Magnetocardiographic differences of ventricular repolarization parameters between Wistar rats and guinea pigs.
The morphology and range of duration of the action potential (AP) in normal Wistar rat's (WR) myocyte markedly differ from those of guinea pigs (GP), whose plateau (phase 2) duration is longer. Thus a clear-cut T wave can be easy defined in GP but not in WR. Aim of this study was to differentiate magnetocardiographic (MCG) ventricular repolarization (VR) parameters of healthy adult WR and GP. 10 female animals (5 Guinea pigs and 5 Wistar rats) were studied with a 36-channel MCG system (sensitivity of 20 fT/square root of Hz1/2) and with one ECG lead, in an unshielded hospital room. To assess VR, HR-corrected, JTpeak, JTend, Tpeak-end and QTend, intervals were measured from both MCG waveforms. Timing was improved by MCG maps analysis. Magnetic field orientation (MFO), its dynamics (MFD) and stability (JTS) during the JT interval, were also automatically computed from MCG maps. All repolarization intervals were significantly shorter in WR than in GP, except the Tpeak-end, which was longer. MFO and MFD also differed. MCG estimate of VR parameters, in adult WR and GP, is precise enough to evidence breed-related differences, consistent with physiological heterogeneity of duration during phases 2 and 3 of the AP, and with an higher degree of transmural dispersion of repolarization in WR.


In vivo and in vitro aging is detrimental to mouse spermatogonial stem cell function.
The development of techniques to maintain the spermatogonial stem cell (SSC) in vivo and in vitro for extended periods essentially allows for the indefinite continuation of an individual germline. Recent evidence indicates that the aging of male reproductive function is due to failure of the SSC niche. SSCs are routinely cultured for 6 mo, and no apparent effect of culture over this period has been observed. To determine the effects of SSC aging, we utilized an in vitro culture system, followed by quantitative transplantation experiments. After culture for 6 mo, SSCs that had been aged in vivo for 1500 days had a slower proliferation rate than SSCs that were aged in vivo to 8 or 300 days. Examination of methylation patterns revealed no apparent difference in DNA methylation between SSCs that were aged 8, 300, or 1500 days before culture. Long-term culture periods resulted in a loss of stem cell potential without an obvious change in the visual appearance of the culture. DNA microarray analysis of in vivo- and in vitro-aged SSCs identified the differential expression of several genes important for SSC function, including B-cell CLL/lymphoma 6, member B (Bcl6b), Lim homeobox protein 1 (Lhx1), and thymus cell antigen 1, theta (Thy1). Collectively, these data indicate that, although both in vitro and in vivo aging are detrimental to SSC function, in vitro aging results in greater loss of function, potentially due to a decrease in core SSC self-renewal gene expression and an increase in germ cell differentiation gene expression.


Volunteer facilitators assist community practices with enhancing cancer control.
To explore the feasibility of recruiting, training, and placing in the field volunteers to assist community practices in enhancing the provision of preventive care. A case series design followed up a cohort of volunteers prospectively as they were recruited, trained, and assigned to practices. The New Hampshire Division of the American Cancer Society recruited and trained the volunteer facilitators. Assistance from the volunteers in implementing a preventive services office system served as the intervention for practices. Volunteers were trained and supported by professional staff and an implementation manual. Recruitment, training, and volunteer experiences in working with practices, as well as the costs of supporting the program, were assessed. Twenty-six volunteers were trained. Of the 15 assigned to practices, 11 had begun to assist their assigned practices to establish a preventive services office system. Extensive planning, patience, and support were required. Volunteers recruited and supported by an intermediary organization can provide assistance to practices in implementing a preventive services office system.


Diagnosis of Acute Heart Failure in the Emergency Department: An Evidence-Based Review.
Heart failure is a common presentation to the emergency department (ED), which can be confused with other clinical conditions. This review provides an evidence-based summary of the current ED evaluation of heart failure. Acute heart failure is the gradual or rapid decompensation of heart failure, resulting from either fluid overload or maldistribution. Typical symptoms can include dyspnea, orthopnea, or systemic edema. The physical examination may reveal pulmonary rales, an S3 heart sound, or extremity edema. However, physical examination findings are often not sensitive or specific. ED assessments may include electrocardiogram, complete blood count, basic metabolic profile, liver function tests, troponin, brain natriuretic peptide, and a chest radiograph. While often used, natriuretic peptides do not significantly change ED treatment, mortality, or readmission rates, although they may decrease hospital length of stay and total cost. Chest radiograph findings are not definitive, and several other conditions may mimic radiograph findings. A more reliable modality is point-of-care ultrasound, which can facilitate the diagnosis by assessing for B-lines, cardiac function, and inferior vena cava size. These modalities, combined with clinical assessment and gestalt, are recommended.


Immune mechanisms in atherosclerosis.
To summarize, it is possible that T cell activation in the plaque has four different effects: a direct inhibition of smooth muscle proliferation mediated by IFN-gamma, an indirect stimulation of smooth muscle proliferation via IFN-induced macrophage activation, an induction of responsiveness to PDGF by induction of PDGF receptor expression, and finally, an up-regulation of HDL receptors. The net effect of T cell activation during the vascular response to injury may, therefore, depend on the balance between these mechanisms in any given situation during lesion development. T cell activation may itself be regulated by apolipoprotein E-containing LDL, which thus could form a direct link between lipoprotein accumulation and immune activation. We have recently tried to assess the effect of T cell activation during the response to experimental arterial injury with the use of a drug model. Cyclosporin A is a drug that specifically inhibits T cell activation. Rats treated with cyclosporin A for a short period had significantly smaller intimal lesions than did controls after balloon injury. This could be due to an inhibition of T cell activation, resulting in an inhibition of monocyte-macrophage activation and thereby loss of an important stimulus for intimal cell proliferation. When interpreting these results, one must, however, bear in mind that cyclosporin A could exert as yet unknown nonimmune vascular effects. It is also worth stressing that cell proliferation in the human atherosclerotic plaque may not be as high as in experimental animal lesions. In fact, cell replication may be a very rare event in the average advanced atherosclerotic plaque. Cell proliferation may, however, be associated with an episodic growth of lesions, and growth factor-mediated responses could, therefore, be important for the eventual clinical outcome in the individual patient. In conclusion, cytokines produced during the immune response affect growth and differentiation of vascular cells and could modulate both the response to injury and the local lipid metabolism in an atherosclerotic plaque There is indirect support for paracrine secretion of several of these factors in the atherosclerotic plaque, and activated T lymphocytes and macrophages are abundant in the plaque. This points to the possibility that specific immune responses are associated with the development of atherosclerosis. It is unknown, however, to what extent such immune responses occur or which antigens may elicit these responses.


NESS002ie: a new fluorinated thiol endopeptidase inhibitor with antinociceptive activity in an animal model of persistent pain.
For the past few decades membrane zinc metallopeptidases have been identified as important therapeutic targets in the control of pain. In particular, neutral endopeptidase (NEP) has been shown to play critical roles in the metabolism of the endogenous peptides Met- and Leu-enkephalins. In this study, we have evaluated the activity of a new fluorinated peptidase inhibitor NESS002ie in both in vitro and in vivo assays. NESS002ie has been compared to the peptidomimetic compound thiorphan and the previously reported NEP selective thiol inhibitor C20. The metallopeptidases inhibitory activity of NESS002ie was tested in vitro using a highly, sensitive, continuous, fluorometric, enzyme assay. Also, the analgesic propriety of NESS002ie, thiorphan and C20 have been evaluated in vivo, by intraplantar, intravenous and intrathecal administration, through nociception assays based on formalin test in mice. Metallopeptidases assays have shown an inhibitory potency of NESS002ie in the nanomolar range for NEP and angiotensin-converting enzyme (ACE). The new fluorinated inhibitor showed higher analgesic activity and bioavailability compared to thiorphan and C20 when administered by both intravenous and intrathecal injections. More significantly, intrathecal injection of NESS002ie reduced both the first and the second phases of the formalin biphasic pain response. In addition, naltrindole and naloxone reversed the analgesic effect of NESS002ie with a diverse profile. This study shows an improvement in relief of inflammation and pain, in vivo, using NESS002ie compared to reference compounds thiorphan and C20. This significant effect could be due to the replacement of isobutyl chain of the thiol C20 with the trifluoromethyl group.


Molecular genetics of C1 inhibitor.
More than 100 different C1 inhibitor gene mutations have been described in hereditary angioedema (HAE) patients. Sixty-nine mutations have been reported in patients with the quantitative C1 inhibitor defect (type 1 HAE) in two recent large-scale studies. These changes were found distributed over all exons and exon/intron boundaries. The molecular defects can be divided as follows: Alu-repeat-mediated deletions or duplications (accounting for 21% of all cases), missense mutations (> 36%), frameshifts (14%), Stop codon mutations (10%), promoter variants (4%), splice site mutations (7-10%), deletions of a few amino acids (less than 3%). Several recent studies indicate that up to 25% of these changes are found in patients without a family history of angioedema and represent de novo mutations. Pathogenic amino acid substitutions were found distributed over the entire length of the coding sequence, except for the 100 amino-acid-long glycosylated amino-terminal extension, whose sequence tolerates extensive variation, as indicated by comparisons across species. Functional studies have been carried out only on a fraction of these amino acid substitutions and indicate that defects affecting intracellular transport are often at the basis of type 1 hereditary angioedema. An interesting promoter variant (a C to T transition at position -103) was found in an exceptional family with recessive transmission of the disease. Regulatory elements in the promoter region and in intron 1 were revealed by their sequence conservation in mouse and man and by functional studies. C1 inhibitor "minigene" constructs directing correct mRNA and protein synthesis in transgenic mice have provided valuable information on hormonal control and cell-type specificity of gene expression.


