Neonatal BCG vaccination and atopic dermatitis before 13 months of age: A randomized clinical trial.

BACKGROUND: Studies have suggested that Bacillus Calmette-Guérin (BCG) vaccination may reduce the risk of allergic diseases, including atopic dermatitis. METHODS: The Danish Calmette Study was conducted 2012-2015. Within 7 days of birth new-borns were randomised 1:1 to BCG or no BCG. Exclusion criteria were gestational age <32 weeks, birth weight <1000 g, known immunodeficiency or no Danish-speaking parent. Data were collected through telephone interviews and clinical examinations until 13 months. RESULTS: Clinical atopic dermatitis was diagnosed in 466/2,052 (22.7%) children in the BCG group and 495/1,952 (25.4%) children in the control group (RR = 0.90 [95% confidence intervals 0.80-1.00]). The effect of neonatal BCG vaccination differed significantly between children with atopic predisposition (RR 0.84 (0.74-0.95)) and children without atopic predisposition (RR 1.09 [0.88-1.37]) (test of no interaction, P = .04). CONCLUSION: Among children with atopic predisposition, the number-needed-to-treat with BCG to prevent one case of atopic dermatitis was 21 (12-76).

Bacillus Calmette-Guérin vaccination at birth and in vitro cytokine responses to non-specific stimulation. A randomized clinical trial.

Several studies have shown increased in vitro cytokine responses to non-related pathogens after Bacillus Calmette-Guérin (BCG) vaccination. A total of 158 infants (80 BCG administered within 7 days of birth; 78 controls) were bled 4 days post-randomization, and at age 3 and 13 months. Geometric mean concentrations of IL-1ß, TNF-α, IL-6 (24 h stimulation) and IFN-γ, IL-10, IL-17, IL-22 (96 h stimulation) in response to in vitro stimulation with RPMI, LPS, PHA, Escherichia coli, Streptococcus pneumoniae, Candida albicans and BCG were compared among BCG vaccinated children and controls. BCG vaccination did not affect in vitro cytokine production, except IFN-γ and IL-22 response to BCG. Stratifying for 'age at randomization' we found a potentiating effect of BCG on cytokine production (TNF-α, IL-6, IL-10) in the 4 days post randomization stimulations, among children who were vaccinated at age 2-7 days versus age 0-1 days. BCG vaccination did not potentiate cytokine production to non-BCG antigens. At 4 days post randomization, BCG was associated with higher cytokine production in the later randomized children.

Methodological quality, completeness of reporting and use of systematic reviews as evidence in clinical practice guidelines for paediatric overweight and obesity.

Paediatric obesity rates remain high despite extensive efforts to prevent and treat obesity in children. We investigated the quality of the methodology and reporting within systematic reviews (SRs) underpinning paediatric content in US clinical practice guidelines (CPGs). In June 2016 we searched guideline clearinghouses and professional organization websites for guidelines published by national or professional organizations in the United States from January 2007 onwards. In our primary, a priori analysis, we used PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) and AMSTAR (A Measurement Tool to Assess Systematic Reviews) instruments to score SRs and meta-analyses that included paediatric populations and were cited by included CPGs. In a secondary, post hoc analysis, we determined the extent to which US CPGs use available, relevant SRs and meta-analyses compared with non-US CPGs. Eight US-based CPGs with 27 references to 22 unique SRs were found. AMSTAR and PRISMA scores were low overall, with only three SRs having 'high' methodological quality. Items dealing with bias assessments and search strategies had especially low scores. US CPGs were also older on average and cited fewer SRs than their international counterparts. Low quality scores and dated guidelines should be a cause for concern among practicing clinicians and a call to action for future guideline developers, publishers and research institutions.

Relationship between [123 I]-FP-CIT SPECT and clinical progression in Parkinson's disease.

