Hypothyroidism in the elderly. When symptoms are not a 'normal' part of aging.
Clinical findings alone may not lead to prompt diagnosis of hypothyroidism in elderly patients. Therefore, routine thyroid function tests may be warranted in older patients, especially women. Serum thyrotropin (TSH) is the most sensitive marker for hypothyroidism, although the test is more costly than that for serum thyroxine (T4). Patients with overt hypothyroidism who have elevated TSH and low T4 levels require replacement therapy. In addition, patients who have a TSH level higher than 20 microU/mL or who have a mildly elevated TSH level and high titers of antithyroid antibodies may benefit from prophylactic treatment. The usual recommended replacement dose is 0.05 to 0.1 mg/day of levothyroxine sodium (Levothroid, Synthroid).


CMV prevention strategies in allogeneic hematopoietic cell transplantation; the role of prophylaxis and pre-emptive monitoring in the era of letermovir.
The preferred strategy for preventing CMV in at-risk populations in alloHCT has undergone a significant practice shift in recent years where the pendulum has swung from a pre-emptive approach to now offering letermovir prophylaxis to all CMV seropositive recipients. Letermovir prophylaxis has resulted in significant reductions in post-transplant clinically significant CMV infection (csCMVi) as well as other important outcomes such as CMV disease, resistant, and refractory CMV infections and nonrelapse mortality. However, prophylactic strategies are not without some limitations, namely delayed onset CMV infections, delayed CMV-specific T cell immune reconstitution, increased drug costs and limited data within pediatric populations. Thus, this review aims to provide an overview of prophylaxis and pre-emptive CMV preventative strategies, and how they are applicable in the current era of letermovir prophylaxis.


BIOCHEMICAL, HEMATOLOGIC AND COAGULATION VALUES IN THE HEREFORD CALF.
Biochemical, hematologic and coagulation values have been determined in normal Hereford calves of both sexes, two to six months of age. These data generally agree with those of others and indicate a similarity in calves irrespective of their age, sex or breed. Prothrombin times are markedly longer in the calf than in man. This has resulted in alterations of anticoagulation regimens after implantation of mechanical circulatory support systems.


The development of language and abstract concepts: the case of natural number.
What are the origins of abstract concepts such as "seven," and what role does language play in their development? These experiments probed the natural number words and concepts of 3-year-old children who can recite number words to ten but who can comprehend only one or two. Children correctly judged that a set labeled eight retains this label if it is unchanged, that it is not also four, and that eight is more than two. In contrast, children failed to judge that a set of 8 objects is better labeled by eight than by four, that eight is more than four, that eight continues to apply to a set whose members are rearranged, or that eight ceases to apply if the set is increased by 1, doubled, or halved. The latter errors contrast with children's correct application of words for the smallest numbers. These findings suggest that children interpret number words by relating them to 2 distinct preverbal systems that capture only limited numerical information. Children construct the system of abstract, natural number concepts from these foundations.


Characterization of messenger ribonucleic acid expression for prostaglandin F2 alpha and luteinizing hormone receptors in various bovine luteal cell types.
LH and prostaglandin F2 alpha (PGF2 alpha) control the life span and function of the corpus luteum (CL). Nevertheless, identification of the various cell types (steroidogenic and nonsteroidogenic) expressing the receptors for these hormones remains controversial. In this study we characterized LH and PGF2 alpha receptor (r) expression in the various luteal cell types using quantitative reverse transcription-polymerase chain reaction. We found, in agreement with previously described functions of PGF2 alpha, that the two steroidogenic cell types, as well as luteal endothelial cells, expressed PGFr. In contrast, LHr was mainly expressed by small luteal cells. A similar pattern of PGFr and LHr expression was observed in steroidogenic cells luteinized in vitro and in cells derived from the mature CL. The expression of these two receptors was inversely affected by increased levels of cAMP (achieved by incubating cells with varying doses of forskolin); LHr expression was down-regulated by 50% in the presence of 10 microM forskolin (p < 0.05), while an increase was observed in PGFr expression. In granulosa-derived luteal cells, maximal expression of PGFr was higher (approximately by 3-fold, p < 0.05) than in the theca-derived luteal cells. PGF2 alpha, mimicking its in vivo effect, markedly down-regulated LHr expression in thecaderived luteal cells, abolishing expression at a concentration of 100 ng/ml. In summary, these studies depict cAMP and PGF2 alpha as major regulators of PGFr and LHr expression in the two steroidogenic cell types. All three major cell types of the CL (steroidogenic and endothelial) express PGFr. LHr mRNA, on the other hand, was detected mainly in small luteal cells. Such broad cellular distribution of PGFr may highlight the significant role played by this prostaglandin in the bovine CL.


Unusual bilateral rim fracture in femoroacetabular impingement.
This is a report of one case of bilateral acetabular rim fracture in association with femoroacetabular impingement (FAI), which was treated with a hip arthroscopic procedure, performing a partial resection, a labral reinsertion, and a subsequential internal fixation with cannulated screws. Up to date, there are in the literature only two reports of rim fracture and "os acetabuli" in association with FAI. In the case we present, the pincer and cam resection were performed without complications; the technique used was published previously. With this technique the head of the screw lays hidden by the reattached labrum. We removed partially the fractured rim fragment and the internal fixation of the remaining portion was achieved with a screw. In the event of a complete resection of the fragment, it would have ended with a LCE angle of 18° and a high probability of hip instability. We believe that this bilateral case helps establish the efficacy and reproducibility of the technique described by Larson.


Left ventricular dysfunction in human immunodeficiency virus infection.
The relationship between human immunodeficiency virus (HIV) infection and cardiovascular disease is still under debate, but it appears that the risk of myocardial infarction in those with HIV infection who are receiving highly active antiretroviral therapy (HAART) is increased. There has been less focus, however, on the effect of HIV and HAART on left ventricular function. Evidence from the past 20 years in both Westernized and developing countries has indicated that subclinical left ventricular dysfunction in HIV-infected individuals with and without well-controlled HIV infection is prevalent and may represent emerging cardiac disease. The specific roles of HIV infection and HAART are unclear, but they may exert independent direct and indirect effects on the myocardium. These effects may include chronic inflammation, metabolic complications (ie, insulin resistance, lipotoxicity, dyslipidemia), and mitochondrial toxicity. The objective of this article is to review the evidence for HIV- and HAART-related left ventricular dysfunction in persons infected with HIV.


Heart defects in connexin43-deficient mice.
Cardiac malformation in connexin43 (CX43)-disrupted mice is restricted to the junction between right ventricle and outflow tract, even though CX43 is also expressed abundantly elsewhere. We analyzed cardiac morphogenesis in immunohistochemically and hybridohistochemically stained and three-dimensionally reconstructed serial sections of CX43-deficient embryos between embryonic day (ED) 10 and birth. The establishment of the D configuration in the ascending loop of CX43-deficient hearts is markedly retarded, so that the right ventricle retains a craniomedial position and is connected with the outflow tract by a more acute bend in ED10 and ED11 embryos. Because of the subsequent growth of the right ventricle, this condition usually evolves into a D loop, but when it persists, a "crisscross" configuration develops, with the atrioventricular cushions rotated 90 degrees, a horizontal muscular ventricular septum, and a parallel course of the endocardial ridges of the outflow tract. After ED12, large intertrabecular pouches develop at the ventricular side of both shelflike myocardial structures that support the endocardial ridges of the outflow tract, ie, at the location that was earlier characterized by the acute bend between the right ventricle and the outflow tract and that subsequently develops into the anterosuperior leaflet of the tricuspid valve. Retarded development of the D configuration in the ascending loop of the embryonic heart predisposes the myocardium at the junction of the right ventricle and outflow tract to excessive development of intertrabecular pouches during subsequent development.


Using vignettes to rethink Latino-white disparities in self-rated health.
Researchers often rely on respondents' self-rated health (SRH) to measure social disparities in health, but recent studies suggest that systematically different reporting styles across groups can yield misleading conclusions about disparities in SRH. In this study, we test whether this finding extends to ethnic differences in self-assessments of health in particular domains. We document differences between US-born whites and four Latino subgroups in respondents' assessments of health in six health domains using data from the second wave of the Los Angeles Family and Neighborhood Survey (N = 1468). We use both conventional methods and an approach that uses vignettes to adjust for differential reporting styles. Our results suggest that despite consistent evidence from the literature that Latinos tend to rate their overall health more poorly than whites, and that Latino immigrants report worse SRH than US-born Latinos, this pattern is not true of self-reports in individual health domains. We find that at the bivariate level, US-born whites (and often US-born Mexicans) have significantly more pessimistic reporting styles than Latino immigrants. After adding controls, we find evidence of significantly different reporting styles for only one domain: US-born Mexicans and whites consistently interpret head pain more severely than the other Latino subgroups. Finally, we find that both before and after adjusting for differences in rating styles across groups, non-Mexican Latino immigrants report better social and physical functioning and less pain than other groups. Our findings underscore the advantages of domain-specific ratings when evaluating ethnic differences in self-assessments of health. We encourage researchers studying social disparities in health to consider respondents' self-assessments in a variety of domains, and to also investigate (when possible) potential biases in their findings due to different reporting styles. The anchoring vignettes approach we use is one potential method for overcoming biases due to different rating styles across groups.


Individual characteristics associated with mismatches between self-reported and accelerometer-measured physical activity.
Accurate assessment tools are required for the surveillance of physical activity (PA) levels and the assessment of the effect of interventions. In addition, increasing awareness of PA is often used as the first step in pragmatic behavioural interventions, as discrepancies between the amount of activity an individual perceives they do and the amount actually undertaken may act as a barrier to change. Previous research has demonstrated differences in the amount of activity individuals report doing, compared to their level of physical activity when measured with an accelerometer. Understanding the characteristics of those whose PA level is ranked differently when measured with either self-report or accelerometry is important as it may inform the choice of instrument for future research. The aim of this project was to determine which individual characteristics are associated with differences between self-reported and accelerometer measured physical activity. Participant data from the 2009 wave of the Commuting and Health in Cambridge study were used. Quartiles of self-reported and accelerometer-measured PA were derived by ranking each measure from lowest to highest. These quartiles were compared to determine whether individuals' physical activity was ranked higher by either method. Multinomial logistic regression models were used to investigate the individual characteristics associated with different categories of mismatch. Data from 486 participants (70% female) were included in the analysis. In adjusted analyses, the physical activity of overweight or obese individuals was significantly more likely to be ranked higher by self-report than by accelerometer than that of normal-weight individuals (OR = 2.07, 95%CI = 1.28-3.34), particularly among women (OR = 3.97, 95%CI = 2.11-7.47). There was a greater likelihood of mismatch between self-reported and accelerometer measured physical activity levels in overweight or obese adults. Future studies in overweight or obese adults should consider employing both methods of measurement.


