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In this study, we report an extensive investigation of the structural language and acoustical specificities of the spontaneous speech of ten three- to five-year-old verbal autistic children. The autistic children were compared to a group of ten typically developing children matched pairwise on chronological age, nonverbal IQ and socioeconomic status, and groupwise on verbal IQ and gender on various measures of structural language (phonetic inventory, lexical diversity and morpho-syntactic complexity) and a series of acoustical measures of speech (mean and range fundamental frequency, a formant dispersion index, syllable duration, jitter and shimmer). Results showed that, overall, the structure and acoustics of the verbal autistic children's speech were highly similar to those of the TD children. Few remaining atypicalities in the speech of autistic children lay in a restricted use of different vocabulary items, a somewhat diminished morpho-syntactic complexity, and a slightly exaggerated syllable duration.
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Sugar-enriched media are used to produce extracellular substances (ECS) by Lactobacillus plantarum WCSF1, with a focus on growing stages and carbon source substrates. Combination of size exclusion chromatography and ATR-FTIR spectroscopy provides physicochemical patterns of bulk ECS produced along culture growing time. Secreted biopolymers present polydisperse and high molecular weight distributions, with significant amounts of carbohydrates and proteins. Results, supported by a multivariate statistical analysis, enable to differentiate the macromolecular content of bacterial ECS along the growing stages regardless of the growing media, highlighting a higher production of proteinaceous materials compared to polysaccharides. At the end of the exponential phase, common exoproteins were present in all the tested sugar-enriched media such as transglycosylases between 20 and 35 kDa, a muropeptidase at 36.9 kDa and a cell wall hydrolase. Additionally, L. plantarum WCFS1 secretes ECS with a greater diversity of proteins, when growing in the sucrose-enriched media. Graphical abstract.
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Proteínas de Bactérias/análise , Meios de Cultura , Lactobacillus plantarum/metabolismo , Substâncias Macromoleculares/análise , Carboidratos/química , Carbono/química , Técnicas de Cultura de Células , Hidrolases/química , Espectrometria de Massas , Peso Molecular , Nitrogênio/química , Polímeros/química , Análise de Componente Principal , Proteômica/métodos , Espectroscopia de Infravermelho com Transformada de Fourier , AçúcaresRESUMO
Whereas a reduced tendency to follow pointing gestures is described as an early sign of autism, the literature on response to joint attention indicates that autistic children perform better when a point is added to other social cues such as eye gaze. The purpose of this study was to explore pointing processing in autism when it is the only available cue and to investigate whether autistic children discriminate intentional pointing gestures from incidental pointing gestures. Eye movements of 58 autistic children (48 male) and 61 typically developing children (36 male) aged 3-5 years were recorded as the children were watching videos of a person uttering a pseudoword and pointing intentionally with one hand and incidentally with the other hand. After 3 s, two different potential referents for the pseudoword gradually emerged in both pointed-at corners. In comparison with typically developing children, autistic children's fixations were significantly farther away from both pointed-at zones. Upon hearing a novel word, typically developing children shifted their visual attention toward the zone pointed intentionally. This trend did not emerge in the group of autistic children regardless of their level of vocabulary. Autistic children, independently of their level of language, pay little attention to pointing when no other social cues are available and fail to discriminate intentional pointing gestures from incidental ones. They seem to grasp neither the spatial nor the social value of pointing.
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Transtorno Autístico , Criança , Sinais (Psicologia) , Tecnologia de Rastreamento Ocular , Fixação Ocular , Gestos , Humanos , MasculinoRESUMO
Acute liver injury (ALI) is a severe disorder resulting from excessive hepatocyte cell death, and frequently caused by acetaminophen intoxication. Clinical management of ALI progression is hampered by the dearth of blood biomarkers available. In this study, a bioinformatics workflow was developed to screen omics databases and identify potential biomarkers for hepatocyte cell death. Then, discovery proteomics was harnessed to select from among these candidates those that were specifically detected in the blood of acetaminophen-induced ALI patients. Among these candidates, the isoenzyme alcohol dehydrogenase 1B (ADH1B) was massively leaked into the blood. To evaluate ADH1B, we developed a targeted proteomics assay and quantified ADH1B in serum samples collected at different times from 17 patients admitted for acetaminophen-induced ALI. Serum ADH1B concentrations increased markedly during the acute phase of the disease, and dropped to undetectable levels during recovery. In contrast to alanine aminotransferase activity, the rapid drop in circulating ADH1B concentrations was followed by an improvement in the international normalized ratio (INR) within 10-48 h, and was associated with favorable outcomes. In conclusion, the combination of omics data exploration and proteomics revealed ADH1B as a new blood biomarker candidate that could be useful for the monitoring of acetaminophen-induced ALI.
