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Human-derived cerebral organoids demonstrate great promise for identifying cell types and cell type specific molecular processes perturbed by genetic variants associated with neuropsychiatric and neurodevelopmental disorders, which are notoriously challenging to study using animal models. However, considerable challenges remain in achieving robust, scalable and generalizable phenotyping of organoids to discover cell types and cell type specific genes. We perform RNA sequencing on 71 samples comprising 1,420 cerebral organoids from 25 donors, and describe a framework (Orgo-Seq) to integrate bulk RNA and single-cell RNA sequence data from human post-mortem brains and cerebral organoids, for the identification of cell types and cell type specific individual genes. We apply Orgo-Seq for two autism-associated loci: 16p11.2 deletions and 15q11-13 duplications, and identify neuroepithelial cells as critical cell types for 16p11.2 deletions, and discover novel and previously reported cell type specific driver genes. Finally, we validated our results that mutations in the KCTD13 gene in the 16p11.2 locus lead to imbalances in the proportion of neuroepithelial cells, using CRISPR/Cas9-edited mosaic organoids. Our work presents a quantitative technological framework to integrate multiple transcriptomics datasets to identify cell types and cell type specific driver genes associated with complex diseases using cerebral organoids.

Burkitt lymphoma is a mature aggressive B-cell lymphoma derived from germinal center B cells(1). Its cytogenetic hallmark is the Burkitt translocation t(8;14)(q24;q32) and its variants, which juxtapose the MYC oncogene with one of the three immunoglobulin loci(2). Consequently, MYC is deregulated, resulting in massive perturbation of gene expression(3). Nevertheless, MYC deregulation alone seems not to be sufficient to drive Burkitt lymphomagenesis. By whole-genome, whole-exome and transcriptome sequencing of four prototypical Burkitt lymphomas with immunoglobulin gene (IG)-MYC translocation, we identified seven recurrently mutated genes. One of these genes, ID3, mapped to a region of focal homozygous loss in Burkitt lymphoma(4). In an extended cohort, 36 of 53 molecularly defined Burkitt lymphomas (68%) carried potentially damaging mutations of ID3. These were strongly enriched at somatic hypermutation motifs. Only 6 of 47 other B-cell lymphomas with the IG-MYC translocation (13%) carried ID3 mutations. These findings suggest that cooperation between ID3 inactivation and IG-MYC translocation is a hallmark of Burkitt lymphomagenesis.

// Alexandra Schrader 1, 2, 10, 11 , Katharina Meyer 3, 6 , Neele Walther 1 , Ailine Stolz 4 , Maren Feist 1, 7 , Elisabeth Hand 1, 6 , Frederike von Bonin 1 , Maurits Evers 3, 6, 13 , Christian Kohler 3, 6 , Katayoon Shirneshan 1 , Martina Vockerodt 5, 8, 10, 12 , Wolfram Klapper 5, 6, 7, 9 , Monika Szczepanowski 5, 6, 7, 9 , Paul G. Murray 8 , Holger Bastians 4 , Lorenz Trumper 1, 2, 5, 7 , Rainer Spang 3, 5, 6, 7 , Dieter Kube 1, 2, 5, 6, 7 1 Department of Haematology and Medical Oncology, University Medical Centre of the Georg-August University Gottingen, Gottingen, Germany 2 GRK1034 of the Deutsche Forschungsgemeinschaft, Georg-August University Gottingen, Gottingen, Germany 3 Department of Statistical Bioinformatics, Institute for Functional Genomics, University of Regensburg, Regensburg, Germany 4 Goettingen Center for Molecular Biosciences (GZMB) and University Medical Center, Institute of Molecular Oncology, Section for Cellular Oncology, Gottingen, Germany 5 Network Molecular Mechanism of Malignant Lymphoma (MMML) of the Deutsche Krebshilfe, Germany 6 BMBF-Network HamatoSys, Germany 7 BMBF-Network Myc-Sys, Germany 8 School of Cancer Sciences, University of Birmingham, Birmingham, UK 9 University-Hospital Schleswig-Holstein, Hematopathology Section and Lymph Node Registry Kiel, Kiel, Germany 10 Department of Anatomy, University Medical Centre of the Georg-August University Gottingen, Gottingen, Germany 11 Present address: Laboratory of Lymphocyte Signaling and Oncoproteome, Department I of Internal Medicine, University Hospital Cologne, Center for Integrated Oncology (CIO) Koln-Bonn, Cologne, Germany 12 Present address: Department of Anatomy, University Medical Centre of the Georg-August University Gottingen, Gottingen, Germany 13 Current address: The John Curtin School of Medical Research the Australian National University Canberra, Australia Correspondence to: Dieter Kube, e-mail: dieter.kube@med.uni-goettingen.de Keywords: lymphoma, B cell receptor signaling, guided clustering, cell cycle delay, chromosomal aberrations Received: September 23, 2015 Accepted: March 31, 2016 Published: May 7, 2016 ABSTRACT To discover new regulatory pathways in B lymphoma cells, we performed a combined analysis of experimental, clinical and global gene expression data. We identified a specific cluster of genes that was coherently expressed in primary lymphoma samples and suppressed by activation of the B cell receptor (BCR) through αIgM treatment of lymphoma cells in vitro . This gene cluster, which we called BCR.1, includes numerous cell cycle regulators. A reduced expression of BCR.1 genes after BCR activation was observed in different cell lines and also in CD10 + germinal center B cells. We found that BCR activation led to a delayed entry to and progression of mitosis and defects in metaphase. Cytogenetic changes were detected upon long-term αIgM treatment. Furthermore, an inverse correlation of BCR.1 genes with c-Myc co-regulated genes in distinct groups of lymphoma patients was observed. Finally, we showed that the BCR.1 index discriminates activated B cell-like and germinal centre B cell-like diffuse large B cell lymphoma supporting the functional relevance of this new regulatory circuit and the power of guided clustering for biomarker discovery.

Catching primal functional changes in early, ‘very far from disease onset’ (VFDO) stages of Huntington’s disease is likely to be the key to a successful therapy. Focusing on VFDO stages, we assessed neuronal microcircuits in premanifest Hdh150 knock-in mice. Employing in vivo two-photon Ca2+ imaging, we revealed an early pattern of circuit dysregulation in the visual cortex - one of the first regions affected in premanifest Huntington’s disease - characterized by an increase in activity, an enhanced synchronicity and hyperactive neurons. These findings are accompanied by aberrations in animal behavior. We furthermore show that the antidiabetic drug metformin diminishes aberrant Huntingtin protein load and fully restores both early network activity patterns and behavioral aberrations. This network-centered approach reveals a critical window of vulnerability far before clinical manifestation and establishes metformin as a promising candidate for a chronic therapy starting early in premanifest Huntington’s disease pathogenesis long before the onset of clinical symptoms.