Evidence-based medicine - engineering the Learning Healthcare System.
Whether for the generation or application of evidence to guide healthcare decisions, the success of evidence-based medicine is grounded in principles common to engineering. In the Learning Healthcare System envisioned by the Institute of Medicine's (IOM) Roundtable on Evidence-Based Medicine, evidence emerges as a natural by-product of care delivery, which is thoroughly documented, pooled for continuous monitoring and analysis, integrated with insights from related studies, and fed back seamlessly to improve the consistency and appropriateness of care decisions by clinicians and their patients. Drawing from lessons shared at the IOM/NAE symposium, Engineering a Learning Healthcare System, this paper provides an overview of the state-of-play in health care today, some of its key challenges, the vision and features of a learning healthcare system, applicable commonalties and principles from engineering, and potential collaborative opportunities moving forward to the benefit of both fields.


XMSF: Structure-preserving noise reduction and pre-segmentation in microscope tomography.
Interpretation of electron tomograms is difficult due to the high noise levels. Thus, denoising techniques are needed to improve the signal-to-noise ratio. XMSF (Microscopy Mean Shift Filtering) is a fast, user-friendly application that succeeds in filtering noise while preserving the structures of interest. It is based on the extension to 3D of a method widely applied in other image processing fields under very different scenarios. XMSF has been tested for a variety of tomograms, showing a great potential to become a state-of-the-art filtering program in electron tomography. Applied iteratively, the algorithm yields pre-segmented volumes facilitating posterior segmentation tasks. Moreover, execution times remain low thanks to parallel computing techniques to exploit current multicore computers. http://sites.google.com/site/xmsfilter/


Nurses' knowledge of heart failure education principles survey: a psychometric study.
To compare the psychometric properties of the original dichotomously scored Nurses' Knowledge of Heart Failure Self-Management Education Principles Survey with a Likert scored version. Nurses must be knowledgeable of HF self-management principles to provide optimal education to patients. Psychometrically strong instruments to measure nurses' knowledge of HF self-management provide important insight into gaps of nurses' knowledge. This psychometric study had a two phase non-experimental design comparing the psychometric characteristics of two versions of an existing survey in addition to test-retest reliability of the revised survey. A convenience sample of 122 nurses was recruited from a healthcare system located in the Southeastern USA. Data were collected from August 2006 through December 2006. Nurses completed the two versions of the survey. The sample was predominantly Caucasian (76%), female (95%) with a mean age of 42 years (SD 11·4). Internal consistency reliability was 0·27 for the dichotomously scored version of the survey and 0·70 for the Likert scored version. The test-retest reliability of the Likert scored survey was r = 0·66. Data from individual Likert scored items indicated nurses were most knowledgeable about the need to continue daily weights even after HF symptoms are gone (mean = 4·43, SD 0·51) and least knowledgeable about how to advise asymptomatic patients to manage a low BP reading (mean = 2·11, SD 0·98). The Likert scored version of the Nurses' Knowledge of Heart Failure Self-Management Principles Survey was stronger psychometrically than the dichotomously scored version. Teaching HF self-management skills to patients is challenging and it is important to identify nurses' gaps in knowledge of self-management principles. The psychometrically sound Likert scored version of the survey potentially may be used to identify nurses' knowledge gaps and inform staff development in-services.


Longevity and survival curves of Rhinella icterica (Anura, Bufonidae) under laboratory conditions.
Life tables and survival curves of tadpoles from Rhinella icterica species were studied in the laboratory, under abiotic conditions controlled by a purification filter, a timer and a chiller. The survival curve for larval stage confirms a great mortality trend in the initial stages, which decreases when reaching the mature morphological condition (r = -0.94). Stages 37, 38, 39, 40 and 41 showed gradual values for their age structures, while stages 42, 43 and 44 presented high variations. Based on the results under laboratory conditions, it can be concluded that the maturity of R. icterica tadpoles development between 37 and 44 stages has a negative correlation and their predicted life expectancy is a logarithmic growth curve (y=-761.96Ln(x)+5298.5).


Increased sensitivity of Treg cells from patients with PBC to low dose IL-12 drives their differentiation into IFN-γ secreting cells.
IL-12 is a pro-inflammatory cytokine that induces the production of interferon-γ (IFNγ) and favours the differentiation of T helper 1 (Th1) cells. In the presence of IL-12 human Treg cells acquire a Th1-like phenotype with reduced suppressive activity in vitro. Primary biliary cholangitis (PBC) is an autoimmune cholestatic liver disease characterised by high Th1 and Th17 infiltrating cells, reduced frequencies of Treg cells, and a genetic association with IL-12 signalling. Herein, we sought to evaluate the IL-12 signalling pathway in PBC pathology, by studying human samples from patients with PBC, alongside those with primary Sjögren's syndrome (pSS)(autoimmune disease with IL-12 signalling gene association), primary sclerosing cholangitis (PSC) (cholestatic liver disease without IL-12 gene association) and healthy individuals. Our data revealed that TLR stimulation of PBC (n = 17) and pSS monocytes (n = 6) resulted in significant induction of IL12A mRNA (p < 0.05, p < 0.01, respectively) compared to PSC monocytes (n = 13) and at similar levels to HC monocytes (n = 8). PSC monocytes expressed significantly less IL-12p70 (108 pg/ml, mean) and IL-23 (358 pg/ml) compared to HC (458 pg/ml and 951 pg/ml, respectively) (p < 0.01, p < 0.05). Treg cells from patients with PBC (n = 16) and pSS (n = 3) but not PSC (n = 10) and HC (n = 8) responded to low dose (10 ng/ml) IL-12 stimulation by significant upregulation of IFNγ (mean 277 and 254 pg/ml, respectively) compared to PSC and HC Treg cells (mean 22 and 77 pg/ml, respectively)(p < 0.05). This effect was mediated by the rapid and strong phosphorylation of STAT4 on Treg cells from patients with PBC and pSS (p < 0.05) but not PSC and HC. In the liver of patients with PBC (n = 7) a significantly higher proportion of IL-12Rβ2+Tregs (16% on average) was detected (p < 0.05) compared to other liver disease controls (5%)(n = 18) which also showed ex vivo high IFNG and TBET expression. CONCLUSION: Our data show an increased sensitivity of PBC and pSS Treg cells to low dose IL-12 stimulation, providing ongoing support for the importance of the IL12-IL-12Rβ2-STAT4 pathway on Treg cells in disease pathogenesis and potentially treatment.


A Case for the Non-Neurologist Telestroke Provider.
Introduction: Telestroke networks have effectively increased the number of ischemic stroke patients who have access to acute stroke therapy. However, the availability of a dedicated group of stroke subspecialists is not always feasible. We hypothesize that rates of tPA recommendation, sensitivity of final diagnosis, and post-tPA hemorrhagic complications do not differ significantly between neurologists and an emergency-medicine physician during telestroke consultations. Methods: Retrospective review of all telestroke consults performed at a comprehensive stroke center over 1 year. Statistical analysis: Chi squared test. Results: Three hundred and three consults were performed among 6 spoke sites. 16% (48/303) were completed by the emergency medicine physician; 25% (76/303) were performed by non-stroke-trained neurologists, and 59% (179/303) were completed by a board-certified Vascular Neurologist. Overall rate of tPA recommendation was 40% (104/255), 38% (18/48), 41% (73/179), and 41% (31/76) among the all neurology-trained, emergency medicine-trained, stroke neurology-trained and other neurology- trained provider groups, respectively (p = 0.427). Sensitivity of final stroke diagnosis was 77% (14/18) and 72% (75/104) in the emergency-medicine trained and neurology-trained provider groups (p = 0.777) No symptomatic hemorrhagic complications following the administration of tPA via telestroke consultation occurred in any group over this time period. One asymptomatic intracerebral hemorrhage was observed (0.96% or 1/104) in the neurology-trained provider group. Discussion/Conclusion: Our results did not illustrate any statistically significant difference between care provided by an emergency medicine-trained physician and neurologists during telestroke consultation. While our study is limited by its relatively low numbers, it suggests that identifying a non-neurologist provider who has requisite clinical experience with acute stroke patients can safely and appropriately provide telestroke consultation. The lack of formerly trained neurologists, therefore, may not need to serve as an impediment to building an effective telestroke network. Future efforts should be focused on illuminating all strategies that facilitate sustainable telestroke implementation.


[Bacteriologic studies of aborted sheep fetuses with special reference to chlamydiae].
A total of 467 tissue specimens recovered from 165 aborted ovine fetuses were examined for chlamydiae microscopically and by means of cell culture. Chlamydiae were detected by microscope in 34 fetuses (20.6%) and by cell culture in 60 fetuses (36.4%). Of the 467 tissue specimens, 51 (10.9%) were microscopically positive, whereas chlamydiae were recovered from 181 specimens (38.8%) using the routine method of BGM cell culture. A total of 87 specimens of fetal organs was tested by the BGM cell culture technique and simultaneously by an enzyme amplified immunoassay to detect chlamydial antigen. Although not as sensitive as cell culture isolation, the ELISA has the advantage of rapidity and of detecting antigen from non-viable chlamydiae in sheep abortion material.