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Álcool Desidrogenase/sangue , Biomarcadores/sangue , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Proteômica/métodos , Acetaminofen/toxicidade , Doença Hepática Induzida por Substâncias e Drogas/patologia , Cromatografia Líquida de Alta Pressão , Biologia Computacional , Humanos , Coeficiente Internacional Normatizado , Limite de Detecção , Espectrometria de Massas em TandemRESUMO
Bone loss and immune dysregulation are among the main adverse outcomes of spaceflight challenging astronauts' health and safety. However, consequences on B-cell development and responses are still under-investigated. To fill this gap, we used advanced proteomics analysis of femur bone and marrow to compare mice flown for 1 mo on board the BION-M1 biosatellite, followed or not by 1 wk of recovery on Earth, to control mice kept on Earth. Our data revealed an adverse effect on B lymphopoiesis 1 wk after landing. This phenomenon was associated with a 41% reduction of B cells in the spleen. These reductions may contribute to explain increased susceptibility to infection even if our data suggest that flown animals can mount a humoral immune response. Future studies should investigate the quality/efficiency of produced antibodies and whether longer missions worsen these immune alterations.-Tascher, G., Gerbaix, M., Maes, P., Chazarin, B., Ghislin, S., Antropova, E., Vassilieva, G., Ouzren-Zarhloul, N., Gauquelin-Koch, G., Vico, L., Frippiat, J.-P., Bertile, F. Analysis of femurs from mice embarked on board BION-M1 biosatellite reveals a decrease in immune cell development, including B cells, after 1 wk of recovery on Earth.
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Linfócitos B/imunologia , Linfócitos B/fisiologia , Fêmur/imunologia , Fêmur/fisiologia , Animais , Medula Óssea/imunologia , Medula Óssea/fisiologia , Células da Medula Óssea/imunologia , Células da Medula Óssea/fisiologia , Diferenciação Celular/imunologia , Diferenciação Celular/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Voo Espacial , Astronave , Baço/imunologia , Baço/fisiologia , Ausência de PesoRESUMO
The cerebellum contains a circadian clock, generating internal temporal signals. The daily oscillations of cerebellar proteins were investigated in mice using a large-scale two-dimensional difference in gel electrophoresis (2D-DIGE). Analysis of 2D-DIGE gels highlighted the rhythmic variation in the intensity of 27/588 protein spots (5%) over 24 h based on cosinor regression. Notably, the rhythmic expression of most abundant cerebellar proteins was clustered in two main phases (i.e., midday and midnight), leading to bimodal distribution. Only six proteins identified here to be rhythmic in the cerebellum are also known to oscillate in the suprachiasmatic nuclei, including two proteins involved in the synapse activity (Synapsin 2 [SYN2] and vesicle-fusing ATPase [NSF]), two others participating in carbohydrate metabolism (triosephosphate isomerase (TPI1] and alpha-enolase [ENO1]), Glutamine synthetase (GLUL), as well as Tubulin alpha (TUBA4A). Most oscillating cerebellar proteins were not previously identified in circadian proteomic analyses of any tissue. Strikingly, the daily accumulation of mitochondrial proteins was clustered to the mid-resting phase, as previously observed for distinct mitochondrial proteins in the liver. Moreover, a number of rhythmic proteins, such as SYN2, NSF and TPI1, were associated with non-rhythmic mRNAs, indicating widespread post-transcriptional control in cerebellar oscillations. Thus, this study highlights extensive rhythmic aspects of the cerebellar proteome.