Impaired mitochondrial energy metabolism contributes to a wide range of pathologic conditions, including neurodegenerative diseases. Mitochondrial apoptosis-inducing factor (AIF) is required for the correct maintenance of mitochondrial electron transport chain. An emerging body of clinical evidence indicates that several mutations in the AIFM1 gene are causally linked to severe forms of mitochondrial disorders. Here we investigate the consequence of WAH-1/AIF deficiency in the survival of the nematode Caenorhabditis elegans. Moreover, we assess the survival of C. elegans strains expressing a disease-associated WAH-1/AIF variant. We demonstrate that wah-1 downregulation compromises the function of the oxidative phosphorylation system and reduces C. elegans lifespan. Notably, the loss of respiratory subunits induces a nuclear-encoded mitochondrial stress response independently of an evident increase of oxidative stress. Overall, our data pinpoint an evolutionarily conserved role of WAH-1/AIF in the maintenance of proper mitochondrial activity.

Burkitt's lymphoma (BL) is a highly malignant, aggressive non-Hodgkin's lymphoma derived from germinal center B cells. Recently, global gene expression profiling of patient samples led to a molecular definition of BL with lymphocyte enhancer-binding factor 1 (LEF1) as a signature gene. Herein, we report the expression of nucleic LEF1 in 15 of 18 patients with BL and the identification of LEF1 target genes. Germinal center B cells were devoid of detectable nuclear LEF1 expression, as were mantle cell lymphoma (0 of 5), marginal zone lymphoma (0 of 6), follicular lymphoma (0 of 12), and diffuse large B-cell lymphoma (1 of 31). Whole-genome gene expression profiling after transient knockdown of LEF1 in BL cell lines identified new LEF1 target genes; these LEF1 targets are enriched with genes associated with cancers. The expression of LEF1 and LEF1-regulated genes in primary BL suggests that LEF1 is not only aberrantly expressed but also transcriptionally active. This study supports a functionally important role for LEF1 and its target genes in BLs.

Injection drug users (IDUs) entering an urban low-threshold opiate detoxification unit in Munich, Germany, between 1991 and 1997 were asked for lifetime suicide attempts and administered a questionnaire, including sociodemographic, IDU-related, and drug user treatment related variables. Among 1,049 participants, 20% had ever tried to commit suicide. In the logistic regression, female gender, older age, drug user treatment experience, lack of drug user counseling, and emergency treatment were associated with attempted suicide. Regular screening for suicidal behavior in IDUs should be undertaken with a special focus to mention risk factors. The study's limitations are noted. This study was partially sponsored by the German Ministry of Health.

Mutations in peroxin (PEX) genes lead to loss of peroxisomes, resulting in the formation of peroxisomal biogenesis disorders (PBDs) and early lethality. Studying PBDs and their animal models has greatly contributed to our current knowledge about peroxisomal functions. Very-long-chain fatty acid (VLCFA) accumulation has long been suggested as a major disease-mediating factor, although the exact pathological consequences are unclear. Here, we show that a Drosophila Pex19 mutant is lethal due to a deficit in medium-chain fatty acids (MCFAs). Increased lipolysis mediated by Lipase 3 (Lip3) leads to accumulation of free fatty acids and lipotoxicity. Administration of MCFAs prevents lipolysis and decreases the free fatty acid load. This drastically increases the survival rate of Pex19 mutants without reducing VLCFA accumulation. We identified a mediator of MCFA-induced lipolysis repression, the ceramide synthase Schlank, which reacts to MCFA supplementation by increasing its repressive action on lip3. This shifts our understanding of the key defects in peroxisome-deficient cells away from elevated VLCFA levels toward elevated lipolysis and shows that loss of this important organelle can be compensated by a dietary adjustment., Author summary Peroxisomes are organelles that contain several enzymes and fatty acids required for many metabolic tasks in the cell, and upon peroxisome loss, their educts accumulate. One example is the accumulation of very-long-chain fatty acids (VLCFAs) with a chain length of more than 20 carbons. These fatty acids cannot be oxidized in mitochondria but are exclusively degraded in peroxisomes. Lowering increased VLCFA levels is sometimes attempted as a treatment option for human disorders with peroxisomal dysfunction, although its effectiveness remains unclear. Here, we have analyzed this process in Drosophila melanogaster and found that peroxisomal loss results not only in VLCFA accumulation but also in a reduction of medium-chain fatty acids (MCFAs). We could show that this is due to a state of high lipolysis and increased mitochondrial activity. By supplementation with MCFAs from coconut oil, we were able to rescue mitochondrial damage and lethality observed in peroxisome-deficient flies. We found that this process is mediated by the ceramide synthase Schlank, which acts as a transcription factor and shuttles between nuclear membrane and endoplasmic reticulum (ER) in response to MCFA availability. We conclude that peroxisome loss triggers the accumulation of free fatty acids and mitochondrial damage in flies and that these effects can be rescued by a diet rich in MCFAs.

This cohort study was conducted to identify risk factors for lifetime emergency room treatment due to overdose in injection drug users. Data of 1049 patients on admission for opioid detoxification were analyzed. More than every third injection drug user (34.7%) experienced emergency room treatment due to an overdose. Using multiple logistic regression not living with a significant other drug user (odds ratio [OR] = 1.78, P = .002), history of suicide attempt (OR = 3.0, P = .000), daily use of barbiturates (OR = 2.17, P = .006) and cannabis (OR = 1.89, P = .001) were independently associated with emergency room treatment, whereas shorter duration of opioid use (OR = 0.23, P = .001) was independently associated with lack of emergency room treatment. Suicidal thoughts and multiple use of central nervous system depressants should be considered in injection drug users entering the emergency room due to an overdose. Emergency rooms should be seen as important places for offering further assistance (e.g., counselling) or referral to an addiction unit to drug users.