Assessment of left ventricular structure and function in rats subjected to pressure-overload hypertrophy in time.
Left ventricular hypertrophy (LVH) is an adaptive response to increased haemodynamic load and an independent risk factor for the development of heart failure. Although the pathophysiological features during the development of cardiac hypertrophy have been extensively studied, the time course of LVH is less clearly defined. To define the time-dependency of the LVH process in vivo and compare the data by necropsy. Using abdominal aortic banding (AAB) in male Wistar strain albino rats we assessed the changes of LV structure and function in short intervals of 5 days for a period of 45 days. We determined the changes by serial echocardiography and confirmed the results in a second echocardiographic experiment and by necropsy. In our model the magnitude of the pressure overload was sufficient to produce significant LVH within a 10-day time frame and further progression on the 15th day after AAB. Interestingly, on the 20th day after banding a short-lasting regression of LVH (and heart weight and LV wall thickness) was found. It was followed by an increase in the next 15 days (till the 35th day), after which LVH was roughly complete (as measured at the 45th day). Following the development of LVH over a relatively long period of time and providing the changes in short intervals, a short lasting regression during ongoing pressure overload was noted. Understanding and targeting the associated signalling underlying this regression may have considerable clinical consequences.


A Monte Carlo analysis of false inference in spatial conflict event studies.
Spatial event data is heavily used in contemporary research on political violence. Such data are oftentimes mapped onto grid-cells or administrative regions to draw inference about the determinants of conflict intensity. This setup can identify geographic determinants of violence, but is also prone to methodological issues. Problems resulting from spatial aggregation and dependence have been raised in methodological studies, but are rarely accounted for in applied research. As a consequence, we know little about the empirical relevance of these general problems and the trustworthiness of a popular research design. We address these questions by simulating conflict events based on spatial covariates from seven high-profile conflicts. We find that standard designs fail to deliver reliable inference even under ideal conditions at alarming rates. We also test a set of statistical remedies which strongly improve the results: Controlling for the geographic area of spatial units eliminates an important source of spurious correlation. In time-series analyses, the same result can be achieved with unit-level fixed effects. Under outcome diffusion, spatial lag models with area controls produce most reliable inference. When those are computationally intractable, geographically larger aggregations lead to similar improvements. Generally, all analyses should be performed at two separate levels of geographic aggregation. To facilitate future research into geographic methods, we release the Simple Conflict Event Generator (SCEG) developed for this analysis.


Assisted standing for Duchenne muscular dystrophy.
Duchenne muscular dystrophy (DMD) is the most common X-linked neuromuscular disorder. When boys with DMD reach the second decade of life, they lose their ability to walk and become wheelchair dependent. Standing devices and orthoses are considered to be an essential component in the therapy management of DMD. Clinical opinion and research from other neurological conditions highlight the proposed benefits of standing device use, however, its effect within this population is currently unknown. A review of the evidence for the use of standing devices and orthoses is necessary to inform all stakeholders, including people with DMD, clinicians, decision makers and funders, and to guide future research. To assess the effects of standing devices and orthoses on musculoskeletal impairments (such as pain, contracture, scoliosis development and bone density) in boys and men with DMD, and secondarily to determine their effect on quality of life, participation in activities, and patient experience (satisfaction). We also considered any adverse events associated with their use. We searched the Cochrane Neuromuscular Specialised Register, CENTRAL, MEDLINE, Embase, AMED, PsycINFO, CINAHL Plus, PEDro, and ProQuest Dissertations & Theses Global up to 5 September 2019. We checked references in identified trials, handsearched journal abstracts, and searched trials registries. We planned to include randomised controlled trials (RCTs) and quasi-RCTs of any model of standing device for use in DMD. The control interventions would have been any other comparison group, including no standing device, a different model of standing device, usual care, or an alternative form of assistive weight bearing. We used standard Cochrane methodological procedures. Although we identified 13 potentially relevant studies, none met the inclusion criteria for this review. Since there were no RCTs or quasi-RCTs available to evaluate the effectiveness of standing devices in people with DMD, studies are needed to investigate the effectiveness of standing devices in this population.


Review of Angiotensin-converting Enzyme Inhibitory Assay: Rapid Method in Drug Discovery of Herbal Plants.
The renin-angiotensin-aldosterone system is a signaling pathway which responsible in the blood pressure regulation. Angiotensin-converting enzyme (ACE) is one of the key elements responsible for the hypertensive mechanism. It converts angiotensin-I to angiotensin-II. The discovery history of the ACE inhibitory activity assay method has been through a long stage for decades and development continues until today. The ACE inhibitory activity has become an effective screening method in the search for new antihypertensive agents from herbal plants. Some of in vitro assay methods were used to examine the activity of ACE inhibitors based on the substrate usage, such as; Cushman and Cheung Method using a substrate hippuryl-histidyl-leucine (HHL), Holmquist method using a substrate furanacryloyl-tripeptide, Elbl and Wagner method using a substrate benzoil-[l-14C] glicyl-L-histidine-L-leucine, Carmel and Yaron method using a substrate o-aminobenzoylglycyl-p-nitrophenylalanilproline, and Lam method using 3-hydroxybutyrylglycyl-glycyl-glycine as substrate. Several different methods to measure the results of enzymatic reactions or separating substrate with products, including spectrophotometric, fluorometric, high-performance liquid chromatography, electrophoresis, and radiochemistry. Application of the test method for screening the ACE inhibitors activity and investigation of active compounds from natural products can be done easily with this method, it is very helpful in research because the results obtained are simple, accurate, and rapid.


Trends in the Use and Characteristics of Chiropractic Services in the Department of Veterans Affairs.
The purpose of this study was to analyze national trends and key features of the Department of Veterans Affairs' (VA's) chiropractic service delivery and chiropractic provider workforce since their initial inception. This was a serial cross-sectional analysis of the VA administrative data sampled from the first record of chiropractic services in VA through September 30, 2015. Data were obtained from VA's Corporate Data Warehouse and analyzed with descriptive statistics. From October 1, 2004, through September 30, 2015, the annual number of patients seen in VA chiropractic clinics increased from 4052 to 37349 (821.7%), and the annual number of chiropractic visits increased from 20072 to 159366 (693.9%). The typical VA chiropractic patient is male, is between the ages of 45 and 64, is seen for low back and/or neck conditions, and receives chiropractic spinal manipulation and evaluation and management services. The total number of VA chiropractic clinics grew from 27 to 65 (9.4% annually), and the number of chiropractor employees grew from 13 to 86 (21.3% annually). The typical VA chiropractor employee is a 45.9-year-old man, has worked in VA for 4.5 years, and receives annual compensation of $97860. VA also purchased care from private sector chiropractors starting in 2000, growing to 159533 chiropractic visits for 19435 patients at a cost of $11155654 annually. Use of chiropractic services and the chiropractic workforce in VA have grown substantially over more than a decade since their introduction.


A new, simple, green, and one-pot four-component synthesis of bare and poly(α,γ, L-glutamic acid)-capped silver nanoparticles.
A simple and green chemical method has been developed to synthesize stable bare and capped silver nanoparticles based on the reduction of silver ions by glucose and capping by poly(α,γ,L-glutamic acid) (PGA). The use of ammonia during synthesis was avoided. PGA has had a dual role in the synthesis and was used as a capping agent to make the silver nanoparticle more biocompatible and to protect the nanoparticles from agglomerating in the liquid medium. The synthesized PGA-capped silver nanoparticles in the size range 5-45 nm were stable over long periods of time, without signs of precipitation. Morphological examination has shown that the silver nanoparticles had a nearly spherical, multiply twinned structure. The effects of the reaction temperature and the reaction time during the synthesis were investigated too. The biocompatibility of the PGA-capped silver nano-particles is discussed in terms of in vitro toxicity with human intestinal Caco-2 cells. The samples were characterized by UV-Visible spectroscopy, field emission scanning electron microscopy, transmission electron microscopy, and zeta potential measurements.


Netting Neutrophils Activate Autoreactive B Cells in Lupus.
Lupus erythematosus (LE) patients develop autoantibodies that form circulating immune complexes (ICs) with extracellular self-nucleic acids. These ICs are deposited into peripheral tissues, where they trigger detrimental organ inflammation. Recent evidence suggests that ICs contain LL37-DNA complexes derived from neutrophil extracellular traps (NETs) and that LE patients develop pathogenic autoantibodies against these structures, including Abs to LL37. However, the mechanism that leads to the generation of these Abs is unknown. In this study, we show that NETs directly trigger Ab production by human memory B cells. This occurs via LL37-DNA complexes present in NETs, which have the unique ability to gain access to endosomal compartments of B cells and to trigger TLR9 activation. In LE patients, NET-derived LL37-DNA complexes trigger polyclonal B cell activation via TLR9, but also specifically expand self-reactive memory B cells producing anti-LL37 Abs in an Ag-dependent manner. These findings suggest a unique link between neutrophils and B cells in which NETs trigger a concerted activation of TLR9 and BCR leading to anti-NET autoantibody production in lupus.


Isolation and immunologic characterization of a human. B-lymphocyte-specific, cell surface antigen.
In addition to HL-A antigens, another cell surface protein complex has been obtained from membranes of the human B-lymphoblast cell line IM-1. This complex which was solubilized with papain, consisted of polypeptides of 23,000 and 30,000 daltons (p23, 30). Rabbit antisera to this material precipitated from [35S]methionine-labeled detergent-solubilized cells, three proteins of 39,000, 34,000, and 29,000 daltons. These antisera were specifically cytotoxic for B lymphocytes of peripheral blood, for B-lymphoblast cell lines, and for EAC rosette receptor-positive surface Ig-negative (Null) lymphocytes. The p23,30 complex was not present on T lymphocytes, EAC rosette receptor-negative Null lymphocytes, or platelets. In addition, the p23,30 complex from several cell lines inhibited alloantisera from multiparous Amish women which had been shown to recognize non-HL-A, B-lymphocyte antigens. Some other properties of the anti-p23,30 sera antisera were described.


