 64.6%-85.6%) had positive TST results, and the same number of subjects had positive QFT-G results. HIV infection was associated with false-negative TST results but not with false-negative QFT-G results. The TST, QFT, and QFT-G have similar sensitivity in persons with culture-confirmed infection. As with the TST, negative QFT and QFT-G results should not be used to exclude the diagnosis of tuberculosis in persons with suggestive signs or symptoms.


Therapeutic efficacy of PUMA for malignant glioma cells regardless of p53 status.
Replacement of the p53 tumor suppressor gene is a rational approach to the management of malignant gliomas because p53 is frequently mutated or inactivated in these cancers. Major weaknesses of this approach are that malignant gliomas are mixtures of cells with wild-type and mutant p53, and that tumor cells exhibiting wildtype p53 are resistant to p53 gene transfer. An effective alternative is needed to overcome these difficulties. p53-upregulated modulator of apoptosis (PUMA) was identified as a p53-inducible proapoptotic molecule. Our purpose was to elucidate a role for PUMA in p53 gene therapy and to investigate whether PUMA is an efficient substitute for p53 in cancer therapy. We demonstrated that PUMA was upregulated in mutant p53 malignant glioma cells (U373-MG and T98G) undergoing apoptosis but was not upregulated in apoptosis-resistant wild-type p53 malignant glioma cells (U87-MG and D54) after adenoviral transfer of p53. Overexpression of PUMA resulted in massive apoptosis associated with mitochondrial damage and caspase-3 activation in all tumor cells tested. Use of the human telomerase reverse transcriptase (hTERT) promoter system induced apoptosis only in malignant glioma cells with telomerase activity, while sparing normal cells lacking telomerase. The ability of PUMA to induce apoptosis was greater than that of caspase-6 or caspase-8 transfer, using the same system. Moreover, exogenous expression of PUMA under the hTERT promoter system significantly suppressed the growth of subcutaneous U87-MG tumors in nude mice and did not induce apoptosis in surrounding nontumor tissues. These results indicate that PUMA, which is regulated under a tumor-specific expression system such as the hTERT promoter, may be better than p53 as a therapeutic tool for malignant gliomas.


Sorting out a problem: A co-production approach to household waste management in Shanghai, China.
Rapid urban population growth in China has resulted in significant challenges for Municipal Solid Waste (MSW) management. To combat these challenges, Chinese authorities implemented a pilot program of MSW sorting in eight Chinese cities including Shanghai in 2000 but is yet to reach a widespread success. This paper uses a qualitative case-study approach to explore the potential of a community-based co-production strategy for household waste sorting as an alternative to the conventional top-down approach. Co-production implies the involvement of citizens, consumers, volunteers and/or community organizations in producing public service. We find two major drivers of the government-volunteer consortium and the peer-pressure effect help realize the potential of waste-management co-production at a lowered cost with improved efficiency and collaborative innovation. In addition, context-specific conditions including policy consistency, strong volunteer effort and compatibility with local culture to promote public participation must be present for further co-production application. We also acknowledge that the co-production approach would be most effective at the initial stage of policy implementation to foster the waste-sorting habit-formation in regions where the waste collection rates remain low.


A review of unmanned vehicles for the detection and monitoring of marine fauna.
Recent technology developments have turned present-day unmanned systems into realistic alternatives to traditional marine animal survey methods. Benefits include longer survey durations, improved mission safety, mission repeatability, and reduced operational costs. We review the present status of unmanned vehicles suitable for marine animal monitoring conducted in relation to industrial offshore activities, highlighting which systems are suitable for three main monitoring types: population, mitigation, and focal animal monitoring. We describe the technical requirements for each of these monitoring types and discuss the operational aspects. The selection of a specific sensor/platform combination depends critically on the target species and its behaviour. The technical specifications of unmanned platforms and sensors also need to be selected based on the surrounding conditions of a particular offshore project, such as the area of interest, the survey requirements and operational constraints.


Relapse and rehospitalization: comparing oral and depot antipsychotics.
A review of studies that compared conventional oral and depot antipsychotic medications highlighted the following points. Mirror-image studies in which patients served as their own controls provided evidence of substantial benefit for depot injectable medications. The randomized clinical trials did not, in general, support the findings of significant decrease in relapse rates between these 2 routes of administration. Across the studies reviewed, the 1-year relapse rate for long-acting depot medication was 27% compared with 42% for patients who received oral medication. The 27% risk of relapse in patients who received guaranteed depot medication suggests that relapse is not always driven by noncompliance. In the only study that lasted for 2 years, the risk of relapse decreased substantially in the depot-treated patients, suggesting that risk of noncompliance may be a more important factor in relapse over extended periods of time. A recent formal meta-analytic review of depot medications concluded that this route of administration resulted in clinical advantages in terms of global outcome.