Background: High sympathetic tone creates a significant risk for ventricular arrhythmias and sudden death, which can especially affect patients after a myocardial infarction (MI) when exercising in a hypoxic environment. Methods and Results: The 16 patients after MI and 10 normal volunteers (NV) underwent a 1-day trip from low altitude (540 m, Bern) to high altitude (3,564 m Jungfraujoch, Swiss Alps). Autonomic function under resting and mental stress conditions at low and high altitude was assessed. MI patients demonstrated a significantly lower stroke volume (P

With lengthening (eccentric) muscle contractions, the magnitude of locomotor-muscle mass and strength increase has been demonstrated to be greater compared with shortening (concentric) muscle contractions. In healthy subjects, energy demand and heart rate responses with eccentric exercise are small relative to the amount of muscle force produced. Thus, eccentric exercise may be an attractive alternative to resistance exercise for patients with limited cardiovascular exercise tolerance.We tested the cardiovascular tolerance of eccentric exercise in 13 coronary patients (ages 40-66) with preserved and/or mild reduced left ventricular function. Patients were randomly assigned to either an eccentric (ECC; N = 7) or a concentric (CON; N = 6) training group and trained for 8 wk. Training workload was increased progressively (from week 1 to 5) to an intensity equivalent to 60% [OV0312]O(2peak).On average, maximum power output achieved with ECC was fourfold compared with CON (357 +/- 96 W vs 97 +/- 21 W; P0.005), whereas measures of oxygen uptake and blood lactate were significantly lower (P0.05 each), and ratings of perceived exertion were similar for ECC and CON. During a 20-min session of ECC and CON, central hemodynamics was measured by means of right heart catheterization. During ECC, responses of mean arterial blood pressure, systemic vascular resistance, pulmonary capillary pressure, cardiac index, and stroke work of the left ventricle on average were in the normal range of values and similar to those observed during CON. Compared with baseline, after 8 wk of training, echocardiographic left ventricular function was unchanged.The results indicate uncoupling of skeletal muscle load and cardiovascular stress during ECC. For low-risk patients with coronary heart disease without angina, inducible ischemia, or left ventricular dysfunction, ECC can be recommended as a safe new approach to perform high-load muscular exercise training with minimal cardiovascular stress.

A review of methods used for exercise training in stable chronic heart failure patients (CHF) shows a lack of standardization to guide prescription. Previous recommendations have been adopted from fitness training or rehabilitation studies. A model for use in CHF patients requires specific guidelines which respect the various manifestations of this illness. Pathology and exercise tolerance of patients with CHF allow only a few selected activities to be performed, such as walking and cycle ergometer training. Although the steady state method has usually been applied for aerobic exercise, the interval method has been shown to cause greater exercise stimuli to peripheral muscle than that obtained during steady state training methods without inducing greater cardiovascular stress. There is no consensus at present as to an optimal parameter for measuring intensity. An intensity of 40-80% peak oxygen consumption (VO(2)) has been applied successfully. A heart rate reserve of 60-80% or 75% of peak heart rate was used as a guide to exercise intensity without consideration of the impaired force-frequency relationship in myocardial performance. Because intensity, duration, and frequency of exercise are closely interrelated, initial exercise should be kept at 40-50% peak VO(2) with exercise duration of > 3-5 min x session performed several times daily. Progression should be followed in this order: duration, then frequency, then intensity. Resistance training can be recommended when small muscle groups are involved, using short bouts of work phases and small numbers of repetitions. To increase respiratory muscle strength and endurance, resistive inspiratory muscle training at intensity 25--35% maximum inspiratory pressure, and performed 20-30 min x d(-1), is recommended. On the basis of currently available research, supervised inpatient training programs should be preferred. Future research should be performed with respect on statistically sufficient, randomized, and controlled long-term studies that compare different training modes, intensities, frequency/duration ratios, and rates of progression.

This study sought to assess the safety of interval exercise training in patients with chronic congestive heart failure (CHF) with respect to left ventricular (LV) function. For effective rehabilitation in CHF, both aerobic capacity and muscle strength need to be improved. We have previously demonstrated in both coronary artery bypass surgery and patients with CHF that interval exercise training (IET) offers advantages over steady-state exercise training (SSET). However, because LV function during IET has not yet been studied, the safety of this method in CHF remains unclear. To assess LV function during IET and SSET, at the same average power output, 11 patients with stable CHF were compared with 9 stable coronary patients with minimal LV dysfunction (control group). Using first-pass radionuclide ventriculography, changes in LV function were assessed during work versus recovery phases, at temporally matched times between the fifth and sixteenth minute of IET and SSET. In CHF during IET, there were no significant variations in the parameters measured during work and/or recovery phases. During the course of both IET and SSET, there was a significant increase in LV ejection fraction (5 vs 4 U; p

Background: The work incapacity of ankylosing spondylitis (AS) ranges between 3% and 50% in Europe. In many countries, work incapacity is difficult to quantify. The work ability index (WAI) is applied to measure the work ability in workers, but it is not well investigated in patients. Aims: To investigate the work incapacity in terms of absence days in patients with AS and to evaluate whether the WAI reflects the absence from work. Hypothesis: Absence days can be estimated based on the WAI and other variables. Design: Cross-sectional design. Setting: In a secondary care centre in Switzerland, the WAI and a questionnaire about work absence were administered in AS patients prior to cardiovascular training. The number of absence days was collected retrospectively. The absence days were estimated using a two-part regression model. Participants: 92 AS patients (58 men (63%)). Inclusion criteria: AS diagnosis, ability to cycle, age between 18 and 65 years. Exclusion criteria: severe heart disease. Primary and secondary outcome measures: Absence days. Results: Of the 92 patients, 14 received a disability pension and 78 were in the working process. The median absence days per year of the 78 patients due to AS alone and including other reasons was 0 days (IQR 0–12.3) and 2.5 days (IQR 0–19), respectively. The WAI score (regression coefficient=−4.66 (p

Melanoma inhibitory activity (MIA), a small soluble secreted protein, is functionally important for progression of malignant melanoma. We recently revealed that p54(nrb) acts as a mediator of MIA action. In this study, we characterize the transcriptional regulation of p54(nrb) by MIA to explain MIA's molecular action. We identified one highly conserved region in the p54(nrb) promoter that is necessary and sufficient for MIA-dependent activation. Functional promoter analysis identified the transcription factor YBX1 as the mediator of MIA activation of p54(nrb) transcription. We screened the genome for further potential MIA-regulated genes carrying the element in their promoter regions. Integrating our sequence data with expression data from human melanomas identified a list of 23 potential MIA-YBX1 targets in melanomas. In summary, we present for the first time effects of MIA on transcriptional regulation. Uncovering new potential downstream effectors working via activation of YBX1 supports the important role of MIA in melanoma.