Regression of dietary copper restriction-induced cardiomyopathy by copper repletion in mice.
Dietary copper deficiency (CuD)(3) leads to cardiac hypertrophy in various animal models. We showed recently that heart failure develops after hypertrophy in FVB mice fed a CuD diet. The present study was undertaken to determine whether CuD-induced cardiac failure is reversible upon copper repletion (CuR). Dams of FVB mice were fed a CuD diet (0.3 mg/kg) starting from d 3 postdelivery; the weanling pups were fed the same diet until CuR with 6.0 mg/kg Cu in the diet at 4 or 5 wk of age. CuR at 4 wk of age prevented the body weight loss; at 5 wk of age, it resulted in the regaining of the lost weight caused by CuD. A significant regression of CuD-induced cardiac hypertrophy was observed in the CuR mice. Histopathological examination revealed that CuR eliminated CuD-caused lipid deposition in the myocardium, and electron microscopy demonstrated that CuD-induced ultrastructural changes such as mitochondrial swelling and organelle structural disarray were all reversed in the CuR mice. Hemodynamic analysis showed that the CuD-depressed systolic and diastolic parameters such as the maximal rate of left ventricular pressure rise (+dP/dt) and decline (-dP/dt), and the contraction and relaxation times were completely recovered in the CuR mice. Furthermore, the CuD-blunted myocardial responses to the beta-adrenergic agonist, isoproterenol, were also restored in the CuR mice. This study thus demonstrates for the first time that CuR results in the regression of heart failure induced by CuD as demonstrated by the reversal of depressed cardiac hemodynamic and contractile function and the restored responsiveness to beta-adrenergic stimulation.


Secretory pathway-dependent localization of the Saccharomyces cerevisiae Rho GTPase-activating protein Rgd1p at growth sites.
Establishment and maintenance of cell polarity in eukaryotes depends upon the regulation of Rho GTPases. In Saccharomyces cerevisiae, the Rho GTPase activating protein (RhoGAP) Rgd1p stimulates the GTPase activities of Rho3p and Rho4p, which are involved in bud growth and cytokinesis, respectively. Consistent with the distribution of Rho3p and Rho4p, Rgd1p is found mostly in areas of polarized growth during cell cycle progression. Rgd1p was mislocalized in mutants specifically altered for Golgi apparatus-based phosphatidylinositol 4-P [PtdIns(4)P] synthesis and for PtdIns(4,5)P(2) production at the plasma membrane. Analysis of Rgd1p distribution in different membrane-trafficking mutants suggested that Rgd1p was delivered to growth sites via the secretory pathway. Rgd1p may associate with post-Golgi vesicles by binding to PtdIns(4)P and then be transported by secretory vesicles to the plasma membrane. In agreement, we show that Rgd1p coimmunoprecipitated and localized with markers specific to secretory vesicles and cofractionated with a plasma membrane marker. Moreover, in vivo imaging revealed that Rgd1p was transported in an anterograde manner from the mother cell to the daughter cell in a vectoral manner. Our data indicate that secretory vesicles are involved in the delivery of RhoGAP Rgd1p to the bud tip and bud neck.


The Antimicrobial Peptide LL-37 as a Possible Adjunct for the Proliferation and Differentiation of Dental Pulp Stem Cells.
This study evaluated the biocompatibility of 5 and 10 μg/mL LL-37 in vitro and its effect on the differentiation of human dental pulp stem cells (DPSCs) into odontoblast-like cells. Cell viability, genotoxicity, nitric oxide production, cell cycle, dentine sialophosphoprotein (DSPP) production, and DSPP gene expression. Concentrations of 5 and 10 μg/mL of LL-37 were not cytotoxic and generally increased cell viability, especially on the third day (P < .05). The tested concentrations did not induce genotoxicity (P < .05). LL-37 did not significantly alter nitrite production at either concentration. Cell cycle analysis revealed that 10 μg/mL of LL-37 arrested cells in G0/G1 (P < .05). The control group exhibited higher numbers of cells in other phases of the cell cycle (P < .05). The expression of the DSPP protein and gene was also higher in the 10 μg/mL of LL-37 group (P < .05). These results demonstrated that LL-37 was biocompatible at these concentrations and increased the number of viable cells, especially during the initial period. The 10 μg/mL concentration arrested the cell cycle and increased expression of the DSPP protein and gene, which indicates that this peptide contributes to odontoblastic differentiation.


Nervous control of the blood pressure in the Atlantic cod, Gadus morhua.
Dorsal (PDA) and ventral aortic blood pressure (PVA) and heart rate (HR) were measured in conscious resting cod, Gadus morhua L., which has been allowed 24 h recovery from surgery. Plasma adrenalin and nonadrenalin concentrations in these fish were 3.4 and 2.2 nmoll-1 respectively, and thus lower than previously reported values from partially recovered cod. Twenty-four hours after treatment with the adrenergic neurone blocking agent bretylium, PDA was significantly reduced by 17% compared to sham-injected controls, although PVA and heart rate were unaltered. Subsequent alpha-adrenoceptor blockade by phentolamine produced no further fall in PDA and no changes in PVA or HR, provided a 5-h period was allowed to overcome the acute toxic side effects of phentolamine. The effectiveness of the bretylium or phentolamine blockade was confirmed by noting the absence of any vasoconstrictor response during sympathetic nerve stimulation in perfused tails from fish used in the in vivo experiments. Bretylium had no significant effect on the sensitivity of the isolated coeliac artery to adrenalin, but effectively blocked the adrenergic innervation of this artery or the vasculature of the tail. Evidence for a non-selective blockade of non-adrenergic nerves to the heart was also obtained. It is concluded that the adrenergic tonus affecting the dorsal aortic blood pressure in resting cod that have recovered for 24 h following surgery is due solely to an adrenergic nervous tone.


Pathology of diencephalic astrocytomas.
Pediatric gliomas span a spectrum of neoplasms ranging from the well-circumscribed, slowly growing lesions that can be totally removed by surgery to highly infiltrating, nonexcisable, rapidly growing tumors that resist any form of postoperative therapy. As part of this symposium, we discuss selected members of this group in the diencephalon, where lesions of both extremes are common in children. The report illustrates and contrasts the clinical, radiographic and pathological features of the two classic regional astrocytomas, pilocytic and fibrillary, as well as two less common but distinctive lesions, pilomyxoid astrocytoma and bithalamic astrocytoma.


Smoking cessation--what works?
Prevalence of tobacco smoking in the Australian community has fallen. However, tobacco smoking remains a major cause of illness and death. General practitioners play an important role in assisting their patients to quit smoking. This article describes evidence based approaches to smoking cessation that can be applied in general practice. Evidence based approaches to smoking cessation include brief counselling, pharmacotherapy, referral to a specialised service such as Quitline, and follow up. The five As approach--Ask, Assess, Advise, Assist and Arrange follow up--provides a structure for intervention. Smoking cessation pharmacotherapies (nicotine replacement therapy and bupropion) have been shown to double quit rates. A new pharmacotherapy (varenicline) has recently become available which evidence to date suggests is more effective. Clinical suitability, the context of the quit attempt, and patient preference are important considerations in choosing a pharmacotherapy. Active follow up helps to reduce the rate of relapse which is otherwise high in what is commonly a chronic relapsing condition.


Modified cardiopulmonary resuscitation (CPR) instruction protocols for emergency medical dispatchers: rationale and recommendations.
International consensus guidelines now support the use of "chest compressions-only" cardiopulmonary resuscitation (CPR) instructions (CCOIs) by emergency medical dispatch (EMD) personnel providing telephone assistance to untrained bystanders at a cardiac arrest scene. These guidelines are based largely on evolving experimental data and a clinical trial conducted in one venue with distinct emergency medical services (EMS) system features. Accordingly, the Council of Standards for the National Academies of Emergency Dispatch was asked to adapt a modified telephone CPR protocol, and specifically one that could be applied more broadly to the spectrum of EMS systems. A group of international EMD specialists, researchers and professional association representatives analyzed available scientific data and considered variations in EMS systems, particularly those in Europe and North America. Several recommendations were established: (1) to avoid confusion, bystanders already providing CPR should continue those previously learned methods; (2) following a sudden collapse unlikely to be of respiratory etiology, CCOIs should be provided when the bystander is not CPR-trained, declining to perform mouth-to-mouth ventilation or unsure of actions to take; (3) following 4 min of CCOIs, ventilations can be provided, but, for now, only at a compression-ventilation ratio of 100:2 until EMS arrives; (4) until more data become available, dispatchers should follow existing compression-ventilation protocols for children and adult cases involving probable respiratory/trauma etiologies; (5) EMD CPR protocols should account for EMS system features and receive quality oversight and expert medical direction.


Aqua-bis-(3-chloro-benzoato-κO)bis-(N,N-di-ethyl-nicotinamide-κN)copper(II).
The title compound, [Cu(C7H4ClO2)2(C10H14N2O)2(H2O)], has twofold symmetry with the Cu(II) cation and the O atom of the coordinating water mol-ecule located on the axis. The Cu(II) cation is coordinated by two carboxyl-ate O atoms of chloro-benzoate (CB) anions, two N atoms of N,N-di-ethyl-nicotinamide (DENA) ligands and one water mol-ecule in a distorted N2O3 square-pyramidal geometry. The benzene and pyridine rings are oriented at a dihedral angle of 82.51 (6)°. In the anionic ligand, the carboxyl-ate group is twisted away from the attached benzene ring by 12.85 (11)°. In the crystal, O-H⋯O hydrogen bonds between the coordinating water mol-ecule and the carboxyl group link the complex mol-ecules into supra-molecular chains running along the c-axis direction.


Autism Detection in Children using Common Spatial Patterns of MEG Signals.
Autism exhibits a wide range of developmental disabilities and is associated with aberrant anatomical and functional neural patterns. To detect autism in young children (4-7 years) in an automatic and non-invasive fashion, we have recorded magnetoencephalogram (MEG) signals from 30 autistic and 30 age-matched typically developing (TD) children. We have used a machine learning classification framework with common spatial pattern (CSP)-based logarithmic band power (LBP) features. When comparing the LBP feature to the conventional logarithmic variance (LV) spatial pattern, CSP + LBP (92.77%) has performed better than CSP + LV (90.66%) in the 1-100 Hz frequency range for distinguishing autistic children from TD children. In frequency band-wise analysis using our proposed method, the high gamma frequency band (50-100 Hz) has shown the highest classification accuracy (97.14%). Our findings reveal that the occipital lobe exhibits the most distinct spatial pattern in autistic children over the whole frequency range. This study shows that spatial brain activation patterns can be utilized as potential biomarkers of autism in young children. The improved performance signifies the clinical relevance of the work for autism detection using MEG signals.