Yoga for depression: the research evidence.
Yoga-based interventions may prove to be an attractive option for the treatment of depression. The aim of this study is to systematically review the research evidence on the effectiveness of yoga for this indication. Searches of the major biomedical databases including MEDLINE, EMBASE, ClNAHL, PsycINFO and the Cochrane Library were conducted. Specialist complementary and alternative medicine (CAM) and the IndMED databases were also searched and efforts made to identify unpublished and ongoing research. Searches were conducted between January and June 2004. Relevant research was categorised by study type and appraised. Clinical commentaries were obtained for studies reporting clinical outcomes. Five randomised controlled trials were located, each of which utilised different forms of yoga interventions and in which the severity of the condition ranged from mild to severe. All trials reported positive findings but methodological details such as method of randomisation, compliance and attrition rates were missing. No adverse effects were reported with the exception of fatigue and breathlessness in participants in one study. No language restrictions were imposed on the searches conducted but no searches of databases in languages other than English were included. Overall, the initial indications are of potentially beneficial effects of yoga interventions on depressive disorders. Variation in interventions, severity and reporting of trial methodology suggests that the findings must be interpreted with caution. Several of the interventions may not be feasible in those with reduced or impaired mobility. Nevertheless, further investigation of yoga as a therapeutic intervention is warranted.


Safety and Efficacy of Combination Antiretroviral Therapy in Human Immunodeficiency Virus-Infected Adults Undergoing Autologous or Allogeneic Hematopoietic Cell Transplantation for Hematologic Malignancies.
The ability to continue combination antiretroviral therapy (cART) in human immunodeficiency virus (HIV)-infected patients undergoing hematopoietic cell transplantation (HCT) for treatment of hematologic malignancies is likely a critical factor in preventing the establishment of an HIV reservoir in transplanted stem cells. Thus, we studied the feasibility of continued antiretroviral therapy in our HIV-infected patients undergoing autologous or allogeneic transplantation. All HIV-infected adults undergoing HCT for hematologic malignancy at Fred Hutchinson Cancer Research Center between 2006 and 2014 were included; most were enrolled in a prospective clinical study to monitor HIV reservoirs after transplantation (NCT00968630 and NCT00112593). Non-nucleotide reverse transcriptase inhibitor or integrase-strand inhibitor-anchored antiretroviral therapy regimens were continued or selected before HCT by infectious disease physicians. Plasma HIV RNA was measured every other day for the first 2 weeks after transplantation and then every 2 weeks. Missed doses of cART and reasons for changing the cART regimen during the post-transplantation hospitalization were documented through review of inpatient pharmacy records. Seven autologous and 8 allogeneic transplantations were performed. In 9 transplantations, the cART regimen was not altered after HCT and no doses were missed. In 2 patients who required alterations in their cART regimen because of development of acute renal failure (n = 1) and small bowel obstruction (n = 1) after HCT, enfuvirtide was used as a bridging component of the regimen. Plasma HIV RNA remained suppressed during the first 28 days in 12 of 15 transplantations, and no patients had a plasma HIV RNA >1000 copies/mL during long-term follow up. Non-nucleotide reverse transcriptase inhibitor- and integrase-strand inhibitor-based cART are safe and effective in HIV-infected persons during the peri-HCT period. Most patients undergoing HCT were able to continue cART without missed doses. Sustained HIV viremia and emergence of resistance were not detected.


An investigation of decision making in anorexia nervosa using the Iowa Gambling Task and skin conductance measurements.
The objective of this study is to determine (a) if decision making ability is impaired in patients with anorexia nervosa (AN) and in people with good recovery from AN and (b) whether any impairment in decision making is associated with alterations in skin conductance responses (SCR). Patients with AN (n = 29), healthy controls comparable in age and IQ (HC, n = 29), and women long term recovered from AN (n = 14), completed the Iowa Gambling Task (IGT) while their SCR were measured. AN patients performed poorly in the IGT compared to the HC and to the recovered AN participants. AN patients had decreased anticipatory SCR prior to choosing cards and reduced SCR after losses compared to HC. IGT performance and the SCR of recovered AN participants did not differ from the HC. Decision making ability is impaired in AN. It is associated with a significantly attenuated SCR. Neither of these features are found in recovered AN. The association between impaired decision making ability and a decreased autonomic response is consistent with the predictions of the Somatic Marker Hypothesis.