Immune cell infiltration varies widely between different glioblastomas (GBMs). The underlying mechanism, however, remains unknown. Here we show that TGF-beta regulates proliferation, migration, and tumorigenicity of mesenchymal GBM cancer stem cells (CSCs) in vivo and in vitro. In contrast, proneural GBM CSCs resisted TGF-beta due to TGFR2 deficiency. In vivo, a substantially increased infiltration of immune cells was observed in mesenchymal GBMs, while immune infiltrates were rare in proneural GBMs. On a functional level, proneural CSC lines caused a significantly stronger TGF-beta-dependent suppression of NKG2D expression on CD8(+) T and NK cells in vitro providing a mechanistic explanation for the reduced immune infiltration of proneural GBMs. Thus, the molecular subtype of CSCs TGF-beta-dependently contributes to the degree of immune infiltration.

Our data suggests that traumatic brain injury (TBI) may account for up to one third of battle-related injuries in today's war. Although the majority of these injuries are classified as mild in severity, service members with severe or penetrating TBI can be faced with many challenges. Injuries sustained on the battlefield require a slightly different approach than the TBI care that is traditionally seen in a civilian setting. This article presents the range of care that occurs beginning on the battlefield and continuing to state-of-the-art rehabilitation within the Department of Defense and Veterans Affairs Polytrauma System of Care.

OBJECTIVE: In Switzerland there is a shortage of population-based information on stroke incidence and case fatalities (CF). The aim of this study was to estimate stroke event rates and both in- and out-of-hospital CF rates. METHODS: Data on stroke diagnoses, coded according to I60-I64 (ICD 10), were taken from the Federal Hospital Discharge Statistics database (HOST) and the Cause of Death database (CoD) for the year 2004. The number of total stroke events and of age- and gender-specific and agestandardised event rates were estimated; overall CF, in-hospital and out-of-hospital, were determined. RESULTS: Among the overall number of 13 996 hospital discharges from stroke (HOST) the number was lower in women (n = 6736) than in men (n = 7260). A total of 3568 deaths (2137 women and 1431 men) due to stroke were recorded in the CoD database. The number of estimated stroke events was 15 733, and higher in women (n = 7933) than in men (n = 7800). Men presented significantly higher age-specific stroke event rates and a higher age-standardised event rate (178.7/100 000 versus 119.7/100 000). Overall CF rates were significantly higher for women (26.9%) than for men (18.4%). The same was true of out-of-hospital CF but not of in-hospital CF rates. CONCLUSION: The data on estimated stroke events obtained indicate that stroke discharge rate underestimates the stroke event rate. Out-of-hospital deaths from stroke accounted for the largest proportion of total stroke deaths. Sex differences in both number of total stroke events and deaths could be explained by the higher proportion of women than men aged 55+ in the Swiss population.

Weight loss affects cardiovascular risk profiles in obese patients. Surgery is not a plausible or viable response to the public health problem of obesity, given that more than 30% of adults are obese in some countries. However, most studies investigating the effects of weight loss on the cardiovascular risk profile are focussed on weight loss and limited to short-term effects. Since newer data show a rebound of cardiovascular risks in studies that complete a short-term follow-up, the question arises whether, and to what extent, long-term treatments offer a more sustained cardiovascular benefit beside the extensive or less marked weight loss. Aims The purpose of this article is to critically review existing data on the long-term cardiovascular effects of weight loss in obese and overweight patients treated with dietary interventions, physical activity programmes, behavioural therapy and pharmacological treatments and their combination. Method Inclusion criteria were peer-reviewed, randomized controlled trials (RCT) in the English language which presented data on cardiovascular effects at a follow-up of at least 18 months during or after weight reduction interventions. The search was limited to adults and the publication years between 1990 and 2007. Studies of patients with diagnoses such as coronary heart disease and cancer, and medically treated diabetes and hypertension were excluded. Results Twenty-three studies measured cardiovascular risk factors after 18 months or more. Mean BMI was 33.9 kg/m2 including 13,733 patients. Mean duration of the studies was 37 months with a dropout rate of 16.1% on average. Regardless of the absolute amount of weight loss a positive effect on cardiovascular risk factors such as blood pressure, lipids and glucose tolerance was found.

Purpose: To evaluate the effects of a comprehensive outpatient rehabilitation program in chronic heart failure (CHF) on quality of life (QoL) in relation to emotional status and clinical severity of disease. Patients and Methods: 25 patients with CHF were included in the 12-week comprehensive rehabilitation program. Initially, and at the end of the program, patients underwent graded cardio-pulmonary exercise testing, echocardiography, and determination of brain natriuretic peptide (BNP) concentration. In addition, they were assessed using: The Minnesota Living with Health Failure Questionnaire [MLHFQ]) for disease Specific QoL, the Hospital Anxiety and Depression Scale [HADS], and the State-Trait Anger Expression Inventory [STAXI]). Results: After 12 weeks of rehabilitation improvements in NYHA class, left ventricular ejection fraction (LVEF), and peak oxygen consumption were found, while mean BNP concentrations did not change. Disease specific QoL demonstrated improvements in physical component and the total score. Relative improvement of psycho-emotional scores correlated positively with relative improvement of disease specific QoL. In patients with LVEF ?30% at baseline, changes in BNP concentration were positively associated with both anxiety and state anger, and with the emotional component score of MLHFQ. Conclusion: Improvements in disease-specific QoL were closely associated to improvements of psycho-emotional status and clinical severity of CHF.

Mitochondrial apoptosis-inducing factor (AIF) influences the oxidative phosphorylation (OXPHOS) system and can be recruited as a mediator of cell death. Pathogenic mutations in the AIFM1 gene cause severe human diseases. Clinical manifestations include inherited peripheral neuropathies, prenatal cerebral abnormalities and progressive mitochondrial encephalomyopathies. In humans, rodents and invertebrates, AIF deficiency results in loss of respiratory complexes and, therefore, impaired OXPHOS. The molecular mechanisms underlying AIF-induced mitochondrial dysfunction remain elusive. Here we show that AIF physically interacts with the oxidoreductase CHCHD4/MIA40. In patient-derived fibroblasts as well as in tissues and glia cells from Harlequin (Hq) mutant mice, AIF deficiency correlates with decreased MIA40 protein levels, without affecting mRNA transcription. Importantly, MIA40 overexpression counteracts loss of respiratory subunits in Hq cells. Together, our findings suggest that MIA40 reduction contributes to the effects of AIF deficiency on OXPHOS, as it may impact on the correct assembly and maintenance of the respiratory subunits. This may be relevant for the development of new therapeutic approaches for AIF-related mitochondrial disorders.

