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The thalamic medial geniculate body (MGB) is uniquely positioned within the neural tinnitus networks. Deep brain stimulation (DBS) of the MGB has been proposed as a possible novel treatment for tinnitus, yet mechanisms remain elusive. The aim of this study was to characterize neurophysiologic hallmarks in the MGB after noise exposure and to assess the neurophysiological effects of electrical stimulation of the MGB. Fourteen male Sprague-Dawley rats were included. Nine subjects were unilaterally exposed to a 16-kHz octave-band noise at 115 dB for 90 min, five received sham exposure. Single units were recorded from the contralateral MGB where spontaneous firing, coefficient of variation, response type, rate-level functions, and thresholds were determined. Local field potentials and electroencephalographical (EEG) recordings were performed before and after high-fre-quency DBS of the MGB. Thalamocortical synchronization and power were analyzed. In total, 214 single units were identified (n = 145 in noise-exposed group, n = 69 in control group). After noise exposure, fast-responding neurons become less respon-sive or nonresponsive without change to their spontaneous rate, whereas sustained-and suppressed-type neurons exhibit enhanced spontaneous activity without change to their stimulus-driven activity. MGB DBS suppressed thalamocortical synchroni-zation in the 13 and c bands, supporting suppression of thalamocortical synchronization as an underlying mechanism of tinnitus suppression by high frequency DBS. These findings contribute to our understanding of the neurophysiologic consequences of noise exposure and the mechanism of potential DBS therapy for tinnitus.NEW & NOTEWORTHY Separate functional classes of MGB neurons might have distinct roles in tinnitus pathophysiology. After noise exposure, fast-responding neurons become less responsive or nonresponsive without change to their spontaneous firing, whereas sustained and suppressed neurons exhibit enhanced spontaneous activity without change to their stimulus-driven activ-ity. Furthermore, results suggest desynchronization of thalamocortical 13 and c oscillations as a mechanism of tinnitus suppres-sion by MGB DBS.

Rationale & Objective In the general population, cognitive impairment is associated with increased mortality, and higher levels of education are associated with lower risks for cognitive impairment and mortality. These associations are not well studied in patients receiving long-term hemodialysis and were the focus of the current investigation. Study Design Prospective cohort study. Setting & Participants Adult hemodialysis patients treated in 20 Italian dialysis clinics. Exposures Patients’ cognitive function across 5 domains (memory, attention, executive function, language, and perceptual-motor function), measured using a neuropsychological assessment comprising 10 tests; and patients’ self-reported years of education. Outcome All-cause mortality. Analytical Approach Nested multivariable Cox regression models were used to examine associations of cognition (any domain impaired, number of domains impaired, and global function score from principal components analysis of unadjusted test scores) and education with mortality and whether there were interactions between them. Results 676 (70.6%) patients participated, with a median age of 70.9 years and including 38.8% women. Cognitive impairment was present in 79.4% (527/664; 95% CI, 76.3%-82.5%). During a median follow-up of 3.3 years (1,874 person-years), 206 deaths occurred. Compared to no cognitive impairment, adjusted HRs for mortality were 1.77 (95% CI, 1.07-2.93) for any impairment, 1.48 (95% CI, 0.82-2.68) for 1 domain impaired, 1.88 (95% CI, 1.01-3.53) for 2 domains, and 2.01 (95% CI, 1.14-3.55) for 3 to 5 domains. The adjusted HR was 0.68 (95% CI, 0.51-0.92) per standard deviation increase in global cognitive function score. Compared with primary or lower education, adjusted HRs were 0.79 (95% CI, 0.53-1.20) for lower secondary and 1.13 (95% CI, 0.80-1.59) for upper secondary or higher. The cognition-by-education interaction was not significant (P = 0.7). Limitations Potential selection bias from nonparticipation and missing data; no data for cognitive decline; associations with education were not adjusted for other socioeconomic factors. Conclusions Cognitive impairment is associated with premature mortality in hemodialysis patients. Education does not appear to be associated with mortality.

Deep brain stimulation of the central auditory pathway is emerging as a promising treatment modality for tinnitus. Within this pathway, the dorsal cochlear nucleus (DCN) plays a key role in the pathophysiology of tinnitus and is believed to be a tinnitus generator. We hypothesized that high-frequency stimulation (HFS) of the DCN would influence tinnitus-related abnormal neuronal activity within the auditory pathway and hereby suppress tinnitus. To this end, we assessed the effect of HFS of the DCN in a noise-induced rat model of tinnitus. The presence of tinnitus was verified using the gap prepulse inhibition of the acoustic startle response paradigm. Hearing thresholds were determined before and after noise trauma by measuring the auditory brainstem responses. In addition, changes in neuronal activity induced by noise trauma and HFS were assessed using c-Fos immunohistochemistry in related structures. Results showed tinnitus development after noise trauma and hearing loss ipsilateral to the side exposed to noise trauma. During HFS of the DCN, tinnitus was suppressed. There was no change in c-Fos expression within the central auditory pathway after HFS. These findings suggest that DCN-HFS changes patterns of activity and results in information lesioning within the network and hereby blocking the relay of abnormal tinnitus-related neuronal activity.

A 4-year-old boy, a β-thalassemia (β-thal) carrier, with an unexplained severe chronic microcytic anemia was referred to us. Sequencing of the α-globin genes revealed a Hb Charlieu [α106(G13)Leu→Pro, HBA1: c.320T>C, p.Leu107Pro] mutation present on both HBA1 genes. Quantitative polymerase chain reaction (qPCR) confirmed αCharlieu mRNA in the proband and his parents, showing that the mutation does not affect mRNA stability. However, we were unable to detect the Hb Charlieu protein by capillary electrophoresis (CE), reverse phase electrophoresis, cation exchange electrophoresis or isoelectric focusing. Mass spectrometry (MS) allowed us to confirm the presence of the Hb Charlieu peptide in erythrocyte progenitors. These findings suggest that the mutation affects the stability of αCharlieu. As hemoglobin (Hb) heat stability tests showed no abnormalities in erythrocytes, we speculated that αCharlieu is already degraded during red blood cell (RBC) development. The clinical severity in the proband and the presence of new methylene blue-stained aggregates in his reticulocytes indicates that incorporation of αCharlieu destabilizes Hb. This, combined with an excess of unstable free α-globins as the result of β-thal minor, results in severely impaired erythropoiesis and, as a consequence, severe and chronic microcytic anemia in the proband.

Objective Secondary education has lifelong implications for well-being. We evaluated associations between the timing and duration of low socioeconomic status (SES) during childhood and academic achievement in secondary school. Methods Cohort design. The structured modeling approach was used to evaluate life-course models for associations between the duration and timing of low SES (across ages 4–5, 6–7, 8–9, 10–11 years) and Grade 7 (median age 12.5 years) reading and numeracy achievement. Linear regressions were fitted for 4 critical period models (each including low SES at 1 age), 1 sensitive period model (including low SES at all ages), and 2 strict accumulation models (including low SES duration in linear/categorical form). Results Of 3734 children, 1718 (46.1%), 1749 (48.6%), 1797 (49.3%), and 1779 (49.8%) experienced low SES at 4 to 5, 6 to 7, 8 to 9, and 10 to 11 years, respectively. For reading, the sensitive period model fitted best. Reading z-score coefficients for low SES (reference: high SES) at 4 to 5, 6 to 7, 8 to 9, and 10 to 11 years were: −0.20, −0.18, −0.02, and −0.22. For numeracy, the categorical strict accumulation model, with SES-by-sex interaction, fitted best. Numeracy z-score coefficients for 1, 2, 3, and 4 periods of low SES (reference: 0 periods) were: −0.38, −0.42, −0.54, and −0.77 for boys, and −0.23, −0.34, −0.42, and −0.54 for girls. Conclusions Low SES at all ages studied except 8 to 9 years has cumulative associations with poorer Grade 7 reading. Longer duration of low SES from 4 to 11 years is associated with poorer Grade 7 numeracy, with stronger associations for boys than girls. Academic interventions should be targeted toward children with persistently low SES.