Nip the bubble in the bud: a guide to avoid gas nucleation in microfluidics.
Gas bubbles are almost a routine occurrence encountered by researchers working in the field of microfluidics. The spontaneous and unexpected nature of gas bubbles represents a major challenge for experimentalists and a stumbling block for the translation of microfluidic concepts to commercial products. This is a startling example of successful scientific results in the field overshadowing the practical hurdles of day-to-day usage. We however believe such hurdles can be overcome with a sound understanding of the underlying conditions that lead to bubble formation. In this tutorial, we focus on the two main conditions that result in bubble nucleation: surface nuclei and gas supersaturation in liquids. Key theoretical concepts such as Henry's law, Laplace pressure, the role of surface properties, nanobubbles and surfactants are presented along with a view of practical implementations that serve as preventive and curative measures. These considerations include not only microfluidic chip design and bubble traps but also often-overlooked conditions that regulate bubble formation, such as gas saturation under pressure or temperature gradients. Scenarios involving electrolysis, laser and acoustic cavitation or T-junction/co-flow geometries are also explored to provide the reader with a broader understanding on the topic. Interestingly, despite their often-disruptive nature, gas bubbles have also been cleverly utilized for certain practical applications, which we briefly review. We hope this tutorial will provide a reference guide in helping to deal with a familiar foe, the "bubble".


Altering the time of vaccination against respiratory pathogens to enhance antibody response and performance of feeder cattle.
Ninety Angus × Hereford calves were ranked by sex, BW, and age and assigned to 1 of 3 vaccination schemes against the bovine respiratory disease complex: 1) vaccination at weaning (d 0) and a booster at feedlot entry (d 30; CON; = 30), 2) vaccination 15 d before weaning (d -15) and a booster 15 d before feedlot entry (d 15; EARLY; = 30), and 3) vaccination 15 d after weaning (d 15) and a booster 15 d after feedlot entry (d 45; DELAYED; = 30). From d -15 to 7, calves were maintained as a single group on pasture. On d 8, calves were placed into 1 of 18 drylot pens (6 pens/treatment; 5 calves/pen) and fed alfalfa-triticale hay. On d 29, calves were transported 1,440 km in a livestock trailer and unloaded on d 30 at the same feed yard with the same pen arrangement used prior to transport. From d 30 to 75, calves were fed a receiving diet based on alfalfa-triticale hay + corn-based concentrate. Calf BW was recorded on 2 consecutive days (d -15, -14, 0, 1, 28, 29, 75, and 76). Blood samples were collected on d -15, 0, 15, 30, 45, 60, and 75. The EARLY calves had less ( ≤ 0.09) ADG before weaning (d -15 to -1); however, they had greater ( ≤ 0.01) ADG during feedlot receiving (d 30 to 75) compared with calves with the other treatments. During preconditioning (d 0 to 29), CON calves had greater ( ≤ 0.04) DMI compared with EARLY and DELAYED calves. During feedlot receiving, no treatment differences were detected ( ≥ 0.17) for hay or concentrate DMI, G:F, and morbidity and mortality rates. There were no treatment effects on calf BW at weaning and at the end of the preconditioning or receiving periods ( ≥ 0.65). Plasma concentrations of antibodies against were greater ( ≤ 0.05) in EARLY calves than in CON and DELAYED calves on d 0, greater ( ≤ 0.04) for CON calves than for EARLY and DELAYED calves on d 15, greater ( ≤ 0.02) in DELAYED and EARLY calves than in CON calves on d 30, and greater ( = 0.03) in EARLY calves than in CON calves on d 75. Plasma concentrations of antibodies against bovine viral diarrhea viruses were greater ( ≤ 0.04) in EARLY calves than in CON and DELAYED calves on d 15 and greater for EARLY and CON calves than for DELAYED calves on d 30 and 45. Collectively, EARLY calves had greater plasma concentrations of antibodies against the evaluated pathogens at feedlot entry and increased ADG during receiving compared with their CON and DELAYED cohorts. Hence, anticipating initial and booster vaccinations against respiratory pathogens to provide both doses prior to feedlot entry appears to be a valid strategy to enhance cattle health and performance during feedlot receiving.