Central giant cell granulomas of the jaws: phenotype and proliferation-associated markers.
Central giant cell granulomas (CGCGs) are jaw tumors of unknown origin that often exhibit an aggressive, though unpredictable, clinical course. The purpose of this study was to determine the immunoprofile of the mononuclear cells that seem to be responsible for the biologic behavior of these tumors. Numbers of cells in cell cycle were also determined and compared in clinically aggressive and non-aggressive CGCGs. Sixteen aggressive and 12 non-aggressive CGCGs were immunohistochemically stained with antibodies to CD34, CD68, factor XIIIa, alpha-smooth muscle actin, prolyl 4-hydroxylase, Ki-67, and p53 protein. Cell populations and numbers of cells in cell cycle were determined through microscopic quantitative assessment. CD34-positive cells were limited to support vessels. CD68-positive mononuclear cells constituted a small population of cells in all tumors. With two exceptions, factor XIIIa-positive cells were rarely seen. Alpha-smooth muscle actin staining was present in approximately half the tumors, and occasionally large numbers of positive cells were seen. Most mononuclear cells were positive for fibroblast-associated antigen. No phenotypic differences were detected between aggressive and non-aggressive tumors. P53 protein did not appear to be overexpressed in CGCGs. Ki-67 staining showed that only mononuclear cells were in cell cycle, and that there were no differences between aggressive and non-aggressive tumors. We conclude that CGCGs are primarily fibroblastic (and myofibroblastic) tumors in which macrophages appear to play a secondary role. Tumor cells show no differentiation toward endothelial cells or macrophage-related dendrocytes (factor XIIIa). Cellular phenotypes and numbers of cells in cell cycle are similar in both aggressive and non-aggressive tumors.


TGFβ2 Induces the Formation of Cross-Linked Actin Networks (CLANs) in Human Trabecular Meshwork Cells Through the Smad and Non-Smad Dependent Pathways.
Increased intraocular pressure results from increased aqueous humor (AH) outflow resistance at the trabecular meshwork (TM) due to pathologic changes including the formation of cross-linked actin networks (CLANs). Transforming growth factor β2 (TGFβ2) is elevated in the AH and TM of primary open angle glaucoma (POAG) patients and induces POAG-associated TM changes, including CLANs. We determined the role of individual TGFβ2 signaling pathways in CLAN formation. Cultured nonglaucomatous human TM (NTM) cells were treated with control or TGFβ2, with or without the inhibitors of TGFβ receptor, Smad3, c-Jun N-terminal kinases (JNK), extracellular signal regulated kinase (ERK), P38, or Rho-associated protein kinase (ROCK). NTM cells were cotreated with TGFβ2 plus inhibitors for 10 days or pretreated with TGFβ2 for 10 days followed by 1-hour inhibitor treatment. NTM cells were immunostained with phalloidin-Alexa-488 and 4',6-diamidino-2-phenylindole (DAPI). Data were analyzed using 1-way ANOVA and Dunnett's post hoc test. TGFβ2 significantly induced CLAN formation (n = 6 to 12, P < 0.05), which was completely inhibited by TGFβ receptor, Smad3, and ERK inhibitors, as well as completely or partially inhibited by JNK, P38, and ROCK inhibitors, depending on cell strains. One-hour exposure to ROCK inhibitor completely resolved formed CLANs (P < 0.05), whereas TGFβ receptor, Smad3 inhibitor, and ERK inhibitors resulted in partial or complete resolution. The JNK and P38 inhibitors showed partial or no resolution. Among these inhibitors, the ROCK inhibitor was the most disruptive to the actin stress fibers, whereas ERK inhibition showed the least disruption. TGFβ2-induced CLANs in NTM cells were prevented and resolved using various pathway inhibitors. Apart from CLAN inhibition, some of these inhibitors also had different effects on actin stress fibers.


What is the correct staging and treatment strategy for locally advanced prostate cancer extending to the bladder?
In locally advanced prostate cancer with bladder invasion, frequently encountered problems such as bleeding, urinary retention, hydronephrosis, and pain create distress for the patients. Therefore patients' quality of life is disrupted and duration of hospitalization is prolonged. Relevant literature about accurate staging and treatment of locally advanced prostate cancer with bladder invasion was investigated. Locally advanced prostate cancer can present as a large-volume aggressive tumor extending beyond boundaries of prostate gland, and involving neighboring structures which can be involved as recurrence(s) following initial local therapy. Survival times of these patients can range between 5 and 8 years. Their common characteristics are adverse and severe local symptoms unfavorably affecting quality of life Control of local symptoms and their effective palliation are independent clinical targets influencing survival outcomes of these patients. The treatment outcomes of locally advanced prostate cancer into the bladder are currently debatable. Although in the current TNM classification, it is defined in T4a, we think that this may be categorized as a subgroup of T3 and thus encourage surgeons for the indication of radical surgeries (radical prostatectomy, radical cystoprostatectomy) in selected patient populations after discussing issues concerning consequences of the treatment alternatives, and expectations with the patients. Cystoprostatectomy followed by immediate androgen deprivation therapy may be a feasible option for selected patients with previously untreated prostate cancer involving the bladder neck because of excellent local control and long term survival.


Orientation and mobility assessment in retinal prosthetic clinical trials.
The purpose of this study was to develop an orientation and mobility (O&M) assessment protocol to implement in subjects who have been implanted with an artificial silicon retina (ASR), and to use this experience to propose a research agenda for O&M with prosthetic vision. A controlled and naturalistic assessment was developed that included walking a prescribed obstacle course and travel to and from a hospital cafeteria. Subjects were tested before and 3 and 6 months after being implanted with the ASR. Outcome measures were walking speed and number of contacts with obstacles. The experiences from this study led to a proposed research agenda in O&M. Eight subjects with retinitis pigmentosa participated in this study. The vision status of the subjects ranged from <20/1600 to 20/80 visual acuity and visual fields of <5 to 40°. Using a repeated-measures analysis of variance, no differences were found in the primary outcome measures. Four subjects were observed to have reduced mobility after implantation. Three subjects self-reported enhancements of travel after implantation, but this enhancement was not observed. This study demonstrates that mobility might not be improved with prosthetic vision. The proposed research agenda emphasizes the importance of developing individualized assessments, identifying specific items of orientation rather than mobility for measuring the effect of prosthetic vision, and to develop and evaluate instructional programs that may be needed to obtain the full benefit of the technology.


Distinct mechanisms determine transposon inheritance and methylation via small interfering RNA and histone modification.
Heritable, but reversible, changes in transposable element activity were first observed in maize by Barbara McClintock in the 1950s. More recently, transposon silencing has been associated with DNA methylation, histone H3 lysine-9 methylation (H3mK9), and RNA interference (RNAi). Using a genetic approach, we have investigated the role of these modifications in the epigenetic regulation and inheritance of six Arabidopsis transposons. Silencing of most of the transposons is relieved in DNA methyltransferase (met1), chromatin remodeling ATPase (ddm1), and histone modification (sil1) mutants. In contrast, only a small subset of the transposons require the H3mK9 methyltransferase KRYPTONITE, the RNAi gene ARGONAUTE1, and the CXG methyltransferase CHROMOMETHYLASE3. In crosses to wild-type plants, epigenetic inheritance of active transposons varied from mutant to mutant, indicating these genes differ in their ability to silence transposons. According to their pattern of transposon regulation, the mutants can be divided into two groups, which suggests that there are distinct, but interacting, complexes or pathways involved in transposon silencing. Furthermore, different transposons tend to be susceptible to different forms of epigenetic regulation.


Leading healthcare in complexity.
Healthcare institutions and providers are in complexity. Networks of interconnections from relationships and technology create conditions in which interdependencies and non-linear dynamics lead to surprising, unpredictable outcomes. Previous effective approaches to leadership, focusing on top-down bureaucratic methods, are no longer effective. Leading in complexity requires leaders to accept the complexity, create an adaptive space in which innovation and creativity can flourish and then integrate the successful practices that emerge into the formal organizational structure. Several methods for doing adaptive space work will be discussed. Readers will be able to contrast traditional leadership approaches with leading in complexity. They will learn new behaviours that are required of complexity leaders, along with challenges they will face, often from other leaders within the organization.


Establishing the tolerability and performance of tamarind seed polysaccharide (TSP) in treating dry eye syndrome: results of a clinical study.
One of the problems arising from available preparations for dry eye syndrome is the limited residence time of products on the ocular surface. In this paper, we look at an innovative new treatment for dry eye, tamarind seed polysaccharide (TSP). TSP possesses mucomimetic, mucoadhesive and pseudoplastic properties. The 'mucin-like' molecular structure of TSP is similar to corneal and conjunctival mucin 1 (MUC1), a transmembrane glycoprotein thought to play an essential role in protecting and wetting the corneal surface and may explain its increased retention on the eye surface. The activity of TSP and hyaluronic acid (HA) in the treatment of dry eye syndrome was compared in an open-label, randomised, single-centre clinical study. Thirty patients were randomised to receive three or more applications per day of either TSP 0.5%, TSP 1% or HA 0.2% (Hyalistiltrade mark) over a period of 90 days. The primary objective of tolerability was assessed by visual analogue scale (VAS), scoring of specific symptoms and the incidence of adverse events. Secondary objectives included improvement in stability of the precorneal tear film, subjective symptoms and corneal and conjunctival staining. TSP 0.5% and 1% were comparable to HA 0.2% with regard to both primary and secondary objective parameters.TSP 1% showed benefits over HA 0.2% for the subjective symptoms; trouble blinking, ocular burning and foreign body sensation. This study suggests that TSP 0.5% and 1% offer at least equivalent relief to HA 0.2% for dry eye syndrome. All treatments demonstrated optimal tolerability and are suitable for frequent use in the therapy of dry eye.TSP 1% produced promising results in terms of improvements in certain patient symptoms and suggests benefits of the TSP formulation. This study paves the way for a larger study to further establish the performance and safety of TSP compared with HA and highlights the need to expand this therapeutic agent to a wider dry eye population.


Nursing theorizing as an ethical endeavor.
This article addresses the ethical dimensions of nursing theorizing. Nursing theorizing, whether it occurs primarily at the outset or emerges during the process of inquiry, is inescapably linked to the theorist's value choices and beliefs about human beings, the environment, and health. These choices are reflected in the conceptual frame of one's research. The normative commitment of the conceptual frame is explored using examples from nursing and nonnursing research. Elements of critical ethical reflection are outlined. It is suggested that the discipline's understanding of what constitutes health and how best to promote health, as well as solutions to ethical dilemmas posed by research, may be enhanced by purposeful ethical inquiry that occurs as an integral component of theorizing activities.