Background Neuromodulation is a promising treatment modality for tinnitus, especially in chronic and severe cases. The auditory thalamus plays a key role in the pathophysiology of tinnitus, as it integrates and processes auditory and limbic information. Objective The effect of high frequency stimulation and low frequency stimulation of the medial geniculate bodies on tinnitus in a noise-induced tinnitus rat model is assessed. Materials and methods Presence of tinnitus was verified using the gap-induced prepulse inhibition of the acoustic startle response paradigm. Hearing thresholds were determined before and after noise trauma with auditory brainstem responses. Anxiety-related side-effects were evaluated in the elevated zero maze and open field. Results Results show tinnitus development after noise exposure and preserved hearing thresholds of the ear that was protected from noise trauma. We found that high frequency stimulation of the medial geniculate bodies suppressed tinnitus. This effect maintained directly after stimulation when the stimulator was turned off. Low frequency stimulation did not have any effects on the gap:no-gap ratio of the acoustic startle response. Conclusion High frequency stimulation of the MGB has a direct and residual suppressing effect on tinnitus in this animal model. Low frequency stimulation of the MGB did not inhibit tinnitus.

ObjectiveThe aim was to compare quality of life (QoL) among children and adolescents with different stages of chronic kidney disease (CKD) and determine factors associated with changes in QoL.DesignCross-sectional.SettingThe Kids with CKD study involved five of eight paediatric nephrology units in Australia and New Zealand.PatientsThere were 375 children and adolescents (aged 6–18 years) with CKD, on dialysis or transplanted, recruited between 2013 and 2016.Main outcome measuresOverall and domain-specific QoL were measured using the Health Utilities Index 3 score, with a scale from −0.36 (worse than dead) to 1 (perfect health). QoL scores were compared between CKD stages using the Mann-Whitney U test. Factors associated with changes in QoL were assessed using multivariable linear and ordinal logistic regression.ResultsQoL for those with CKD stages 1–2 (n=106, median 0.88, IQR 0.63–0.96) was higher than those on dialysis (n=43, median 0.67, IQR 0.39–0.91, pConclusionsThe overall QoL and domains such as pain and emotion are substantially worse in children on dialysis compared with earlier stage CKD and those with kidney transplants.

Background Computerized Dynamic Posturography (CDP)was developed by the American space program to assess imbalance in astronauts, and eventually evolved into a clinical diagnostic tool. However it is not a specific measure of vestibular function. Vestibular Evoked Myogenic Potential testing (VEMPs) is a new clinical tool which is sensitive and specific for measuring otolithic pathology, especially in the atypical vestibular patient. Research question As posturography measures ability to maintain balance, and VEMP testing measures the structures responsible for this, we wondered if CDP results would correlate with VEMP abnormalities in the clinical setting. Methods We analysed 180 patients sequentially referred to our unit for vestibular complaints. All patients had a full battery of vestibular assessments. We correlated VEMP results with CDP results to look for abnormality patterns and correlations. An occasional patient’s only abnormality was on CDP Results There was a high rate of VEMP abnormalities seen, which correlates with the fact that our referral base consists of patients with chronic vestibular complaints. The rate of VEMP abnormalities was the same in patients with normal CDP and those with abnormal CDP. Significance Our results do not suggest that CDP is unnecessary, but we feel that they emphasize the idea that these tests are measuring two different aspects of balance control. In some patients, all assessments are abnormal, but in some patients only one assessment is abnormal, suggesting that these modalities measure different things and are all important in the diagnostic armamentarium. Hopefully in the near future, the use of virtual reality will reduce the cost of CDP to the point where it can be made widely accessible to patients and clinicians.

Due to the COVID- 19 outbreak in the Netherlands (March 2020) and the associated social distancing measures, families were enforced to stay at home as much as possible. Adolescents and their families may be particularly affected by this enforced proximity, as adolescents strive to become more independent. Yet, whether these measures impact emotional well-being in families with adolescents has not been examined. In this ecological momentary assessment study, we investigated if the COVID-19 pandemic affected positive and negative affect of parents and adolescents and parenting behaviors (warmth and criticism). Additionally, we examined possible explanations for the hypothesized changes in affect and parenting. To do so, we compared daily reports on affect and parenting that were gathered during two periods of 14 consecutive days, once before the COVID-19 pandemic (2018–2019) and once during the COVID-19 pandemic. Multilevel analyses showed that only parents’ negative affect increased as compared to the period before the pandemic, whereas this was not the case for adolescents’ negative affect, positive affect and parenting behaviors (from both the adolescent and parent perspective). In general, intolerance of uncertainty was linked to adolescents’ and parents’ negative affect and adolescents’ positive affect. However, Intolerance of uncertainty, nor any pandemic related characteristics (i.e. living surface, income, relatives with COVID-19, hours of working at home, helping children with school and contact with COVID-19 patients at work) were linked to the increase of parents’ negative affect during COVID-19. It can be concluded that on average, our sample (consisting of relatively healthy parents and adolescents) seems to deal fairly well with the circumstances. The substantial heterogeneity in the data however, also suggest that whether or not parents and adolescents experience (emotional) problems can vary from household to household. Implications for researchers, mental health care professionals and policy makers are discussed.

Background Chronic kidney disease (CKD) is a devastating illness associated with increased mortality, reduced quality of life, impaired growth, neurocognitive impairment and psychosocial maladjustment in children. There is growing evidence of socioeconomic disparities in health outcomes among children with CKD. Patient navigators are trained non-medical personnel who assist patients with chronic conditions journey through the continuum of care and transit across different care settings. They help vulnerable and underserved populations to better understand their diagnosis, treatment options, and available resources, guide them through complex medical systems, and help them to overcome barriers to health care access. Given the complexity and chronicity of the disease process and concerns that current models of care may not adequately support the provision of high-level care in children with CKD from socioeconomically disadvantaged backgrounds, a patient navigator program may improve the provision of care and overall health of children with CKD. Methods The NAV-KIDS2 trial is a multi-centre, staggered entry, waitlisted randomised controlled trial assessing the health benefits and costs of a patient navigator program in children with CKD (stages 3–5, on dialysis, and with kidney transplants), who are of low socioeconomic backgrounds. Across 5 sites, 210 patients aged from 3 to 17 years will be randomised to immediate receipt of a patient navigator intervention for 24 weeks or waitlisting with standard care until receipt of a patient navigator at 24 weeks. The primary outcome is child self-rated health (SRH) 6-months after completion of the intervention. Other outcomes include utility-based quality of life, caregiver SRH, satisfaction with healthcare, progression of kidney dysfunction, other biomarkers, missed school days, hospitalisations and mortality. The trial also includes an economic evaluation and process evaluation, which will assess the cost-effectiveness, fidelity and barriers and enablers of implementing a patient navigator program in this setting. Discussion This study will provide clear evidence on the effectiveness and cost-effectiveness of a new intervention aiming to improve overall health and well-being for children with CKD from socioeconomically disadvantaged backgrounds, through a high quality, well-powered clinical trial. Trial registration Prospectively registered (12/07/2018) on the Australian New Zealand Clinical Trials Registry (ACTRN12618001152213).

Objective Here, we present a 7-year-old patient suffering from severe haemolytic anaemia. The most common cause of chronic hereditary non-spherocytic haemolytic anaemia is red blood cell pyruvate kinase (PK-R) deficiency. Because red blood cells rely solely on glycolysis to generate ATP, PK-R deficiency can severely impact energy supply and cause reduction in red blood cell lifespan. We determined the underlying cause of the anaemia and investigated how erythroid precursors in the patient survive. Methods PK activity assays, Western blot and Sanger sequencing were employed to determine the underlying cause of the anaemia. Patient erythroblasts were cultured and reticulocytes were isolated to determine PK-R and PKM2 contribution to glycolytic activity during erythrocyte development. Results We found a novel homozygous mutation (c.583G>A) in the PK-R coding gene (PKLR). Although this mutation did not influence PKLR mRNA production, no PK-R protein could be detected in the red blood cells nor in its precursors. In spite of the absence of PK-R, the reticulocytes of the patient exhibited 20% PK activity compared with control. Western blotting revealed that patient erythroid precursors, like controls, express residual PKM2. Conclusions We conclude that PKM2 rescues glycolysis in PK-R-deficient erythroid precursors.