BACKGROUND: The demonstration of presynaptic dopaminergic deficiency on [123 I]-FP-CIT SPECT imaging is a useful ancillary tool in the diagnosis of Parkinson's disease (PD). Whilst there is evidence of a cross-sectional relationship between the degree of dopaminergic deficiency and severity of bradykinesia and rigidity, longitudinal studies are rare. Moreover, the relationship between motor subtypes and their dopaminergic deficient state is not well characterized. AIM: Our primary aim was to assess the correlations between dopaminergic deficiency on baseline [123 I]-FP-CIT SPECT imaging with the progression of motor severity in patients classified by motor subtype, and the development of motor complications. Our secondary aim was to assess the correlation between UPDRS-III subscores and the time to onset of motor complications. METHODS: 42 PD patients with abnormal baseline [123 I]-FP-CIT SPECT scans and at least 3 years of clinical follow-up were classified by motor subtype: akinetic-rigid, tremor-dominant or mixed. UPDRS-III scores at baseline and at 3-year follow-up, and time to onset of motor complications were recorded. RESULTS: [123 I]-FP-CIT uptake ratios were inversely correlated with UPDRS-III scores at 3 years only in akinetic-rigid patients (r=-.51, P=.04). Time to onset of motor complications was inversely correlated with UPDRS-III subscores for bradykinesia and rigidity at baseline (r=-.52, P=.02) and at 3 years (r=-.54, P=.01). CONCLUSION: The degree of dopaminergic deficiency on baseline [123 I]-FP-CIT SPECT inversely correlates with motor severity at 3-year follow-up in akinetic-rigid patients only. Furthermore, UPDRS-III subscores for bradykinesia and rigidity at baseline show an inverse correlation with time to onset of motor complications across all PD subtypes.

Rapid balloon distension as a tool to study cortical processing of visceral sensations and pain.

BACKGROUND: The processing of discomfort and pain in the central nervous system is normally studied with experimental methods, but it is mandatory that they are reliable over time to ensure that any interventions will result in valid results. We investigated reliability of rapid balloon distension in the rectum to elicit cortical evoked potentials (CEPs) to study the reliability of central processing of visceral sensation and discomfort/pain. METHODS: Eighteen healthy volunteers had two series of rectal balloon distensions performed on two separate days. Individualized balloon pressure, corresponding to pain detection threshold or to the maximum possible distension (30 psi), was used. Within- and between days reliability was measured in terms of amplitudes and latencies of the CEP global field power, topography and underlying brain networks. KEY RESULTS: There were two prominent peaks in the CEP recordings at mean latencies of 157 and 322 ms. There were no differences in latencies or amplitudes (p = 0.3) and they passed the Bland-Altman test for reproducibility. There were no differences in topographies (p > 0.7). Brain source connectivity revealed the cingulate-operculum network as the most consistent network within and between subjects. There were no differences in the location of brain sources in this network (p = 0.9) and the source coordinates were reproducible. Finally, the cingulate source generally had higher strength than operculum source (p < 0.001). CONCLUSIONS & INFERENCES: A reliable method to study central mechanisms underlying visceral sensation and pain was established. The method may improve our understanding of visceral pain and could be an objective method for studying efficacy of analgesics on visceral pain.

Cortical evoked potentials in response to rapid balloon distension of the rectum and anal canal.

BACKGROUND: Neurophysiological evaluation of anorectal sensory function is hampered by a paucity of methods. Rapid balloon distension (RBD) has been introduced to describe the cerebral response to rectal distension, but it has not successfully been applied to the anal canal. METHODS: Nineteen healthy women received 30 RBDs in the rectum and the anal canal at intensities corresponding to sensory and unpleasantness thresholds, and response was recorded as cortical evoked potentials (CEPs) in 64-channels. The anal canal stimulations at unpleasantness level were repeated after 4 min to test the within-day reproducibility. CEPs were averaged, and to overcome latency variation related to jitter the spectral content of single sweeps was also computed. KEY RESULTS: Repeated stimulation of the anal canal generated CEPs with similar latencies but smaller amplitudes compared to those from the rectum. Due to latency jitter, reproducibility of averaged CEPs was lower than what was found in the rectum. The most reproducible feature was N2P2 peak-to-peak amplitude with intra-class correlation coefficient (ICC) of 0.7 and coefficient of variation (CV) of 18%. Spectral content of the single sweeps showed reproducibility with ICCs for all bands >0.8 and corresponding CVs <7%. CONCLUSIONS & INFERENCES: Cortical potentials evoked from the anal canal are challenged by latency jitter likely related to variability in muscle tone due to the distensions. Using single-sweep analysis, anal CEPs proved to be reproducible and should be used in future evaluation of the anal function.

Reproducibility of psychophysics and electroencephalography during offset analgesia.