Atrazine in public water supplies and risk of ovarian cancer among postmenopausal women in the Iowa Women's Health Study.
Few studies have evaluated environmental chemical exposures in relation to ovarian cancer. We previously found an increased risk of ovarian cancer among postmenopausal women in Iowa associated with higher nitrate levels in public water supplies (PWS). However, elevated nitrate levels may reflect the presence of other agricultural chemicals, such as atrazine, one of the most commonly detected pesticides in Iowa PWS. We evaluated the association between atrazine in drinking water and incident ovarian cancer (N=145, 1986-2010) among 13 041 postmenopausal women in the Iowa Women's Health Study who used their PWS for ≥11 years as reported in 1989. Average levels of atrazine (1986-1987), nitrate-nitrogen (NO3-N, 1955-1988) and estimated levels of total trihalomethanes (TTHM, 1955-1988) from PWS monitoring data were linked to the participants' cities of residence. We computed HRs and 95% CIs by categories of the average atrazine level (not detected, ≤ or >0.37 parts per billion=median) using Cox proportional hazards regression adjusting for ovarian cancer risk factors. Atrazine was detected in water samples from 69 cities where 4155 women (32%) lived and levels were moderately correlated with NO3-N (ρ=0.35) and TTHM (ρ=0.24). Atrazine levels were not associated with ovarian cancer risk with or without adjusting for NO3-N and TTHM levels (p-trend=0.50 and 0.81, respectively). Further, there was no evidence for effect modification of the atrazine association by NO3-N or TTHM levels. In our study with low atrazine detection rates, we found no association between atrazine in PWS and postmenopausal ovarian cancer risk.


The Success and Evolution of a Urological "Boot Camp" for Newly Appointed UK Urology Registrars: Incorporating Simulation, Nontechnical Skills and Assessment.
Urological training has dramatically changed in recent years. Training durations are shorter and a drive toward consultant led care has reduced trainees experience. Within the UK, approximately 50 registrars annually embark on a 5-year Urology training programme, with variable levels of basic urological experience. To describe a simulation programme aimed at delivering the knowledge and skills necessary to safely and effectively start working as a registrar in Urology by intensive training with a 1:1 faculty to delegate ratio. Our course content mirrors the UK training syllabus for junior Urology registrars. We delivered 8 modules over a 4-day programme with a fifth day of assessments. Delegates level of urological knowledge, operative competency and confidence pre-, immediately post-training and at 3-months postcourse were assessed. Objective delegate and faculty feedback was also collected. Technical skills modules include; inguinoscrotal surgery, ureteroscopy, transurethral resection, urodynamics, and Botox administration as well as basic reconstructive and laparoscopic operative skills. "Nontechnical" skills included simulated ward round, out-patient, and emergency scenarios. Feedback from delegates and faculty members has been overwhelmingly positive. We have used this feedback to tailor the content of the course for following years. An increased knowledge level (based on mean examination scores [precourse 55.5%, postcourse 70.1%]) and operative competency was observed in all skills assessed (transurethral resection of the prostate, transurethral resection of bladder tumor, Ureteroscopy, laparoscopic skills, and instrument assembly). Operative confidence was increased immediately and at 3-months postcourse. Our "boot camp" course provides a realistic introduction and foundation to begin Urological practice. Being delivered at the beginning of the training scheme, prior to intensive patient exposure, registrars are in an optimum position to develop their newly acquired knowledge and skills to enhance training and intends to improve patient safety and satisfaction.


The Reversal of the Gender Gap in Education and Trends in Marital Dissolution.
The reversal of the gender gap in education has potentially far-reaching consequences for marriage markets, family formation, and relationship outcomes. One possible consequence of this is the growing number of marriages in which wives have more education than their husbands. Previous studies have found this type of union to be at higher risk of dissolution. Using data on marriages formed between 1950 and 2004 in the United States, we evaluate whether this association has persisted as the prevalence of this relationship type has increased. Our results show a large shift in the association between spouses' relative education and marital dissolution. In particular, we confirm that marriages in which wives have the educational advantage were once more likely to dissolve, but we show that this association has disappeared in more recent marriage cohorts. Another key finding is that the relative stability of marriages between educational equals has increased. These results are consistent with a shift away from rigid gender specialization toward more flexible, egalitarian partnerships and provide an important counterpoint to claims that progress toward gender equality in heterosexual relationships has stalled.