To determine the association of socioeconomic disadvantage and parent-rated health in children with chronic kidney disease (CKD). A total of 377 children (aged 6–18 years) with CKD stages I–V (n = 199), on dialysis (n = 43), or with a kidney transplant (n = 135) were recruited from 2012 to 2016 in Australia and New Zealand. Associations of five socioeconomic status (SES) components and the global SES index with parent-rated health of the child were examined using adjusted logistic regression. The median age of participants was 12.6 years (interquartile range (IQR) 8.9–15.5). In the entire cohort, the adjusted odds ratios (ORs) and 95% confidence intervals (CIs) for poor parent-rated health were 1.85 (1.13–3.03) for lower household income, 1.78 (1.08–2.96) for families that did not own their own home, 2.50 (1.50–4.16) for caregivers who rated their financial status as poor, 0.84 (0.51–1.38) for lower educational attainment, and 1.68 (1.04–2.72) for children whose primary caregivers were unemployed. With reference to the highest global SES index quartile, adjusted ORs for poor parent-rated health in descending order were 1.49 (0.69–3.21), 2.11 (1.06–4.20), and 2.20 (1.09–4.46), respectively. The association between low SES and poor parent-rated health was modified by CKD stage, where lower global SES index was independently associated with poor parent-rated health in children with CKD stages I–V, but not children on dialysis or with kidney transplants (p = 0.04). Low SES is associated with poor parent-rated health in children with CKD stages I–V, but not children on dialysis and with kidney transplants.

Background The efficacy of novel targeted therapies is often tested at the time of tumor recurrence. However, for glioblastoma (GBM) patients, surgical resections at recurrence are performed only in a minority of patients; therefore, molecular data are predominantly derived from the initial tumor. Molecular data of the initial tumor for patient selection into personalized medicine trials can therefore be used only when the specific genetic change is retained in the recurrent tumor. Methods In this study we determined whether EGFR amplification and expression of the most common mutation in GBMs (EGFRvIII) is retained at tumor recurrence. Because retention of genetic changes may be dependent on the initial treatment, we only used a cohort of GBM samples that were uniformly treated according to the current standard of care (ie, chemo-irradiation with temozolomide). Results Our data show that, in spite of some quantitative differences, the EGFR amplification status remains stable in the majority (84%) of tumors evaluated. EGFRvIII expression remained similar in 79% of GBMs. However, within the tumors expressing EGFRvIII at initial diagnosis, approximately one-half lose their EGFRvIII expression at tumor recurrence. Conclusions The relative stability of EGFR amplification indicates that molecular data obtained in the primary tumor can be used to predict the EGFR status of the recurrent tumor, but care should be taken in extrapolating EGFRvIII expression from the primary tumor, particularly when expressed at first diagnosis.

The Sydney rock oyster, Saccostrea glomerata, has been demonstrated as a useful biomonitor of estrogenic compounds following laboratory exposures, yet its utility in the assessment of estrogenic exposure and effects under field conditions requires investigation. To achieve this aim, S. glomerata were deployed in Newcastle, Australia in the effluent receiving marine waters of Burwood Beach WWTP (Burwood Beach "near"

Introduction: Newborn screening (NBS) techniques have been developed for several lysosomal storage disorders (LSDs), including Mucopolysaccharidosis type I (MPS I). MPS I is an LSD with a wide phenotypic spectrum that ranges from the severe Hurler phenotype to the attenuated Scheie phenotype. To improve the ethical discussion about NBS for MPS I, we performed an interview study to explore the experiences of MPS I patients and their parents with the timings of their diagnoses. Methods: We used a qualitative research approach consisting of 17 interviews with the parents of patients with all MPS I phenotypes and with patients with attenuated forms of MPS I. The interviews were audio-recorded, transcribed and subsequently analyzed to identify the main themes identified by the participants. Results: Five important themes, focusing on the experienced disadvantages of delayed diagnosis and the advantages and disadvantages of a hypothetical earlier diagnosis, were identified in our group of participants: 1) delayed diagnosis causing parental frustration, 2) delayed diagnosis causing patient frustration, 3) early diagnosis enabling reproductive decision-making, 4) early diagnosis enabling focusing on the diagnosis, and 5) early diagnosis enabling timely initiation of treatment. There was a remarkable similarity in the experiences with timing of diagnosis between parents of patients with the severe and the attenuated forms. Conclusion: This was the first study to explore the personal experiences of MPS I patients and their parents with diagnostic timing. Our study identified five important themes that are highly relevant to the ethical discussion on expanding NBS programs for MPS I. (C) 2012 Elsevier Inc. All rights reserved

Objectives To examine the effect of age on associations between household income and overall health from birth to adolescence, and whether age patterns vary by country. It is uncertain whether income-related health inequalities remain stable, widen, or narrow as children age, which impacts optimal timing of equity-focused interventions. Study design Systematic review (CRD42016038583) of MEDLINE, Embase, PsycINFO, CINAHL, SocINDEX (full-text), and EconLit (full-text) to April 2017. We included observational studies and trials in children and adolescents (0-18 years of age), examining age differences in associations between income and overall health (self-rated, clinician-rated, proxy-rated). One reviewer extracted data; 2 evaluated risk of bias. Results Thirty-eight articles containing 43 studies (30 cross-sectional, 13 cohort) were identified, from high-income (n = 39) and middle-income (n = 4) countries. In the US (n = 21), positive income-health associations emerged in early childhood, and these inequalities typically widened progressively into adolescence. Relative to 0- to 3-year-olds, ratios of income-health coefficients ranged from 1.10-3.71 for 4-8 years of age, 1.26-3.86 for 9-12 years of age, 1.36-6.71 for 13-17 years. In the United Kingdom and Ireland (n = 8), inequalities emerged in early-to-mid childhood, but age patterns were less consistent. In other high-income countries (Australia, Canada, France, Germany, Japan, Republic of Korea), inequalities mostly persisted or widened with age. In middle-income countries, inequalities appeared to narrow (Indonesia n = 2) or persist (Brazil n = 2) with age. Limitations are unclear/high risk of bias and dataset overlap for some studies. Conclusions In many countries, income-related health status inequalities persist or widen as children age. Interventions that improve health equity early in the life-course are needed.

To investigate in a randomised controlled trial the effect of basic elements of developmental care (incubator covers and positioning aids) on growth and neurodevelopment in infants born at32 weeks.Infants were randomised within 48 h of birth to a developmental care (DC) or standard care (C) group. Outcome measures at 1 and 2 years corrected age were growth, standardised neurological examinations, and mental (MDI) and psychomotor (PDI) development (Dutch version of the Bayley Scales of Infant Development II).192 infants were recruited (DC = 98; C = 94). Thirteen infants (DC = 7, C = 6) were excluded because they were admitted for5 days or died within the first 5 days. In total, 179 infants met the inclusion criteria. In-hospital mortality was 12/91 (13.2%) in the DC group and 8/88 (9.1%) in the C group. Assessments were carried out on 147 children (DC = 74, C = 73) at 1 year and 142 children (DC = 72, C = 70) at 2 years. No significant difference in growth, neurological outcomes or MDI was found. A positive trend in PDI at 1 year (p = 0.05) did not continue once the children reached 2 years. There was no difference found when neurological and developmental scores were combined.Basic developmental care has no positive effect on neurological and mental development or growth at 1 and 2 years of age in infants born at32 weeks. A positive effect on psychomotor development at 1 year did not continue at 2 years of age.

Background. In general practice, depression is often not recognized. As treatment of depression is effective, screening has been proposed as one solution to combat this 'hidden morbidity'. The results of screening programmes for depression, however, are inconsistent and most studies do not show a positive effect on patient outcomes. Patients do not always accept this diagnosis and hence do not receive proper treatment. Nothing is known about the tendency of those patients who screen positive for depression to accept treatment for their 'disclosed' disorder. Objective. In this study, we aimed to better understand the views of patients who screened positive in a screening programme for depression. Methods. We performed a qualitative study with semi-structured in-depth interviews with 17 patients. These adult patients (nine females), all suffering from major depressive disorder, were disclosed by a screening programme for depression performed within 11 Dutch general practices. The transcripts were independently analysed by two researchers using MAXqda2. Results. All patients appreciated the active way in which they were approached for screening. Fifteen of the 17 patients recognized the depressive symptoms but nine of them did not accept the diagnosis. The first explanation for resistance to the diagnosis of depression is fear of stigmatization and scepticism about the usefulness of labelling. Secondly, patients experienced their depressive symptoms as a normal and transitory reaction to adversity. Thirdly, patients had doubts about the necessity and effectiveness of treatment. Depressive symptoms, such as feelings of guilt, self-depreciation and fatigue, hamper help-seeking behaviour. Conclusions. We conclude that some patients with undisclosed depression, who took the trouble of going through a complete screening programme, felt aversion to being diagnosed as having depression. In the context of screening for depression, we recommend that the patients' view on depression be elicited before diagnosing and offering treatment

