The role of complexity metrics in a multi-institutional dosimetry audit of VMAT

OBJECTIVE: To demonstrate the benefit of complexity metrics such as the modulation complexity score (MCS) and monitor units (MUs) in multi-institutional audits of volumetric-modulated arc therapy (VMAT) delivery.

METHODS: 39 VMAT treatment plans were analysed using MCS and MU. A virtual phantom planning exercise was planned and independently measured using the PTW Octavius(®) phantom and seven29(®) 2D array (PTW-Freiburg GmbH, Freiburg, Germany). MCS and MU were compared with the median gamma index pass rates (2%/2 and 3%/3 mm) and plan quality. The treatment planning systems (TPS) were grouped by VMAT modelling being specifically designed for the linear accelerator manufacturer's own treatment delivery system (Type 1) or independent of vendor for VMAT delivery (Type 2). Differences in plan complexity (MCS and MU) between TPS types were compared.

RESULTS: For Varian(®) linear accelerators (Varian(®) Medical Systems, Inc., Palo Alto, CA), MCS and MU were significantly correlated with gamma pass rates. Type 2 TPS created poorer quality, more complex plans with significantly higher MUs and MCS than Type 1 TPS. Plan quality was significantly correlated with MU for Type 2 plans. A statistically significant correlation was observed between MU and MCS for all plans (R = -0.84, p < 0.01).

CONCLUSION: MU and MCS have a role in assessing plan complexity in audits along with plan quality metrics. Plan complexity metrics give some indication of plan deliverability but should be analysed with plan quality.

ADVANCES IN KNOWLEDGE: Complexity metrics were investigated for a national rotational audit involving 34 institutions and they showed value. The metrics found that more complex plans were created for planning systems which were independent of vendor for VMAT delivery.

Plasma levels of intermedin (adrenomedullin-2) in healthy human volunteers and patients with heart failure

Intermedin/adrenomedullin-2 (IMD) is a member of the adrenomedullin/CGRP peptide family. Less is known about the distribution of IMD than for other family members within the mammalian cardiovascular system, particularly in humans. The aim was to evaluate plasma IMD levels in healthy subjects and patients with chronic heart failure. IMD and its precursor fragments, preproIMD25–56 and preproIMD57–92, were measured by radioimmunoassay in 75 healthy subjects and levels of IMD were also compared to those of adrenomedullin (AM) and mid-region proadrenomedullin45–92 (MRproAM45–92) in 19 patients with systolic heart failure (LVEF < 45%). In healthy subjects, plasma levels (mean + SE) of IMD (6.3 + 0.6 pg ml−1) were lower than, but correlated with those of AM (25.8 + 1.8 pg ml−1; r = 0.49, p < 0.001). Plasma preproIMD25–56 (39.6 + 3.1 pg ml−1), preproIMD57–92 (25.9 + 3.8 pg ml−1) and MRproAM45–92 (200.2 + 6.7 pg ml−1) were greater than their respective bioactive peptides. IMD levels correlated positively with BMI but not age, and were elevated in heart failure (9.8 + 1.3 pg ml−1, p < 0.05), similarly to MRproAM45–92 (329.5 + 41.9 pg ml−1, p < 0.001) and AM (56.8 + 10.9 pg ml−1, p < 0.01). IMD levels were greater in heart failure patients with concomitant renal impairment (11.3 + 1.8 pg ml−1) than those without (6.5 + 1.0 pg ml−1; p < 0.05). IMD and AM were greater in patients receiving submaximal compared with maximal heart failure drug therapy and were decreased after 6 months of cardiac resynchronization therapy. In conclusion, IMD is present in the plasma of healthy subjects less abundantly than AM, but is similarly correlated weakly with BMI. IMD levels are elevated in heart failure, especially with concomitant renal impairment, and tend to be reduced by high intensity drug or pacing therapy.