Added Benefit of Stent Retriever Technology for Acute Ischemic Stroke: A Pooled Analysis of the NINDS tPA, SWIFT, and STAR Trials.
Endovascular treatment is increasingly being used in acute stroke care. However, although stent retrievers show improved flow restoration rates, their clinical benefits have been uncertain. To assess the incremental effect of using stent retrievers compared with intravenous tissue plasminogen activator (IV tPA; alteplase) alone or placebo/control. We conducted a pooled analysis of 4 studies using stent retrievers (Solitaire), IV tPA, or placebo/control. We applied the ischemic stroke risk score (www.sorcan.ca/iscore) to each participant to adjust for differences in baseline characteristics. We used a shift analysis to account for the potential benefits across the entire modified Rankin scale score at 90 days, adjusting for time-to-treatment, baseline Alberta Stroke Program Early CT score, and ischemic stroke risk score. Of the 915 participants in this analysis, 312 (34.1%) patients received placebo, 312 (34.1%) received tPA alone, 131 (14.4%) received stent retrievers alone, and 160 (17.5) received combined therapy (IV tPA plus stent retrievers). The shift analysis revealed that more patients remained independent at 90 days if receiving stent retrievers alone (number needed to treat 3.5) or combined with tPA (number needed to treat 3.1) compared with tPA alone. After adjustment, participants receiving stent retrievers alone (odds ratio, 2.95; 95% confidence interval, 1.48-5.89) or combined with tPA (odds ratio, 4.45; 95% confidence interval, 2.40-8.27) were more likely to be independent at 90 days compared with tPA alone. Patients with acute ischemic stroke who received IV tPA or revascularization therapies had a higher likelihood of achieving independence at 3 months. Stent retriever technology combined with tPA was associated with the greatest benefit compared with placebo, tPA alone, or endovascular therapy alone.


Evaluation of selective prostaglandin E2 (PGE2) receptor agonists as therapeutic agents for the treatment of asthma.
Prostaglandin E2 (PGE2) released in asthmatic airways has bronchodilator properties and inhibits allergen-induced bronchoconstriction and release of inflammatory mediators. Although considered as a potential treatment for asthma, PGE2 also has some proinflammatory properties. PGE2 acts through four different receptor subtypes (EP1, EP2, EP3, and EP4) that may explain some of PGE2's diverse effects. In a mouse model of allergic inflammation in which the four receptors were individually deleted, only EP3(-/-) mice showed an enhancement of inflammation, whereas an EP3 agonist was inhibitory, with PGE2 being inactive. Thus, EP3 agonists may lead to a new approach for the treatment of asthma. However, other PGE2 receptor subtypes may also have beneficial effects, and a greater understanding of the signaling pathways of these receptor subtypes will help to clarify the role of these receptors in asthma.


People and guns involved in denied and completed handgun sales.
Denial of handgun purchases by prohibited people and knowledge of the structure of gun commerce have helped to deter and prevent firearm violence. The authors hypothesize that handguns involved in a denied purchase would more closely resemble those used in crime compared with handguns sold. Cross sectional. Denied and completed handgun sales in California, 1998-2000. Handgun and purchaser characteristics of denied and completed sales were compared. In particular, handgun characteristics most closely associated with crime guns (type, caliber, barrel length, price) were examined. Compared with handguns sold, handguns in denied sales were somewhat more likely to be semiautomatic pistols (74.6% v 69.4%), to have short barrels (25.9% v 22.2%), and be of medium caliber (48.9% v 37.3%). Ten percent of the handguns in denied sales and 3.4% of handguns sold were identified as inexpensive. The characteristics of denied handguns are similar to those seen among crime guns. Both groups of guns may reflect the desirability for criminal purposes of pistols, which have larger ammunition capacities than other handguns, and short barrels, which increase their ability to be concealed.