Purpose: Although exercise training is established as an integrated part of treatment regimes in both patients with transmural myocardial infarction (MI) and chronic congestive heart failure (CHF), there is no consensus yet on the appropriateness of water exercises and swimming. One reason is the lack of information concerning both central hemodynamic volume and pressure responses during immersion in these patients. Methods: This paper presents explorative studies on changes in cardiac dimensions and central hemodynamics during graded immersion and swimming in patients with moderate and/or severe MI and in patients with moderate and/or compensated severe CHF. For comparison purposes, healthy subjects were assessed. Measurements were performed by using Swan-Ganz right heart catheterization, subxiphoidal echocardiography, and Doppler-echocardiography. Results: The major findings were: 1) Indicators of an increase in preload were seen in patients with moderate and severe MI. In both patient groups, upright immersion to the neck and supine body position at rest in the water resulted in abnormal mean pulmonary artery pressure (PAm) and mean pulmonary capillary pressures (PCPm), respectively. During low-speed swimming (20–25 m·min−1), the PAm and/or PCPm were higher than during supine cycle ergometry at a load of 100 W. 2) Left ventricular overload and decrease and/or no change in stroke volume occurred in patients with severe CHF who were immersed up to the neck. 3) Patient’s well-being was maintained despite hemodynamic deterioration. Conclusion: The acute responses during immersion and swimming suggest the need for additional studies on long-term changes in cardiac dimensions and central hemodynamic in both patients with severe MI and severe CHF who undergo a swimming program, compared with nonswimming patients with MI and CHF of similar etiology and severity of disease.

This evaluation was performed to assess the effects of a new, comprehensive outpatient rehabilitation program on generic and disease-specific quality of life related to exercise tolerance in stable chronic heart failure patients. Fifty-one patients (aged 59+/-11 years; 84% men) were treated for 12 weeks. Patients underwent optimized drug treatment, exercise training, and counseling and education. At baseline and at the end of the program, functional status, exercise capacity, and quality of life were assessed using the Medical Outcomes Study 36-item Short-Form Health Survey and the Minnesota Living with Heart Failure Questionnaire. Left ventricular ejection fraction and New York Heart Association functional class, as well as measures of physical fitness and walking distance covered in 6 minutes, improved significantly (by 11%-20% and by 58% on average, respectively). Physical functioning (effect size, 0.38; p

This retrospective study aims to determine whether there is a difference in the additional consumption of alcohol between addicts treated with methadone or dihydrocodeine (DHC) and untreated addicts injecting heroin.1,685 patients admitted for opioid withdrawal between 1991 and 1997 were reviewed. Cross-reference tables and multiple logistic regression analyses were carried out. 28% of patients take more than 40 g of alcohol daily (on average 176 g). We found that patients who are treated with methadone or DHC drink alcohol significantly more often daily than the heroin-dependent patients (p < 0.01). Using multiple regression analyses, the results were confirmed. Additionally, we found that co-abuse of alcohol was predicted by male gender, longer duration of drug use, additional daily consumption of tetrahydrocannabinol and daily consumption of benzodiazepines. Alcohol consumption by opioid-addicted patients treated with methadone or DHC presents a serious medical problem. Co-abuse of alcohol will receive more attention.

To increase muscle mass and strength in patients with chronic congestive heart failure (CHF), there is a need for implementing resistance exercises in exercise training programs. This study sought to assess the safety of rhythmic strength exercise with respect to left ventricular function in 9 patients with stable CHF, compared with 6 stable coronary patients with mild left ventricular dysfunction (control group). With use of right-sided catheterization, changes in left ventricular function were assessed during double leg press exercise at loads of 60% and 80% of maximum voluntary contraction. The exercise sessions lasted 14 minutes each, divided into work and recovery phases of 60/120 seconds. In CHF, during exercise at a 60% load, there was a significant increase in heart rate (mean +/- SEM 90 +/- 4 beats/min; p0.05), mean arterial blood pressure (95 +/- 3 mm Hg; p0.01), diastolic pulmonary artery pressure (20.2 +/- 2.7 mm Hg; p0.01), and cardiac index (3 +/- 0.3 L/m2/min; p0.05). Additionally, during leg press exercise at an 80% load, there was a significant decrease in systemic vascular resistance (1,086 +/- 80 dynes x s x cm(-5); p0.001), an increased cardiac index (3.4 +/- 0.1; p0.001), and left ventricular stroke work index (75 +/- 5 g x m/m2; p0.01), suggesting enhanced left ventricular function. Compared with controls, in CHF the magnitude of changes in hemodynamic parameters during exercise, demonstrated at a 60% load, was significantly smaller (systemic vascular resistance: [mean] 1,613 --1000 vs 1472 --1,247 dynes x s x cm(-5); cardiac index: 2.4 --3 vs 2.8 --4.4 L/m2/min, and stroke work index: 60 --69 vs 114 --155 g x m/m2; p0.05 each). Nevertheless, changes indicated an enhanced contractile function of the left ventricle in CHF. This study demonstrates stability of left ventricular function during resistance exercise in well-compensated CHF patients with optimal drug therapy, as well as the appropriateness of the chosen mode and intensity applied as these factors relate to cardiovascular stress. This conclusion cannot be extrapolated to patients with less well-compensated heart failure, or to more protracted resistance training.

Measures of left ventricular function during exercise provide information that is more accurate than the exercise ECG in the diagnosis of coronary artery disease, supportive of the data provided by myocardial perfusion studies, and of great prognostic significance. We review basic methods for evaluating left ventricular function during exercise and responses to various types of exercise, including incremental exercise and exercise training conditions. Additionally, we review changes in both incremental exercise test responses and responses to training in various pathological conditions. Case reports are included to illustrate the utility of measuring left ventricular function during exercise.