Development of a Novel Assay for the Detection of Active Neutrophil Elastase in Patients with Chronic Obstructive Pulmonary Disease

Neutrophil elastase (NE), a biomarker of infection and inflammation, correlates with the severity of several respiratory diseases including chronic obstructive pulmonary disease (COPD). However, it’s detection and quantification in biological samples is confounded by a lack of reliable and robust methodologies. Standard assays using chromogenic or fluorogenic substrates are not specific when added to complex clinical samples containing multiple proteolytic and hydrolytic enzymes which have the ability to hydrolyse the substrate, thereby resulting in an over-estimation of the target protease. Furthermore, ELISA systems measure total protease levels which can be a mixture of latent, active and protease-inhibitor complexes. Therefore, we have developed a novel immunoassay (ProteaseTag™ Active NE Immunoassay) which is selective and specific for the capture of active NE in sputum and Bronchoalveolar Lavage (BAL) in patients with COPD. The objective of this study was to clinically validate ProteaseTag™ Active NE Ultra Immunoassay for the detection of NE in sputum from COPD patients. 20 matched sputum sol samples were collected from 10 COPD patients (M=6, F=4; 73 ± 6 years) during stable and exacerbation phases. Samples were assayed for NE activity utilising both ProteaseTag™ Active NE Ultra Immunoassay and a fluorogenic substrate-based kinetic activity assay. Both assays detected elevated levels of NE in the majority of patients (n=7) during an exacerbation (mean=217.2 μg/ml ±296.6) compared to their stable phase (mean=92.37 μg/ml ±259.8). However, statistical analysis did not show this difference to be significant (p=0.07, ProteaseTag™ Active NE Ultra Immunoassay; p=0.06 kinetic assay), most likely due to the low study number. A highly significant correlation was found between the 2 assay types (p≤0.0001, r=0.996). NE as a primary efficacy endpoint in clinical trials or as a marker of inflammation within the clinic has been hampered by the lack of a robust and simple to use assay. ProteaseTag™ Active NE Immunoassay specifically measures only active NE in clinical samples, is quick and easy to use (< 3 hours) and has no dependency on a kinetic readout. ProteaseTag™ technology is currently being transferred to a lateral flow device for use at Point of Care.

Utilisation of a novel ProteaseTag™ activity immunoassay for the specific measurement of neutrophil elastase in BAL from children with cystic fibrosis

Introduction: Neutrophil elastase (NE) is a serine protease implicated in the pathogenesis of several respiratory diseases including cystic fibrosis (CF). The presence of free NE in BAL is a predictor of subsequent bronchiectasis in children with CF (Sly et al, 2013, NEJM 368: 1963-1970). Furthermore, children with higher levels of sputum NE activity (NEa) tend to experience a more rapid decline in FEV1 over time even after adjusting for age, gender and baseline FEV1 (Sagel et al, 2012, AJRCCM 186: 857-865). Its detection and quantification in biological samples is however confounded by a lack of robust methodologies. Standard assays using chromogenic or fluorogenic substrates are not specific when added to complex samples containing multiple proteolytic and hydrolytic enzymes. ELISA systems measure total protein levels which can be a mixture of latent, active and protease-inhibitor complexes. We have therefore developed a novel assay (ProteaseTag™ Active NE Immunoassay), which couples an activity dependent NE-Tag with a specific antibody step, resulting in an assay which is both selective and specific for NEa. Aims: To clinically validate ProteaseTag™ Active NE for the detection of free NEa in BAL from children with CF. Methods: A total of 95 paediatric BAL samples [CF (n=76; 44M, 32F) non-CF (n=19; 12M, 7F)] collected through the Study of Host Immunity and Early Lung Disease in CF (SHIELD CF) were analysed for NEa using ProteaseTag™ Active NE (ProAxsis Ltd) and a fluorogenic substrate-based assay utilising Suc-AAPV-AMC (Sigma). IL-8 was measured by ELISA (R&D Systems). Results were analysed to show comparisons in free NEa between CF and non-CF samples alongside correlations with a range of clinical parameters. Results: NEa measured by ProteaseTag™ Active NE correlated significantly with age (r=0.3, p=0.01) and highly significantly with both IL-8 (r=0.4, p=<0.0001) and the absolute neutrophil count (ANC) (r=0.4, p=<0.0001). These correlations were not observed when NEa was measured by the substrate assay even though a significant correlation was found between the two assays (r=0.8, p<0.0001). A trend towards significance was found between NEa in the CF and non-CF groups when measured by ProteaseTag™ Active NE (p=0.07). Highly significant differences were found with the other inflammatory parameters between the 2 groups (IL-8: p=0.0002 and ANC: p=0.006). Conclusion: NEa as a primary efficacy endpoint in clinical trials or as a marker of inflammation within the clinic has been hampered by the lack of a robust and simple to use assay. ProteaseTag™ Active NE has been shown to be a specific and superior tool in the measurement of NEa in paediatric CF BAL samples (supporting data from previous studies using adult CF expectorated samples). The technology is currently being transferred to a lateral flow device for use at Point of Care. Acknowledgements: This work was supported by the National Children’s Research Centre, Dublin (SHIELD CF) and grants from the Medical Research Council and Cystic Fibrosis Foundation Therapeutics.