Continuous glucose monitoring in the ICU: clinical considerations and consensus.
Glucose management in intensive care unit (ICU) patients has been a matter of debate for almost two decades. Compared to intermittent monitoring systems, continuous glucose monitoring (CGM) can offer benefit in the prevention of severe hyperglycemia and hypoglycemia by enabling insulin infusions to be adjusted more rapidly and potentially more accurately because trends in glucose concentrations can be more readily identified. Increasingly, it is apparent that a single glucose target/range may not be optimal for all patients at all times and, as with many other aspects of critical care patient management, a personalized approach to glucose control may be more appropriate. Here we consider some of the evidence supporting different glucose targets in various groups of patients, focusing on those with and without diabetes and neurological ICU patients. We also discuss some of the reasons why, despite evidence of benefit, CGM devices are still not widely employed in the ICU and propose areas of research needed to help move CGM from the research arena to routine clinical use.


Identification of genome-wide targets of Olig2 in the adult mouse spinal cord using ChIP-Seq.
In jawed vertebrates, oligodendrocytes (OLs) are the myelin-producing glial cells responsible for ensheathment of axons within the central nervous system and are also crucial for remyelination following injury or disease. Olig2 is a crucial factor in the specification and differentiation of oligodendrocyte precursor cells (OPCs) that give rise to mature, myelin-producing OLs in the developing and postnatal CNS; however, its role in adulthood is less well understood. To investigate the role Olig2 plays in regulating gene expression in the adult OL lineage in a physiologically-relevant context, we performed chromatin immunoprecipitation followed by next generation sequencing analysis (ChIP-Seq) using whole spinal cord tissue harvested from adult mice. We found that many of the Olig2-bound sites were associated with genes with biological processes corresponding to OL differentiation (Nkx2.2, Nkx6.2, and Sip1), myelination and ensheathment (Mbp, Cldn11, and Mobp), as well as cell cycle and cytoskeletal regulation. This suggests Olig2 continues to play a critical role in processes related to OL differentiation and myelination well into adulthood.


Progress in Cancer Control Leads to Significant Number of Cancer Deaths Avoided in Canada.
It is currently not known how many more cancer deaths would have occurred among Canadians if cancer mortality rates were unchanged due to various modern human interventions. The objective of this study was to estimate the number of cancer deaths that have been avoided in Canada since the age-standardized cancer mortality rate peaked in 1988. We applied the age-specific cancer mortality rates from 1988 to the Canadian population for all subsequent years to estimate the number of expected deaths. Avoided cancer deaths were estimated as the difference between the observed and expected number of cancer deaths for each year. Since 1988, there have been 372,584 (SMR=0.77) and 120,045 (SMR=0.90) avoided cancer deaths in males and females, respectively (492,629 total). There have been nearly half a million cancer deaths avoided in Canada since the overall cancer mortality rate peaked. This demonstrates the exceptional progress made in modern cancer control in Canada.


[Programmed cell death protein 1 and its ligands regulate immune balance in allergic rhinitis].
Objective: To explore the expression and significance in regulating immune balance of programmed cell death protein 1 (PD-1) and its ligands PD-L1, PD-L2 in allergic rhinitis (AR). Methods: Eighty-two patients who received outpatient treatment due to high nasal reaction symptoms or were hospitalized due to nasal septum deviation and underwent nasal septum correction surgery in Department of Otorhinolaryngology Head and Neck Surgery of Renmin Hospital of Wuhan University from May 2018 to May 2019 were enrolled, including 42 males and 40 females, with the age ranging from 14 to 38 years old. Blood, inferior turbinate nasal mucosal tissue and relevant clinical data were collected. Patients were divided into AR group and control group due to clinical manifestation, skin prick test and detection of specific IgE (sIgE) in serum. Immunohistochemistry was used to detect the expression of PD-1 and its ligands in nasal mucosa of the two groups. Flow cytometry was used to detect the proportions of PD-1(+)CD4(+)T cells, PD-L1(+) myeloid dendritic cells (mDCs), PD-L2(+)mDCs and Th2 cells in peripheral blood of the two groups. The expression levels of total IgE, sPD-1, sPD-L1 and sPD-L2 in serum of the two groups were detected by ELISA. The measurement data of normal distribution or normal distribution after the logarithm conversion to Ln were compared by t test. Pearson correlation or Spearman correlation was used to analyze the correlation among the indicators. P<0.05 was considered statistically significant. Results: The expression of PD-1 and its ligands on the surface of immune cells in the nasal mucosa of the AR group was significantly higher than that of the control group. The ratio of PD-1(+)CD4(+)T cells, PD-L1(+)mDCs and Th2 cells in peripheral blood of AR group was significantly higher than that of the control group ((15.24±6.45)% vs (8.71±5.33)%, (8.79±2.01)% vs (5.74±2.90)%, (7.89±1.95)% vs (2.52±1.34)%, all P<0.05). There was no significant difference in the ratio of PD-L2(+)mDCs between the two groups. Correlation analysis found that the proportion of PD-1(+)CD4(+) T cells was positively correlated with the Visual Analogue Scale (VAS) score of AR, total IgE concentration and the serum sIgE concentration (r value was 0.501, 0.541, 0.608, respectively, all P<0.05). The proportion of PD-L1(+)mDCs was positively correlated with the VAS score of AR and the serum sIgE concentration (r value was 0.604, 0.563, respectively, all P<0.05). The proportion of Th2 cells in peripheral blood was positively correlated with the proportion of PD-L1(+)mDCs and PD-1(+)CD4(+)T cells (r value was 0.538, 0.623, respectively, all P<0.05). Serum total IgE, sPD-1 and sPD-L1 in the AR group were significantly higher than those in the control group ((6.34±1.38) ng/ml vs (4.89±1.10) ng/ml, (4.40±1.01) pg/ml vs (3.79±1.21) pg/ml, (3.88±0.25) pg/ml vs (3.57±0.23) pg/ml, all P<0.05), and there was no significant difference in sPD-L2 levels between the two groups. Correlation analysis showed that sPD-L1 was positively correlated with total IgE and sIgE concentration (r values was 0.32, 0.45, respectively, all P<0.05). Conclusions: PD-1 and PD-L1 are highly expressed on the surface of immune cells in peripheral blood and nasal mucosa of AR patients, and sPD-1 and sPD-L1 expression levels in peripheral blood of AR patients are increased. The PD-1/PD-L1 signaling pathway promote AR inflammatory response by inducing Th2 type immune response.


The use of subcutaneous infusion in medication administration.
The subcutaneous administration of medications is an area that receives little attention compared with other types of parenteral therapy. Parenteral administration is used by many thousands of patients who self-administer their medication on a daily basis-for example, those using insulin to manage diabetes, recipients of some types of hormone therapy and so on. It is also an effective route for the continuous administration of medication(s) in individuals who are terminally ill. Patients approaching the end of their life may be unable to tolerate the administration of oral medication to control their symptoms and make them more comfortable. This paper will discuss how subcutaneous infusion can be used to deliver these medications, but at the same time how important the selection of the most appropriate subcutaneous infusion device is to the overall comfort of the patient, and to reduce the potential for sharps-related injuries to healthcare workers. Appropriate device selection, together with its management, is an important contributing factor to patient safety and comfort. It will diminish the potential for premature device loss, which can lead to repeated insertion procedures for the patient, as well as delaying their medication. There is also a resource implication for the NHS, as the replacement of any device involves the use of additional equipment and staff time. Additionally, the use of any infusion device poses a risk to healthcare workers of acquiring a bloodborne infection should they experience a percutaneous injury. Knowledge of what equipment is available will reduce the potential risk to these staff.


Engineering Quasi-Vivo in vitro organ models.
Cell culture is the workhorse of biologists, toxicologists, tissue engineers and a whole host of research fields in both academia and industry. Having explored individual molecular mechanisms inside cells for decades using traditional cell culture techniques, researchers have only just begun to appreciate that the intricate interconnectivity between cells and cellular networks as well as with the external environment is far more important to cellular orchestration than are single molecular events inside the cell. For example many questions regarding cell, tissue, organ and system response to drugs, environmental toxins, stress and nutrients cannot possibly be answered by concentrating on the minutiae of what goes on in the deepest recesses of single cells. New models are required to investigate cellular cross-talk between different cell types and to construct complex in-vitro models to properly study tissue, organ and system interaction without resorting to animal experiments. This chapter describes how tissue and organ models can be developed using the Quasi-Vivo system and discusses how they may be used in drug toxicity studies.


Lifespan extension by dietary intervention in a mouse model of Cockayne syndrome uncouples early postnatal development from segmental progeria.
Cockayne syndrome (CS) is a rare autosomal recessive segmental progeria characterized by growth failure, lipodystrophy, neurological abnormalities, and photosensitivity, but without skin cancer predisposition. Cockayne syndrome life expectancy ranges from 5 to 16 years for the two most severe forms (types II and I, respectively). Mouse models of CS have thus far been of limited value due to either very mild phenotypes, or premature death during postnatal development prior to weaning. The cause of death in severe CS models is unknown, but has been attributed to extremely rapid aging. Here, we found that providing mutant pups with soft food from as late as postnatal day 14 allowed survival past weaning with high penetrance independent of dietary macronutrient balance in a novel CS model (Csa(-/-) | Xpa(-/-)). Survival past weaning revealed a number of CS-like symptoms including small size, progressive loss of adiposity, and neurological symptoms, with a maximum lifespan of 19 weeks. Our results caution against interpretation of death before weaning as premature aging, and at the same time provide a valuable new tool for understanding mechanisms of progressive CS-related progeroid symptoms including lipodystrophy and neurodysfunction.


Testing for treatment related trend with partially exchangeable clustered data.
When testing for treatment related effect, it is common to focus on data that describe some primary response, even though an experiment might yield data on secondary, less conspicuous effects. An example of such an experiment can be found in developmental toxicity studies, where a primary response might be the presence of skeletal malformation in a fetus, and a secondary effect might be reduced weight. In such studies, weight reduction may occur not only among malformed fetuses, but also among normal fetuses. In this paper, we use a likelihood ratio procedure to construct treatment related trend tests for developmental experiments with such outcomes. We assume that within each cluster, data are partially exchangeable and we use the presence or absence of malformations as categorical covariates. Two models are considered for the covariance structures: In the general model, partially exchangeable covariance matrices are used for each cluster within dose groups. In the homogeneous model, a common exchangeable covariance matrix is used for all clusters. Maximum likelihood estimates (MLEs) of unknown parameters in the general model are obtained by directly maximizing the log-likelihood function through careful numerical consideration, while MLEs of parameters of teh homogeneous model are obtained through an iterative procedure. Two applications to developmental toxicity data are described.