BACKGROUND: Offset analgesia (OA) is a pain-inhibiting mechanism, defined as a disproportionately large decrease in pain perception in response to a discrete decrease in noxious stimulus intensity. Hence, the aims were (1) to investigate whether psychophysics and electroencephalography (EEG) can be assessed simultaneously during OA and (2) to assess whether OA is reproducible within the same day as well as between different days. METHODS: Two separate studies investigated OA: Study I (13 healthy volunteers; seven men; 25.5 ± 0.65 years) aimed at determining the feasibility of recording psychophysics and EEG simultaneously during OA. Study II (18 healthy volunteers; 12 men; 34 ± 3.15 years) assessed reproducibility of OA in terms of psychophysics and EEG. Subjects were presented to a 30-s OA heat stimulus paradigm on the volar forearm and psychophysics, and EEG recordings were obtained throughout the procedure. Reproducibility was assessed within the same day and between different days, using intraclass correlation coefficients (ICCs). Additionally, the reproducible psychophysical parameters were correlated to relevant EEG frequency bands. RESULTS: Simultaneous recording of psychophysics and EEG affects the frequency distribution in terms of alpha suppression. Reproducibility was proven for the psychophysics and EEG frequency bands both within the same day (all ICCs > 0.62) and between different days (all ICCs > 0.66, except for the delta band). Correlations between psychophysics and EEG were found in the theta (4-8 Hz), alpha (8-12 Hz) and gamma (32-80 Hz) bands (all p < 0.01). CONCLUSION: OA is a robust and reproducible model for experimental pain research, making it suitable for future research.

Baseline [(123) I]FP-CIT SPECT (DaTSCAN) severity correlates with medication use at 3 years in Parkinson's disease.

BACKGROUND: Presynaptic dopaminergic deficiency on dopamine transporter imaging supports a clinical diagnosis of Parkinson's disease and correlates with the severity of rigidity and bradykinesia. Baseline dopaminergic deficiency predicts clinical severity, but the relationship with subsequent medication use has not been reported. METHODS: A randomly selected cross section of 83 Parkinson's disease (PD) patients who had [(123) I] FP-CIT SPECT at the time of clinical diagnosis was identified. Dopaminergic deficiency was graded 1, 2 or 3 with increasing severity using visual assessment and by semiquantitative analysis of putamen and caudate uptake. Antiparkinson medication usage and clinical severity by Hoehn and Yahr were noted annually to 3 years. RESULTS: In 83 patients (66% male, median age 65.0 years, IQ 55.4-71.8), [(123) I]FP-CIT SPECT was grade 1 in 20 (24%), grade 2 in 53 (64%) and grade 3 in 10 patients (12%). Dopamine transporter uptake ratios were inversely associated with antiparkinson medication usage (r = -0.26, P = 0.0201) and Hoehn Yahr stage (r = -0.32, P = 0.0029) at 3 years from baseline, but there was considerable variation in drug usage in individual patients. At 3 years, patients with grade 1 scans at baseline received a median dose of 325 levodopa equivalent units (LEU) (interquartile range 175-433); grade 2 scan patients 400 LEU (interquartile range 300-635); and grade 3 scan patients 460 LEU (interquartile range 252-658). CONCLUSION: The degree of reduction in presynaptic dopaminergic uptake at baseline is associated with higher antiparkinson drug dosage at follow-up, but the wide variation means that the baseline FP-CIT SPECT does not reliably predict drug use in individual cases.

Duration of L-dopa and dopamine agonist monotherapy in Parkinson's disease.

The expected duration of initial antiparkinson monotherapy before the need for supplementation is not clearly defined for routine practice. The aim of this study was to define the length of L-dopa (L-3, 4-dihydrophenylalanine) and dopamine agonist monotherapy. The duration of monotherapy and discontinuation rates were investigated in a natural observational setting by plotting Kaplan-Meier survival curves. Out of 345 patients, 180 (52.2%) received L-dopa and 165 (47.8%) received a dopamine agonist as initial monotherapy. Half of the patients starting L-dopa received supplementary therapy with- in 3.6 years (95% confidence interval, 3.2-4.6), significantly longer than for dopamine agonist monotherapy (half required a second agent at 2.3 years [2.0-2.9]; P = 0.00017). Discontinuation of L-dopa therapy was 1%. Dopamine agonists were stopped (due to side-effects like impulse control disorders [6%], somnolence [4%] and light-headedness [3%]) in 20% over four years. The duration and tolerability of L-dopa and dopamine agonists as initial Parkinson's disease monotherapy are defined in this study; this may form part of the information exchange with patients.

Colorectal distension-evoked potentials in awake rats: a novel method for studies of visceral sensitivity.