Adult Saccostrea glomerata were exposed to environmentally relevant concentrations of 4-nonylphenol (1microg/L and 100microg/L) and 17alpha-ethynylestradiol (5ng/L and 50ng/L) in seawater over 8 weeks. Exposures were performed to assess effects on vitellogenin induction and gonadal development during reproductive conditioning. Chronic direct estrogenicity within gonadal tissue was assessed via an estrogen receptor-mediated, chemical-activated luciferase reporter gene-expression assay (ER-CALUX). Estradiol equivalents (EEQ) were greatest in the 100microg/L 4-nonylphenol exposure (28.7+/-2.3ng/g tissue EEQ) while 17alpha-ethynylestradiol at concentrations of 50ng/L were 2.2+/-1.5ng/g tissue EEQ. Results suggest 4-nonylphenol may be accumulated in tissue and is partly resistant to biotransformation; maintaining its potential for chronic estrogenic action, while 17alpha-ethynylestradiol, although exhibiting greater estrogenic potency on biological endpoints possibly exerts its estrogenic action before being rapidly metabolised and/or excreted. A novel methodology was developed to assess vitellogenin using high-performance liquid chromatography (HPLC). Exposure to both 17alpha-ethynylestradiol (50ng/L) and 4-nonylphenol (100microg/L) produced increases in vitellogenin for females, whereas males exhibited increases in vitellogenin when exposed to 50ng/L 17alpha-ethynylestradiol only. Females exhibited greater vitellogenin responses than males at 50ng/L 17alpha-ethynylestradiol only. Histological examination of gonads revealed a number of individuals exhibiting intersex (ovotestis) in 50ng/L 17alpha-ethynylestradiol exposures. Male individuals in 1microg/L and 100microg/L 4-nonylphenol exposures and 5ng/L 17alpha-ethynylestradiol were at earlier stages of spermatogenic development than corresponding controls.

The observation that the use of antidepressants has strongly increased during the past decade implies that on a micro level doctors and patients more often decide that antidepressants are the appropriate treatment. Therefore, it is important to increase insight into patients' decision making regarding the use of antidepressants. The decision making model used in the present study was based on A. Bandura's (1986) social cognitive theory. Two cohorts of patients were recruited and followed for 9 months. Among patients who use antidepressants (N = 166), the stronger pros and weaker cons of discontinuation and self-efficacy predicted more proximal goal intentions. Goal intentions predicted discontinuation after 9 months. Among patients who had used antidepressants in the past (N = 73), stronger pros of discontinuation and the weaker perceived functions of antidepressants predicted a more positive evaluation of their present state, compared with when they still used antidepressants. These temporal comparisons, in turn, predicted renewed use of antidepressants after 9 months. The results provide a framework for supporting and influencing decision making with regard to the use of antidepressants.

Congenital dyserythropoietic anaemia type II (CDAII) is a rare autosomal recessive disease characterized by ineffective erythropoiesis, haemolysis, erythroblast morphological abnormalities, hypoglycosylation of some red blood cell membrane proteins, particularly band 3, and mutations in the SEC23B gene. We report the analysis of 101 patients from 91 families with a median follow-up of 23 years (range 0-65); 68 patients are newly reported. Clinical and haematological parameters were separately analysed in early infancy and thereafter, when feasible. Molecular analysis of the SEC23B gene confirmed the high heterogeneity of the defect, leading to the identification of 54 different mutations, 24 of which are newly described. To evaluate the genotype-phenotype correlation, patients were grouped according to their genotype (two missense mutations vs. one missense/one drastic mutation) and assigned to two different severity gradings based on laboratory data and on therapeutic needs; by this approach only a weak genotype-phenotype correlation was observed in the analysed groups.

Objective: There is concern that patients included in trials do not represent the true patient population and women in particular may selectively be excluded. We looked at trial data submitted to the European Medicines Agency (EMEA) by drug companies to achieve marketing authorization in Europe between 2000 and 2003. Methods: We reviewed the EMEA database and included the main studies for the risk/benefit assessment (pivotal trials) submitted between 2000 and 2003. Results: In pivotal trials submitted to the EMEA there was no, or generally clinically negligible, evidence for gender bias; however, women were underrepresented in hypertension, diabetes and hepatitis B trials, and overrepresented in rheumatoid arthritis and allergic conjunctivitis. Conclusions: In trials submitted for marketing authorization to the EMEA gender bias was not a serious problem.

OBJECTIVE. The aim of our study was to examine long-term effects of nephrocalcinosis in prematurely born children. PATIENTS AND METHODS. Preterm neonates (gestational age RESULTS. Blood pressure did not differ in ex-preterm infants with and without nephrocalcinosis but was significantly higher than expected for healthy children. In comparison to healthy children, more ex-preterm infants with neonatal nephrocalcinosis had (mild) chronic renal insufficiency (glomerular filtration rate: CONCLUSIONS. Nephrocalcinosis in preterm neonates can have long-term sequelae for glomerular and tubular function. Furthermore, prematurity per se is associated with high blood pressure, relatively small kidneys, and (distal) tubular dysfunction. Long-term follow-up of blood pressure and renal glomerular and tubular function of preterm neonates, especially with neonatal nephrocalcinosis, seems warranted.

Both hypertension and congestive heart failure are characterized by a reduced vasodilatory capacity. In both conditions, the impairment of endothelial function is mainly the result of a reduced nitric oxide availability. The highly beta1-selective third-generation adrenoceptor blocker nebivolol displays additional endothelium-dependent vasodilating actions in humans as well as in animal models. In this study, we investigated whether these vasodilating properties of nebivolol are preserved in conditions with endothelial dysfunction. The vasodilatory effects of nebivolol were compared with those of the muscarinic agonist methacholine in isolated aortic rings obtained from spontaneous hypertensive rats and rabbits with experimental heart failure. The methacholine-induced responses were attenuated in aortic rings from both spontaneous hypertensive rats and congestive heart failture rabbits (42+/-6% and 25+/-3% vs. 89+/-3% and 54+/-7% for controls, respectively; P 0.05, n=6-13). These results implicate that the favorable hemodynamic profile of nebivolol may be preserved in patients with hypertension or congestive heart failure despite an impaired endothelial function

Familial hemophagocytic lymphohistiocytosis type 5 (FHL-5) is a rare genetic disorder caused by mutations in STXBP2/Munc18-2. Munc18-2 plays a role in the degranulation machinery of natural killer cells and cytotoxic T lymphocytes. Mutations in STXBP2/Munc18-2 lead to impaired killing of target cells by natural killer cells and cytotoxic T lymphocytes, which in turn results in elevated levels of the inflammatory cytokine interferon gamma, macrophage activation, and hemophagocytosis. Even though patients with FHL-5 present with anemia and hemolysis, no link between the disease and the erythroid lineage has been established. Here we report that red blood cells express Munc18-2 and that erythroid cells from patients with FHL-5 exhibit intrinsic defects caused by STXBP2/Munc18-2 mutations. Red blood cells from patients with FHL-5 expose less phosphatidylserine on their surface upon Ca2+ ionophore ionomycin treatment. Furthermore, cultured erythroblasts from patients with FHL-5 display defective erythropoiesis characterized by decreased CD235a expression and aberrant cell morphology. Copyright (C) 2015 ISEH - International Society for Experimental Hematology. Published by Elsevier Inc

PURPOSE: The aim of this study was to explore the perceived need for supportive care including healthy lifestyle programs among cancer survivors, their attitude towards self-management and eHealth, and its association with several sociodemographic and clinical variables and quality of life.METHODS: A questionnaire on the perceived need for supportive care and attitude towards self-management and eHealth was completed by 212 cancer survivors from an online panel.RESULTS: Highest needs were reported regarding physical care (66 %), followed by healthy lifestyle programs (54 %), social care (43 %), psychological care (38 %), and life question-related programs (24 %). In general, cancer survivors had a positive attitude towards self-management and eHealth. Supportive care needs were associated with male gender, lower age, treatment with chemotherapy or (chemo)radiation (versus surgery alone), hematological cancer (versus skin cancer, breast cancer, and other types of cancer), and lower quality of life. A positive attitude towards self-management was associated with lower age. A more positive attitude towards eHealth was associated with lower age, higher education, higher income, currently being under treatment (versus treatment in the last year), treatment with chemotherapy or (chemo)radiation (versus surgery alone), prostate and testicular cancer (versus hematological, skin, gynecological, and breast cancer and other types of cancer), and lower quality of life.CONCLUSIONS: The perceived need for supportive care including healthy lifestyle programs was high, and in general, cancer survivors had a positive attitude towards self-management and eHealth. Need and attitude were associated with sociodemographic and clinical variables and quality of life. Therefore, a tailored approach seems to be warranted to improve and innovate supportive care targeting cancer survivors.