PURPOSE: Kinetics of VO2 at onset of constant work rate exercise was previously shown to be slowed in patients with chronic heart failure (CHF) compared with that in healthy normals. Because bicycle ergometry with ramp protocol is usually used for exercise testing with CHF patients, it would be of practical importance if it can be shown that a delay in the time interval of linear increase of VO2 (TILIV) to work rate occurs after beginning ramp exercise. Data of central hemodynamics (CHF) and noninvasive cardiopulmonary parameters (CHF, normals) should also correlate with VO2 delay time if this parameter is related to cardiopulmonary exercise capacity. METHODS: Fifteen males with CHF (mean +/- SEM: age 52 +/- 2 yr; ejection fraction 32 +/- 4%; peak cardiac index 3.9 +/- 0.3 L x m(-2) x min(-1)) and 28 healthy males (50 +/- 1 yr) were assessed. During ramp bicycle ergometry (3 min unloaded, work rate increments of 12.5 W x min(-1)), VO2 was measured breath by breath. RESULTS: After the onset of ramp exercise, there was a difference in the TILIV between patients and normals (83.7 +/- 3.6 vs 66.8 +/- 2.9 s; P < 0.001). Significant differences between both groups were also found for VO2 at ventilatory threshold (VT) (10.1 +/- 0.1 vs 15.2 +/- 0.7 mL x kg(-1) x min(-1); P < 0.0001), VO2 at VT relative to predicted VT (58 +/- 4 vs 97 +/- 4%; P < 0.0001), peak VO2 (13.2 +/- 1.0 vs 34 +/- 1.4 mL x kg(-1) x min(-1), P < 0.001), and increase of systolic blood pressure (36 +/- 7 vs 71 +/- 5 mm Hg; P < 0.0001). In CHF, the TILIV correlated significantly with peak cardiac index and VO2 at VT (r = -0.71; P < 0.005 each), relative value of VO2/kg at VT (r = -0.61; P < 0.03), peak VO2/kg (r = -0.63; P < 0.01), and increase of systolic blood pressure (r = -0.52; P < 0.02). In the normals only VO2/kg at VT correlated significantly with TILIV (r = -0.41; P < 0.03). In patients, stepwise regression analysis identified three predictors which could explain 79% of the variance of TILIV: VO2/kg at VT (r2 = 0.51), peak cardiac index (r2 = 0.20), and peak VO2/kg (r2 = 0.08). CONCLUSION: TILIV, determined at the onset of ramp exercise, is prolonged in CHF patients compared with that in normals and reflects severity of functional impairment because of reduced cardiac index and aerobic capacity. TILIV can provide information about changes in cardiopulmonary exercise capacity and thus can be used for follow-up and treatment studies in CHF.

Aqua therapy for patients with chronic heart failure may be dangerous Aqua exercise and swimming are traditionally recommended for low risk cardiac patients. In patients with severe myocardial infarction (MI) and compensated severe chronic heart failure (CHF), physicians may have reservations about water exercises and swimming. Concerns become understandable when central haemodynamic responses in healthy subjects during immersion and swimming are considered. During neck deep water immersion, a 100 cm column of water exerts a pressure of 76 mm Hg on the body surface. During swimming, a pressure of 40–60 mm Hg can be assumed. This pressure compresses superficial veins, particularly of the lower extremities and abdomen, resulting in a blood volume shift to the thorax and heart. On immersion up to the iliac crest, the blood volume shift is not significant, but, on immersion up to the neck, the central blood volume has been shown to be increased by about 700 ml; 180–240 ml of this was allotted to the heart volume, with enlargement of all four chambers.1 Planimetry of the diastolic poster-anterior area of the heart has shown an average increase in heart size of 30% within six seconds.2 From a central haemodynamic point of view, crucial immersion starts at the diaphragm/xiphoid level. At this level, both the buoyancy effect3 and the external hydrostatic pressure result in a blood volume shift, which ( a ) corresponds to up to 85% of …

We prospectively assessed whether baseline central hemodynamics and exercise capacity can predict improvement of VO2 at ventilatory threshold (VT) after exercise training in patients with severe chronic congestive heart failure. Eighteen patients (mean +/- SEM; age 52 +/- 2 years), half of them listed for transplant, underwent 3 weeks of exercise training (interval cycle and treadmill walking; 5 x/week) and 3 weeks of activity restriction in a random-order crossover trial. Baseline data were not significantly different for groups with exercise training first and activity restriction first: cardiac index at rest (2.1 +/- 0.1 L/m2/min), maximum cardiac index (3.1 +/- 0.2 L/m2/min) (Fick), and echocardiographic ejection fraction (21 +/- 1%). The same was true for cardiopulmonary exercise data (cycle ergometry; up 12.5 W/min): VO2 at VT (9.3 +/- 0.4 ml/kg/min), maximum VO2 (12.2 +/- 0.7 ml/kg/min), VT in percentage of predicted maximum VO2 (31 +/- 2%), heart rate at VT (95 +/- 4 beats/min), and decrease of dead space-to-tidal volume ratio from rest to VT (33 +/- 1 --29 +/- 1). Improvement of VO2 at VT after training (2.2 +/- 0.4 ml/kg/min; p0.001) was not related to baseline central hemodynamics (r =0.10 for each), but was greater in patients with a lower baseline VO2 at VT (r = -0.65; p0.01), peak VO2 (r = -0.66; p0.01), VT in percentage of predicted maximum VO2 (r = -0.74; p0.001), heart rate at VT (r = -0.63; p0.01), and smaller decrease of dead space-to-tidal volume ratio from rest to VT (r = 0.65; p0.01). Ejection fraction after exercise training (24 +/- 2%) and activity restriction (23 +/- 2%) did not differ significantly compared with baseline, and patient status (heart failure and cardiac rhythm) remained stable. Three parameters accounted for 84% of the variance of improvement in VO2 at VT: VO2 at VT in percent predicted maximum VO2, decrease of dead space-to-tidal volume ratio, and heart rate at VT. The findings suggest that there was a greater increase in VO2 at VT after exercise training in patients with greater peripheral deconditioning at baseline. The improvement was unrelated to central hemodynamics. Clinically stable patients with severe chronic congestive heart failure, potential heart transplant candidates, and those awaiting transplantation may benefit from involvement in a short-term exercise training program.