BACKGROUND: Quantification of the visceromotor response induced by colorectal distension (CRD) in rodents is commonly used for preclinical studies of visceral pain. The model is well established but does not fully assess the central response to stimulation. The aim of this study was to establish a novel model assessing cerebral evoked potentials (CEPs) in response to CRD in awake rats. METHODS: Epidural recording electrodes were chronically implanted in the skull of female Sprague-Dawley rats. Colorectal distension-induced CEPs were recorded using either rapid balloon distensions (100 ms, 20-80 mmHg) or electric stimulation (1 ms, 1-4 mA) using stimulation probes placed in the distal colon. KEY RESULTS: Colorectal distension-induced CEPs were separated in three partly temporally overlapping components consisting of five prominent peaks. Peak latencies at 80 mmHg were (P1, N1) 23 ± 1 and 55 ± 4 ms, (N2, P2a, P2b) 91 ± 3, 143 ± 5 and 174 ± 3 ms, and (P3) 297 ± 3 ms. Amplitudes and latencies were, except for the early component, intensity dependent. Intrarectal administration of lidocaine significantly reduced the amplitude of N2 (by 42 ± 6%, P < 0.001) and P2 (by 34 ± 6%, P < 0.001). Electrically induced CEPs were intensity dependent and had similar topography and latencies as the mechanical evoked potentials (P1: 26 ± 2 ms; N1: 61 ± 1 ms; P2: 84 ± 6 ms; N2: 154 ± 6 ms; P3: 326 ± 10 ms), but there were large variations in amplitudes in between repeated electrical stimulations. CONCLUSIONS & INFERENCES: Colorectal distension-induced CEPs can be recorded reliably in awake rats and may serve as a surrogate marker of colonic sensation and be a useful parameter in studies of visceral sensitivity.

Multimodal sensory testing of the rectum and rectosigmoid: development and reproducibility of a new method.

Evaluation of rectal and rectosigmoid sensation is important in basic, clinical and pharmacological studies. New methods to evoke and assess multimodal (electrical, thermal and mechanical) experimental pain of the upper gut activate distinct pathways and mimics clinical pain. The aims of the current study were to characterize the sensory response and reproducibility to multimodal stimulation of rectum and the rectosigmoid. A multimodal rectal probe was developed. Mucosal electrostimulation was delivered at the recto-sigmoid junction. In Rectum, impedance planimetry was used for measurement of cross-sectional area (CSA) during distension. Circulation of water within the bag at either 4 or 60 degrees C was applied for thermal stimulation. The method was tested in 12 healthy volunteers (six men mean age 32 years) on two subsequent days. Mechanical and sensory responses and referred pain areas were assessed. Stimulation with electrical, thermal and mechanical modalities resulted in different sensory perceptions. The relationship between stimulus intensity and sensory response was linear for all modalities. Sensory response to different modalities did not differ between investigation days (all P-values > 0.1). Approximately 75% of subjects felt referred pain in distinct skin locations. Between-days reproducibility was good for all modalities [intra-class correlation (ICC) > or = 0.6]. At sensory threshold, CSA showed best reproducibility (ICC > or = 0.9). At pain detection threshold stretch ratio, CSA and electrostimulation showed best reproducibility (ICC = 1.0; 0.9; 0.9). The present model was easily implemented, robust and showed good reproducibility. It can be used to study pathophysiology or pharmacological interventions in healthy controls and in patients with diseases involving the distal hindgut.

Danish stable schools for experiential common learning in groups of organic dairy farmers.

The farmer field school (FFS) is a concept for farmers' learning, knowledge exchange, and empowerment that has been developed and used in developing countries. In Denmark, a research project focusing on explicit non-antibiotic strategies involves farmers who have actively expressed an interest in phasing out antibiotics from their herds through promotion of animal health. One way of reaching this goal was to form participatory focused farmer groups in an FFS approach, which was adapted to Danish conditions and named "stable schools." Four stable schools were established and went through a 1-yr cycle with 2 visits at each of the 5 or 6 farms connected to each group. A facilitator was connected to each group whose role was to write the meeting agenda together with the host farmer, direct the meeting, and write the minutes to send to the group members after the meeting. Through group focus interviews and individual semistructured qualitative interviews of all participants, the approach of the farmers' goal-directed work toward a common goal was judged to be very valuable and fruitful and based on a common learning process. Complex farming situations were the focus of all groups and in this context, problems were identified and solutions proposed based on each farmer's individual goals. In this article, we describe the experiences of 4 stable school groups (each comprising farmers and a facilitator), and the common process of building a concept that is suitable for Danish organic dairy farming.