Objective: To determine whether the results from placebo-controlled studies conducted in patients with manic episode can be generalised to a routine population of hospitalised acute manic patients. Methods: A list of four most prevalent inclusion and the nine most prevalent exclusion criteria was constructed for participation in previous randomised-controlled trials (RCTs). On the basis of this list, a consecutive series of 68 patients with 74 episodes of acute mania who had been referred for routine treatment were retrospectively assessed to determine their eligibility for a hypothetical but representative randomised controlled trial. Results: Only 16% of the manic episodes would qualify for the hypothetical trial (male episodes 28%, female episodes 10%), whereas 37%, 20% and 27% of the manic episodes would have to be excluded because they did no fulfil one, two or at least three of the inclusion or exclusion criteria. The most common exclusion criterion was "no use of contraceptives". If this criterion was not taken into account, 28% of the male episodes and 33% of the female episodes would qualify for inclusion in the hypothetical study. Apart from the use of contraceptives, no significant differences between male and female episodes were observed in the reasons for exclusion: 11% suicidal ideation, 29% prior mood stabilising medication, 1% depot medication, 22% other axis I diagnosis, 27% internal disease somatic disease, 5% neurological disorder, 15% alcohol use disorder and 10% drug use disorder. Conclusion: Only a small percentage acute manic episodes in a routine mental hospital seem to qualify for a standard placebo-controlled RCT. It could be argued, however, that certain exclusion criteria (e.g. no use of contraceptives) are not very likely to reduce the external validity of a standard RCT. In contrast, some other exclusion criteria (e.g. comorbid alcohol and drug use disorders) may have resulted in an overestimation of the efficacy of anti-manic medications. These notions should be taken into account when evaluating the results of RCTs in bipolar patients with an acute manic episode. (C) 2003 Elsevier B.V./ECNP. All rights reserved

Sympathetic neurotransmission in tissues with intact sympathetic nerve arborization is extensively dependent on calcium influx via N-type calcium-channels. It was the objective of the present study to assess and compare the claimed sympatholytic effect of the 1,4-dihydropyridine compound cilnidipine with other voltage-dependent calcium-channel (VDCC) antagonists. We studied these compounds by means of three different models. In the rabbit isolated thoracic aorta, the alleged sympatholytic properties displayed by these compounds were evaluated in the noradrenaline spillover model. Additionally, the influence of cilnidipine on stimulation-induced constrictor responses was studied in the rat isolated tail artery (male Wistar rats, 250-300 g) in addition to its effect on noradrenaline-induced contractions. Finally, we studied the influence of cilnidipine and other calcium-channel blockers on stimulation-induced chronotropic responses, in order to address N- or L-type selectivity, in the pithed rat model (male Wistar rats, 260-320 g). Furthermore, we evaluated their effect on noradrenaline-induced tachycardia. In the isolated rabbit thoracic aorta preparation omega-conotoxin GVIA (0.1 microM) nearly abolished the sympathetic outflow caused by stimulation, whereas nifedipine (0.1 microM) and amlodipine (1 microM) did not influence the evoked noradrenaline release. Cilnidipine (1 microM) significantly attenuated the response by nearly 18% and mibefradil (1 microM) by c. 42%. The stimulation-induced constrictor response (prejunctional effect) in the rat isolated tail artery could be blocked by omega-conotoxin GVIA (0.5 and 1 microM). Cilnidipine (10 nm and 0.1 microM) significantly attenuated responses to stimulation by maximally 20%, whereas it did not influence the constrictor response to noradrenaline (postjunctional effect). The mean heart rate in the pithed rat model amounted to 309.3 +/- 3.6 beats/min (bpm). Electrical stimulation of the cardio-accelerator nerves (C7-Th1) resulted in an increase by 106.7 +/- 2.2 bpm. All antagonists studied, except for nifedipine, attenuated the chronotropic response to stimulation (P < 0.05). The rank order of sympatholytic efficacy was: omega-conotoxin GVIA (84.8%), mibefradil (75.1%), cilnidipine (43.0%) and amlodipine (34.8%). Noradrenaline (10 nmol/kg) increased the heart rate by 117.8 +/- 2.7 bpm. This chronotropic response was influenced equally well by the calcium-channels blockers as observed in the stimulation (prejunctional) experiment. In conclusion, the N-type channel blocking properties and thus sympatholytic effect of cilnidipine could be demonstrated in some (vascular) but not all (cardiac) models studied. At the level of the vasculature cilnidipine reduced the neurotransmitter release to electrical stimulation in both the noradrenaline spillover model and in the model of the rat isolated tail artery, respectively. For amlodipine and nifedipine no sympatholytic activity could be demonstrated. In the pithed rat model, we were unable to demonstrate a selective N-type blocking effect for the VDCC-antagonists.

Objective It was the objective of this study to investigate whether a facilitatory role of vasopressin (AVID) on sympathetic nerve activity can be demonstrated in humans at the peripheral level. Methods Eight subjects (32 +/- 2.3 years) participated in this study. Forearm blood flow (FABF) was measured using the venous occlusion plethysmography model. Each session was performed in the presence of a continuous infusion (into the brachial artery) of AVID in sub-pressor dosage of 0.008 ng/kg per min, or NaCl 0.9%. Using lower-body negative pressure (LBNP) (-10, -20 and -30 mmHg) the combined pre- and postsynaptic action of AVP on the sympathetic nervous system was investigated. This was followed by a second protocol in which the possible postsynaptic effects of AVID were evaluated with intra-arterial infused norepinephrine (NE). Results The baseline FABF was 5.2 +/- 0.6 ml/100 ml per min. After infusion of AVP (0.008 ng/kg per min), the FABF remained unchanged at a flow of 5.5 +/- 0.6 ml/100 ml per min (P = 0.26). LBNP caused a pressure-dependent decrease in FABF (25.6 +/- 4.4, 29.0 +/- 6.1 and 38.6 +/- 6.9%, for -10, -20 and -30 mmHg, respectively). AVP significantly enhanced the FABF responses to lower-body negative pressures (38.0 +/- 8.6, 49.3 +/- 5.1 and 58.9 +/- 6.3%, respectively (P = 0.014). NE caused a dose-dependent vasoconstriction by 3.1 +/- 4.6, 17.0 +/- 4.3 and 23.2 +/- 4.9%, at dosages of 10, 20 and 40 pg/min, respectively, unaffected by AVID (P = 0.91). Conclusions We conclude that AVID can facilitate vasoconstriction mediated by the peripheral sympathetic nervous system at the presynaptic level in humans. (C) 2004 Lippincott Williams Wilkins

The beta-blocker nebivolol is a racemic mixture of D- and L- enantiomers that displays negative inotropic as well as direct vasorelaxant activity. In addition, it has been proposed that nebivolol exerts endothelium-protective effects caused by its antioxidant properties. In the present study we investigated the effect of D-, L-, and d/l-nebivolol on reactive oxygen species (ROS)-induced endothelial damage and compared it with carvedilol and metoprolol. Isolated rat aortic rings were exposed to ROS by electrolysis of the organ bath medium. Before and after electrolysis, endothelial function was measured by preconstricting the vessels with phenylephrine followed by the addition of methacholine. Carvedilol and nebivolol protected against ROS-induced endothelial damage, whereas metoprolol did not. The protective effect of nebivolol proved not to be stereoselective. Furthermore, we attempted to determine whether nebivolol acts a scavenger itself or whether another mechanism is involved. By means of HPLC measurements it was shown that nebivolol concentrations were decreased after exposure to electrolysis-induced ROS, thus indicating that nebivolol is degraded by its reaction with ROS. Functional experiments, in the rat aorta, demonstrated that exposure of nebivolol to ROS also affects its vasodilator activity. In conclusion, the present study demonstrates that nebivolol alleviates ROS-induced impairment of endothelium-dependent vasorelaxation. This protective effect is very likely the result of a direct ROS-scavenging action by the nebivolol molecule itself.

In recent years it has been shown that angiotensin II (Ang II) stimulates formation of reactive oxygen species (ROS), presumably by activation of NAD(P)H oxidase. This ROS formation has been primarily associated with cellular growth regulation by Ang II. The objective of the present study was to investigate whether these ROS contribute to Ang II-induced vasoconstriction. Experiments were performed in isolated rat thoracic aorta. Concentration response curves were constructed for Ang II in the absence and presence of the NAD(P)H oxidase inhibitor DPI, and ROS scavengers catalase and EUK-8. Inhibition of NAD(P)H oxidase as well as scavenging of ROS, decreased the contractile response to Ang II. Administration of NADPH, a substrate for NAD(P)H oxidase, produced vasoconstriction that proved to be sensitive for DPI, catalase, and EUK-8. Exposure of the vessels to exogenous ROS, induced by electrolysis of the organ bath medium, also resulted in a contractile response that was decreased by ROS scavenging. The results suggest that ROS play a role in Ang II-induced vasoconstriction via the activation of NAD(P)H oxidase.