Validity of model devices used to assess canine temperament in behavioral tests

Temperament tests are widely accepted as instruments for profiling behavioral variability in dogs, and they are applied in numerous areas of investigation (e.g. suitability for adoption or for breeding). During testing, to elicit a dog's reaction toward novel stimuli and predict its behavior in everyday life, model devices such as a child-like doll, or a fake dog, are often employed. However, the reliability of these devices to accurately stimulate dogs' reactions to children or dogs, is unknown and perhaps overestimated. This may be a particular concern in the case of aggressive behavior toward humans, a significant public health issue. The aim of this study was to: (1) evaluate the correlation between dogs' reactions to these devices, and owners' reports of their dog's aggression history (using the C-BARQ ??); (2) compare reactions toward the devices of dogs with and without histories of aggression. Subjects were selected among those visiting for behavioral consultation at the Veterinary Hospital of the University of Pennsylvania, and previously categorized as aggressive toward unfamiliar children, conspecifics, or as non-aggressive dogs (control). The test consisted of different components: an unfamiliar female tester approaching the dog; the presentation of a child-like doll, an ambiguous object, and a fake plastic dog. All tests were videotaped and durations of behaviors were later analyzed on the basis of a specified ethogram. Dogs' reactions were compared to C-BARQ scores, and interesting correlations emerged for 'dog-directed aggression/fear' (R = 0.48, P = 0.004), and 'stranger-directed aggression' (R = 0.58, P <0.001) factors. Dogs differed in their reactions toward the devices: the child-like doll and the fake dog elicited more social behaviors than the ambiguous object used as a control stimulus. Issues concerning the reliability of these tools to assess canine temperament are discussed. ?? 2012 Elsevier B.V. All rights reserved.

Serum Cytokines and Adipokines in Periodontitis

Objective: To investigate whether there was an association between serum levels of pro-inflammatory cytokines, adipokines and periodontitis in 60-70 year-old men in Northern Ireland. Methods: Test subjects (n=61) with established periodontitis, categorised by loss of periodontal attachment (PAL) and pocketing of at least 5mm, were matched for age, smoking and BMI with controls (n=60) who were periodontitis resistant (no PAL or pocketing over 3mm). Serum levels of IL 1 alpha, TNF alpha, IL 6, adiponectin, resistin, leptin and C-reactive protein (CRP) were measured from serum using a multiplex array (Randox, UK). Mann Whitney analysis was used with the level of significance set at p<0.05. Results: There was considerable inter-individual variability in all the analytes measured. There was a significantly higher level of IL 6 in men with periodontitis (3.25 pg/ml, SD 2.29) than in those with no periodontitis (2.38 pg/ml, SD1.62), p=0.0041 (after Bonferroni correction for multiple testing p=0.029). Serum IL 6 was strongly correlated with CRP: r=0.52 (95% CI 0.38-0.64), p<0.0001. There was a lower serum level of adiponectin in men with periodontitis (3662.1 ng/ml, SD 2383.4) compared with those who were periodontitis resistant (4265.9 ng/ml, SD 2266.0), which just escaped statistical significance at p=0.068. There were no significant differences in the serum levels of any of the other analytes measured in relation to periodontal status. Conclusions: There was evidence in the 60-70 year-old male population investigated that periodontitis was associated with an increased serum level of IL 6.