A mucosal antibody response following systemic Haemophilus influenzae type B infection in children.
The possibility that mucosal antibody is produced as a host response to Haemophilus influenzae type b (Hib) infection was examined in this study. 17 of 18 prospectively evaluated children ranging in age from 2 mo to 7 yr developed a detectable level of anticapsular antibody in their nasopharyngeal secretions after systemic Hib infection. The mean concentration of nasal anti-capsular antibody of the 18 children was 554 ng/mg IgA (SD = 35-8,863) during the acute phase of illness and declined to 224 ng/mg IgA (SD = 19-2,688) in convalescence. Some children had mucosal antibody detectable at least 10 mo after infection. The mucosal antibody levels were not affected by the length of illness before diagnosis, type of disease, age of the patient, sex, or presence of detectable capsular antigen or viable bacteria in the nasopharynx. The mucosal antibody was predominantly of the IgA class and occurred independent of the serum antibody. Six of the children aged less than 1 yr who did not produce and/or sustain a serum antibody level correlated with protection demonstrated a persistent mucosal antibody response. These findings suggest that the mucosal immune system may have the ability to respond at an earlier age than the serum immune system and lead us to postulate that protective secretory antibodies to prevent systemic Hib disease may be inducible in young infants in spite of the poor serum antibody response occurring at this age.


A survey on regenerative surgery performed by Swiss specialists in periodontology with special emphasis on the application of enamel matrix derivatives in infrabony defects.
This survey aimed to evaluate the common practice of regenerative periodontal surgery with special regard to the use of enamel matrix derivatives (EMD, Emdogain® ) by board-certified specialists in periodontology and non-certified, but active members of the Swiss Society of Periodontology (SSP). A cross-sectional postal survey of 533 dentists, representing all members of the SSP practising in Switzerland, was conducted. The questionnaire consisted of three sections, assessing: 1) general personal information regarding the practice setting and education, 2) general questions regarding periodontal surgery practices and 3) specific questions regarding the use of EMD. The information obtained was compared and differences between specialists and non-specialists were calculated. P-values smaller than 5% were considered significant. Sixty-nine percent of the specialists answered the questionnaire, compared to only 37.4% of the non-specialists (overall: 42.4%). In general, specialists performed surgeries more frequently, and presented a significantly higher percentage of EMD users than the non-specialists. The application guidelines were followed in general. Some differences were observed in application and selection criteria. The subjective perception of clinical success varied greatly among clinicians. Residual pockets were reported to be present in approximately one third of the defects after therapy. In conclusion, this survey revealed that EMD was used on a regular basis by dentists performing periodontal therapy. In addition, the answers by both groups generally corresponded well with the current available literature.


Evaluating high-throughput ab initio gene finders to discover proteins encoded in eukaryotic pathogen genomes missed by laboratory techniques.
Next generation sequencing technology is advancing genome sequencing at an unprecedented level. By unravelling the code within a pathogen's genome, every possible protein (prior to post-translational modifications) can theoretically be discovered, irrespective of life cycle stages and environmental stimuli. Now more than ever there is a great need for high-throughput ab initio gene finding. Ab initio gene finders use statistical models to predict genes and their exon-intron structures from the genome sequence alone. This paper evaluates whether existing ab initio gene finders can effectively predict genes to deduce proteins that have presently missed capture by laboratory techniques. An aim here is to identify possible patterns of prediction inaccuracies for gene finders as a whole irrespective of the target pathogen. All currently available ab initio gene finders are considered in the evaluation but only four fulfil high-throughput capability: AUGUSTUS, GeneMark_hmm, GlimmerHMM, and SNAP. These gene finders require training data specific to a target pathogen and consequently the evaluation results are inextricably linked to the availability and quality of the data. The pathogen, Toxoplasma gondii, is used to illustrate the evaluation methods. The results support current opinion that predicted exons by ab initio gene finders are inaccurate in the absence of experimental evidence. However, the results reveal some patterns of inaccuracy that are common to all gene finders and these inaccuracies may provide a focus area for future gene finder developers.


Elevated serum S100B protein in drug-free bipolar patients during first manic episode: a pilot study.
S100B protein is a calcium-binding protein mostly derived from glial cells, which exerts trophic or toxic effects on neural cells depending on its concentration. Since serum S100B levels has been tested as a potential marker in neuropsychiatric disorders, and structural abnormalities on glial cells have been recently associated with bipolar disorder patients, we conducted this preliminary study to examine if S100B serum levels are altered during first manic episode. We quantitated S100B in serum of 40 subjects (20 unmedicated patients during manic episode and 20 healthy matched controls). The mean+/-S.D. values for S100B for bipolar subjects were 0.065+/-0.068 microg/l and 0.018+/-0.029 microg/l for healthy controls. Increased levels of S100B in bipolar mania was statistically significant (Wilcoxon signed ranks test, Z=-2.45, P=0.01). These preliminary findings suggest that mania may increase the levels of S100B in serum of bipolar disorder patients, which could be related to adaptative neural mechanisms in bipolar mania.


Photoprotective capacity of non-photochemical quenching in plants acclimated to different light intensities.
Arabidopsis plants grown at low light were exposed to a gradually increasing actinic light routine. This method allows for the discerning of the photoprotective component of NPQ, pNPQ and photoinhibition. They exhibited lower values of Photosystem II (PSII) yield in comparison to high-light grown plants, and higher calculated dark fluorescence level (F'o calc.) than the measured one (F'o act.). As a result, in low-light grown plants, the values of qP measured in the dark appeared higher than 1. Normally, F'o act. and F'o calc. match well at moderate light intensities but F'o act. becomes higher at increasing intensities due to reaction centre (RCII) damage; this indicates the onset of photoinhibition. To explain the unusual increase of qP in the dark in low-light grown plants, we have undertaken an analysis of PSII antenna size using biochemical and spectroscopic approaches. Sucrose gradient separation of thylakoid membrane complexes and fast fluorescence induction experiments illustrated that the relative PSII cross section does not increase appreciably with the rise in PSII antenna size in the low-light grown plants. This suggests that part of the increased LHCII antenna is less efficiently coupled to the RCII. A model based upon the existence of an uncoupled population LHCII is proposed to explain the discrepancies in calculated and measured values of F'o.


Comparing uni-modal and multi-modal therapies for improving writing in acquired dysgraphia after stroke.
Writing therapy studies have been predominantly uni-modal in nature; i.e., their central therapy task has typically been either writing to dictation or copying and recalling words. There has not yet been a study that has compared the effects of a uni-modal to a multi-modal writing therapy in terms of improvements to spelling accuracy. A multiple-case study with eight participants aimed to compare the effects of a uni-modal and a multi-modal therapy on the spelling accuracy of treated and untreated target words at immediate and follow-up assessment points. A cross-over design was used and within each therapy a matched set of words was targeted. These words and a matched control set were assessed before as well as immediately after each therapy and six weeks following therapy. The two approaches did not differ in their effects on spelling accuracy of treated or untreated items or degree of maintenance. All participants made significant improvements on treated and control items; however, not all improvements were maintained at follow-up. The findings suggested that multi-modal therapy did not have an advantage over uni-modal therapy for the participants in this study. Performance differences were instead driven by participant variables.


Should living wills be legalized?
Living wills allow patients to state their wish to die and not be kept alive through the use of medications, artificial means or "heroic" measures. They have been made legal documents in 38 states in the United States. Living wills permit advance expression of a patient's wishes, promote effective communication and demonstrate respect for the patient as a person. Problems with legal recognition of such wills include the need for agreement on fundamental terms, possible restriction of patients' rights, limitation of options in decision-making and possible negative effects on the physician-patient relationship. Before legislation is enacted, public and professional attitudes toward the care of terminally ill patients should be assessed. All health care professionals should receive better education in this area, and palliative care services should be made more widely available. Only if these measures fail should living wills be made legal documents in Canada.


Firing rates of motor units in human vastus lateralis muscle during fatiguing isometric contractions.
We investigated the firing rate of motor units in the vastus lateralis muscle in five healthy young men (mean = 21.4 yr, SD = 0.9) during a sequence of isometric constant-torque contractions repeated to exhaustion. The contractions were sustained at 20% of the maximal voluntary level, measured at the beginning of the test sequence. Electromyographic (EMG) signals were recorded via quadrifilar fine-wire electrodes and subsequently decomposed into their constituent motor unit action potentials to obtain the motor unit firing times. In addition, we measured the whole muscle mechanical properties during the fatigue task using electrical stimulation. The firing rate of motor units first decreased within the first 10-20% of the endurance time of the contractions and then increased. The firing rate increase was accompanied by recruitment of additional motor units as the force output remained constant. The elicited twitch and tetanic torque responses first increased and then decreased. The two processes modulated in a complementary fashion at the same time. Our data suggest that, when the vastus lateralis muscle is activated to maintain a constant torque output, its motoneuron pool receives a net excitatory drive that first decreases to compensate for the short-lived potentiation of the muscle force twitch and then increases to compensate for the diminution of the force twitch. The underlying inverse relationship between the firing rate and the recruitment threshold that has been reported for nonfatigued contractions is maintained. We, therefore, conclude that the central nervous system control of vastus lateralis motor units remains invariant during fatigue in submaximal isometric isotonic contractions.


Biological and clinical significance of KRAS mutations in lung cancer: an oncogenic driver that contrasts with EGFR mutation.
KRAS and epidermal growth factor receptor (EGFR) are the two most frequently mutated proto-oncogenes in adenocarcinoma of the lung. The occurrence of these two oncogenic mutations is mutually exclusive, and they exhibit many contrasting characteristics such as clinical background, pathological features of patients harboring each mutation, and prognostic or predictive implications. Lung cancers harboring the EGFR mutations are remarkably sensitive to EGFR tyrosine kinase inhibitors such as gefitinib or erlotinib. This discovery has dramatically changed the clinical treatment of lung cancer in that it almost doubled the duration of survival for lung cancer patients with an EGFR mutation. In this review, we describe the features of KRAS mutations in lung cancer and contrast these with the features of EGFR mutations. Recent strategies to combat lung cancer harboring KRAS mutations are also reviewed.