Early dialogue between the developers of new technologies and pricing and reimbursement agencies: a pilot study.
It is common practice for developers of new health care technologies to engage in early dialogue with the major regulatory agencies; such discussions frequently center around the proposed clinical trial designs to support the registration of new interventions and suggestions on their improvement. Pricing and reimbursement agencies are increasingly using the results from health technology assessments to inform their decision making for new technologies. Such assessments are invariably underpinned by the phase 3 clinical trial evidence which may not provide answers to the key questions. Technology developers are beginning to realize that direct, early dialogue on the evidence requirements of the major pricing and reimbursement agencies, before phase 3 clinical trial designs for their key development compounds have been finalized, may be beneficial. This article reports on the pioneering efforts of one technology developer in seeking early dialogue with seven pricing and reimbursement agencies in five countries globally in 2007-2008 on their likely evidence requirements for a new oral treatment for patients with chronic plaque psoriasis. The pilot project demonstrated that a feasible process of early dialogue could be established, through a face-to-face meeting with prior circulation of a briefing book. Although there was some variation in the advice the similarities far outweighed the differences. More experience of early dialogue needs to be accumulated, involving a wider range of pricing and reimbursement agencies and compounds. The conclusion of this study, however, was that early dialogue can be a worthwhile process for all parties and can lead to a common understanding about evidence development for market access.


Neonatal morphometric similarity mapping for predicting brain age and characterizing neuroanatomic variation associated with preterm birth.
Multi-contrast MRI captures information about brain macro- and micro-structure which can be combined in an integrated model to obtain a detailed "fingerprint" of the anatomical properties of an individual's brain. Inter-regional similarities between features derived from structural and diffusion MRI, including regional volumes, diffusion tensor metrics, neurite orientation dispersion and density imaging measures, can be modelled as morphometric similarity networks (MSNs). Here, individual MSNs were derived from 105 neonates (59 preterm and 46 term) who were scanned between 38 and 45 weeks postmenstrual age (PMA). Inter-regional similarities were used as predictors in a regression model of age at the time of scanning and in a classification model to discriminate between preterm and term infant brains. When tested on unseen data, the regression model predicted PMA at scan with a mean absolute error of 0.70 ± 0.56 weeks, and the classification model achieved 92% accuracy. We conclude that MSNs predict chronological brain age accurately; and they provide a data-driven approach to identify networks that characterise typical maturation and those that contribute most to neuroanatomic variation associated with preterm birth.


Posterior short-segment fixation with or without fusion for thoracolumbar burst fractures. a five to seven-year prospective randomized study.
The impact of fusion as a supplement to short-segment instrumentation for the treatment of thoracolumbar burst fractures is unclear. We conducted a controlled clinical trial to define the effect of fusion on lumbar spine and patient-related functional outcomes. From 2000 to 2002, seventy-three consecutive patients with a single-level Denis type-B burst fracture involving the thoracolumbar spine and a load-sharing score of <or=6 were managed with posterior pedicle screw instrumentation. The patients were randomly assigned to treatment with posterolateral fusion (fusion group, n = 37) or without posterolateral fusion (nonfusion group, n = 36). The patients were followed for at least five years after surgery and were assessed with regard to clinical and radiographic outcomes. Clinical outcomes were evaluated with use of the Frankel scale, the motor score of the American Spinal Injury Association, a visual analog scale, and the Short Form-36 (SF-36) questionnaire. Radiographic outcomes were assessed on the basis of the local kyphosis angle and loss of kyphosis correction. No significant difference in radiographic or clinical outcomes was noted between the patients managed with the two techniques. Both operative time and blood loss were significantly less in the nonfusion group compared with the fusion group (p < 0.05). Twenty-five of the thirty-seven patients in the fusion group still had some degree of donor-site pain at the time of the latest examination. Posterolateral bone-grafting is not necessary when a Denis type-B thoracolumbar burst fracture associated with a load-sharing score of <or=6 is treated with short-segment pedicle screw fixation.


A TLR7 antagonist restricts interferon-dependent and -independent immunopathology in a mouse model of severe influenza.
Cytokine-mediated immune-cell recruitment and inflammation contribute to protection in respiratory virus infection. However, uncontrolled inflammation and the "cytokine storm" are hallmarks of immunopathology in severe infection. Cytokine storm is a broad term for a phenomenon with diverse characteristics and drivers, depending on host genetics, age, and other factors. Taking advantage of the differential use of virus-sensing systems by different cell types, we test the hypothesis that specifically blocking TLR7-dependent, immune cell-produced cytokines reduces influenza-related immunopathology. In a mouse model of severe influenza characterized by a type I interferon (IFN-I)-driven cytokine storm, TLR7 antagonist treatment leaves epithelial antiviral responses unaltered but acts through pDCs and monocytes to reduce IFN-I and other cytokines in the lung, thus ameliorating inflammation and severity. Moreover, even in the absence of IFN-I signaling, TLR7 antagonism reduces inflammation and mortality driven by monocyte-produced chemoattractants and neutrophil recruitment into the infected lung. Hence, TLR7 antagonism reduces diverse types of cytokine storm in severe influenza.


Speech intelligibility enhancement after maxillary denture treatment and its impact on quality of life.
Tooth loss and its prosthetic rehabilitation significantly affect speech intelligibility. However, little is known about the influence of speech deficiencies on oral health-related quality of life (OHRQoL). The aim of this study was to investigate whether speech intelligibility enhancement through prosthetic rehabilitation significantly influences OHRQoL in patients wearing complete maxillary dentures. Speech intelligibility by means of an automatic speech recognition system (ASR) was prospectively evaluated and compared with subjectively assessed Oral Health Impact Profile (OHIP) scores. Speech was recorded in 28 edentulous patients 1 week prior to the fabrication of new complete maxillary dentures and 6 months thereafter. Speech intelligibility was computed based on the word accuracy (WA) by means of an ASR and compared with a matched control group. One week before and 6 months after rehabilitation, patients assessed themselves for OHRQoL. Speech intelligibility improved significantly after 6 months. Subjects reported a significantly higher OHRQoL after maxillary rehabilitation with complete dentures. No significant correlation was found between the OHIP sum score or its subscales to the WA. Speech intelligibility enhancement achieved through the fabrication of new complete maxillary dentures might not be in the forefront of the patients' perception of their quality of life. For the improvement of OHRQoL in patients wearing complete maxillary dentures, food intake and mastication as well as freedom from pain play a more prominent role.


Dopaminergic medication boosts action-effect binding in Parkinson's disease.
Parkinson's disease (PD) is a neurodegenerative disorder affecting voluntary motor control. However, little is known about the experience of voluntary action in PD patients. A key component of action experience is the feeling of controlling one's own actions, and through them, external events. In healthy individuals this sense of agency (SoA) is associated with a subjective compression of time, such that actions and their effects are perceived as bound together across time. This action-effect binding provides an indirect measure of SoA. Nine PD patients and age-matched controls judged the time of voluntary actions and of an auditory effect (a tone) of the action. The pattern of results resembled previous studies, with the perceived time of actions showing a shift towards the subsequent tone, relative to a baseline condition involving actions without tones. Similarly, the perceived times of tones showed a shift towards the preceding action that caused the tone, relative to a baseline condition involving tones only. The patients were tested both on and off dopaminergic medication. PD patients off medication showed no significant change in action-effect binding relative to controls. Conversely, PD patients on medication showed a significant increase in action-effect binding relative to their own performance off medication. Increased availability of dopamine strengthened the experience of association between actions and external events, enhancing the sense of agency. These results shed light on the contribution of dopamine to the experience of instrumental action, and also on impulse control disorders and psychosis in medicated PD patients.


Identification of Spen as a Crucial Factor for Xist Function through Forward Genetic Screening in Haploid Embryonic Stem Cells.
In mammals, the noncoding Xist RNA triggers transcriptional silencing of one of the two X chromosomes in female cells. Here, we report a genetic screen for silencing factors in X chromosome inactivation using haploid mouse embryonic stem cells (ESCs) that carry an engineered selectable reporter system. This system was able to identify several candidate factors that are genetically required for chromosomal repression by Xist. Among the list of candidates, we identify the RNA-binding protein Spen, the homolog of split ends. Independent validation through gene deletion in ESCs confirms that Spen is required for gene repression by Xist. However, Spen is not required for Xist RNA localization and the recruitment of chromatin modifications, including Polycomb protein Ezh2. The identification of Spen opens avenues for further investigation into the gene-silencing pathway of Xist and shows the usefulness of haploid ESCs for genetic screening of epigenetic pathways.


Altered macrophage function contributes to colitis in mice defective in the phosphoinositide-3 kinase subunit p110δ.
Innate immune responses are crucial for host defense against pathogens but need to be tightly regulated to prevent chronic inflammation. Initial characterization of mice with a targeted inactivating mutation in the p110δ subunit of phosphoinositide 3-kinase (PI3K p110δ(D910A/D910A)) revealed defects in B- and T-cell signaling and chronic colitis. Here, we further characterize features of inflammatory bowel diseases in these mice and investigate underlying innate immune defects. Colons and macrophages from PI3K p110δ(D910A/D910A) mice were evaluated for colonic inflammation and innate immune dysfunction. Colonic p110δ messenger RNA expression was examined in interleukin (IL)-10(-/-) and wild-type germ-free mice during transition to a conventional microbiota. To assess polygenic impact on development of colitis, p110δ(D910A/D910A) mice were backcrossed to IL-10(-/-) mice. A mild spontaneous colitis was shown in PI3K p110δ(D910A/D910A) mice at 8 weeks, with inflammation increasing with age. An inflammatory mucosal and systemic cytokine profile was characterized by expression of IL-12/23. In PI3K p110δ(D910A/D910A) macrophages, augmented toll-like receptor signaling and defective bactericidal activity were observed. Consistent with an important homeostatic role for PI3K p110δ, wild-type mice raised in a germ-free environment markedly up-regulated colonic PI3K p110δ expression with the introduction of the enteric microbiota; however, colitis-prone IL-10(-/-) mice did not. Moreover, PI3K p110δ(D910A/D910A) mice crossed to IL-10(-/-) mice developed severe colitis at an early age. This study describes a novel model of experimental colitis that highlights the importance of PI3K p110δ in maintaining mucosal homeostasis and could provide insight into the pathogenesis of human inflammatory bowel disease.