Co-expression of NPY and VIP in human dental pulp

Introduction: The regulation of pulpal haemodynamics in health and disease involves sympathetic and parasympathetic mechanisms in which both neuropeptide Y (NPY; a sympathetic vasoconstrictor) and vasoactive intestinal polypeptide (VIP; a parasympathetic vasodilator) may play potential pathophysiological roles. We have previously investigated the levels of NPY or VIP present in human dental pulp tissue and shown that their expression is up-regulated in caries induced pulpal inflammation. Objectives: The aim of this study was to investigate the potential correlation between NPY and VIP levels measured in the same dental pulp samples using radioimmunoassay (RIA). Methods: Pulp tissue was obtained from extracted teeth, classified as follows; healthy (n=22), moderately carious (n=20) and grossly carious (n=26). Samples were processed for RIA by boiling in acetic acid as previously described. The levels of NPY and VIP, measured by RIA, were expressed as ng/gram of pulp tissue. The nature of the relationship between NPY and VIP levels in human pulp tissue was tested by calculating Pearson's product moment correlation coefficient using the linear regression test. Results: Calculation of Pearson product moment correlation coefficient showed a significant negative correlation between NPY and VIP levels in pulp tissue samples from non-carious teeth (p = 0.02, r = -.48). This negative correlation in non-carious teeth changed to a significant positive correlation in carious teeth when the levels of NPY and VIP were compared (p = 0.03, r= 0.311). Conclusions: In non-carious teeth, the negative correlation between NPY and VIP levels is in keeping with the previously described modulatory influence of cholinergic nerves on sympathetic function which may be perturbed as caries develops.

Influence of corneal structure, corneal responsiveness, and other ocular parameters on tonometric measurement of intraocular pressure.

PURPOSE: To estimate the relationships between ocular parameters and tonometrically measured intraocular pressure (IOP), to determine the influence of ocular parameters on different instrument measurements of IOP, and to evaluate the association of ocular parameters with a parameter called hysteresis. METHODS: Patients presenting at a glaucoma clinic were recruited for this study. Subjects underwent IOP measurement with the Goldmann applanation tonometer (GAT), the TonoPen, and the Reichert Ocular Response Analyzer (ORA), and also measurements of central corneal thickness (CCT), axial length, corneal curvature, corneal astigmatism, central visual acuity, and refractive error. Chart information was reviewed to determine glaucoma treatment history. The ORA instrument provided a measurement called corneal hysteresis. The association between measured IOP and the other ocular characteristics was estimated using generalized estimating equations. RESULTS: Among 230 patients, IOP measurements from the TonoPen read lowest, and ORA read highest, and GAT measurements were closest to the mean IOP of the 3 instruments. In a multiple regression model adjusting for age, sex, race, and other ocular characteristics, a 10 microm increase in CCT was associated with an increase of 0.79 mm Hg measured IOP in untreated eyes (P<0.0001). Of the 3 tonometers, GAT was the least affected by CCT (0.66 mm Hg/10 mum, P<0.0001). Hysteresis was significantly correlated with CCT with a modest correlation coefficient (r=0.20, P<0.0007). CONCLUSIONS: Among parameters related to measured IOP, features in addition to CCT, such as hysteresis and corneal curvature, may also be important. Tonometric instruments seem to be affected differently by various physiologic characteristics.

Dark room provocative test and extent of angle closure: an anterior segment OCT study.