Clinical efficacy of intravesical electrical stimulation on detrusor underactivity: 8 Years of experience from a single center.
The aim of this study was to retrospectively evaluate the effectiveness of intravesical electrical stimulation (IVES) on detrusor underactivity (DU).From 2009 to 2016, a total of 105 patients with symptoms of DU who were treated with IVES were included in this retrospective study. The medical records, physical examination findings, urine culture results, and video-urodynamic studies were reviewed. Changes in post-void residual urine (PVR) and voiding efficiency (VE) were included for evaluation of efficacy. Patients achieving a >50% reduction in the PVR were regarded as responders. A >80% reduction in the PVR was considered obvious improvement. A questionnaire was administered to patients with bladder sensation.Of the 105 patients, the information of residual urine volume and voiding volume was obtained in 89 patients, and detailed pre- and post-IVES bladder sensation information was available on 96 patients. Of the 89 patients, 47.2% (42/89) were responders and achieved a >50% reduction in the PVR. Obvious improvement in the PVR, defined as a >80% reduction, occurred in 27% (24/89) of the patients. VE developed in 76.4% (68/89) of the patients, and 30.3% (27/89) of the patients increased >50%. Significant improvements in the PVR and VE were observed during IVES treatment (P < .05). Based on the questionnaire, bladder sensation developed and was sustained in 44.8% (43/96) of the patients.IVES provides a promising method for improving the PVR and VE in a majority of patients with DU. Thus, IVES is worth to further study and carry out.


Marked improvements in survival of patients with rectal cancer in the Netherlands following changes in therapy, 1989-2006.
Since the 1990s, treatment of patients with rectal cancer has changed in the Netherlands. Aim of this study was to describe these changes in treatment over time and to evaluate their effects on survival. All patients in the Netherlands Cancer Registry with invasive primary rectal cancer diagnosed during the period 1989-2006 were selected. The Cochran-Armitage trend test was used to analyse trends in treatment over time. Multivariate relative survival analyses were performed to estimate relative excess risk (RER) of dying. In total, 40,888 patients were diagnosed with rectal cancer during the period 1989-2006. The proportion of patients with stages II and III disease receiving preoperative radiotherapy increased from 1% in the period 1989-1992 to 68% in the period 2004-2006 for younger patients (<75 years) and from 1% to 51% for older patients (>or=75 years), whereas the use of postoperative radiotherapy decreased. Administration of chemotherapy to patients with stage IV disease increased over time from 21% to 66% for patients younger than 75 years. Both males and females exhibited an increase in five-year relative survival from 53% to 60%. The highest increase in survival was found for patients with stage III disease. In the multivariate analyses survival improved over time for patients with stages II-IV disease. After adjustment for treatment variables, this improvement remained significant for patients with stages III and IV disease. The changes in therapy for rectal cancer have led to a markedly increased survival. Patients with stage III disease experienced the greatest improvement in survival.


Effects of imidacloprid and thiamethoxam on the development and reproduction of the soybean aphid Aphis glycines.
The soybean aphid Aphis glycines Matsumura (Hemiptera: Aphididae) is a primary pest of soybeans and poses a serious threat to soybean production. Our studies were conducted to understand the effects of different concentrations of insecticides (imidacloprid and thiamethoxam) on A. glycines and provided critical information for its effective management. Here, we found that the mean generation time and adult and total pre-nymphiposition periods of the LC50 imidacloprid- and thiamethoxam-treatment groups were significantly longer than those of the control group, although the adult pre-nymphiposition period in LC30 imidacloprid and thiamethoxam treatment groups was significantly shorter than that of the control group. Additionally, the mean fecundity per female adult, net reproductive rate, intrinsic rate of increase, and finite rate of increase of the LC30 imidacloprid-treatment group were significantly lower than those of the control group and higher than those of the LC50 imidacloprid-treatment group (P < 0.05). Moreover, both insecticides exerted stress effects on A. glycines, and specimens treated with the two insecticides at the LC50 showed a significant decrease in their growth rates relative to those treated with the insecticides at LC30. These results provide a reference for exploring the effects of imidacloprid and thiamethoxam on A. glycines population dynamics in the field and offer insight to agricultural producers on the potential of low-lethal concentrations of insecticides to stimulate insect reproduction during insecticide application.


Immunologic Insights on the Membrane Proximal External Region: A Major Human Immunodeficiency Virus Type-1 Vaccine Target.
Broadly neutralizing antibodies (bNAbs) targeting conserved regions within the human immunodeficiency virus type-1 (HIV-1) envelope glycoprotein (Env) can be generated by the human immune system and their elicitation by vaccination will be a key point to protect against the wide range of viral diversity. The membrane proximal external region (MPER) is a highly conserved region within the Env gp41 subunit, plays a major role in membrane fusion and is targeted by naturally induced bNAbs. Therefore, the MPER is considered as an attractive vaccine target. However, despite many attempts to design MPER-based immunogens, further study is still needed to understand its structural complexity, its amphiphilic feature, and its limited accessibility by steric hindrance. These particular features compromise the development of MPER-specific neutralizing responses during natural infection and limit the number of bNAbs isolated against this region, as compared with other HIV-1 vulnerability sites, and represent additional hurdles for immunogen development. Nevertheless, the analysis of MPER humoral responses elicited during natural infection as well as the MPER bNAbs isolated to date highlight that the human immune system is capable of generating MPER protective antibodies. Here, we discuss the recent advances describing the immunologic and biochemical features that make the MPER a unique HIV-1 vulnerability site, the different strategies to generate MPER-neutralizing antibodies in immunization protocols and point the importance of extending our knowledge toward new MPER epitopes by the isolation of novel monoclonal antibodies. This will be crucial for the redesign of immunogens able to skip non-neutralizing MPER determinants.


Math anxiety and math motivation in online learning during stress: The role of fearful and avoidance temperament and implications for STEM education.
Students' math motivation can predict engagement, achievement, and career interest in science, technology, engineering, and mathematics (STEM). However, it is not well understood how personality traits and math anxiety may be linked to different types or qualities of math motivation, particularly during high-stress times such as the COVID-19 pandemic. In this study, we examined how fearful or avoidant temperaments contribute to math anxiety and math motivations for college students during the COVID-19 pandemic. Ninety-six undergraduate students from a large public university were assessed on temperamental fear, math anxiety, and math motivation in an online math course. Results showed that higher levels of temperamental fear are directly linked to higher levels of math anxiety. In addition, temperamental fear is indirectly linked to higher levels of autonomous motivation (i.e., intrinsic motivation and identified regulation) and lower levels of controlled motivation (i.e., external regulation) through math anxiety. Results have implications for helping students at high risk for both high math anxiety and for low motivation to engage in math learning.


Clinical course of patients with familial early-onset Alzheimer's disease potentially lacking senile plaques bearing the E693Δ mutation in amyloid precursor protein.
Oligomeric amyloid β (Aβ) is currently considered to induce Alzheimer's disease (AD). We examined 2 patients with familial AD who possessed the Osaka (E693Δ) mutation in amyloid precursor protein. To the best of our knowledge, these patients are the first AD cases presumably affected with Aβ oligomers in the absence of senile plaques, and they support the Aβ oligomer hypothesis. We evaluated the clinical course, neuropsychological data, cerebrospinal fluid biomarker levels, magnetic resonance imaging (MRI) scans, fluorodeoxyglucose-positron emission tomography (PET) scans, and Pittsburgh compound B (PiB)-PET images of these patients. In the early stages, these patients developed memory disturbances in a similar rate to patients with sporadic AD. Despite their memory disturbances, both patients showed only limited brain atrophy on MRI and little amyloid accumulation on PiB-PET. Subsequent to the development of memory disturbances, both patients suffered from motor dysfunction, probably due to cerebellar ataxia, and, within a few years, the patients fell into an apallic state. Familial AD patients with Osaka (E693Δ) mutation show severe dementia, cerebellar ataxia, and gait disturbances.


Predicting the graft diameter of the peroneus longus tendon for anterior cruciate ligament reconstruction.
The aim of this study was to evaluate the correlation between various anthropometric parameters and the graft diameter of the peroneus longus tendon (PLT).We retrospectively analyzed the data of 156 patients who underwent anterior cruciate ligament reconstruction (ACLR) with the PLT graft at our institution. Anthropometric parameters, including height, weight, gender, age, duration of injury, and preinjury activity levels, were recorded. t tests, correlation coefficients (Pearson r), and a multiple linear regression analysis were used to evaluate the influence of these anthropometric variables on the diameter of the graft obtained.The mean PLT graft diameter was 8.3 mm, and 21 patients (13.5%) had a diameter less than 8 mm; 85 patients (54.5%) had a diameter between 8 and 9 mm, and 50 patients (32.0%) had a diameter greater than or equal to 9 mm. The correlation analysis showed that height (P < .001), weight (P < .001), and duration of injury (P = .012) were significantly related to graft diameter. On the basis of these 3 predictors, the following regression equation was obtained: Diameter = 2.28 + 0.028*height (cm) + 0.013*weight (kg) + 0.289*duration of injury (0 or 1). Patients who were short and shin were more likely to own smaller graft diameters (<8 mm), especially the one ruptured his or her anterior cruciate ligament (ACL) over 3 months.Height, weight, and duration of injury were associated with the diameter of PLT. They are important preoperative information for surgeon about the size of PLT and can be used for alternative graft source planning and patient counseling.Level of evidence: IV.


The kinetics of biological phase transitions manifested by sigmoid time curves: a review of approaches.
Experimental biological data showing a time course of sigmoid shape are customarily interpreted in terms of mass-action chemical kinetic theory of reactions in free solution. But the reactive sites in biological systems are often located in solid or structured cellular components through which cooperative interactions may occur, resulting in phase transitions. In inorganic systems, the time course of phase transitions often shows experimental curves of sigmoid shape, and this may be predicted theoretically. Therefore it is suggested that when sigmoid time curves are observed in biological systems containing solid or structured components, mathematical analysis based on cooperative interactions and phase transitions is more likely to be valid than analysis based on mass-action concepts.