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Cerebelo/metabolismo , Relógios Circadianos , Regulação da Expressão Gênica , Proteoma/análise , Proteoma/genética , Animais , Cerebelo/química , Ritmo Circadiano , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteômica , RNA Mensageiro/análise , RNA Mensageiro/genética , Eletroforese em Gel Diferencial BidimensionalRESUMO
The safety of space flight is challenged by a severe loss of skeletal muscle mass, strength, and endurance that may compromise the health and performance of astronauts. The molecular mechanisms underpinning muscle atrophy and decreased performance have been studied mostly after short duration flights and are still not fully elucidated. By deciphering the muscle proteome changes elicited in mice after a full month aboard the BION-M1 biosatellite, we observed that the antigravity soleus incurred the greatest changes compared with locomotor muscles. Proteomics data notably suggested mitochondrial dysfunction, metabolic and fiber type switching toward glycolytic type II fibers, structural alterations, and calcium signaling-related defects to be the main causes for decreased muscle performance in flown mice. Alterations of the protein balance, mTOR pathway, myogenesis, and apoptosis were expected to contribute to muscle atrophy. Moreover, several signs reflecting alteration of telomere maintenance, oxidative stress, and insulin resistance were found as possible additional deleterious effects. Finally, 8 days of recovery post flight were not sufficient to restore completely flight-induced changes. Thus in-depth proteomics analysis unraveled the complex and multifactorial remodeling of skeletal muscle structure and function during long-term space flight, which should help define combined sets of countermeasures before, during, and after the flight.
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Mitocôndrias/metabolismo , Músculo Esquelético/metabolismo , Atrofia Muscular/genética , Proteoma/genética , Ausência de Peso/efeitos adversos , Animais , Apoptose/genética , Sinalização do Cálcio , Regulação da Expressão Gênica , Resistência à Insulina , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Mitocôndrias/patologia , Desenvolvimento Muscular/genética , Músculo Esquelético/patologia , Atrofia Muscular/etiologia , Atrofia Muscular/metabolismo , Atrofia Muscular/patologia , Estresse Oxidativo , Proteoma/metabolismo , Voo Espacial , Serina-Treonina Quinases TOR/genética , Serina-Treonina Quinases TOR/metabolismo , Telômero/metabolismo , Telômero/patologiaRESUMO
Background and aims: Nongenerative speech is the rote repetition of words or phrases heard from others or oneself. The most common manifestations of nongenerative speech are immediate and delayed echolalia, which are a well-attested clinical feature and a salient aspect of atypical language use in autism. However, there are no current estimates of the frequency of nongenerative speech, and the individual characteristics associated with nongenerative speech use in individuals across the autistic spectrum are poorly understood. In this study, we aim to measure and characterize spontaneous and nongenerative speech use in minimally verbal and verbally fluent autistic children and adolescents. Methods: Participants were 50 minimally verbal and 50 verbally fluent autistic individuals aged 6 to 21 years. Spontaneous and nongenerative speech samples were derived from SALT transcripts of ADOS-2 assessments. Participants' intelligible speech utterances were categorized as spontaneous or nongenerative. Spontaneous versus nongenerative utterances were compared between language subgroups on frequency of use and linguistic structure. Associations between nongenerative speech use and a series of individual characteristics (ADOS-2 subscale scores, nonverbal IQ, receptive vocabulary, and chronological age) were investigated over the whole sample and for each language subgroup independently. Results: Almost all participants produced some nongenerative speech. Minimally verbal individuals produced significantly more nongenerative than spontaneous utterances, and more nongenerative utterances compared to verbally fluent individuals. Verbally fluent individuals produced limited rates of nongenerative utterances, in comparison to their much higher rates of spontaneous utterances. Across the sample, nongenerative utterance rates were associated with nonverbal IQ and receptive vocabulary, but not separately for the two language subgroups. In verbally fluent individuals, only age was significantly inversely associated with nongenerative speech use such that older individuals produced fewer nongenerative utterances. In minimally verbal individuals, there were no associations between any of the individual characteristics and nongenerative speech use. In terms of linguistic structure, the lexical diversity of nongenerative and spontaneous utterances of both language subgroups was comparable. Morphosyntactic complexity was higher for spontaneous compared to nongenerative utterances in verbally fluent individuals, while no differences emerged between the two utterance types in minimally verbal individuals. Conclusions: Nongenerative speech presents differently in minimally verbal and verbally fluent autistic individuals. Although present in verbally fluent individuals, nongenerative speech appears to be a major feature of spoken language in minimally verbal children and adolescents. Implications: Our results advocate for more research on the expressive language profiles of autistic children and adolescents who remain minimally verbal and for further investigations of nongenerative speech, which is usually excluded from language samples. Given its prevalence in the spoken language of minimally verbal individuals, nongenerative speech could be used as a way to engage in and maintain communication with this subgroup of autistic individuals.