PURPOSE: To clarify the risk parameters measured by anterior segment optical coherence tomography (AS-OCT) for elevated intraocular pressures (IOP) provoked by the darkroom test and to provide recommendations for its clinical usage. METHODS: Subjects aged >40 years and whose peripheral anterior chambers were ≤1/4 corneal thickness were recruited. The anterior segment of the eye was imaged in sitting position and under both light and dark conditions and biometry was performed using anterior segment optical coherence tomography. The analyzed parameters were: (1) central anterior chamber depth (ACD); (2) anterior chamber width; (3) pupil diameter; (4) iris curvature; (5) lens thickness; and (6) number of meridians with closed angles (NCA). Then the darkroom test was performed and a positive provocative test result was defined as a rise in IOP ≥8 mm Hg after the test. Statistical analyses included: (1) the difference in parameters between positive and negative eyes; (2) the association between posttest IOP and the parameters; and (3) the difference in parameters between the 2 eyes in subjects with the asymmetric results. RESULTS: A total of 70 subjects were recruited. ACD (P=0.022), NCA in light (P<0.001), and NCA in dark (P<0.001) were different significantly between eyes with positive and negative results. There was a strong association between NCA in dark (r=0.755, P<0.001) and the posttest IOP. Among subjects with asymmetric results between the 2 eyes, the ACD was shallower and the lens thickness was larger in the positive eye. CONCLUSIONS: The posttest IOP is determined by the extent of functionally closed angles in the dark. The test may be useful in the early diagnosis of primary angle closure. At the same time, angle configuration should be evaluated to remove false positive result.

Height, stunting, and refractive error among rural Chinese schoolchildren: the See Well to Learn Well project.

PURPOSE: To evaluate the hypothesis that changes in nutritional status could be partly responsible for observed increases in myopia prevalence among Chinese children. DESIGN: Cross-sectional cohort study. METHODS: Rural Chinese secondary school children participating in a study of interventions to promote spectacle use were randomly sampled (20% of children with uncorrected vision >6/12 bilaterally, and 100% of remaining children) and underwent cycloplegic refraction with subjective refinement and measurement of height and weight. Stunting was defined according to the World Health Organization standard population. RESULTS: Among 3226 children in the sample, 2905 (90.0%) took part. Among 1477 children undergoing refraction, 1371 (92.8%) had height and weight measurements. These children had a mean age of 14.5 +/- 1.4 years, 59.8% were girls, and mean spherical equivalent refraction was -1.93 +/- 1.82 diopters. Stunting was present in 87 children (6.4%). While height was inversely associated with refractive error (RE) (taller children were more myopic) among boys (r = -0.147, P = .001), this disappeared when adjusting for age, and no such association was observed among girls. Neither girls nor boys with stunting differed significantly in refraction from children without stunting, and neither stunting nor height was associated with RE when adjusting for age, height, and parental education. The power of this study to have detected a 0.75 diopters difference in RE between children with and without stunting was 0.96. CONCLUSION: Results from this cross-sectional study are not consistent with the hypothesis that nutritional status is a determinant of RE in this setting.

Corneal biomechanics, refractive error, and axial length in Chinese primary school children.

PURPOSE: Low corneal hysteresis is associated with longer axial length in Chinese secondary school children. The authors sought to explore this association in primary school children. METHODS: LogMAR presenting visual acuity, cycloplegic refractive error, ocular biometry, central corneal thickness (CCT), and corneal hysteresis (CH) was assessed for children in grades 1 to 3 at an academically competitive urban school in Shantou, China. RESULTS: Among 872 eligible children (mean age, 8.6 ± 2.1 years), 651 (74.7%) completed the examination. Among 1299 examined eyes, 111 (8.5%) had uncorrected vision ≤6/12. Mean spherical equivalent refractive error for all eyes was +0.26 ± 1.41 D, and axial length (AL) was 22.7 ± 0.90 mm. CH for the lowest (mean AL, 21.7 ± 0.39 mm), two middle (mean AL, 22.4 ± 0.15 and 22.9 ± 0.15 mm), and highest quartiles (mean AL, 23.7 ± 0.74 mm) of AL were 10.6 ± 2.1 mm Hg, 10.4 ± 2.1 mm Hg, 10.3 ± 2.3 mm Hg, and 10.2 ± 2.3 mm Hg respectively (age- and gender-adjusted Pearson's correlation coefficient r = -0.052; P = 0.001). In generalized estimating equation models adjusting for age, gender, and CCT, lower CH was significantly associated with longer AL (P < 0.001) and more myopic refractive error (P = 0.001). CONCLUSIONS: CH measurement is practical in young children because this is when myopia undergoes its most rapid progression. Prospective follow-up of this cohort at high risk for myopia is under way to determine whether low CH is predictive, or a consequence, of long AL.

Comet 322P/SOHO 1: An asteroid with the smallest perihelion distance?