Diagnostic schemes for reducing epidemic size of African viral hemorrhagic fever outbreaks.
Viral hemorrhagic fever (VHF) outbreaks, with high mortality rates, have often been amplified in African health institutions due to person-to-person transmission via infected body fluids. By collating and analyzing epidemiological data from documented outbreaks, we observed that diagnostic delay contributes to epidemic size for Ebola and Marburg hemorrhagic fever outbreaks. We used a susceptible-exposed-infectious-removed (SEIR) model and data from the 1995 outbreak in Kikwit, Democratic Republic of Congo, to simulate Ebola hemorrhagic fever epidemics. Our model allows us to describe the dynamics for hospital staff separately from that for the general population, and to implement health worker-specific interventions. The model illustrates that implementing World Health Organization/US Centers for Disease Control and Prevention guidelines of isolating patients who do not respond to antimalarial and antibacterial chemotherapy reduces total outbreak size, from a median of 236, by 90% or more. Routinely employing diagnostic testing in post-mortems of patients that died of refractory fevers reduces the median outbreak size by a further 60%. Even greater reductions in outbreak size were seen when all febrile patients were tested for endemic infections or when febrile health-care workers were tested. The effect of testing strategies was not impaired by the 1-3 day delay that would occur if testing were performed by a reference laboratory. In addition to improving the quality of care for common causes of febrile infections, increased and strategic use of laboratory diagnostics for fever could reduce the chance of hospital amplification of VHFs in resource-limited African health systems.


Affective-associative two-process theory: a neurocomputational account of partial reinforcement extinction effects.
The partial reinforcement extinction effect (PREE) is an experimentally established phenomenon: behavioural response to a given stimulus is more persistent when previously inconsistently rewarded than when consistently rewarded. This phenomenon is, however, controversial in animal/human learning theory. Contradictory findings exist regarding when the PREE occurs. One body of research has found a within-subjects PREE, while another has found a within-subjects reversed PREE (RPREE). These opposing findings constitute what is considered the most important problem of PREE for theoreticians to explain. Here, we provide a neurocomputational account of the PREE, which helps to reconcile these seemingly contradictory findings of within-subjects experimental conditions. The performance of our model demonstrates how omission expectancy, learned according to low probability reward, comes to control response choice following discontinuation of reward presentation (extinction). We find that a PREE will occur when multiple responses become controlled by omission expectation in extinction, but not when only one omission-mediated response is available. Our model exploits the affective states of reward acquisition and reward omission expectancy in order to differentially classify stimuli and differentially mediate response choice. We demonstrate that stimulus-response (retrospective) and stimulus-expectation-response (prospective) routes are required to provide a necessary and sufficient explanation of the PREE versus RPREE data and that Omission representation is key for explaining the nonlinear nature of extinction data.


Garenoxacin treatment of experimental endocarditis caused by viridans group streptococci.
The activity of garenoxacin was compared to that of levofloxacin or penicillin in a rabbit model of Streptococcus mitis group (penicillin MIC, 0.125 microg/ml) and Streptococcus sanguinis group (penicillin MIC, 0.25 microg/ml) endocarditis. Garenoxacin and levofloxacin had MICs of 0.125 and 0.5 microg/ml, respectively, for both study isolates. Rabbits with catheter-induced aortic valve endocarditis were given no treatment, penicillin at 1.2x10(6) IU/8 h intramuscularly, garenoxacin at 20 mg/kg of body weight/12 h intravenously, or levofloxacin at 40 mg/kg/12 h intravenously. For both isolates tested, garenoxacin area under the curve (AUC)/MIC and maximum concentration of drug in serum (Cmax)/MIC ratios were 368 and 91, respectively. Rabbits were sacrificed after 3 days of treatment; cardiac valve vegetations were aseptically removed and quantitatively cultured. For S. mitis group experimental endocarditis, all studied antimicrobial agents were more active than no treatment (P<0.001), whereas for S. sanguinis group endocarditis, no studied antimicrobial agents were more active than no treatment. We conclude that AUC/MIC and Cmax/MIC ratios may not predict activity of some quinolones in experimental viridans group endocarditis and that garenoxacin and levofloxacin may not be ideal choices for serious infections caused by some quinolone-susceptible viridans group streptococci.


Systematic review and meta-analysis of Transurethral Needle Ablation in symptomatic Benign Prostatic Hyperplasia.
Benign prostatic hyperplasia (BPH) constitutes a major clinical problem. Minimally invasive therapies for the treatment of symptomatic BPH include Transurethral Needle Ablation (TUNA), but it is unclear what impact this technique has on the disease and its role among other currently available therapeutic options. The objective of this study is to ascertain the efficacy and safety of TUNA in the treatment of BPH. Systematic review of the literature until January 2005 and meta-analysis of clinical studies assessing TUNA in symptomatic BPH. Studies were critically appraised. Estimates of effect were calculated according to the random-effects model. 35 studies (9 comparative, 26 non-comparative) were included. Although evidence was limited by methodological issues, the analysis of relevant outcomes indicates that while TUNA significantly improves BPH parameters with respect to baseline, it does not reach the same level of efficacy as TURP in respect to all subjective and objective variables. Further, its efficacy declines in the long-term with a rate of secondary-treatment significantly higher than of TURP [OR: 7.44 (2.47, 22.43)]. Conversely, TUNA seems to be a relatively safe technique and shows a lower rate of complications than TURP [OR:0.14 (0.05, 0.14)] with differences being particularly noteworthy in terms of postoperative bleeding and sexual disorders. Likewise, TUNA has fewer anesthetic requirements and generates a shorter hospital stay than TURP [WMD: -1.9 days (-2.75, -1.05)]. Scarce data and lack of replication of comparisons hinder the assessment of TUNA vs. other local therapies. No comparisons with medical treatment were found. The body of evidence on which TUNA has been introduced into clinical practice is of only moderate-low quality. Available evidence suggest that TUNA is a relatively effective and safe technique that may eventually prove to have a role in selected patients with symptomatic BPH. TUNA significantly improves BPH parameters with respect to baseline values, but it does not reach the same level of efficacy and long-lasting success as TURP. On the other hand, TUNA seems to be superior to TURP in terms of associated morbidity, anesthetic requirements and length of hospital stay. With respect to the role of TUNA vis-à-vis other minimally invasive therapies, the results of this review indicate that there are insufficient data to define this with any degree of accuracy. Overall cost-effectiveness and the role of TUNA versus medical treatment need further evaluation.


Complications in ambulatory pediatric patients with nonidiopathic spinal deformity undergoing fusion to the pelvis using the sacral-alar-iliac technique within 2 years of surgery.
Significant investigation in the adult population has generated a body of research regarding proximal junctional kyphosis (PJK) and proximal junctional failure (PJF) following long fusions to the sacrum and pelvis. However, much less is known regarding early complications, including PJK and PJF, in the ambulatory pediatric patient. As such, the objective of this study was to address the minimal literature on early complications after ambulatory pediatric patients underwent fusion to the sacrum with instrumentation to the pelvis in the era of sacral-alar-iliac (S2AI) instrumentation. The authors performed a retrospective review of pediatric patients with nonidiopathic spinal deformity < 18 years of age with ambulatory capacity who underwent fusion to the pelvis at a multisurgeon pediatric academic spine practice from 2016 to 2018. All surgeries were posterior-only approaches with S2AI screws as the primary technique for sacropelvic fixation. Descriptive, outcome, and radiographic data were obtained. The definition of PJF included symptomatic PJK presenting as fracture, screw pullout, or disruption of the posterior osseoligamentous complex. Twenty-five patients were included in this study. Nine patients (36.0%) had 15 complications for an overall complication rate of 60.0%. Unplanned return to the operating room occurred 8 times in 6 patients (24.0%). Four patients (16.0%) had wound issues (3 with deep wound infection and 1 with wound breakdown) requiring reoperation. Three patients (12.0%) had PJF, all requiring reoperation. A 16-year-old female patient with syndromic scoliosis underwent extension of fusion due to posterior tension band failure at 6 months. A 17-year-old male patient with neuromuscular scoliosis underwent extension of fusion due to proximal screw pullout at 5 months. A 10-year-old female patient with congenital scoliosis underwent extension for PJF at 5 months following posterior tension band failure. One patient had pseudarthrosis requiring reoperation 20 months postoperatively. Fixation to the pelvis enables significant deformity correction, but with rather high rates of complications and unexpected returns to the operating room. Considerations of sagittal plane dynamics for PJK and PJF should be strongly analyzed when performing fixation to the pelvis in ambulatory pediatric patients.


Image quality improvement of parallel four-step phase-shifting digital holography by using the algorithm of parallel two-step phase-shifting digital holography.
We propose an algorithm that can improve the quality of the reconstructed image from the single hologram recorded by the optical system of the parallel four-step phase-shifting digital holography. The proposed algorithm applies the image-reconstruction algorithm of parallel two-step phase-shifting digital holography to the hologram so as to reduce errors in the reconstructed image and eliminate ghosts. We numerically and experimentally confirmed that the proposed algorithm decreased 25% in terms of root mean square error in amplitude, and eliminated the ghosts, respectively.


Sustained delivery of salbutamol and beclometasone from spray-dried double emulsions.
The sustained delivery of multiple agents to the lung offers potential benefits to patients. This study explores the preparation of highly respirable dual-loaded spray-dried double emulsions. Spray-dried powders were produced from water-in-oil-in-water (w/o/w) double emulsions, containing salbutamol sulphate and/or beclometasone dipropionate in varying phases. The double emulsions contained the drug release modifier polylactide co-glycolide (PLGA 50 : 50) in the intermediate organic phase of the original micro-emulsion and low molecular weight chitosan (Mw<190 kDa: emulsion stabilizer) and leucine (aerosolization enhancer) in the tertiary aqueous phase. Following spray-drying resultant powders were physically characterized: with in vitro aerosolization performance and drug release investigated using a Multi-Stage Liquid Impinger and modified USP II dissolution apparatus, respectively. Powders generated were of a respirable size exhibiting emitted doses of over 95% and fine particle fractions of up to 60% of the total loaded dose. Sustained drug release profiles were observed during dissolution for powders containing agents in the primary aqueous and secondary organic phases of the original micro-emulsion; the burst release of agents was witnessed from the tertiary aqueous phase. The novel spray-dried emulsions from this study would be expected to deposit and display sustained release character in the lung.