Angiotensin (AT) 11 is known to enhance responses to electrical field stimulation (EFS) via AT(1) receptors located on sympathetic nerve terminals. Differences in potency exist between AT(1) receptor antagonists regarding the inhibition of the prejunctional and postjunctional AT(1) receptors. It is hypothesized that prejunctional AT(1) receptors might belong to the AT(1B) receptor subtype. Accordingly, the authors investigated whether AT(1B) receptor inhibition by high concentrations of PD123319 could suppress ATII-augmented noradrenergic transmission (prejunctional) in the rabbit thoracic aorta by means of a noradrenaline spillover model. Additionally, the influence of PD123319 on ATII-enhanced constrictor responses to electrical field stimulation was investigated in the isolated rabbit mesenteric artery. Furthermore; the authors investigated whether PD123319 could influence the constrictor responses (postjunctional) to ATII in both preparations. In the thoracic aorta, ATII (10 nM) caused a significant enhancement of EFS-evoked [H-3]-noradrenaline release by a factor of 2.0 +/- 0.1. This reinforcement could be inhibited by PD123319 (0.1, 1, and 10 muM). The constrictor response to ATII was unaffected by PD123319. In the mesenteric artery, ATII (0.5 nM) caused a significant enhancement of constrictor responses to EFS by factors of 2.9 +/- 0.3, 2.3 +/- 0.3, and 1.6 +/- 0.1 at 1, 2, and 4 Hz, respectively. This enhancement could be attenuated by PD123319 (I and 10 muM. The constrictor response to ATII was unaffected by PD123319. It is concluded that the prejunctional AT(1) receptors belong to the AT(1B) subtype whereas postjunctional AT(1) receptors do not

The present study was designed to investigate the influence of maturation (young versus adult) on the angiotensin II-mediated facilitation of sympathetic nerve traffic (prejunctional AT(1)-receptor) as well as on the angiotensin II-mediated vasoconstriction (postjunctional AT(1)-receptor). Additionally, we investigated the inhibitory effect of the selective AT(1)-receptor antagonist eprosartan on angiotensin II-mediated responses at both sites during maturation. Male New Zealand White rabbits, aged 12 to 14 and 35 to 38 weeks (young versus adult, respectively), were used. To study angiotensin II at the neuronal AT(1)-receptor we investigated its influence on electrical field stimulation (EFS)-evoked sympathetic neurotransmission in the isolated thoracic aorta in a noradrenaline spillover model. To study the effects of angiotensin II at the level of the vasculature concentration-response curves for angiotensin II were constructed. In both models the influence of eprosartan on angiotensin II-mediated responses was studied. Angiotensin II (0.01 nM-0.1 muM) concentration-dependently enhanced the EFS-evoked noradrenaline release in both groups. No differences concerning the relative (approximately 100%, P > 0.05) and absolute facilitation were observed between groups, although concentrations required in adult rabbits exceeded those in young animals by 1 unity log M increment. Eprosartan concentration-dependently attenuated the angiotensin II-enhanced (10 nM) sympathetic outflow. The inhibitory potency differed approximately by a factor ten between both groups (young-pIO(50) 7.91 +/- 0.12 and adult; pIC(50) 8.81 +/- 0.3 1, respectively, P

Nebivolol is a highly selective beta(1) adrenoceptor blocker with additional vasodilating properties. Although it has been shown that the nebivolol-induced vasorelaxation is nitric oxide (NO) and cGMP dependent, the receptor that mediates these actions remains controversial, and serotonergic as well as beta-adrenergic pathways may be involved. Therefore, functional experiments investigating the receptor involved in nebivolol-induced vasorelaxation were performed in the rat aorta. Isolated aortic rings were exposed to cumulative concentrations of nebivolol. Nebivolol concentrations of 3 micromol/L and higher caused vasorelaxation, which was inhibited by the presence of the NO synthase inhibitor l-NNA (100 micromol/L), or by mechanical removal of the endothelium. Exposure of the vessel rings to the selective 5-HT(1A) antagonist NAN-190 (1 micromol/L) or the 5-HT(1/2) antagonist methysergide (1 micromol/L) did not influence nebivolol-induced vasorelaxation. Similarly, the incubation with the beta(2)-adrenoceptor antagonist butoxamine (50 micromol/L) did not prevent vasorelaxation. The selective beta(3)-adrenoceptor antagonist S-(-)-cyanopindolol (1 micromol/L), however, significantly counteracted the nebivolol-induced vasorelaxation. Furthermore, exposure of the aortic rings to cumulative concentrations of the beta(3) selective adrenoceptor agonist BRL37344 caused, like nebivolol, NO-dependent vasorelaxation that was antagonized by S-(-)-cyanopindolol. The results suggest that nebivolol-induced NO-dependent vasorelaxation is, at least in part, caused by a beta(3)-adrenoceptor agonistic effect.

Angiotensin II (Ang II) enhances sympathetic neurotransmission via AT1-receptors located on sympathetic nerve terminals. We recently demonstrated that inhibition of Ang II-mediated facilitation in the pithed rat by irbesartan resulted in a U-shaped dose response curve, which was not observed when PD 123319, at a concentration that selectively blocks the AT2-receptor, was co-administered. Hence, the irbesartan-mediated `upstroke' might be explained by the involvement of the AT2-receptor after AT1 blockade with high-dose irbesartan. In the present study, we further investigated the possible role of the AT2-receptor in Ang II-mediated facilitation in vitro. We studied the effect of the AT2-receptor antagonist PD 123319 (10 nM) on Ang II-enhanced sympathetic outflow evoked by electrical field stimulation (EFS) in the rat isolated inferior vena cava. Additionally, we investigated the effect of the AT1-receptor blocker irbesartan (0.1 nM—1 µM) on the sequelae of Ang II-enhanced, EFS-evoked sympathetic nerve traffic in the presence or absence of PD 123319 (10 nM). PD 123319 did not influence Ang II-enhanced sympathetic outflow. Irbesartan dose-dependently attenuate Ang II-augmented transmitter release (pIC50 7.99±0.03), whereas no U-shaped concentrationresponse relationship for irbesartan was observed. Co-administration of PD 123319 with irbesartan proved unable to influence Ang II-mediated facilitation differently compared with irbesartan alone. The experimental observations indicate that the AT2-receptor is not involved in Ang II-mediated enhancement of sympathetic nerve traffic in the present in vitro study.

Both in human and in experimental heart failure (HF), the renin-angiotensin system and the sympathetic nervous system are activated. In a previous study a facilitatory action of angiotensin II (Ang II) was shown in the rabbit mesenteric artery, which was mediated via prejunctionally located Ang II type I (AT(1)) receptors. Very little is known about the effects of Ang II on sympathetic neurotransmission at the peripheral level in congestive heart failure (CFH). Accordingly, in the isolated mesenteric arteries obtained from rabbits with experimentally induced CHF, as well as in age-matched control rabbits, the effect of Ang II on contractions provoked by electrical field stimulation was investigated in the presence and absence of the AT(1) receptor antagonist eprosartan. Additionally, to investigate a possible postjunctional facilitation, the effects of Ang II on alpha-adrenoceptor-mediated responses were studied using noradrenaline (NA). Lastly, the vasoconstrictor effects of Ang II were compared between HF rabbits and controls, by constructing concentration-response curves to Ang II. In control rabbits, Ang II 0.5 nM caused an enhancement of stimulation-induced responses by a factor 3.2 +/- 0.5, 2.4 +/- 0.3, and 1.5 +/- 0.08, at 1, 2, and 4 Hz, respectively (P