Development and daily management of an explicit strategy of nonuse of antimicrobial drugs in twelve Danish organic dairy herds.

Promotion of animal health and well-being at the individual animal and herd level is an important goal in organic farming. At the same time, chemical products affecting the natural balance among living organisms are prohibited in all areas of the organic farm. From an animal welfare point of view, however, no animal must suffer. Therefore, veterinary drugs are allowed under the European Union's regulations for organic farming, despite the fact that they are powerful cell toxins affecting both pathogenic and necessary bacteria, and as such in organic terminology, are regarded as "chemical" or "artificial" products. In this article, we present and discuss interviews with 12 Danish organic dairy producers who claim that minimized use or nonuse of antimicrobial drugs is an explicit goal. The dairy producers were at different levels with regard to reduced antimicrobial treatment. An explicit strategy of no antimicrobial treatments is based primarily on a long-term effort to improve herd health, and secondarily, on finding alternative treatments for diseased animals. Improved hygiene, outdoor access, use of nursing cows, and blinding of chronic mastitis quarters were the main techniques in developing a strategy of not using antimicrobial treatments in the herd by dairy producers. Producers' perception of disease changed from something unavoidable to a disturbing break in the daily rhythm that often could have been avoided. Change toward a nonantimicrobial strategy was gradual and stepwise. All dairy producers in this study desired to preserve the possibility of using antimicrobial drugs in emergencies.

[Tools for the diagnosis of attention-deficit/hyperactivity disorder in adults. Self-rating behaviour questionnaire and diagnostic checklist]. / Instrumente zur Diagnostik der Aufmerksamkeitsdefizit-/Hyperaktivitätsstörung (ADHS) im ErwachsenenalterSelbstbeurteilungsskala (ADHS-SB) und Diagnosecheckliste (ADHS-DC).

We report on the development of a German self-rating behaviour questionnaire (ADHD-SR) and diagnostic checklist (ADHD-DC) for the diagnosis of attention-deficit/hyperactivity disorder in adults according to DSM IV and ICD 10 research criteria. When comparing self-rating with expert rating, we found good concordance measured by intraclass coefficients on the level of single symptoms and syndrome scores. High retest reliability of the ADHD-SR demonstrated the ability to assess time-stable behaviour traits. Evaluation of the psychometric properties revealed good internal consistency and adequate convergent and divergent validity measured by the "big five" derived from the NEO-FFI and the constructs impulsivity, venturesomeness, and empathy of Eysenck's impulsiveness questionnaire. We detected a remarkable correlation with the Wender Utah Rating Scale, which targets the detection of childhood ADHD symptoms. Diagnostic sensitivity for different cutoff points was calculated by ROC analysis at 65--88%. Specificity was 67% to 92%.

Expression of a cytoplasmic transhydrogenase in Saccharomyces cerevisiae results in formation of 2-oxoglutarate due to depletion of the NADPH pool.

The intracellular redox state of a cell is to a large extent defined by the concentration ratios of the two pyridine nucleotide systems NADH/NAD(+) and NADPH/NADP(+) and has a significant influence on product formation in microorganisms. The enzyme pyridine nucleotide transhydrogenase, which can catalyse transfer of reducing equivalents between the two nucleotide systems, occurs in several organisms, but not in yeasts. The purpose of this work was to analyse how metabolism during anaerobic growth of Saccharomyces cerevisiae might be altered when transfer of reducing equivalents between the two systems is made possible by expression of a cytoplasmic transhydrogenase from Azotobacter vinelandii. We therefore cloned sth, encoding this enzyme, and expressed it under the control of a S. cerevisiae promoter in a strain derived from the industrial model strain S. cerevisiae CBS8066. Anaerobic batch cultivations in high-performance bioreactors were carried out in order to allow quantitative analysis of the effect of transhydrogenase expression on product formation and on the intracellular concentrations of NADH, NAD(+), NADPH and NADP(+). A specific transhydrogenase activity of 4.53 U/mg protein was measured in the extracts from the strain expressing the sth gene from A. vinelandii, while no transhydrogenase activity could be detected in control strains without the gene. Production of the transhydrogenase caused a significant increase in formation of glycerol and 2-oxoglutarate. Since NADPH is used to convert 2-oxoglutarate to glutamate while glycerol formation increases when excess NADH is formed, this suggested that transhydrogenase converted NADH and NADP(+) to NAD(+) and NADPH. This was further supported by measurements of the intracellular nucleotide concentrations. Thus, the (NADPH/NADP(+)):(NADH/NAD(+)) ratio was reduced from 35 to 17 by the transhydrogenase. The increased formation of 2-oxoglutarate was accompanied by a two-fold decrease in the maximal specific growth rate. Also the biomass and ethanol yields were significantly lowered by the transhydrogenase.