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LAY ABSTRACT: For most autistic children, spoken language emergence and development happen after the age of 3. Once they start developing and using spoken language, some eventually manage to reach typical levels of language abilities, while others remain minimally speaking into adulthood. It is therefore difficult to consider young autistic preschoolers as a homogeneous group in terms of spoken language levels. In our study, we breakdown a representative and inclusive group of children on the spectrum aged from 3 to 5 into five subgroups that correspond to different linguistic profiles. To do so, we qualitatively described children's (pre)verbal productions elicited during interactions with a parent and with an experimenter. We then used a type of statistical analysis called cluster analysis to group together the children that had a similar expressive (pre)linguistic behavior. Using this analysis, we were able to delineate five linguistic profiles with qualitatively different patterns of vocal production. Two of these profiles are composed of speaking children; the three others are composed of non- or minimally speaking children. Our findings show that traditional binary division of speaking versus nonspeaking autistic children is not precise enough to describe the heterogeneity of early spoken language in young autistic children. They also support the use of qualitative descriptions of vocal productions and speech to accurately document children's level of language, which could, in turn, help design very finely tailored language intervention specific to each child.
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Transtorno do Espectro Autista , Transtorno Autístico , Transtornos do Desenvolvimento da Linguagem , Humanos , Criança , Idoso , Pré-Escolar , Linguística , Análise por ConglomeradosRESUMO
BACKGROUND: In the spring of 2020, Belgian authorities enforced a full lockdown period to contain the spread of the SARS-CoV-2 virus. This lockdown drastically disrupted the daily life of autistic individuals' and that of their families. In the midst of these extraordinary circumstances, we assessed the impact of social restrictions on autistic individuals' behavior and their parents' or caregivers' quality of life; we also sought to identify individual characteristics that may influence such changes. METHODS: We designed an online survey targeting caregivers living with an autistic child or adult. The questionnaire included 125 five-point Likert questions which targeted changes in families' quality of life and in autistic individuals' behavior, as well as factors likely to influence the extent and direction of these changes. RESULTS: We collected data from 209 French-speaking Belgian respondents. Respondents reported that the lockdown brought about a higher frequency of nonfunctional socio-communicative behaviors, as well as a decrease in families' quality of life. Parents who had less access to respite care experienced a steeper decrease in their quality of life. Autistic individuals with comorbidities, and whose parents had less access to respite care and implemented fewer rules at home during lockdown were more likely to display nonfunctional socio-communicative behaviors. CONCLUSION: COVID-19 lockdown restrictions had a negative impact on both autistic individuals and their parents.
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Transtorno Autístico , COVID-19 , Adulto , Transtorno Autístico/epidemiologia , Bélgica/epidemiologia , COVID-19/epidemiologia , Criança , Controle de Doenças Transmissíveis , Humanos , Pais , Qualidade de Vida , SARS-CoV-2RESUMO
The cerebellum harbors a circadian clock that can be shifted by scheduled mealtime and participates in behavioral anticipation of food access. Large-scale two-dimensional difference gel electrophoresis (2D-DIGE) combined with mass spectrometry was used to identify day-night variations in the cerebellar proteome of mice fed either during daytime or nighttime. Experimental conditions led to modified expression of 89 cerebellar proteins contained in 63 protein spots. Five and 33 spots were changed respectively by time-of-day or feeding conditions. Strikingly, several proteins of the heat-shock protein family (i.e., Hsp90aa1, 90ab1, 90b1, and Hspa2, 4, 5, 8, 9) were down-regulated in the cerebellum of daytime food-restricted mice. This was also the case for brain fatty acid protein (Fabp7) and enzymes involved in oxidative phosphorylation (Ndufs1) or folate metabolism (Aldh1l1). In contrast, aldolase C (Aldoc or zebrin II) and pyruvate carboxylase (Pc), two enzymes involved in carbohydrate metabolism, and vesicle-fusing ATPase (Nsf) were up-regulated during daytime restricted feeding, possibly reflecting increased neuronal activity. Significant feeding × time-of-day interactions were found for changes in the intensity of 20 spots. Guanine nucleotide-binding protein G(o) subunit alpha (Gnao1) was more expressed in the cerebellum before food access. Neuronal calcium-sensor proteins [i.e., parvalbumin (Pvalb) and visinin-like protein 1 (Vsnl1)] were inversely regulated in daytime food-restricted mice, compared to control mice fed at night. Furthermore, expression of three enzymes modulating the circadian clockwork, namely heterogeneous nuclear ribonucleoprotein K (Hnrnpk), serine/threonine-protein phosphatases 1 (Ppp1cc and Ppp1cb subunits) and 5 (Ppp5), was differentially altered by daytime restricted feeding. Besides cerebellar proteins affected only by feeding conditions or daily cues, specific changes in in protein abundance before food access may be related to behavioral anticipation of food access and/or feeding-induced shift of the cerebellar clockwork.