The aim of the present study was to determine the inhibitory potency of two selective angiotensin AT, receptor antagonists, eprosartan and candesartan, at the level of the sympathetic nerve terminal and the vascular smooth muscle. Male New Zealand White rabbits, weighing 2100-2550 g, were used. To study eprosartan and candesartan at the neuronal angiotensin AT, receptor, we investigated their influence on the angiotensin II-enhanced, electrical field stimulation-evoked sympathetic transmission in the rabbit isolated thoracic aorta in a noradrenaline spillover model. To study both antagonists at the vascular angiotensin AT(1) receptor, concentration-response curves for angiotensin II were constructed in the presence or absence of the two angiotensin AT(1) receptor antagonists. Angiotensin II (10 nM) caused a significant increase by 107 +/- 11.1% of the stimulation-evoked sympathetic outflow, which was concentration-dependently inhibited by both eprosartan (pIC(50) 7.91 +/- 0.12) and candesartan (pIC(50) 10.76 +/- 0.13). Angiotensin II (1 nM-0.3 muM) caused a concentration-dependent increase in contractile force (E-max 20.62 +/- 2.24 mN, pD(2) 8.16 +/- 0.04). Both eprosartan (pA(2) 8.90 +/- 0.11, pIC(50) 8.87 +/- 0.12 (10 nM angiotensin II)) and candesartan (pD(2)' 10.80 +/- 0.13) counteracted the contractions evoked by cumulative concentrations of angiotensin II. Candesartan proved a more potent antagonist than eprosartan at both the pre- and postjunctional angiotensin AT(1) receptor. For eprosartan, vascular inhibitory concentrations were 10-fold lower than sympatho-inhibitory concentrations, whereas for candesartan, inhibitory concentrations at both sites were similar. The results may be explained by differences between the pre- and postjunctional angiotensin AT(1) receptor subtype. (C) 2003 Elsevier Science B.V All rights reserved

Summary 1. Reactive oxygen species (ROS) are known to be involved in the progression of various cardiovascular diseases. One source of ROS is activated neutrophils, which can release superoxide anion radicals and hydrogen peroxide by membrane-bound NAD(P)H oxidases. These ROS not only destroy bacteria, but may also affect mammalian tissue. In addition, hydrogen peroxide serves as a substrate for myeloperoxidase, an enzyme that is released by activated neutrophils during inflammatory processes, as seen, for instance, in reperfusion injury and atherosclerosis. Myeloperoxidase catalyses the oxidation of chloride by hydrogen peroxide, yielding hypochlorite, an extremely potent oxidant. 2. The purpose of the present study was to evaluate the effects of hypochlorite on a variety of receptor-dependent processes in rat isolated left atria and rat thoracic aorta and to compare these results with the phenomena observed after incubation with hydrogen peroxide. 3. In the presence of hypochlorite (300 µmol/L), the positive inotropic response of α1-adrenoceptor stimulation by methoxamine (300 µmol/L) was converted into a negative inotropic response. In contrast, the positive inotropic effects of the β1/β2-adrenoceptor agonist isoprenaline (3 µmol/L) and endothelin (ET)-1 (100 nmol/L) remained largely unaffected. 4. The inversion of α1-adrenoceptor-mediated inotropy was not obtained in the presence of hydrogen peroxide (500 µmol/L). Hydrogen peroxide did not affect the positive inotropic response of isoprenaline, but it completely abolished the inotropic effect of ET-1. 5. The effect of cardiac M2-receptor stimulation was studied in the presence of hypochlorite and hydrogen peroxide. The negative inotropic response to acetylcholine (ACh) was significantly enhanced after hypochlorite incubation compared with control. 6. In the rat thoracic aorta, endothelial function, evaluated by means of ACh-induced vasodilation, was completely abolished in the presence of hypochlorite (100 µmol/L), but remained unaffected by treatment with the same concentration of hydrogen peroxide. 7. From these data, we conclude that hypochlorite exerts more toxic properties than its precursor hydrogen peroxide, leading to substantial physiological alterations in cardiac and vascular tissue.

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In a family with mild dominant spherocytosis, affected members showed partial band 3 deficiency. The index patient showed more severe clinical symptoms than his relatives, and his red blood cells displayed concomitant low pyruvate kinase activity. We investigated the contribution of partial PK deficiency to the phenotypic expression of mutant band 3 in this family. Pyruvate kinase deficiency and band 3 deficiency were characterized by DNA analysis. Results of red cell osmotic fragility testing, the results of cell deformability obtained by the Automated Rheoscope and Cell Analyzer and the results obtained by Osmotic Gradient Ektacytometry, which is a combination of these tests, were related to the red cell ATP content. Spherocytosis in this family was due to a novel heterozygous mutation in SLC4A1, the gene for band 3. Reduced PK activity of the index patient was attributed to a novel mutation in PKLR inherited from his mother, who was without clinical symptoms. Partial PK deficiency was associated with decreased red cell ATP content and markedly increased osmotic fragility. This suggests an aggravating effect of low ATP levels on the phenotypic expression of band 3 deficiency.

Background Haemoglobin (Hb) variants are well-known factors interfering with accurate HbA1c testing. This report describes two novel Hb variants leading to inappropriate quantification of HbA1c by ion-exchange chromatography. Methods Glycated forms of novel Hb variants were recognised in the blood of two patients with diabetes mellitus screened by HbA1c ion-exchange chromatography. Dedicated high-resolution cation-exchange chromatography and subsequent DNA sequencing revealed the exact nature of the variants. Other common techniques for quantifying HbA1c were applied on both samples and haematological parameters were determined to judge possible pathology associated with the novel Hb variants. Results A fraction of 15% of abnormal Hb was observed in a 37-year-old female. DNA sequencing revealed a heterozygous mutation in the α1-globin gene, resulting in a leucine-to-phenylalanine amino-acid substitution (HBA1: c.301C>T, p.Leu101Phe). We named this variant Hb Weesp. The other novel variant, Hb Haelen, presented as a 40% fraction in a 63-year-old male and resulted from a heterozygous amino acid substitution in the β-globin gene (HBB: c.335T>C, p.Val112Gly). The presence of both Hb variants resulted in aberrant separation of the Hb components, leading to an inadequate quantification of HbA1c. Conclusions Close examination of HbA1c chromatograms revealed two novel, clinically silent Hb variants that interfere with HbA1c quantification. Healthcare providers need to be aware of the potential of such Hb variants when interpreting HbA1c results.

Male rats were treated with a muscarinic receptor antagonist at 3, 10, and 100 mg/kg/day for 4 weeks prior to mating with untreated females and their reproductive status was determined on gestation days (GD) 15–17. Treatment-related decreases in the pregnancy rate were observed at 100 mg/kg/day without any effects on mating performance. Impairment of male fertility by this compound was also observed after treatment for 1 week, but there were no effects after a 1-week withdrawal period suggesting reversibility of the effect. There were no treatment-related effects on sperm production or motility, or testicular histopathology in any group. In order to determine whether the reduced fertility was a class effect of muscarinic receptor antagonists, atropine was examined. Males received atropine for 1 week at 62.5 and 125 mg/kg/day and were mated with untreated females. A low pregnancy rate associated with a decrease in the number of implantations was observed at 125 mg/kg/day. The effect on implantation was also observed at 62.5 mg/kg/day. These findings suggest that the impairment of fertility in male rats induced by muscarinic receptor antagonists is a class effect, and has a relatively short onset of effect and is quickly reversible.

Effects of angiotensin H type I (AT(1)) receptor antagonists eprosartan and candesartan and AT(2) receptor antagonist PD123319 on Ana II-induced facilitation of noradrenergic neurotransmission were investigated in isolated rabbit mesenteric artery under isometric conditions. Sympathoinhibitory potency of AT(1) blockers was compared with their potency concerning inhibition of direct vasoconstrictor effect of Ana II. To investigate blockade of presynaptic AT(1) and AT(2) receptors, effects of Ang II on electrical field stimulation (EFS)-induced contractions in presence or absence of eprosartan, candesartan. or PD123319 were studied. To investigate blockade of postsynaptic AT, receptors, effects of either eprosartan or candesartan on concentration-response curves of Ana II were studied. In addition, effect of Ana 11 on postsynaptic alpha-adrenoceptor-mediated responses was studied using noradrenaline. EFS (1, 2, and 4 Hz) caused an increase of contractile force. At stimulation frequencies of 1, 2, and 4 Hz, a subpressor concentration of Ana II (0.5 nM) increased stimulation-induced vasoconstrictor responses by 2.8 +/- 0.5, 2.4 +/- 0.4. and 1.6 +/- 0.1 of control values, respectively (p

Summary1 It was shown recently that stimulation of cardiac muscarinic M2-receptors revealed an enhanced negative inotropic response in isolated rat left atria after exposure to hypochlorite-induced oxidative stress. This phenomenon was not observed after stimulation of the cardiac A1-receptor, which like the M2-receptor is coupled to Gi-proteins. Since even the contractile response to M3-receptor stimulation was not amplified in the rat portal vein, we hypothesized a M2-receptor specificity of this hypochlorite-induced enhancement. 2 The present study was performed in order to investigate whether the sympathoinhibitory response to presynaptically located M2-receptor stimulation would also be modified after exposure to hypochlorite in the rat tail artery. We applied electrical field stimulation (EFS) in order to mimic sympathetic neurotransmission. 3 EFS increased the vascular tone frequency-dependently (0.3–4 Hz). EFS-induced vasoconstriction could be attenuated by acetylcholine (30 nm−1 μm) in a concentration-dependent manner. Hypochlorite (10 and 100 μm) did not affect the sympathoinhibitory effect of acetylcholine (100 nm). 4 In conclusion, in contrast to cardiac M2-receptors, hypochlorite did not amplify the sympathoinhibitory effects of presynaptic M2-receptors. The different responsiveness between neuronal and cardiac M2-receptors to hypochlorite may be explained by the different G-protein subunits involved in the activation of the underlying signalling cascade.