Network topologies and dynamics leading to endotoxin tolerance and priming in innate immune cells.
The innate immune system, acting as the first line of host defense, senses and adapts to foreign challenges through complex intracellular and intercellular signaling networks. Endotoxin tolerance and priming elicited by macrophages are classic examples of the complex adaptation of innate immune cells. Upon repetitive exposures to different doses of bacterial endotoxin (lipopolysaccharide) or other stimulants, macrophages show either suppressed or augmented inflammatory responses compared to a single exposure to the stimulant. Endotoxin tolerance and priming are critically involved in both immune homeostasis and the pathogenesis of diverse inflammatory diseases. However, the underlying molecular mechanisms are not well understood. By means of a computational search through the parameter space of a coarse-grained three-node network with a two-stage Metropolis sampling approach, we enumerated all the network topologies that can generate priming or tolerance. We discovered three major mechanisms for priming (pathway synergy, suppressor deactivation, activator induction) and one for tolerance (inhibitor persistence). These results not only explain existing experimental observations, but also reveal intriguing test scenarios for future experimental studies to clarify mechanisms of endotoxin priming and tolerance.


White pine blister rust resistance in limber pine: evidence for a major gene.
Limber pine (Pinus flexilis) is being threatened by the lethal disease white pine blister rust caused by the non-native pathogen Cronartium ribicola. The types and frequencies of genetic resistance to the rust will likely determine the potential success of restoration or proactive measures. These first extensive inoculation trials using individual tree seed collections from >100 limber pine trees confirm that genetic segregation of a stem symptom-free trait to blister rust is consistent with inheritance by a single dominant resistance (R) gene, and the resistance allele appears to be distinct from the R allele in western white pine. Following previous conventions, we are naming the R gene for limber pine "Cr4." The frequency of the Cr4 allele across healthy and recently invaded populations in the Southern Rocky Mountains was unexpectedly high (5.0%, ranging from 0 to 13.9%). Cr4 is in equilibrium, suggesting that it is not a product of a recent mutation and may have other adaptive significance within the species, possibly related to other abiotic or biotic stress factors. The identification of Cr4 in native populations of limber pine early in the invasion progress in this region provides useful information for predicting near-term impacts and structuring long-term management strategies.


In vivo evaluation of periodontal microcirculatory changes associated with endodontic treatment.
The purpose of this study was to investigate in vivo the gingival microcirculatory changes associated with endodontic treatment using the continuous wave of condensation technique. Twenty necrotic one canal roots of 20 cooperative patients of both sexes, aged between 20 and 43 years, were selected. All patients were examined by capillaroscopy before, immediately after endodontic treatment, and after 7 days. The last examination was carried out by the same operator, and repeated twice for each examined area: masticatory, buccal and labial mucosa corresponding to the endodontically treated root. All canals were prepared using a simultaneous technique with Ni-Ti files (MTwo files). The images of the masticatory mucosa after root canal obturation showed evident micro-areas of extravasation, with significant bleeding and angio-morphological alterations due to heat. One hour after the endodontic treatment evident extravasation was observed, but a decrease of all altered parameters, was present. After seven days from treatment, in the periodontal tissues, a complete healing was observed. The in vivo evaluation of the vascular pattern during root canal obturation with System B showed that the high temperature in the canal determines visible effects on the vasculature of adjacent sites. It was found that microangiotectonic alterations decrease up to a complete healing after 7 days from treatment. All the changes in microcirculation, due to thermal shock of periodontal tissues, are reversible.


Body mass and prostatic cancer: a prospective study.
Previous studies have suggested that increased body mass is associated with an increased risk of prostatic cancer, but these studies have been limited by the fact that they were based on a few simple measurements such as height and weight. Similar results were found in a prospective study of the incidence of prostatic cancer in a cohort of Japanese men born in 1900-19 and living in Hawaii. Further evaluation of the extensive anthropomorphic measurements made in this cohort suggested that the association between measures of body mass and prostatic cancer might be accounted for more by lean tissue than by fat tissue. There was a significant positive association of the risk of prostatic cancer with area of muscle in the arm but not with area of fat in the arm. Further research is needed on the biological mechanisms of carcinogenesis that may be related to both lean and fat tissue and the development of prostatic cancer.


Predicting the infarct-related artery in STEMI from the surface ECG: independent validation of proposed criteria.
This study independently evaluated the diagnostic performance of electrocardiographic (ECG) criteria to predict the infarct-related artery (IRA) in patients with an acute ST-segment elevation myocardial infarction (STEMI). While a number of ECG criteria have been proposed to predict the IRA in STEMI, many of these "rules" came from modestly sized populations and did not undergo external validation. Therefore, we aimed to evaluate popular criteria from the literature in an independent cohort. All acute STEMI cases over a 10-year period from a single hospital were retrospectively identified. We excluded patients with a missing pre-intervention ECG, irretrievable angiographic films, prior coronary artery bypass grafting, left bundle branch block, ventricular pacing, or not meeting strict STEMI criteria. After review of the angiograms for the IRA, cases with either no or multiple culprits were excluded. We included 480 subjects meeting STEMI criteria in inferior leads (192, 40%), anterior leads (184, 38%), both anterior and inferior leads (88, 18%), isolated lateral leads (nine, 2%), or a posterior pattern (seven, 1%). Notably, every pattern except isolated lateral STEMI included an IRA in both the right and left coronary arteries. Existing ECG criteria to predict the IRA in STEMI have modest diagnostic performance when externally validated, and lower than in the original reports. Distinguishing the level of obstruction in the left anterior descending artery remains especially challenging. Hence, their use should be pragmatic when selecting an initial catheter for treating STEMI, since discordances will occur when compared to the actual angiogram.


Adding Value to Fruit Processing Waste: Innovative Ways to Incorporate Fibers from Berry Pomace in Baked and Extruded Cereal-based Foods-A SUSFOOD Project.
This article communicates the set-up of BERRYPOM, a European research project established in the second call of the SUStainable FOOD Production and Consumption (SUSFOOD) network. The project deals with the by-product from berry processing, which is frequently recycled as animal feed, composted or utilized for biogas production. With BERRYPOM it is proposed to analyze the value of berry pomace, to optimize the recovery of bioactive compounds from pomace material, and to incorporate processed berry pomace in cereal-based foods to take advantage of nutritional benefits that originate from its fiber and the content of bioactive substances. Additionally, extraction methods will be evaluated to obtain products rich in phytochemicals, and the influence of processing steps on the antioxidant capacity of pomace will be analyzed. The fiber extracts will then also be utilized in different cereal-based foods and extruded products. As project outcome we expect a substantial increase of knowledge concerning fiber and phytochemicals extraction from berry pomace, its suitability for enhancing nutritional and sensory properties of cereal-based foods, and its effects on the sustainability of the food chain.


[Reconstruction of mutism. Studies of subjective experiences of former patients with the modified Landfield categories].
The subjective experience of catatonic patients was investigated with the repertory grid-technique after recovery from the acute state of disease and compared with non-catatonic schizoaffective and depressive patients. The sample consists of 18 catatonic patients, 22 schizoaffective, 16 unipolar depressive, 8 unipolar manic and 23 patients with bipolar affective disorder as well as of 32 psychiatrically healthy orthopaedic controls. Interviews were held well before discharge. The contents of personal constructs was analyzed by means of modified Landfield categories. "Self" of catatonic patients did not differ significantly from the other groups. The "Idealself" of catatonic patients differed significantly in the categories "high self esteem" (from the controls and the non-catatonic group) and "high empathy" (from the controls). The "disease-self" ("In the acute catatonic state") was construed as isolated (lack of social contact). The results underline the importance of an active therapeutical interaction in the acute catatonic state. The modified Landfield Categories proved to be a useful psychodiagnostic instrument for the idiographic assessment of personality in particular, and for psychiatric therapy research in general.


Experiences of Older Adults During the 2020 COVID-19 Pandemic in the United States: An Initial Exploration of Nationally Representative Data at the Intersection of Gender and Race.
Little is known about the overall experiences and feelings of diverse older populations during the 2020 COVID-19 pandemic. To provide the baseline information for future research and policy, this study analyzed the 2020 Health and Retirement Study COVID-19 project data (n = 1782). More than 70% of older adults reported the following activities: watching TV (98%), reading (90%), using a computer and the internet (83%), gardening (82%), walking (75%), baking and cooking (73%), and praying (73%). Volunteering and attending community groups, which are known to benefit well-being, were unpopular (less than 8%). During the pandemic, older adults were generally satisfied with their lives, but more than half of them were concerned about their own health, family's health, and future prospects. Our study also showed the differences in the experiences and feelings by gender and race as well as the intersection of gender and race in the United States.


Health status of Asian Americans: United States, 1992-94.
This report compares the health status of selected Asian national origin groups. 1992-94 National Health Interview Survey (NHIS) data were analyzed for six Asian national origin groups (Chinese, Filipino, Asian Indian, Japanese, Vietnamese, and Korean), the Asian and Pacific Islander (API) population as a whole, and the non-Hispanic white population. Unadjusted and age-adjusted estimates and standard errors of health indicators and sociodemographic characteristics were generated. A broad range of health issues was studied including respondent-assessed health status, activity limitation, physician contacts, restricted activity days, hospital episodes, smoking status, and knowledge of acquired immunodeficiency syndrome (AIDS). A greater age-adjusted percent of Vietnamese (17.2 percent) and Korean (12.8 percent) persons had fair or poor respondent-assessed health status than persons of Chinese, Filipino, and Japanese descent (6.1-7.4 percent). A lower age-adjusted percent of Chinese persons (6.5 percent) experienced activity limitation compared with Filipino, Japanese, and Vietnamese persons (9.4-13.2 percent). Japanese persons (4.9 contacts) had a greater average annual number of physician contacts than Chinese persons (3.1 contacts) after age adjusting the data. When the data were age adjusted, a higher percent of Korean adults (22.5 percent) were current smokers than Chinese (10.0 percent) and Asian Indian adults (8.7 percent). A higher age-adjusted percent of Vietnamese (21.2 percent) and Asian Indian (18.0 percent) adults reported knowing nothing about AIDS compared with Japanese adults (5.1 percent). A greater proportion of Vietnamese adults (91.6 percent) had not been tested for the AIDS virus infection compared with Chinese, Filipino, Asian Indian, and Japanese adults (72.6-78.5 percent) after age adjusting the data. Differences in health emerge when data on the API population are analyzed by national origin group. Estimates of health presented for the API population as a whole mask differences among subgroups.