Fracture load of milled polymeric fixed dental prostheses as a function of connector cross-sectional areas.
Polymeric computer-aided design/computer-aided manufacturing (CAD/CAM) blocks exhibit various advantages in contrast to conventionally processed resin restorations. However, the influence of connector dimensions on resin fixed dental prostheses (FDPs) has not yet been investigated. This study evaluated the impact of connector cross-sectional area (CSA) on the fracture load of 3-unit CAD/CAM FDPs and compared this with conventionally fabricated ones. Anatomically shaped 3-unit FDPs with the CSAs of 6, 9, 12, and 16 mm(2) (N=240, n=15 per material and per CSA) were fabricated from the following CAD/CAM materials: artBloc Temp (AT), TelioCAD (TC), CAD-Temp (CT), and one conventional resin material, CronMix K (CMK) as the control group. The fracture load was tested and the data were analyzed with 2-way ANOVA, 1-way ANOVA, the Scheffé post hoc test, and Weibull statistics (α=05). CMK showed the significant lowest values for all CSAs followed by CT, except for the 12 mm(2) connector (P<.001). The CAD/CAM FDPs exhibited a significant increase in fracture load with the increase of CSA (P<.001). Conventionally fabricated FDPs presented an increase of values up to the CSA of 12 mm(2). For TC, the shape increased with a larger CSA. The other materials exhibited no tendencies in this regard. Among the 12 mm(2) groups, AT exhibited the highest shape value (19.1), and among the 16 mm(2) groups, TC showed the highest shape value (17.0). The CMK FDPs with a CSA of 16 mm(2) showed almost half as high a shape parameter (6.4) than the other three CSAs. CAD/CAM resin FDPs revealed significantly higher fracture load values than conventionally fabricated FDPs and showed a significant increase in fracture load with the increase of the CSA.


Statin myalgia is not associated with reduced muscle strength, mass or protein turnover in older male volunteers, but is allied with a slowing of time to peak power output, insulin resistance and differential muscle mRNA expression.
Statins are associated with muscle myalgia and myopathy, which probably reduce habitual physical activity. This is particularly relevant to older people who are less active, sarcopaenic and at increased risk of statin myalgia. We hypothesised that statin myalgia would be allied to impaired strength and work capacity in older people, and determined whether differences aligned with divergences in lean mass, protein turnover, insulin sensitivity and the molecular regulation of these processes. Knee extensor strength and work output during 30 maximal isokinetic contractions were assessed in healthy male volunteers, nine with no statin use (control 70.4 ± 0.7 years) and nine with statin myalgia (71.5 ± 0.9 years). Whole body and leg glucose disposal, muscle myofibrillar protein synthesis (MPS) and leg protein breakdown (LPB) were measured during fasting (≈5 mU l(-1) insulin) and fed (≈40 mU l(-1) insulin + hyperaminoacidaemia) euglyceamic clamps. Muscle biopsies were taken before and after each clamp. Lean mass, MPS, LPB and strength were not different but work output during the initial three isokinetic contractions was 19% lower (P < 0.05) in statin myalgic subjects due to a delay in time to reach peak power output. Statin myalgic subjects had reduced whole body (P = 0.05) and leg (P < 0.01) glucose disposal, greater abdominal adiposity (P < 0.05) and differential expression of 33 muscle mRNAs (5% false discovery rate (FDR)), six of which, linked to mitochondrial dysfunction and apoptosis, increased at 1% FDR. Statin myalgia was associated with impaired muscle function, increased abdominal adiposity, whole body and leg insulin resistance, and evidence of mitochondrial dysfunction and apoptosis.


The primate neocortex in comparative perspective using magnetic resonance imaging.
In this study we use neuroanatomic data from living anthropoid primate subjects to test the following three hypotheses: (1) that the human neocortex is significantly larger than expected for a primate of our brain size, (2) that the human prefrontal cortex is significantly more convoluted than expected for our brain size, and (3) that increases in cerebral white matter volume outpace increases in neocortical gray matter volume among anthropoid primates. Whole brain MRI scans were obtained from 44 living primate subjects from 11 different species. Image analysis software was used to calculate total brain volume, neocortical gray matter volume, cerebral white matter volume, and the cross sectional area of the spinal cord in each scan. Allometric regression analyses were used to compare the relative size of these brain structures across species, with an emphasis on determining whether human brain proportions correspond with predictions based on nonhuman primate allometric trajectories. All three hypotheses were supported by our analysis. The results of this study provide additional insights into human brain evolution beyond the important observation that brain volume approximately tripled in the hominid lineage by demonstrating that the neocortex was uniquely modified throughout hominid evolution. These modifications may constitute part of the neurobiological substrate that supports some of our species most distinctive cognitive abilities.