Objective Several studies have shown that arginine vasopressin (AVP) potentiates the sympathetic nervous transmission in isolated vessels. The present study investigates such a potentiation in the pithed rat model. Methods Male Wistar rats weighing 270-310 g were used. Spinal-cord stimulation was applied, with frequencies of 0.25-4 Hz, in the presence or absence of a subpressor dose of intravenous (i.v.) AVP (1 pmol/kg per min). In addition, the effect of AVP on postsynaptic alpha-adrenoceptor-mediated responses was studied using exogenously administered noradrenaline (NA). For this purpose dose-response curves (DRCs) for NA (i.v.) were constructed. Results In the pithed rat model endogenously generated angiotensin II facilitates neurally mediated increments in vascular resistance. Without the administration of the angiotensin II type 1 (AT(1)) antagonist irbesartan, the facilitating effect of AVP was not visible. However, after the administration of the AT(1) antagonist irbesartan, the facilitating effect of AVP became apparent. The stimulation-induced rise in diastolic blood pressure (DBP) was enhanced in the presence of AVP from 63.7 +/- 4.5 to 78.6 +/- 4.2 mmHg, at a stimulation frequency of 4 Hz. The vasopressin receptor V-1 antagonist, SR-49059, completely inhibited this AVP-inDuced facilitation, whereas the V-2 antagonist, SR-1211463B, or the V-2 agonist desmopressin, did not. The DRC of exogenously administered NA was not influenced by AVP. Conclusion The stimulating effect of AVP on sympathetic neurotransmission is completely dependent on the stimulation of presynaptically located V-2 receptors. The facilitating effect of angiotensin II on the sympathetic nervous system (SNS) in the pithed rat model masks the facilitating effect of AVP in this preparation. (C) 2002 Lippincott Williams Wilkins

Angiotensin II (Ang II) causes facilitation of sympathetic neurotransmission via prejunctionallylocated AT1-receptors. The pithed rat is a suitable model to study the interactions between endogenously produced Ang II and the sympathetic nervous system at the peripheral level. Previously, we demonstrated that inhibition of the facilitatory actions of Ang II is a class effect of all AT1-receptor blockers (ARB). However, all ARBs caused less than maximal inhibition after the highest dose, thus causing a U-shaped dose-response curve with respect to sympatho-inhibition. In the present study, we investigated whether the AT2-receptor is involved in this `upturn' of the dose-response relationship. Accordingly, we studied the effect of the ARB, irbesartan (1—60 mg/kg), on the sequelae of electric stimulation of the thoraco-lumbar sympathetic outflow in the presence and absence of the AT2blocker, PD 123319 (0.5 mg/kg +50 µg/kg/min). Additionally, the effect of the combined (non-selective) AT1/AT2-receptor antagonist saralasin (0.001, 0.003, 0.01 or 0.03 mg/kg/min), on stimulation-induced responses was studied. In addition, we measured PRA-levels after administration of irbesartan, in this model. The stimulation-induced increase in diastolic blood pressure (DBP) could be dose-dependently reduced by irbesartan. Co-infusion with PD 123319 increased the sympatho-inhibitory potency of irbesartan, possibly through displacement of irbesartan from plasma protein binding sites. The U-shaped dose-response relationship observed with irbesartan, which is illustrative for other ARBs in this model, was not observed when PD 123319 was co-administered with irbesartan, nor with the non-selective AT1/AT2blocker, saralasin. PRA-levels increased from 111.0±17.8 to 198.7±22.2 ng/ml/hour after administration of irbesartan. PRA-levels did not differ when measured after the three highest doses of irbesartan. Conclusions The present findings indicate a facilitatory role for the AT2-receptor, which is unmasked by the highest dose of irbesartan. Different plasma Ang II-levels are unlikely to have caused the less than maximal inhibition after the highest dose of irbesartan.

Introduction We investigated whether a group of pathologists could reproducibly apply the International Association for the Study of Lung Cancer/American Thoracic Society/European Respiratory Society (IASLC/ATS/ERS) classification for lung adenocarcinoma to a cohort of stage 1 tumors and whether this architectural classification and/or other parameters could demonstrate survival advantage. Methods A total of 145 cases of 7 th edition of tumor, node, metastasis stage 1 adenocarcinoma were retrospectively reviewed for predominant architectural pattern, including cribriform pattern, nuclear grade, mitotic index, and necrosis. The parameters were assessed for reproducibility and survival and using multivariate analysis, compared with stage, age, and sex. Results The majority of tumors had a mixed architecture with the acinar pattern being the most common predominant architecture. Micropapillary and cribriform architecture were the least frequent patterns. This study demonstrated that a group of five pathologists could reproducibly apply the IASLC/ATS/ERS classification. Although there were insufficient cribriform-predominant adenocarcinomas for assessment, when the percentage of all cribriform was combined with other architectures, it was associated with a worse prognosis. The majority of the parameters assessed demonstrated significance with univariate analysis but only mitotic index, as assessed by the highest count/10 high-power fields remained significant with multivariate analysis. Conclusion In this study of resected stage 1 primary lung adenocarcinoma, we found mitotic index to be the only independent prognostic marker. It was more closely associated with outcome than either pathologic T stage or IASLC/ATS/ERS architecture-based classification. Further validation of concordance and reproducibility in reporting mitotic index, as well as validation of prognostic significance, needs to be undertaken in independent data sets.

In a family with mild dominant spherocytosis, affected members showed partial band 3 deficiency. The index patient showed more severe clinical symptoms than his relatives, and his red blood cells displayed concomitant low pyruvate kinase activity. We investigated the contribution of partial PK deficiency to the phenotypic expression of mutant band 3 in this family. Pyruvate kinase deficiency and band 3 deficiency were characterized by DNA analysis. Results of red cell osmotic fragility testing, the results of cell deformability obtained by the Automated Rheoscope and Cell Analyzer and the results obtained by Osmotic Gradient Ektacytometry, which is a combination of these tests, were related to the red cell ATP content. Spherocytosis in this family was due to a novel heterozygous mutation in SLC4A1, the gene for band 3. Reduced PK activity of the index patient was attributed to a novel mutation in PKLR inherited from his mother, who was without clinical symptoms. Partial PK deficiency was associated with decreased red cell ATP content and markedly increased osmotic fragility. This suggests an aggravating effect of low ATP levels on the phenotypic expression of band 3 deficiency.

A 41-year-old man had a palpable mass on the volar side of his hand. X-ray, ultrasound and MRI showed an inhomogeneous tumour with calcification. Histologic examination ruled out the suspicion of a liposarcoma. The surgeon successfully excised the ossifying lipoma, which is a rare variant of a common lipoma and can clinically and radiologically mimic malignant neoplasms.

Aim: Atomic force microscopy nanoindentation of myofibers was used to assess and quantitatively diagnose muscular dystrophies from human patients. Materials & methods: Myofibers were probed from fresh or frozen muscle biopsies from human dystrophic patients and healthy volunteers, as well as mice models, and Young’s modulus stiffness values were determined. Results: Fibers displaying abnormally low mechanical stability were detected in biopsies from patients affected by 11 distinct muscle diseases, and Young’s modulus values were commensurate to the severity of the disease. Abnormal myofiber resistance was also observed from consulting patients whose muscle condition could not be detected or unambiguously diagnosed otherwise. Discussion & conclusion: This study provides a proof-of-concept that atomic force microscopy yields a quantitative read-out of human muscle function from clinical biopsies, and that it may thereby complement current muscular dystrophy diagnosis.