Ethanol produces corticotropin-releasing factor receptor-dependent enhancement of spontaneous glutamatergic transmission in the mouse central amygdala.
Ethanol (EtOH) modulation of central amygdala (CeA) neurocircuitry plays a key role in the development of alcoholism via activation of the corticotropin-releasing factor (CRF) receptor (CRFR) system. Previous work has predominantly focused on EtOH × CRF interactions on the CeA GABA circuitry; however, our laboratory recently showed that CRF enhances CeA glutamatergic transmission. Therefore, this study sought to determine whether EtOH modulates CeA glutamate transmission via activation of CRF signaling. The effects of EtOH on spontaneous excitatory postsynaptic currents (sEPSCs) and basal resting membrane potentials were examined via standard electrophysiology methods in adult male C57BL/6J mice. Local ablation of CeA CRF neurons (CRF(CeAhDTR) ) was achieved by targeting the human diphtheria toxin receptor (hDTR) to CeA CRF neurons with an adeno-associated virus. Ablation was quantified post hoc with confocal microscopy. Genetic targeting of the diphtheria toxin active subunit to CRF neurons (CRF(DTA) mice) ablated CRF neurons throughout the central nervous system, as assessed by quantitative reverse transcriptase polymerase chain reaction quantification of CRF mRNA. Acute bath application of EtOH significantly increased sEPSC frequency in a concentration-dependent manner in CeA neurons, and this effect was blocked by pretreatment of co-applied CRFR1 and CRFR2 antagonists. In experiments utilizing a CRF-tomato reporter mouse, EtOH did not significantly alter the basal membrane potential of CeA CRF neurons. The ability of EtOH to enhance CeA sEPSC frequency was not altered in CRF(CeAhDTR) mice despite a ~78% reduction in CeA CRF cell counts. The ability of EtOH to enhance CeA sEPSC frequency was also not altered in the CRF(DTA) mice despite a 3-fold reduction in CRF mRNA levels. These findings demonstrate that EtOH enhances spontaneous glutamatergic transmission in the CeA via a CRFR-dependent mechanism. Surprisingly, our data suggest that this action may not require endogenous CRF.


Transcriptional profiling of degraded RNA in cryopreserved and fixed tissue samples obtained at autopsy.
Traditional multiplexed gene expression methods require well preserved, intact RNA. Such specimens are difficult to acquire in clinical practice where formalin fixation is the standard procedure for processing tissue. Even when special handling methods are used to obtain frozen tissue, there may be RNA degradation; for example autopsy samples where degradation occurs both pre-mortem and during the interval between death and cryopreservation. Although specimens with partially degraded RNA can be analyzed by qRT-PCR, these analyses can only be done individually or at low levels of multiplexing and are laborious and expensive to run for large numbers of RNA targets. We evaluated the ability of the cDNA-mediated Annealing, Selection, extension, and Ligation (DASL) assay to provide highly multiplexed analyses of cryopreserved and formalin fixed, paraffin embedded (FFPE) tissues obtained at autopsy. Each assay provides data on 1536 targets, and can be performed on specimens with RNA fragments as small as 60 bp. The DASL performed accurately and consistently with cryopreserved RNA obtained at autopsy as well as with RNA extracted from formalin-fixed paraffin embedded tissue that had a cryopreserved mirror image specimen with high quality RNA. In FFPE tissue where the cryopreserved mirror image specimen was of low quality the assay performed reproducibly on some but not all specimens. The DASL assay provides reproducible results from cryopreserved specimens and many FFPE specimens obtained at autopsy. Gene expression analyses of these specimens may be especially valuable for the study of non-cancer endpoints, where surgical specimens are rarely available.


Evaluation of leukocyte adherence inhibition (LAI) test in primary and secondary hepatic malignancies.
Leukocyte adherence inhibition (LAI) has been demonstrated to be a simple, rapid, and reliable technique in the diagnosis of various malignancies including hepatocellular carcinoma. The LAI test was carried out employing modified tube LAI technique in patients with primary hepatocellular carcinoma (HCC) and secondary carcinomas of liver. Positive LAI response to HCC antigen was obtained in all six (100%) cases of hepatocellular carcinoma tested. None of the control cases, which included 8 healthy subjects and 16 cases of benign liver diseases, gave positive LAI response. Two out of 19 cases of secondary carcinoma gave a positive LAI reaction to HCC antigen. In secondary carcinomas, 19 out of 21 cases (90.48%) gave positive LAI reaction to secondary adenocarcinoma antigen. There were two false positives in controls (1 each of cirrhosis and amebic liver abscess), and 1 out of 8 cases of HCC also gave positive response to secondary carcinoma antigen. Thus, LAI test was found to be useful in the diagnosis of hepatic malignancies.


Bowel disturbances are the most important risk factors for late onset fecal incontinence: a population-based case-control study in women.
Age, diarrhea, and certain chronic illnesses are risk factors for fecal incontinence (FI). However, the contribution of obstetric injury to the development of FI later in life is unclear. We sought to better understand the risk factors for FI. Through the Rochester Epidemiology Project, a nested case-control study of 176 randomly selected women with FI (cases; mean age, 58 years) and 176 age-matched community controls was conducted in a population-based cohort from Olmsted County, Minnesota. Risk factors for FI were evaluated by reviewing inpatient and outpatient medical (including original obstetric) records. Analyses focused on conditions that preceded the index date (incidence date of FI for case in each matched pair). In 88% of cases, FI began at age ≥40 years; severity was mild (37%), moderate (58%), or severe (5%). By multivariable analysis, current smoking (odds ratio [OR], 4.7; 95% confidence interval [CI], 1.4-15), body mass index (OR per unit, 1.1; 95% CI, 1.004-1.1), diarrhea (OR, 53; 95% CI, 6.1-471), irritable bowel syndrome (OR, 4.8; 95% CI, 1.6-14), cholecystectomy (OR, 4.2; 95% CI, 1.2-15), rectocele (OR, 4.9; 95% CI, 1.3-19), and stress urinary incontinence (OR, 3.1; 95% CI, 1.4-6.5), but not obstetric events, were independent risk factors for FI. Bowel disturbances rather than prior obstetric injury are the main risk factors for FI. Measures to ameliorate bowel disturbances and other potentially reversible risk factors should be implemented before anal imaging is performed on women with FI.


Antiretroviral initiation is associated with increased skeletal muscle area and fat content.
A greater burden of physical function impairment occurs in HIV-infected adults; the impact of antiretroviral therapy (ART) initiation on muscle density (less dense = more fat), a measure of muscle quality, is unknown. AIDS Clinical Trials Group Study A5260s, a cardiometabolic substudy of A5257, randomized HIV-infected, ART-naive adults to ritonavir-boosted atazanavir, darunavir, or raltegravir with tenofovir/emtricitabine backbone. Single-slice abdominal computed tomography scans from baseline and week 96 were reanalyzed for total and lean muscle area and density. Two-sample t-tests described the differences between baseline and week 96 variables. Linear regression analysis was used to explore the role of a priori identified variables and potential confounders. Participants (n = 235) were mostly men (90%); 31% were Black non-Hispanic; 21% were Hispanic. Over 96 weeks, small but significant increases were seen in oblique/transverse abdominal, rectus, and psoas muscle total area (range 0.21-0.83 cm; P < 0.05) but not the lean muscle component (all P ≥ 0.33). Significant decreases in overall density, consistent with increases in fat, were seen in all muscle groups (range -0.87 to -2.4 HU; P < 0.01); for the lean muscle component, only decreases in oblique/transverse abdominal and rectus reached statistical significance (P < 0.05). In multivariable analyses, Black race was associated with increased muscle density and female sex with decreased density; treatment arm was not associated with changes in mass or density. The ART-associated increase in muscle area, regardless of regimen, is likely a reflection of increased fat within the muscle. The consequences of fatty infiltration of muscle on subsequent muscle function require further investigation.


Expression of the Acidothermus cellulolyticus E1 endoglucanase in Caldicellulosiruptor bescii enhances its ability to deconstruct crystalline cellulose.
The Caldicellulosiruptor bescii genome encodes a potent set of carbohydrate-active enzymes (CAZymes), found primarily as multi-domain enzymes that exhibit high cellulolytic and hemicellulolytic activity on and allow utilization of a broad range of substrates, including plant biomass without conventional pretreatment. CelA, the most abundant cellulase in the C. bescii secretome, uniquely combines a GH9 endoglucanase and a GH48 exoglucanase in one protein. The most effective commercial enzyme cocktails used in vitro to pretreat biomass are derived from fungal cellulases (cellobiohydrolases, endoglucanases and a β-d-glucosidases) that act synergistically to release sugars for microbial conversion. The C. bescii genome contains six GH5 domains in five different open reading frames. Four exist in multi-domain proteins and two as single catalytic domains. E1 is a GH5 endoglucanase reported to have high specific activity and simple architecture and is active at the growth temperature of C. bescii. E1 is an endo-1,4-β-glucanase linked to a family 2 carbohydrate-binding module shown to bind primarily to cellulosic substrates. We tested if the addition of this protein to the C. bescii secretome would improve its cellulolytic activity. In vitro analysis of E1 and CelA shows synergistic interaction. The E1 gene from Acidothermus cellulolyticus was cloned and expressed in C. bescii under the transcriptional control of the C. bescii S-layer promoter, and secretion was directed by the addition of the C. bescii CelA signal peptide sequence. The vector was integrated into the C. bescii chromosome at a site previously showing no detectable detrimental consequence. Increased activity of the secretome of the strain containing E1 was observed on both carboxymethylcellulose (CMC) and Avicel. Activity against CMC increased on average 10.8 % at 65 °C and 12.6 % at 75 °C. Activity against Avicel increased on average 17.5 % at 65 °C and 16.4 % at 75 °C. Expression and secretion of E1 in C. bescii enhanced the cellulolytic ability of its secretome. These data agree with in vitro evidence that E1 acts synergistically with CelA to digest cellulose and offer the possibility of engineering additional enzymes for improved biomass deconstruction with the knowledge that C. bescii can express a gene from Acidothermus, and perhaps other heterologous genes, effectively.