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BACKGROUND: With the overarching objective to gain better insights into social attention in autistic adults, the present study addresses three outstanding issues about face processing in autism. First, do autistic adults display a preference for mouths over eyes; second, do they avoid direct gaze; third, is atypical visual exploration of faces in autism mediated by gender, social anxiety or alexithymia? METHODS: We used a novel reinforced preferential looking paradigm with a group of autistic adults (n = 43, 23 women) pairwise matched on age with neurotypical participants (n = 43, 21 women). Participants watched 28 different pairs of 5 s video recordings of a speaking person: the two videos, simultaneously displayed on the screen, were identical except that gaze was directed at the camera in one video and averted in the other. After a 680 ms transition phase, a short reinforcement animation appeared on the side that had displayed the direct gaze. RESULTS: None of the groups showed a preference for mouths over eyes. However, neurotypical participants fixated significantly more the stimuli with direct gaze, while no such preference emerged in autistic participants. As the experiment progressed, neurotypical participants also increasingly anticipated the appearance of the reinforcement, based on the location of the stimulus with the direct gaze, while no such anticipation emerged in autistic participants. LIMITATIONS: Our autistic participants scored higher on the social anxiety and alexithymia questionnaires than neurotypicals. Future studies should match neurotypical and autistic participants on social anxiety and alexithymia and complement questionnaires with physiological measures of anxiety. CONCLUSIONS: The absence of preference for direct versus averted gaze in the autistic group is probably due to difficulties in distinguishing eye gaze direction, potentially linked to a reduced spontaneous exploration or avoidance of the eye region. Social attention and preference for direct versus averted gaze correlated with alexithymia and social anxiety scores, but not gender.
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Transtorno Autístico/fisiopatologia , Comportamento de Escolha , Fixação Ocular/fisiologia , Reforço Psicológico , Adulto , Sintomas Afetivos/complicações , Sintomas Afetivos/fisiopatologia , Ansiedade/complicações , Ansiedade/fisiopatologia , Transtorno Autístico/complicações , Tecnologia de Rastreamento Ocular , Feminino , Humanos , Masculino , Estimulação Luminosa , Comportamento SocialRESUMO
The oral mucosal pellicle is a thin lubricating layer generated by the binding of saliva proteins on epithelial oral cells. The protein composition of this biological structure has been to date studied by targeted analyses of specific salivary proteins. In order to perform a more exhaustive proteome characterization of pellicles, we used TR146 cells expressing or not the transmembrane mucin MUC1 and generated pellicles by incubation with human saliva and washing to remove unbound proteins. A suitable method was established for the in vitro isolation of the mucosal pellicle by "shaving" it from the cells using trypsin. The extracts, the washing solutions and the saliva used to constitute the pellicles were analyzed by LC MS/MS (data are available via ProteomeXchange with identifier PXD017268). Comparison of pellicle and saliva compositions evidenced the adsorption of proteins not previously reported as pellicle constituents such as proteins of the PLUNC family. Pellicles formed on TR146 and TR146/MUC1 were also analyzed and compared by protein label-free quantification. The two types of samples appeared as distinct clusters in multivariate analyses, but the discriminant proteins (Welch test p < .05, FDR < 0.1) were cellular rather than salivary proteins. SIGNIFICANCE: The oral mucosal pellicle is made of salivary proteins tightly bound to oral epithelial cells. It is essential to oral health, with biological functions depending largely on its protein constituents. Characterizing its proteome is difficult due to the intimate association of this protein layer to cell membranes. In this work, we report a trypsin "shaving" protocol which enabled to sample the pellicle formed on an in vitro cellular model of oral epithelium. Analyzing such samples by high-resolution mass spectrometry provided novel information on the mucosal pellicle composition. This work is therefore a good starting point for further characterization of this biological structure.