Evolution in interacting species alters predator life-history traits, behaviour and morphology in experimental microbial communities.
Predator-prey interactions heavily influence the dynamics of many ecosystems. An increasing body of evidence suggests that rapid evolution and coevolution can alter these interactions, with important ecological implications, by acting on traits determining fitness, including reproduction, anti-predatory defence and foraging efficiency. However, most studies to date have focused only on evolution in the prey species, and the predator traits in (co)evolving systems remain poorly understood. Here, we investigated changes in predator traits after approximately 600 generations in a predator-prey (ciliate-bacteria) evolutionary experiment. Predators independently evolved on seven different prey species, allowing generalization of the predator's evolutionary response. We used highly resolved automated image analysis to quantify changes in predator life history, morphology and behaviour. Consistent with previous studies, we found that prey evolution impaired growth of the predator, although the effect depended on the prey species. By contrast, predator evolution did not cause a clear increase in predator growth when feeding on ancestral prey. However, predator evolution affected morphology and behaviour, increasing size, speed and directionality of movement, which have all been linked to higher prey search efficiency. These results show that in (co)evolving systems, predator adaptation can occur in traits relevant to foraging efficiency without translating into an increased ability of the predator to grow on the ancestral prey type.


An homozygous mutation in KCNK3 is associated with an aggressive form of hereditary pulmonary arterial hypertension.
Pulmonary arterial hypertension (PAH) is a rare devastating disease characterized by a high genetic heterogeneity with several related genes recently described, including BMPR2,TBX4 and KCNK3. The association between KCNK3 and PAH has been recently identified, but the prognosis and phenotype associated with these mutations have been poorly described. We studied a series of 136 idiopathic and hereditary PAH Spanish patients for BMPR2, TBX4 and KCNK3 mutations. We report the results of KCNK3 in which we were able to describe two new mutations (p.Gly106Arg and p.Leu214Arg) in three patients. The first one was found in a patient belonging to a consanguineous Romani family, who carried a homozygous mutation in KCNK3 and developed a severe and early form of the disease. To the best of our knowledge, this is the first time that a homozygous mutation in KCNK3 is reported in a PAH patient. The second one was found in a patient who presented at the young adult age a severe form of the disease. The present report supports the contribution of KCNK3 mutations to the genetic etiology of PAH and strongly suggests that mutations in KCNK3 follow incomplete dominance with worsening of the clinical features in homozygous patients.


Risk of cerebral hemorrhage in mild traumatic brain injury and antithrombotic treatment.
The observation time in mild traumatic brain injury (mTBI) is controversial. Our aim was to assess the risk of neurological complications in mTBI with and without antithrombotic treatment. We retrospectively evaluated patients with mTBI seen in the emergency room for 3 years. We considered MTBI those with Glasgow ≥13 at admission. A cranial CT was performed in all cases with ≥1 risk factor at admission and at 24 h in those with neurological impairment or initial pathological cranial CT. Complications in the following 3 months were retrospectively reviewed. We evaluated 907 patients with a mean age of 73 ± 19 years. Ninety-one percent presented risk factors, with 60% on antithrombotic treatment. We detected 11% of initial brain hemorrhage, 0.4% at 24 h, and no cases at 3 months. Antithrombotic treatment was not associated with an increased risk of brain hemorrhage (9.9% with vs 11.9% without treatment, p = 0.3). 39% of the hemorrhages presented neurological symptoms (18% post-traumatic amnesia, 12% headache, 8% vomiting, 1% seizures), with 78.4% having mild symptoms. Of the 4 hemorrhages detected at 24 h, 3 were asymptomatic and one case that worsened the initial headache. No asymptomatic patient without lesion on initial clinical cranial CT presented at 24 h. Our study suggests that patients with asymptomatic mTBI, without a lesion on the initial cranial CT, would not require the observation period or CT control regardless of antithrombotic treatment or INR level.


CpGassoc: an R function for analysis of DNA methylation microarray data.
With the increasing availability of high-density methylation microarrays, there has been growing interest in analysis of DNA methylation data. We have developed CpGassoc, an R package that can efficiently perform the statistical analysis needed for increasingly large methylation datasets. CpGassoc is a modular, expandable package with functions to perform rapid analyses of DNA methylation data via fixed or mixed effects models, to perform basic quality control, to carry out permutation tests, and to display results via an array of publication-quality plots. CpGassoc is implemented in R and is freely available at http://genetics.emory.edu/conneely; we are in the process of submitting it to CRAN.


Cardiac Arrest Following Drug Abuse with Intravenous Tapentadol: Case Report and Literature Review.
BACKGROUND Tapentadol is a centrally acting opioid analgesic, with a dual mode of action, as a norepinephrine reuptake inhibitor and an agonist of the μ-opioid receptor (MOR). Tapentadol is used for the management of musculoskeletal pain, and neuropathic pain associated with diabetic peripheral neuropathy. CASE REPORT A 32-year-old woman attended hospital for evaluation of an intractable headache. Computed tomography and magnetic resonance imaging of the brain were negative. She was found unresponsive in the bathroom on the day following hospital admission, and despite resuscitative measures, the patient died following cardiac arrest. Autopsy toxicology revealed significantly elevated levels of tapentadol, and bedside evidence suggested that the patient had self-administered this medication intravenously before her death. CONCLUSIONS We report a rare adverse effect of tapentadol causing respiratory depression leading to cardiac arrest. Medical examiners and forensic toxicologists should be aware of the toxicity of this novel opiate drug.


Gold nanoparticle-fluorophore complexes: sensitive and discerning "noses" for biosystems sensing.
Gold nanoparticles (NPs) efficiently quench adsorbed fluorophores. Upon disruption of such complexes by an analyte, fluorescence turn-on is observed. By judicious choice of the functionalized NP and the fluorophore, these complexes display different responses to analytes, thus leading to versatile yet simple array-based sensor platforms. Using this strategy, we can identify proteins in buffer and serum, distinguish between both different species and different strains of bacteria, and differentiate between healthy, cancerous, and metastatic human and murine cells.


[18F-FDG PET/CT Metabolic Parameters and Circulating Tumour DNA Mutation Abundance in Diffuse Large B-Cell Lymphoma: Correlation and Survival Analysis].
To investigate the correlation between 18Fluoro-deoxyglucose positron emission tomography/computed tomography (18F-FDG PET/CT) metabolic parameters and peripheral blood circulating tumour DNA (ctDNA) in patients with diffuse large B-cell lymphoma (DLBCL), and the prognostic value of these two types of parameters in predicting progression-free survival (PFS). Clinical, PET/CT and ctDNA data of DLBCL patients who underwent peripheral blood ctDNA testing and corresponding PET/CT scans during the same period were retrospectively analyzed. At the time of ctDNA sampling and PET scan, patients were divided into baseline and relapsed/refractory (R/R) groups according to different disease conditions. CtDNA mutation abundance was expressed as variant allele frequency (VAF), including maximum VAF (maxVAF) and mean VAF (meanVAF). Total metabolic tumour volume (TMTV) and total lesion glycolysis (TLG) were obtained by the 41% maximum normalized uptake value method, and the distance between the two farthest lesions (Dmax) was used to assess the correlation between PET parameters and ctDNA mutation abundance using Spearman correlation analysis. The receiver operating characteristic (ROC) curves were used to obtain the optical cut-off values of those parameters in predicting PFS in the baseline and R/R groups, respectively. Survival curves were outlined using the Kaplan-Meier method and log-rank test was performed to compare survival differences. A total of 67 DLBCL patients [28 males and 39 females, median age 56.0(46.0, 67.0) years] were included and divided into baseline group (29 cases) and R/R group (38 cases). Among these PET parameters, baseline TMTV, TLG, and Dmax were significantly correlated with baseline ctDNA mutation abundance, except for maximum standardized uptake value (SUVmax) (maxVAF vs TMTV: r=0.711; maxVAF vs TLG: r=0.709; maxVAF vs Dmax: r=0.672; meanVAF vs TMTV: r=0.682; meanVAF vs TLG: r=0.677; meanVAF vs Dmax: r=0.646). While in all patients, these correlations became weaker significantly. Among R/R patients, only TMTV had a weak correlation with meanVAF (r=0.376). ROC analysis showed that, the specificity of TMTV, TLG and Dmax in predicting PFS was better than mutation abundance, while the sensitivity of ctDNA mutation abundance was better. Except R/R patients, TMTV, TLG, Dmax, and VAF were significantly different at normal/elevated lactate dehydrogenase in baseline group and all patients (all P<0.05). Survival curves indicated that high TMTV (>109.5 cm3), high TLG (>2 141.3), high Dmax (>33.1 cm) and high VAF (maxVAF>7.74%, meanVAF>4.39%) were risk factors for poor PFS in baseline patients, while only high VAF in R/R patients (both maxVAF and meanVAF >0.61%) was a risk factor for PFS. PET-derived parameters correlate well with ctDNA mutation abundance, especially in baseline patients. VAF of ctDNA predicts PFS more sensitively than PET metabolic parameters, while PET metabolic tumour burden with better specificity. TMTV, TLG and VAF all have good prognostic value for PFS. PET/CT combined with ctDNA has potential for further studies in prognostic assessment and personalized treatment.


MUC1 alters oncogenic events and transcription in human breast cancer cells.
MUC1 is an oncoprotein whose overexpression correlates with aggressiveness of tumors and poor survival of cancer patients. Many of the oncogenic effects of MUC1 are believed to occur through interaction of its cytoplasmic tail with signaling molecules. As expected for a protein with oncogenic functions, MUC1 is linked to regulation of proliferation, apoptosis, invasion, and transcription. To clarify the role of MUC1 in cancer, we transfected two breast cancer cell lines (MDA-MB-468 and BT-20) with small interfering (si)RNA directed against MUC1 and analyzed transcriptional responses and oncogenic events (proliferation, apoptosis and invasion). Transcription of several genes was altered after transfection of MUC1 siRNA, including decreased MAP2K1 (MEK1), JUN, PDGFA, CDC25A, VEGF and ITGAV (integrin alphav), and increased TNF, RAF1, and MMP2. Additional changes were seen at the protein level, such as increased expression of c-Myc, heightened phosphorylation of AKT, and decreased activation of MEK1/2 and ERK1/2. These were correlated with cellular events, as MUC1 siRNA in the MDA-MB-468 line decreased proliferation and invasion, and increased stress-induced apoptosis. Intriguingly, BT-20 cells displayed similar levels of apoptosis regardless of siRNA, and actually increased proliferation after MUC1 siRNA. These results further the growing knowledge of the role of MUC1 in transcription, and suggest that the regulation of MUC1 in breast cancer may be more complex than previously appreciated. The differences between these two cell lines emphasize the importance of understanding the context of cell-specific signaling events when analyzing the oncogenic functions of MUC1, and caution against generalizing the results of individual cell lines without adequate confirmation in intact biological systems.