Modified vaccinia Ankara-expressing Ag85A, a novel tuberculosis vaccine, is safe in adolescents and children, and induces polyfunctional CD4+ T cells.
Modified vaccinia Ankara-expressing Ag85A (MVA85A) is a new tuberculosis (TB) vaccine aimed at enhancing immunity induced by BCG. We investigated the safety and immunogenicity of MVA85A in healthy adolescents and children from a TB endemic region, who received BCG at birth. Twelve adolescents and 24 children were vaccinated and followed up for 12 or 6 months, respectively. Adverse events were documented and vaccine-induced immune responses assessed by IFN-gamma ELISpot and intracellular cytokine staining. The vaccine was well tolerated and there were no vaccine-related serious adverse events. MVA85A induced potent and durable T-cell responses. Multiple CD4+ T-cell subsets, based on expression of IFN-gamma, TNF-alpha, IL-2, IL-17 and GM-CSF, were induced. Polyfunctional CD4+ T cells co-expressing IFN-gamma, TNF-alpha and IL-2 dominated the response in both age groups. A novel CD4+ cell subset co-expressing these three Th1 cytokines and IL-17 was induced in adolescents, while a novel CD4+ T-cell subset co-expressing Th1 cytokines and GM-CSF was induced in children. Ag-specific CD8+ T cells were not detected. We conclude that in adolescents and children MVA85A safely induces the type of immunity thought to be important in protection against TB. This includes induction of novel Th1-cell populations that have not been previously described in humans.


Validation of sun exposure and protection index (SEPI) for estimation of sun habits.
In both Sweden and Australia high incidence rates of skin cancer have become a major health problem. In prevention and risk communication, it is important to have reliable ways for identifying people with risky sun habits. In this study the validity and reliability of the questionnaire Sun Exposure Protection Index (SEPI), developed to assess individual's sun habits and their propensity to increase sun protection during routine, often brief, clinical encounters, has been evaluated. The aim of our study was to evaluate validity and reliability of the proposed SEPI scoring instrument, in two countries with markedly different ultraviolet radiation environments (Sweden and Australia). Two subpopulations in Sweden and Australia respectively were asked to fill out the SEPI together with the previously evaluated Readiness to Alter Sun Protective Behaviour questionnaire (RASP-B) and the associated Sun-protective Behaviours Questionnaire. To test reliability, the SEPI was again filled out by the subjects one month later. Comparison between SEPI and the questions in the Sun-protective Behaviours Questionnaire, analyzed with Spearman's Rho, showed good correlations regarding sun habits. Comparison between SEPI and RASP-B regarding propensity to increase sun protection showed concurrently lower SEPI mean scores for action stage, but no difference between precontemplation and contemplation stages. The SEPI test-retest analysis indicated stability over time. Internal consistency of the SEPI, assessed with Cronbach's alpha estimation showed values marginally lower than the desired >0.70 coefficient value generally recommended, and was somewhat negatively affected by the question on sunscreen use, likely related to the classic "sunscreen paradox". There were some differences in the performance of the SEPI between the Swedish and Australian samples, possibly due to the influence of "available" sunlight and differing attitudes to behaviour and protection "at home" and on vacation. SEPI appears to be a stable instrument with an overall acceptable validity and reliability, applicable for use in populations exposed to different UVR environments, in order to evaluate individual sun exposure and protection.


Genomic analysis of natural selection and phenotypic variation in high-altitude mongolians.
Deedu (DU) Mongolians, who migrated from the Mongolian steppes to the Qinghai-Tibetan Plateau approximately 500 years ago, are challenged by environmental conditions similar to native Tibetan highlanders. Identification of adaptive genetic factors in this population could provide insight into coordinated physiological responses to this environment. Here we examine genomic and phenotypic variation in this unique population and present the first complete analysis of a Mongolian whole-genome sequence. High-density SNP array data demonstrate that DU Mongolians share genetic ancestry with other Mongolian as well as Tibetan populations, specifically in genomic regions related with adaptation to high altitude. Several selection candidate genes identified in DU Mongolians are shared with other Asian groups (e.g., EDAR), neighboring Tibetan populations (including high-altitude candidates EPAS1, PKLR, and CYP2E1), as well as genes previously hypothesized to be associated with metabolic adaptation (e.g., PPARG). Hemoglobin concentration, a trait associated with high-altitude adaptation in Tibetans, is at an intermediate level in DU Mongolians compared to Tibetans and Han Chinese at comparable altitude. Whole-genome sequence from a DU Mongolian (Tianjiao1) shows that about 2% of the genomic variants, including more than 300 protein-coding changes, are specific to this individual. Our analyses of DU Mongolians and the first Mongolian genome provide valuable insight into genetic adaptation to extreme environments.


[Local coordination of health care in North Rhine-Westphalia Interim review of the North Rhine-Westphalian model].
In January 1998, the Public Health Service Act (OGDG) was put into force by North Rhine-Westphalia's state parliament as Article 3 of the Act on the Strengthening of the Efficiency of Districts, Cities and Municipalities. The objective of the Public Health Service Act is the performance of functions through the public health service in a way which corresponds to problems, ambitions and priorities at the local level. The effects of the Act have been documented over a period of 5 years. The authors describe the method of evaluation and the most important results. As well as describing traditional tasks, they focus in particular on process and structural data of the local health conferences. These have been introduced as a consequence of the Public Health Service Act in all 54 districts and self-administered cities in North Rhine-Westphalia. Another task which has become obligatory as a result of the Public Health Service Act is health reporting. Here, the authors report on the state of affairs with regard to implementation, major topics and differences in concept. Based on the state of development at the end of 2002, the authors describe present developments and prospects. They expect continuous need for support, particularly in the fields of health conferences and health reporting.


[Lumbar spinal stenosis: diagnosis and conservative treatment].
Lumbar Spinal Stenosis is a typical disease of the elderly patient that mainly originates in degenerative multisegmental changes of the lumbar vertebral column. The classical symptom of pain irradiation into the legs whilst walking and relief with standing is similar to peripheral arterial disease presentation but differs in the sense that symptoms can be triggered through lumbar extension and relieved with lumbar flexion whereas arterial disease is correlated with pathological arteriovascular findings. Diagnosis is usually confirmed through magnetic resonance imaging (MRI) and response to conservative treatment (analgetics, physiotherapy, epidural injections) is usually good in the majority of cases. Only a minority of about 20% of all cases show progressive disease and may necessitate surgical interventions.


Hybrid visibility compositing and masking for illustrative rendering.
In this paper, we introduce a novel framework for the compositing of interactively rendered 3D layers tailored to the needs of scientific illustration. Currently, traditional scientific illustrations are produced in a series of composition stages, combining different pictorial elements using 2D digital layering. Our approach extends the layer metaphor into 3D without giving up the advantages of 2D methods. The new compositing approach allows for effects such as selective transparency, occlusion overrides, and soft depth buffering. Furthermore, we show how common manipulation techniques such as masking can be integrated into this concept. These tools behave just like in 2D, but their influence extends beyond a single viewpoint. Since the presented approach makes no assumptions about the underlying rendering algorithms, layers can be generated based on polygonal geometry, volumetric data, point-based representations, or others. Our implementation exploits current graphics hardware and permits real-time interaction and rendering.


Conjugated linoleic acid stimulates an anti-tumorigenic protein NAG-1 in an isomer specific manner.
Conjugated linoleic acids (CLAs), naturally occurring fatty acids in ruminant food products, have anti-tumorigenic and pro-apoptotic properties in animal as well as in vitro models of cancer. However, the cellular mechanism has not been fully understood. NAG-1 (non-steroidal anti-inflammatory drug-activated gene-1) is induced by several dietary compounds and belongs to a TGF-beta superfamily gene associated with pro-apoptotic and anti-tumorigenic activities. The present study was performed to elucidate the molecular mechanism by which CLA stimulates anti-tumorigenic activity in human colorectal cancer (CRC) cells. The trans-10, cis-12-CLA (t10,c12-CLA) repressed cell proliferation and induced apoptosis, whereas linoleic acid or c9,t11-CLA showed no effect on cell proliferation and apoptosis. We also found that t10,c12-CLA induced the expression of a pro-apoptotic gene, NAG-1, in human CRC cells. Inhibition of NAG-1 expression by small interference RNA (siRNA) results in repression of t10,c12-CLA-induced apoptosis. Microarray analysis using t10,c12-CLA-treated HCT-116 cells revealed that activating transcription factor 3 (ATF3) was induced and its expression was confirmed by western analysis. The t10,c12-CLA treatment followed by the overexpression of ATF3 increased NAG-1 promoter activity in HCT-116 cells. We further provide the evidence that t10,c12-CLA inhibited the phosphorylation of AKT and the blockage of GSK-3 by siRNA abolished t10,c12-CLA-induced ATF3 and NAG-1 expression. The current study demonstrates that t10,c12-CLA stimulates ATF3/NAG-1 expression and subsequently induces apoptosis in an isomer specific manner. These effects may be through inhibition of AKT/GSK-3beta pathway in human CRC cells.


Ex vivo and in vitro Monocyte Responses Do Not Reflect in vivo Immune Responses and Tolerance.
Cytokine production by ex vivo (EV)-stimulated leukocytes is commonly used to gauge immune function and frequently proposed to guide immunomodulatory therapy. However, whether EV cytokine production capacity accurately reflects the in vivo (IV) immune status is largely unknown. We investigated relationships between EV monocyte cytokine responses and IV cytokine responses in a large cohort of healthy volunteers using a highly standardized IV model of short-lived LPS-induced systemic inflammation, which captures hallmarks of both hyperinflammation and immunological tolerance. Therefore, 110 healthy volunteers were intravenously challenged with 1 ng/kg LPS twice: on day 0 to determine the extent of the IV (hyper)inflammatory response and on day 7 to determine the degree of IV endotoxin tolerance. Baseline EV monocyte cytokine production capacity was assessed prior to LPS administration. Short-term and long-term EV tolerance was assessed in monocytes isolated 4 h and 7 days after LPS administration, respectively. No robust correlations were observed between baseline EV cytokine production capacity and IV cytokine responses following LPS administration. However, highly robust inverse correlations were observed between IV cytokine responses and EV cytokine responses of monocytes isolated 4 h after IV LPS administration. No correlations between IV and EV tolerance were found. In conclusion, attenuated EV cytokine production capacity reflects ongoing IV inflammation rather than immune suppression. Results of EV assays should be interpreted with caution at the risk of improper use of immuno-stimulatory drugs.