We observed comet 322P/SOHO 1 (P/1999 R1) from the ground and with the Spitzer Space Telescope when it was between 2.2 and 1.2 AU from the Sun. These are the first observations of any SOHO-discovered periodic comet by a non-solar observatory, and allow us to investigate its behavior under typical cometary circumstances. 322P appeared inactive in all images. Its lightcurve suggests a rotation period of 2.8+/-0.3 hr and has an amplitude greater than ~0.3 mag, implying a density of at least 1000 kg m$^{-3}$, considerably higher than that of any known comet. It has average colors of g'-r' = 0.52+/-0.04 and r'-i' = 0.04+/-0.09. We converted these to Johnson colors and found that the V-R color is consistent with average cometary colors, but R-I is somewhat bluer; these colors are most similar to V- and Q-type asteroids. Modeling of the optical and IR photometry suggests it has a diameter of 150-320 m and a geometric albedo of 0.09-0.42, with diameter and albedo inversely related. Our upper limits to any undetected coma are still consistent with a sublimation lifetime shorter than the typical dynamical lifetimes for Jupiter Family Comets. These results suggest that it may be of asteroidal origin and only active in the SOHO fields of view via processes different from the volatile-driven activity of traditional comets. If so, it has the smallest perihelion distance of any known asteroid.

Avaliação do potencial reprodutivo de touros através do exame andrológico

Este trabalho refere-se às atividades do estágio de Mestrado Integrado em Medicina Veterinária, apresentando-se casuística no âmbito dos planos sanitários e atividade clínica acompanhados. Como tema para discussão optou-se pela avaliação de touros através do exame andrológico. Após revisão bibliográfica analisaram-se dados obtidos pela VetAl (2008- 2012) caracterizando a população dos touros de carne no Sul de Portugal. Dos 184 touros avaliados foram aprovados 72,28 %, aumentando a probabilidade de reprovação tendencialmente com idade. Em parâmetros reprodutivos importantes como perímetro testicular existe influência de: idade (p<0,001), raça (p<0,05) e Pontuação da Condição Corporal (PCC) (p<0,05). Encontraram-se correlações significativas entre parâmetros tais como perímetro testicular e idade (p<0,001; r=0,52), PCC e parâmetros seminais microscópicos (p<0,05) e dos vários parâmetros entre si. O exame andrológico é essencial para estimar o potencial reprodutivo dos touros e importa fomentar a sua realização em Portugal para melhorar os níveis de fertilidade e rentabilidade das explorações; Abstract Rating of bulls’ reproductive potential through breeding soundness evaluation This document reports the activities of the traineeship for the Master in Veterinary Medicine, including the presentation of the number of cases attended during herd health plans and clinical activity. The theme chosen for discussion was bull fertility through breeding soundness evaluation. Data obtained by Vetal (2008-2012) were analyzed, according to the state of the art, with the aim of characterizing beef bulls’ population in Southern Portugal. Out of 184 bulls evaluated, 72,28 % were approved, unsuccessful outcome apparently increases with age. Scrotal circumference was significantly influenced by: age (p<0,001), breed (p<0,05) and Body Condition Scoring (BCS) (p<0,05). Correlations were significant between scrotal circumference and age (p<0,001; r=0,52), between BCS and microscopic semen evaluation parameters (p<0,05) and between these semen evaluation parameters. The breeding soundness evaluation is essential to assess bulls’ reproductive potential and routine performance of this exam should be encouraged in Portugal to improve fertility and herd profitability.

Medication regimen complexity in institutionalized elderly people in an aging society

Background Complex medication regimens may adversely affect compliance and treatment outcomes. Complexity can be assessed with the medication regimen complexity index (MRCI), which has proved to be a valid, reliable tool, with potential uses in both practice and research. Objective To use the MRCI to assess medication regimen complexity in institutionalized elderly people. Setting Five nursing homes in mainland Portugal. Methods A descriptive, cross-sectional study of institutionalized elderly people (n = 415) was performed from March to June 2009, including all inpatients aged 65 and over taking at least one medication per day. Main outcome measure Medication regimen complexity index. Results The mean age of the sample was 83.9 years (±6.6 years), and 60.2 % were women. The elderly patients were taking a large number of drugs, with 76.6 % taking more than five medications per day. The average medication regimen complexity was 18.2 (±SD = 9.6), and was higher in the females (p < 0.001). The most decisive factors contributing to the complexity were the number of drugs and dosage frequency. In regimens with the same number of medications, schedule was the most relevant factor in the final score (r = 0.922), followed by pharmaceutical forms (r = 0.768) and additional instructions (r = 0.742). Conclusion Medication regimen complexity proved to be high. There is certainly potential for the pharmacist’s intervention to reduce it as part as the medication review routine in all the patients.