Utilization of a murine model to investigate the molecular process of transphyseal bone formation.

The purpose of this investigation was to establish a model that allows efficient and broad-based molecular studies of bony bridge formation and to correlate basic histologic findings and novel in situ hybridization assays with previous studies. We disrupted the hindlimb tibial physis of skeletally immature mice. Histologic examination of the damaged limbs at weeks 1, 3, and 5 using the HBQ method and in situ hybridization for col2, col1O, Indian hedgehog (ihh), and vascular endothelial growth factor (VEGF) were consistent with previous reports in non-murine models documenting initial evidence of ossification at 1 week post-injury and a fully ossified bony bridge at 3 weeks. In situ hybridization assays documented the absence of ihh and VEGF in the disrupted region, suggesting that bone formation is not occurring via the process of endochondral ossification. We conclude that a murine model provides an excellent opportunity for further examination of the molecular mechanism of induced transphyseal ossification.

[Drug therapy of patients with emotionally unstable personality]. / Medikamentell behandling av personer med emosjonelt ustabil personlighet.

BACKGROUND: Most patients with borderline personality disorders are treated with psychotropic medication. There are, however, no clear clinical guidelines concerning optimal treatment of the disorder. MATERIAL AND METHODS: This article reviews the data on the efficacy of treatment with different classes of psychopharmacological drugs. The emphasis is on clinical trials published after 1981, as most placebo-controlled double-blind studies have been done in the last two decades. Furthermore, studies conducted before 1980 are more difficult to interpret because of changes in clinical diagnostic criteria. RESULTS: Neuroleptics have been studied most extensively followed by antidepressants. Neuroleptics have a modest, but broad therapeutic effect on symptoms in all domains. Doses are lower than those used for treating schizophrenia. Antidepressants have a more inconsistent effect. Tricyclics have been the least successful, whereas irreversible MAO inhibitors and selective serotonin reuptake inhibitors (SSRIs) have been effective in treating mood symptoms and impulsivity. Lithium has a possible effect in diminishing anger and suicidal symptoms. INTERPRETATION: As there is no "drug of choice" for the treatment of borderline personality disorder, a more rational clinical approach might be to treat different symptom clusters (cognitive/schizotypal, affective, impulsive) rather than the disorder itself. Finally, some practical guidelines for drug treatment of the disorder are proposed.

Optimization of ethanol production in Saccharomyces cerevisiae by metabolic engineering of the ammonium assimilation.

Ethanol is still one of the most important products originating from the biotechnological industry with respect to both value and amount. In addition to ethanol, a number of byproducts are formed during an anaerobic fermentation of Saccharomyces cerevisiae. One of the most important of these compounds, glycerol, is produced by yeast to reoxidize NADH, formed in synthesis of biomass and secondary fermentation products, to NAD+. The purpose of this study was to evaluate whether a reduced formation of surplus NADH and an increased consumption of ATP in biosynthesis would result in a decreased glycerol yield and an increased ethanol yield in anaerobic cultivations of S. cerevisiae. A yeast strain was constructed in which GLN1, encoding glutamine synthetase, and GLT1, encoding glutamate synthase, were overexpressed, and GDH1, encoding the NADPH-dependent glutamate dehydrogenase, was deleted. Hereby the normal NADPH-consuming synthesis of glutamate from ammonium and 2-oxoglutarate was substituted by a new pathway in which ATP and NADH were consumed. The resulting strain TN19 (gdh1-A1 PGK1p-GLT1 PGK1p-GLN1) had a 10% higher ethanol yield and a 38% lower glycerol yield compared to the wild type in anaerobic batch fermentations. The maximum specific growth rate of strain TN19 was slightly lower than the wild-type value, but earlier results suggest that this can be circumvented by increasing the specific activities of Gln1p and Glt1p even more. Thus, the results verify the proposed concept of increasing the ethanol yield in S. cerevisiae by metabolic engineering of pathways involved in biomass synthesis.