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Proteômica , Espectrometria de Massas em Tandem , Película Dentária , Epitélio , Humanos , Saliva , Proteínas e Peptídeos SalivaresRESUMO
We hypothesized that skeletal muscle contraction produces a cellular stress signal, triggering adipose tissue lipolysis to sustain fuel availability during exercise. The present study aimed at identifying exercise-regulated myokines, also known as exerkines, able to promote lipolysis. Human primary myotubes from lean healthy volunteers were submitted to electrical pulse stimulation (EPS) to mimic either acute intense or chronic moderate exercise. Conditioned media (CM) experiments with human adipocytes were performed. CM and human plasma samples were analyzed using unbiased proteomic screening and/or ELISA. Real-time qPCR was performed in cultured myotubes and muscle biopsy samples. CM from both acute intense and chronic moderate exercise increased basal lipolysis in human adipocytes. Growth and differentiation factor 15 (GDF15) gene expression and secretion increased rapidly upon skeletal muscle contraction. GDF15 protein was upregulated in CM from both acute and chronic exercise-stimulated myotubes. We further showed that physiological concentrations of recombinant GDF15 protein increased lipolysis in human adipose tissue, while blocking GDF15 with a neutralizing antibody abrogated EPS CM-mediated lipolysis. We herein provide the first evidence to our knowledge that GDF15 is a potentially novel exerkine produced by skeletal muscle contraction and able to target human adipose tissue to promote lipolysis.
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Exercício Físico/fisiologia , Fator 15 de Diferenciação de Crescimento/metabolismo , Lipólise/fisiologia , Músculo Esquelético/metabolismo , Adulto , Humanos , MasculinoRESUMO
A major class of clinical biomarkers is constituted of intracellular proteins which are leaking into the blood following ischemia, exposure to toxic xenobiotics or mechanical aggression. Their ectopic presence in plasma/serum is an indicator of tissue damage and raises a warning signal. These proteins, referred to as cytolysis biomarkers, are generally of cytoplasmic origin and as such, are devoid of glycosylation. In contrast, most plasma/serum proteins originate from the hepatic secretory pathway and are heavily glycosylated (at the exception of albumin). Recent advances in targeted proteomics have supported the parallelized evaluation of new blood biomarkers. However, these analytical methods must be combined with prefractionation strategies that reduce the complexity of plasma/serum matrix. In this article, we present the glycodepletion method, which reverses the hydrazide-based glycocapture concept to remove plasma/serum glycoproteins from plasma/serum matrix and facilitates the detection of cytolysis biomarkers. Glycodepletion was integrated to a targeted proteomics pipeline to evaluate 4 liver cytolysis biomarker candidates in the context of acetaminophen-induced acute hepatitis.
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Proteínas Sanguíneas/isolamento & purificação , Glicoproteínas/isolamento & purificação , Proteínas/análise , Proteômica/métodos , Sequência de Aminoácidos , Biomarcadores/análise , Biomarcadores/sangue , Fracionamento Químico/métodos , Cromatografia Líquida/métodos , Glicoproteínas/sangue , Glicosilação , Hepatite/sangue , Humanos , Espectrometria de Massas em Tandem/métodosRESUMO
While safety of fasting therapy is debated in humans, extended fasting occurs routinely and safely in wild animals. To do so, food deprived animals like breeding penguins anticipate the critical limit of fasting by resuming feeding. To date, however, no molecular indices of the physiological state that links spontaneous refeeding behaviour with fasting limits had been identified. Blood proteomics and physiological data reveal here that fasting-induced body protein depletion is not unsafe "per se". Indeed, incubating penguins only abandon their chick/egg to refeed when this state is associated with metabolic defects in glucose homeostasis/fatty acid utilization, insulin production and action, and possible renal dysfunctions. Our data illustrate how the field investigation of "exotic" models can be a unique source of information, with possible biomedical interest.
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Proteínas Sanguíneas/metabolismo , Jejum/sangue , Segurança , Spheniscidae/sangue , Animais , HumanosRESUMO
¹H Nuclear magnetic resonance (NMR) spectroscopy (400 MHz) was used in the context of food surveillance to develop a reliable analytical tool to differentiate brands of cola beverages and to quantify selected constituents of the soft drinks. The preparation of the samples required only degassing and addition of 0.1% of TSP in D2O for locking and referencing followed by adjustment of pH to 4.5. The NMR spectra obtained can be considered as "fingerprints" and were analyzed by principal component analysis (PCA). Clusters from colas of the same brand were observed, and significant differences between premium and discount brands were found. The quantification of caffeine, acesulfame-K, aspartame, cyclamate, benzoate, hydroxymethylfurfural (HMF), sulfite ammonia caramel (E 150D), and vanillin was simultaneously possible using external calibration curves and applying TSP as internal standard. Limits of detection for caffeine, aspartame, acesulfame-K, and benzoate were 1.7, 3.5, 0.8, and 1.0 mg/L, respectively. Hence, NMR spectroscopy combined with chemometrics is an efficient tool for simultaneous identification of soft drinks and quantification of selected constituents.