Eleven men with severe chronic heart failure (peak cardiac index 4.0 ± 0.2 L/m 2 /min), six on a heart transplantation waiting list, were prospectively assessed. To determine reproducibility of cardiopulmonary and hemodynamic variables for clinical purposes during ramp bicycle ergometry, the patients underwent two ramp bicycle ergometer tests (3 minutes unloaded, work rate increments of 12.5 W/min) with a 1-week interval between tests. Oxygen uptake (v O 2 ) carbon dioxide production (v CO 2 ), and ventilation were measured breath by breath, and calculations were performed to determine gas exchange ratio, oxygen pulse, ventilatory equivalents of oxygen and carbon dioxide, and end-tidal partial pressure for oxygen and carbon dioxide. Additionally, heart rate, blood pressure, and lactate levels were assessed. Measurements were performed at submaximum work rate levels of 25 W, 50 W, and 75 W at ventilatory threshold and at peak work rate. At all measurement points, the coefficient of variation for cardiopulmonary variables was between 1.4% and 7.1% for submaximum work rate levels, between 1.2% and 4.4% at ventilatory threshold, and between 2.4% and 7.1% at peak work rate. For heart rate, blood pressure, and lactate levels, coefficient of variation was between 2.7% and 5.7% for submaximum work rate levels, between 1.4% and 6.1% at ventilatory threshold, and between 1.2% and 5.5% at peak work rate. The data suggest high reproducibility for duplicate measurements of cardiopulmonary and hemodynamic variables during ramp bicycle ergometry in patients with severe chronic heart failure. The results may be used to determine whether any variable in a single patient is significantly different from that obtained in a previous exercise test or if the change is within experimental error. (Am Heart J 1997;134: 20-6.)

The effect of a 3-week exercise programme on performance and economy of walking was analysed in 16 male patients with chronic heart failure [mean age 51.8 (SD 6.9) years, height 174.9 (SD 6.3) cm, body mass 75.3 (SD 11.5) kg, ejection fraction 20.8 (SD 5.0)%]. They were submitted to a cardiopulmonary exercise test on a cycle ergometer and a 6-min walking test on a treadmill before and after the period of exercise training. The training programme consisted of interval cycle (five times a week for 15 min), and treadmill ergometer training (three times a week for 10 min) at approximately 70% cycling peak oxygen uptake (VO2peak) and supplementary exercises (three times a week for 20 min). Compared to the pre values cycling VO2peak [11.9 (SD 2.9) vs 14.0 (SD 2.3) ml. kg-1.min-1], maximal self paced walking speed [0.68 (SD 0.33) vs 1.16 (SD 0.30) m.s-1], and net walking power [2.16 (SD 0.89) vs 2.73 (SD 0.91) W.kg-1] had increased (P < 0.01) while net energy cost [3.31 (SD 0.66) vs 2.33 (SD 0.38) J.kg-1. m-1] had decreased (P < 0.001) after the training period. Approximately 42% of the increase of walking speed resulted from a higher walking power output, whereas approximately 58% corresponded to a positive effect on walking economy. The improvement in walking economy was a function of an increase in walking velocity itself and a result of a more efficient walking technique. These results would indicate that in patients with marked exercise intolerance, adequate exercise training programmes could contribute to favourable metabolic changes with positive effects on the economy of motion.

Previous exercise training studies in patients with chronic congestive heart failure (CHF) were performed for periods lasting > 2 months, and effects of activity restriction on exercise induced-benefits were not systematically assessed. With one exception study, patients were not reported to be transplant candidates. In this random-order crossover study, effects of 3 weeks of exercise training and 3 weeks of activity restriction on functional capacity in 18 hospitalized patients with severe CHF [(mean +/- SEM) age 52 +/- 2 years; ejection fraction 21 +/- 1%; half of them on a transplant waiting list] were assessed. The training program consisted of interval exercise with bicycle ergometer (15 minutes) 5 times weekly, interval treadmill walking (10 minutes), and exercises (20 minutes), each 3 times weekly. With training, the onset of ventilatory threshold was delayed (p < 0.001), with increased work rate by 57% (p < 0.001) and oxygen uptake by 23.7% (p < 0.001). On average, there was a 14.6% decrease in slope of ventilation/carbon dioxide production before the onset of ventilatory threshold (p < 0.05), and ventilatory equivalent of carbon dioxide production by 10.3% (p < 0.01). At the highest comparable work rate (56 +/- 5 W) the following variables were decreased: heart rate (7.3%; p < 0.05), lactate (26.6%; p < 0.001), and ratings of perceived leg fatigue and dyspnea (14.5% and 16.5%; p < 0.001 each). At peak exercise, oxygen uptake was increased by 19.7% (p < 0.01) and oxygen pulse by 14.2% (p < 0.01). There was a correlation of baseline peak oxygen uptake and increase of peak oxygen uptake due to training (r = -0.75; p < 0.004). Independently of the random order, data after activity restriction did not differ significantly from data measured at baseline. Patients with stable, severe CHF can achieve significant improvements in aerobic and ventilatory capacity and symptomology by short-term exercise training using interval exercise methods. Impairments due to activity restriction suggest the need for long-term exercise training.

Classes I/II and III of the classification systems of the New York Heart Association (NYHA), Canadian Cardiovascular Society (CCS) and American Medical Association (AMA) were compared with each other and with the Weber classification (O2 uptake, VO2/kg during treadmill walking) in 35 male patients with severe left ventricular dysfunction. Measured end points were ventilatory threshold (VT) and peak exercise. Also investigated was whether the CCS and AMA scales, due to their more stringent differentiation, are more precise than the NYHA system in determining a limited physical capacity and whether there are other differentiating factors useful in classification which may be derived from cardiopulmonary exercise testing. At the VT, the mean VO2/kg did not differ significantly in any classification system between classes I/II and III (12.8 +/- 2.5 vs. 11.1 +/- 2.3 ml/kg/min) and corresponded to Weber class B. At peak exercise, the mean VO2/kg only differed significantly within the NYHA classification; classes I/II (16.3 +/- 3.1 ml/kg/min) corresponded to Weber class B, and class III (13 +/- 3 ml/kg/min) to Weber class C. The individual values displayed a large scatter. Factors differing in classes I/II and III of all three systems at peak exercise were the ventilatory equivalent of O2 and CO2 as well as end-tidal partial pressure for O2 and CO2. At VT these factors showed a separating character only in the AMA classification. It is not possible to determine objective functional impairment by use of the NYHA, CCS and AMA systems because they are not analogous to the Weber system. Nevertheless, these classification systems can be used for clinical assessment and follow-up.