Metabonomic studies to assess beer quality and stability

The work reported in this thesis aimed at applying the methodology known as metabonomics to the detailed study of a particular type of beer and its quality control, with basis on the use of multivariate analysis (MVA) to extract meaningful information from given analytical data sets. In Chapter 1, a detailed description of beer is given considering the brewing process, main characteristics and typical composition of beer, beer stability and the commonly used analytical techniques for beer analysis. The fundamentals of the analytical methods employed here, namely nuclear magnetic resonance (NMR) spectroscopy, gas-chromatography-mass spectrometry (GC-MS) and mid-infrared (MIR) spectroscopy, together with the description of the metabonomics methodology are described shortly in Chapter 2. In Chapter 3, the application of high resolution NMR to characterize the chemical composition of a lager beer is described. The 1H NMR spectrum obtained by direct analysis of beer show a high degree of complexity, confirming the great potential of NMR spectroscopy for the detection of a wide variety of families of compounds, in a single run. Spectral assignment was carried out by 2D NMR, resulting in the identification of about 40 compounds, including alcohols, amino acids, organic acids, nucleosides and sugars. In a second part of Chapter 3, the compositional variability of beer was assessed. For that purpose, metabonomics was applied to 1H NMR data (NMR/MVA) to evaluate beer variability between beers from the same brand (lager), produced nationally but differing in brewing site and date of production. Differences between brewing sites and/or dates were observed, reflecting compositional differences related to particular processing steps, including mashing, fermentation and maturation. Chapter 4 describes the quantification of organic acids in beer by NMR, using different quantitative methods: direct integration of NMR signals (vs. internal reference or vs. an external electronic reference, ERETIC method) and by quantitative statistical methods (using the partial least squares (PLS) regression) were developed and compared. PLS1 regression models were built using different quantitative methods as reference: capillary electrophoresis with direct and indirect detection and enzymatic essays. It was found that NMR integration results generally agree with those obtained by the best performance PLS models, although some overestimation for malic and pyruvic acids and an apparent underestimation for citric acid were observed. Finally, Chapter 5 describes metabonomic studies performed to better understand the forced aging (18 days, at 45 ºC) beer process. The aging process of lager beer was followed by i) NMR, ii) GC-MS, and iii) MIR spectroscopy. MVA methods of each analytical data set revealed clear separation between different aging days for both NMR and GC-MS data, enabling the identification of compounds closely related with the aging process: 5-hydroxymethylfurfural (5-HMF), organic acids, γ-amino butyric acid (GABA), proline and the ratio linear/branched dextrins (NMR domain) and 5-HMF, furfural, diethyl succinate and phenylacetaldehyde (known aging markers) and, for the first time, 2,3-dihydro-3,5-dihydroxy-6-methyl-4(H)-pyran-4-one xii (DDMP) and maltoxazine (by GC-MS domain). For MIR/MVA, no aging trend could be measured, the results reflecting the need of further experimental optimizations. Data correlation between NMR and GC-MS data was performed by outer product analysis (OPA) and statistical heterospectroscopy (SHY) methodologies, enabling the identification of further compounds (11 compounds, 5 of each are still unassigned) highly related with the aging process. Data correlation between sensory characteristics and NMR and GC-MS was also assessed through PLS1 regression models using the sensory response as reference. The results obtained showed good relationships between analytical data response and sensory response, particularly for the aromatic region of the NMR spectra and for GC-MS data (r > 0.89). However, the prediction power of all built PLS1 regression models was relatively low, possibly reflecting the low number of samples/tasters employed, an aspect to improve in future studies.