Metabolic Syndrome, Gamma-Glutamyl Transferase, and Risk of Sudden Cardiac Death.
Background: Metabolic syndrome is associated with a significantly increased risk of sudden cardiac death (SCD). However, whether temporal changes in the metabolic syndrome status are associated with SCD is unknown. We aimed to determine whether metabolic syndrome and gamma-glutamyl transferase (ɣ-GTP), including their temporal changes, are associated with the risk of SCD. Methods: We performed a nationwide population-based analysis using the Korean National Health Insurance Service. People who underwent a national health check-up in 2009 and 2011 were enrolled. The influence of metabolic syndrome and ɣ-GTP on SCD risk was evaluated. Results: In 2009, 4,056,423 (848,498 with metabolic syndrome) people underwent health screenings, 2,706,788 of whom underwent follow-up health screenings in 2011. Metabolic syndrome was associated with a 50.7% increased SCD risk (adjusted hazard ratio (aHR) = 1.507; p < 0.001). The SCD risk increased linearly as the metabolic syndrome diagnostic criteria increased. The ɣ-GTP significantly impacted the SCD risk; the highest quartile had a 51.9% increased risk versus the lowest quartile (aHR = 1.519; p < 0.001). A temporal change in the metabolic syndrome status and ɣ-GTP between 2009 and 2011 was significantly correlated with the SCD risk. Having metabolic syndrome in 2009 or 2011 indicated a lower SCD risk than having metabolic syndrome in 2009 and 2011 but a higher risk than having no metabolic syndrome. People with a ≥20-unit increase in ɣ-GTP between 2009 and 2011 had an 81.0% increased SCD risk versus those with a change ≤5 units (aHR = 1.810; p < 0.001). Conclusions: Metabolic syndrome and ɣ-GTP significantly correlated with an increased SCD risk. SCD was also influenced by temporal changes in the metabolic syndrome status and ɣ-GTP, suggesting that appropriate medical treatment and lifestyle modifications may reduce future SCD risk.


One year of burns at a role 3 Medical Treatment Facility in Afghanistan.
Historically, burns have formed a significant proportion of the casualties of war. The management of burns in recent conflict has been found to be a resource-heavy undertaking, though its impact on both personnel and resources in current conflicts is unclear. A case analysis has been carried out in order to quantify the logistical impact of the management of burns on Role 3 Medical Treatment Facility (MTF) infrastructure and to examine if and how the cause and management of burns have evolved in early 21st century conflict. All casualties treated for burns at a Role 3 MTF over one calendar year were identified and scanned copies of their notes obtained from the UK Joint Theatre Trauma Registry and retrospectively analysed. 88 of the 1461 (6.0%) trauma patients presenting to the Role 3 MTF over the year were treated for burns of whom 52.3% were combat troops and 45.4% civilians. Half of the burns were caused by non-conflict related mechanisms; the two commonest mechanisms were flame burns in 38/88 mostly non-conflict related cases and blast in 30/88 cases most of which were conflict related. The management of burns at war is a complex process. It is further confounded by the management of civilians with non-conflict related burns, which places a predictable strain on Role 3 MTF resources: theatre time, nursing time, dressing resources and bed space. This must be planned for in current and future deployed operations.


Purification and general properties of the DNA-binding protein (P16) from rat liver mitochondria.
The mitochondrial DNA-binding protein P16 was isolated from rat liver mitochondrial lysates by affinity chromatography on single strand DNA agarose and separated from DNA in the preparation by alkaline CsCl isopycnic gradients. The top fraction of the gradients contained a single polypeptide species (Mr approximately equal to 15,200) based upon SDS PAGE. Digestion of single strand DNA-bound P16 with proteinase K produced a protease-insensitive, DNA-binding fragment (Mr approximately equal to 6,000) that has been purified by essentially the same procedures used for intact P16. The partial amino acid compositions for P16 and the DNA-binding fragment were obtained by conventional methods. Analysis of subcellular fractions revealed that nearly all of the cellular P16 was located in the mitochondria and that only trace amounts of protein of comparable electrophoretic mobility could be isolated from the nuclear or cytoplasmic fractions. The labeling of P16 with [35S]methionine in primary rat hepatocyte cultures was inhibited by more than 90% by the cytoplasmic translation inhibitor cycloheximide, but unaffected by the mitochondrial-specific agent chloramphenicol. These results indicate that P16 is synthesized on cytoplasmic ribosomes and imported into the mitochondria. The addition of purified P16 to deproteinized mitochondrial DNA resulted in the complete protection of the labeled nascent strands of displacement loops against branch migrational loss during cleavage of parental DNA with SstI, thus providing strong evidence that P16 is the single entity required for this in vitro function. Incubation of P16 with single strand phi X174 DNA, double strand (RF) phi X174 DNA, or Escherichia coli ribosomal RNA and subsequent analysis of the nucleic acid species for bound protein indicated a strong preference of P16 for single strand DNA and no detectable affinity for RNA or double strand DNA. Examination of P16-single strand phi X174 DNA complexes by direct electron microscopy revealed thickened, irregular fibers characteristic of protein-associated single strand DNA.


Efficient targeted mutagenesis in Borrelia burgdorferi.
Genetic studies in Borrelia burgdorferi have been hindered by the lack of a nonborrelial selectable marker. Currently, the only selectable marker is gyrB(r), a mutated form of the chromosomal gyrB gene that encodes the B subunit of DNA gyrase and confers resistance to the antibiotic coumermycin A(1). The utility of the coumermycin-resistant gyrB(r) gene for targeted gene disruption is limited by a high frequency of recombination with the endogenous gyrB gene. A kanamycin resistance gene (kan) was introduced into B. burgdorferi, and its use as a selectable marker was explored in an effort to improve the genetic manipulation of this pathogen. B. burgdorferi transformants with the kan gene expressed from its native promoter were susceptible to kanamycin. In striking contrast, transformants with the kan gene expressed from either the B. burgdorferi flaB or flgB promoter were resistant to high levels of kanamycin. The kanamycin resistance marker allows efficient direct selection of mutants in B. burgdorferi and hence is a significant improvement in the ability to construct isogenic mutant strains in this pathogen.


Molecular genetic description of the cryptic wheat-Aegilops geniculata introgression carrying rust resistance genes Lr57 and Yr40 using wheat ESTs and synteny with rice.
The cryptic wheat-alien translocation T5DL.5DS-5MgS(0.95), with leaf rust and stripe rust resistance genes Lr57 and Yr40 transferred from Aegilops geniculata (UgMg) into common wheat, was further analyzed. Molecular genetic analysis using physically mapped ESTs showed that the alien segment in T5DL.5DS-5MgS(0.95) represented only a fraction of the wheat deletion bin 5DS2-0.78-1.00 and was less than 3.3 cM in length in the diploid wheat genetic map. Comparative genomic analysis indicated a high level of colinearity between the distal region of the long arm of chromosome 12 of rice and the genomic region spanning the Lr57 and Yr40 genes in wheat. The alien segment with genes Lr57 and Yr40 corresponds to fewer than four overlapping BAC or PAC clones of the syntenic rice chromosome arm 12L. The wheat-alien translocation breakpoint in T5DL.5DS-5MgS(0.95) was further localized to a single BAC clone of the syntenic rice genomic sequence. The small size of the terminal wheat-alien translocation, as established precisely with respect to Chinese Spring deletion bins and the syntenic rice genomic sequence, further confirmed the escaping nature of cryptic wheat-alien translocations in introgressive breeding. The molecular genetic resources and information developed in the present study will facilitate further fine-scale physical mapping and map-based cloning of the Lr57 and Yr40 genes.


Angiostatic effects of Brazilian green propolis and its chemical constituents.
Propolis, a resinous substance collected by honeybees from various plant sources, has several pharmacological actions, such as anti-tumor and anti-inflammatory effects. The aim of this study was to evaluate the anti-angiogenic effects of a water extract of Brazilian green propolis (WEP) and its constituents, caffeoylquinic acid derivatives, against angiogenic processes in human umbilical vein endothelial cells (HUVECs) in vitro. We also examined the anti-angiogenic effects of WEP against retinal neovascularization in a murine oxygen-induced retinopathy model in vivo. WEP and its constituents significantly suppressed vascular endothelial growth factor (VEGF)-induced HUVEC proliferation, migration, and tube formation in vitro. WEP and its caffeoylquinic acid derivatives suppressed VEGF-stimulated phosphorylation of mitogen-activated protein kinase in HUVECs (versus VEGF alone). Moreover, WEP (300 mg/kg/day, subcutaneously for 5 days) significantly suppressed retinal neovascularization in the murine oxygen-induced retinopathy model. These data indicate that (i) WEP has angiostatic effects against angiogenic processes in vitro and in an in vivo model of murine oxygen-induced retinopathy and (ii) the inhibitory effects of WEP against in vitro angiogenesis are chiefly derived from its caffeoylquinic acid derivatives. Judging from these findings, WEP and its caffeoylquinic acid derivatives may represent candidates for preventive or therapeutic agents against diseases caused by angiogenesis.


The competitive nature of signal transducer and activator of transcription complex formation drives phenotype switching of T cells.
Signal transducers and activators of transcription (STATs) are key molecular determinants of T-cell fate and effector function. Several inflammatory diseases are characterized by an altered balance of T-cell phenotypes and cytokine secretion. STATs, therefore, represent viable therapeutic targets in numerous pathologies. However, the underlying mechanisms by which the same STAT proteins regulate both the development of different T-cell phenotypes and their plasticity during changes in extracellular conditions remain unclear. In this study, we investigated the STAT-mediated regulation of T-cell phenotype formation and plasticity using mathematical modelling and experimental data for intracellular STAT signalling proteins. The close fit of our model predictions to the experimental data allows us to propose a potential mechanism for T-cell switching. According to this mechanism, T-cell phenotype switching is the result of the relative redistribution of STAT dimer complexes caused by the extracellular cytokine-dependent STAT competition effects. The developed model predicts that the balance between the intracellular STAT species defines the amount of the produced cytokines and thereby T-cell phenotypes. The model predictions are consistent with the experimentally observed interferon-γ to interleukin-10 switching that regulates human T helper type 1/type 1 regulatory T-cell responses. The proposed model is applicable to a number of STAT signalling circuits.