This study compares hemodynamic, metabolic, and gas exchange responses, catecholamine levels, and symptoms in 35 male patients with chronic heart failure (CHF) ([mean ± SD] age 53 ± 11 yr ; ejection fraction 24 ± 11%) during three differently graded exercise test protocols. On three consecutive days patients performed cycle ergometry supine, with prolonged steps (prol BE) and right heart catheterization, ramplike cycle ergometry sitting (ramp BE), and ramplike treadmill walking (TMW). As in routine clinical practice, the prol BE was terminated when pathologic central hemodynamics and/or increased symptomology occurred, and ramp BE and TMW due to increased symptomology and/or physician's decision. During prol BE at ventilatory threshold (VT) the VO 2 (8.6 ± 1.8 ml kg -1 .min -1 ) was lower than during ramp BE (9.3 ± 2.1 ml.kg -1 .min -1 ) (P < 0.017) and TMW (11.8 ± 2.3 ml.kg -1 .min -1 ) (P < 0.0001). Prol BE, ramp BE, and TMW also differed significantly with respect to ventilation (22 ± 7 l.min -1 ; 26 ± 6 l.min -1 ; 29 ± 7 l.min -1 ; P < 0.01) and heart rate (100 ± 15 beats.min -1 ; 103 ± 18 beats.min -1 ; 110 ± 16 beats.min -1 ; P < 0.017). No differences were found in lactate levels, catecholamine levels, and ratings of leg fatigue and dyspnea. At test termination, the peak VO 2 during prol BE (10.8 ± 3.3 ml.kg -1 .min -1 ) was lower than during ramp BE (13.3 ± 4.1 ml.kg -1 .min -1 ) (P < 0.0001) and TMW (14.7 ± 3.4 ml.kg -1 .min -1 ) (P < 0.0001). Peak norepinephrine value during ramp BE (4.531 ± 2.788 nmol.l -1 ) was higher than during prol BE (3.707 ± 2.262 nmol.l -1 ) (P < 0.001). Among the three tests, no significant differences were found for peak values of heart rate, lactate, and ratings of dyspnea. Although the VO 2 .kg -1 at VT was significantly higher during ramp BE and TMW compared to prol BE (P < 0.001), the values expressed as a percent of peak VO 2 .kg -1 were significantly lower (70 ± 4% ; 72 ± 6% ; 79 ± 3% ; P < 0.017). A systematic effect on aerobic capacity with reduced peak values during ramp BE and TMW was demonstrated when test termination was based primarily on pathological findings of central hemodynamics from prol BE.

In 309 postmyocardial infarction patients (age 40 +/- 7.7 years) without persistent ischemia there is only a weak correlation between ejection fraction (EF) and exercise tolerance (r = 0.45, p0.01), and between EF and maximum cardiac output (CO; r = 0.41, p0.01) and maximum pulmonary capillary wedge pressure (PCWP; r = -0.32, p0.001). The same was true for exercise tolerance and maximum PCWP (r = -0.53, p0.001). A big scattering of individual values could be observed. Although we found a high positive correlation between maximum CO and exercise tolerance (r = 0.80, p0.001), in individual cases a low CO could be related to high exercise tolerance and vice versa. In the multivariate analysis, only the heart volume/kg body weight and maximum PCWP could be shown to be of independent prognostic importance for survival and/or mortality in the following years (chi 2 = 5.9, p0.015 and chi 2 = 7.2, p0.007, respectively).

AtT20 (pituitary corticotroph) cells were transfected with either the native or a mutant [AspB10]rat insulin II gene, using a plasmid containing the insulin gene and a neomycin resistance gene under the control of independent constitutive promoters. The cellular immunoreactive insulin (IRI) content ranged from 0.8-440 ng/10(6) cells, with the highest value similar to that found for a rat insulinoma cell line (RIN) and corresponding to approximately 1% that of native pancreatic B-cells. There was a direct correlation between insulin mRNA levels and IRI content and no correlation between mRNA levels and rat insulin II gene copy number. Furthermore, in some lines the insulin II transgene was lost even though the gene encoding neomycin resistance was retained. IRI release was stimulated up to 4-fold by isobutylmethylxanthine in all lines transfected with the native rat insulin II gene, and HPLC analysis showed most IRI as fully processed insulin, with less than 5% as proinsulin. These cells, thus, directed most proinsulin to secretory granules for conversion and regulated release regardless of the absolute amount of IRI expressed. One of the lines transfected with the AspB10 mutant gene (line AA9) released nearly 50% of IRI as proinsulin under basal conditions, with stimulation of insulin, but not proinsulin, release by isobutylmethylxanthine. This confirmed our previous finding of partial diversion of this mutant proinsulin from the regulated to the constitutive pathway. A second line (IC6) expressing the same mutant gene at much higher levels appeared to direct all mutant proinsulin to the regulated pathway, suggesting that for this particular mutant proinsulin, the secretory pathway employed by the transfected cells can be affected by the amount of proinsulin synthesized.

With water immersion, gravity is partly eliminated, and the water exerts a pressure on the body surface. Consequently there is a blood volume shift from the periphery to the central circulation, resulting in marked volume loading of the thorax and heart. This paper presents a selection of published literature on water immersion, balneotherapy, aqua exercises, and swimming, in patients with left ventricular dysfunction (LVD) and/or stable chronic heart failure (CHF). Based on exploratory studies, central hemodynamic and neurohumoral responses of aquatic therapies will be illustrated. Major findings are: 1. In LVD and CHF, a positive effect of therapeutic warm-water tub bathing has been observed, which is assumed to be from afterload reduction due to peripheral vasodilatation caused by the warm water. 2. In coronary patients with LVD, at low-level water cycling the heart is working more efficiently than at lowlevel cycling outside of water. 3. In patients with previous extensive myocardial infarction, upright immersion to the neck resulted in temporary pathological increases in mean pulmonary artery pressure (mPAP) and mean pulmonary capillary pressures (mPCP). 4. Additionally, during slow swimming (20-25m/min) the mPAP and/or PCP were higher than during supine cycling outside water at a 100W load. 5. In CHF patients, neck- deep immersion resulted in a decrease or no change in stroke volume. 6. Although patients are hemodynamically compromised, they usually maintain a feeling of well-being during aquatic therapy. Based on these findings, clinical indications for aquatic therapies are proposed and ideas are presented to provoke further research.