Isolation of Acetylated and Unmodified Protein N-Terminal Peptides by Strong Cation Exchange Chromatographic Separation of TrypN-Digested Peptides.
We developed a simple and rapid method to enrich protein N-terminal peptides, in which the protease TrypN is first employed to generate protein N-terminal peptides without Lys or Arg and internal peptides with two positive charges at their N termini, and then, the N-terminal peptides with or without N-acetylation are separated from the internal peptides by strong cation exchange chromatography according to a retention model based on the charge/orientation of peptides. This approach was applied to 20 μg of human HEK293T cell lysate proteins to profile the N-terminal proteome. On average, 1550 acetylated and 200 unmodified protein N-terminal peptides were successfully identified in a single LC/MS/MS run with less than 3% contamination with internal peptides, even when we accepted only canonical protein N termini registered in the Swiss-Prot database. Because this method involves only two steps, protein digestion and chromatographic separation, without the need for tedious chemical reactions, it should be useful for comprehensive profiling of protein N termini, including proteoforms with neo-N termini.


Transcriptional activation by the Kaposi's sarcoma-associated herpesvirus latency-associated nuclear antigen is facilitated by an N-terminal chromatin-binding motif.
In immunocompromised patients, infection with Kaposi's sarcoma-associated herpesvirus (KSHV) can give rise to Kaposi's sarcoma and several lymphoproliferative disorders. In these tumors, KSHV establishes a latent infection in many of the rapidly proliferating and morphologically abnormal cells. Only a few viral gene products are expressed by the latent virus, and one of the best characterized is the latency-associated nuclear antigen (LANA), a nuclear protein required for the maintenance of viral episomal DNA in the dividing host cell. LANA can also activate or repress an assortment of cellular and viral promoters and may contribute to pathogenesis by allowing the proliferation and survival of host cells. Here we show that activation of the human E2F1 and cyclin-dependent kinase-2 (CDK2) promoters requires elements from both the N- and C-terminal regions of LANA. Deletion of the first 22 amino acids, which are necessary for episome tethering, does not affect nuclear localization but significantly reduces transactivation. Within the deleted peptide, we have identified a short sequence, termed the chromatin-binding motif (CBM), that binds tightly to interphase and mitotic chromatin. A second chromatin-binding activity resides in the C terminus but is not sufficient for optimal transactivation. Alanine substitutions within the CBM reveal a close correlation between the transactivation and chromatin binding activities, implying a mechanistic link. In contrast to promoter activation, we find that the 223 amino acids of the LANA C terminus are sufficient to inhibit p53-mediated activation of the human BAX promoter, indicating that the CBM is not required for all transcription-related functions.


Telomerase inhibitor Imetelstat (GRN163L) limits the lifespan of human pancreatic cancer cells.
Telomerase is required for the unlimited lifespan of cancer cells. The vast majority of pancreatic adenocarcinomas overexpress telomerase activity and blocking telomerase could limit their lifespan. GRN163L (Imetelstat) is a lipid-conjugated N3'→P5' thio-phosphoramidate oligonucleotide that blocks the template region of telomerase. The aim of this study was to define the effects of long-term GRN163L exposure on the maintenance of telomeres and lifespan of pancreatic cancer cells. Telomere size, telomerase activity, and telomerase inhibition response to GRN163L were measured in a panel of 10 pancreatic cancer cell lines. The cell lines exhibited large differences in levels of telomerase activity (46-fold variation), but most lines had very short telomeres (2-3 kb in size). GRN163L inhibited telomerase in all 10 pancreatic cancer cell lines, with IC50 ranging from 50 nM to 200 nM. Continuous GRN163L exposure of CAPAN1 (IC50 = 75 nM) and CD18 cells (IC50 = 204 nM) resulted in an initial rapid shortening of the telomeres followed by the maintenance of extremely short but stable telomeres. Continuous exposure to the drug eventually led to crisis and to a complete loss of viability after 47 (CAPAN1) and 69 (CD18) doublings. Crisis In these cells was accompanied by activation of a DNA damage response (γ-H2AX) and evidence of both senescence (SA-β-galactosidase activity) and apoptosis (sub-G1 DNA content, PARP cleavage). Removal of the drug after long-term GRN163L exposure led to a reactivation of telomerase and re-elongation of telomeres in the third week of cultivation without GRN163L. These findings show that the lifespan of pancreatic cancer cells can be limited by continuous telomerase inhibition. These results should facilitate the design of future clinical trials of GRN163L in patients with pancreatic cancer.


Inter-relationship between autoregulation of glomerular filtration rate, tubuloglomerular feedback and juxtaglomerular renin activity in normotensive and hypertensive rats.
1. Reduction of renal perfusion pressure from 133 mmHg to 117 mmHg in control rats did not induce a significant change of kidney glomerular filtration rate (GFR) or nephron GFR determined in distal tubules. In contrast, nephron GFR measured in proximal tubular segments (NGFR-P) fell significantly. 2. Qualitatively the same response of filtration rate to changes of arterial blood pressure was found in the chronically clipped kidneys of Goldblatt hypertensive rats after acute removal of the clip. 3. In contrast, autoregulation of kidney GFR, NGFR-D and NGFR-P was abolished in the contralateral kidneys of Goldblatt hypertensive rats. 4. Microperfusion studies showed that tubuloglomerular feedback regulation of NGFR was present in the renin-rich ischaemic kidneys of Goldblatt rats after removal of the constricting clip, but greatly attenuated in the renin-depleted contralateral kidneys. 5. These data indicate that tubuloglomerular feedback participates in establishing renal autoregulation, possibly by mediation of the renin-angiotensin system.


Five-year follow-up study of tardive dyskinesia.
A 5-year follow-up investigation of TD was carried out in 85 Hungarian schizophrenic out-patients. No substantial changes occurred during the 5 years in pharmacotherapy: most patients continued on neuroleptics and antiparkinson drugs. Comparison of 1978 and 1983 ratings revealed increased dyskinesia scores, which was statistically significant on the DRS Choreo-athetosis cluster. There was an increase in the number of patients with "moderately" severe TD from 4 in 1978 to 6 in 1981 to 10 (9 "Moderate" and 1 "Severe" TD) in 1983. The total number of TD cases as defined by research diagnostic criteria increased by only 3. The long-term trend in these neuroleptic-treated schizophrenics is for dyskinesia to plateau after reaching mild intensity, while in a minority of cases, the dyskinesia tends to extend from the oro-facial region to other body regions.


Adolescent caffeine consumption and aggressive behavior: A longitudinal assessment.
Background: Adolescent caffeine consumption has been linked to aggressive behaviors, although no longitudinal tests have been reported to date. The purpose of this study was to test the longitudinal relations between daily adolescent caffeine consumption and aggressive behaviors. Methods: Two waves of survey data collected 12 months apart in the spring of 2018 and 2019, from the 2004 birth cohort in the Icelandic LIFECOURSE study, were analyzed using structural equation modeling (N = 2,278). Both direct and mediated models were employed. Results: Caffeine use at time 1 (T1) was associated with aggressive behavior at time 2 (T2) (β = .12, p < .001) independent of aggressive behavior at T1. A considerable added relation was observed between caffeine at T1 and aggressive behavior at T2 via indirect (i.e., mediated) effects of aggressive behavior at T1 (standardized β = .20, p < .001). Over 64% of the standardized total effect (β = .31) observed between caffeine at T1 and aggressive behavior at T2 was due to mediation. Conclusion: Adolescent caffeine consumption forecasts aggressive behaviors. Caffeine use at T1 increased the likelihood of self-reported aggressive behaviors 12 months later irrespective of level of reported aggressive behavior at T1 while controlling for common confounders.


TNF alpha--a pivotal role in rheumatoid arthritis?
This review seeks to describe data which support the concept that TNF alpha has a pivotal role in the pathogenesis of RA. There is now compelling evidence from in vitro studies and in animal models that this is the case. The in vitro studies suggest that the direct pathogenic effects of TNF alpha are amplified by its potential to act as a potent paracrine molecule, by inducing other pro-inflammatory molecules such as IL-1, and GM-CSF. It has yet to be established whether similar paracine effects of TNF-alpha are observed in the collagen type II mouse, or in human TNF alpha transgenic mice, where (as discussed in this review), TNF alpha undoubtedly has a pathogenic involvement. During the ongoing inflammation in RA, TNF alpha (and other mediators?) induce the production of soluble TNF-R which can act as natural inhibitors. This process is, however, inadequate within the inflamed synovium, due to the fact that cytokine interactions are very local, through autocrine and paracrine mechanisms. In this environment there is a continuous competition between the interactions of surface or soluble receptor with TNF alpha. It may well be that soluble TNF-Rs play a more important role in the periphery where they act as an essential clearance mechanism for the ligand. If, as the available data suggest, TNF alpha does indeed have a pivotal role in the pathogenesis of RA, this needs to be verified by demonstrating that removal of TNF alpha locally in the synovial joint of RA patients has a significant effect on clinical parameters of disease, and the progression of the disease process itself.