Salivary molecular spectroscopy : a sustainable, rapid and non-invasive monitoring tool for diabetes mellitus during insulin treatment

Monitoring of blood glucose is an invasive, painful and costly practice in diabetes. Consequently, the search for a more cost-effective (reagent-free), non-invasive and specific diabetes monitoring method is of great interest. Attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy has been used in diagnosis of several diseases, however, applications in the monitoring of diabetic treatment are just beginning to emerge. Here, we used ATR-FTIR spectroscopy to evaluate saliva of non-diabetic (ND), diabetic (D) and insulin-treated diabetic (D+I) rats to identify potential salivary biomarkers related to glucose monitoring. The spectrum of saliva of ND, D and D+I rats displayed several unique vibrational modes and from these, two vibrational modes were pre-validated as potential diagnostic biomarkers by ROC curve analysis with significant correlation with glycemia. Compared to the ND and D+I rats, classification of D rats was achieved with a sensitivity of 100%, and an average specificity of 93.33% and 100% using bands 1452 cm-1 and 836 cm-1, respectively. Moreover, 1452 cm-1 and 836 cm-1 spectral bands proved to be robust spectral biomarkers and highly correlated with glycemia (R2 of 0.801 and 0.788, P < 0.01, respectively). Both PCA-LDA and HCA classifications achieved an accuracy of 95.2%. Spectral salivary biomarkers discovered using univariate and multivariate analysis may provide a novel robust alternative for diabetes monitoring using a non-invasive and green technology

The influence of a hot environment on physiological stress responses in exercise until exhaustion

Exhaustive exercise in a hot environment can impair performance. Higher epinephrine plasma levels occur during exercise in heat, indicating greater sympathetic activity. This study examined the influence of exercise in the heat on stress levels. Nine young healthy men performed a maximal progressive test on a cycle ergometer at two different environmental conditions: hot (40 degrees C) and normal (22 degrees C), both between 40% and 50% relative humidity. Venous blood and saliva samples were collected pre-test and post-test. Before exercise there were no significant changes in salivary biomarkers (salivary IgA: p = 0.12; alpha-amylase: p = 0.66; cortisol: p = 0.95; nitric oxide: p = 0.13; total proteins: p = 0.07) or blood lactate (p = 0.14) between the two thermal environments. Following exercise, there were significant increases in all variables (salivary IgA 22 degrees C: p = 0.04, 40 degrees C: p = 0.0002; alpha-amylase 22 degrees C: p = 0.0002, 40 degrees C: p = 0.0002; cortisol 22 degrees C: p = 0.02, 40 degrees C: p = 0.0002; nitric oxide 22 degrees C: p = 0.0005, 40 degrees C: p = 0.0003, total proteins 22 degrees C: p<0.0001, 40 degrees C: p<0.0001 and; blood lactate 22 degrees C: p<0.0001, 40 degrees C: p<0.0001) both at 22 degrees C and 40 degrees C. There was no significant adjustment regarding IgA levels between the two thermal environments (p = 0.74), however the levels of alpha-amylase (p = 0.02), cortisol (p<0.0001), nitric oxide (p = 0.02) and total proteins (p = 0.01) in saliva were higher in the hotter conditions. Blood lactate was lower under the hot environment (p = 0.01). In conclusion, enduring hot temperature intensified stressful responses elicited by exercise. This study advocates that hot temperature deteriorates exercise performance under exhaustive stress and effort conditions

Study of Melipona quadrifasciata brain under operant learning using proteomic and phosphoproteomic analysis

Abstract Learning to anticipate events based on the predictive relationship between an action and an outcome (operant conditioning) is a form of associative learning shared by humans and most of other living beings, including invertebrates. Several behavioral studies on the mechanisms of operant conditioning have included Melipona quadrifasciata, a honey bee that is easily manipulated due to lack of sting. In this work, brain proteomes of Melipona bees trained using operant conditioning and untrained (control) bees were compared by two-dimensional gel electrophoresis analysis within pI range of 3-10 and 4–7; in order to find proteins specifically related to this type of associative learning.One protein was detected with differential protein abundance in the brains of trained bees, when compared to not trained ones, through computational gel imaging and statistical analysis. This protein was identified by peptide mass fingerprinting and MS/MS peptide fragmentation using a MALDI-TOF/TOF mass spectrometer as one isoform of arginine kinase monomer, apparently dephosphorylated. Brain protein maps were obtained by 2-DE (Two-dimensional gel electrophoresis) from a total proteins and phosphoproteins extract of the bee Melipona quadrifasciata. One isoform of arginine kinase, probably a dephosphorylated isoform, was significantly more abundant in the brain of trained bees using operant conditioning. Arginine kinase has been reported as an important enzyme of the energy releasing process in the visual system of the bee, but it may carry out additional and unexpected functions in the bee brain for learning process

The use of Open Reading frame ESTs (ORESTES) for analysis of the honey bee transcriptome

BACKGROUND: The ongoing efforts to sequence the honey bee genome require additional initiatives to define its transcriptome. Towards this end, we employed the Open Reading frame ESTs (ORESTES) strategy to generate profiles for the life cycle of Apis mellifera workers. RESULTS: Of the 5,021 ORESTES, 35.2% matched with previously deposited Apis ESTs. The analysis of the remaining sequences defined a set of putative orthologs whose majority had their best-match hits with Anopheles and Drosophila genes. CAP3 assembly of the Apis ORESTES with the already existing 15,500 Apis ESTs generated 3,408 contigs. BLASTX comparison of these contigs with protein sets of organisms representing distinct phylogenetic clades revealed a total of 1,629 contigs that Apis mellifera shares with different taxa. Most (41%) represent genes that are in common to all taxa, another 21% are shared between metazoans (Bilateria), and 16% are shared only within the Insecta clade. A set of 23 putative genes presented a best match with human genes, many of which encode factors related to cell signaling/signal transduction. 1,779 contigs (52%) did not match any known sequence. Applying a correction factor deduced from a parallel analysis performed with Drosophila melanogaster ORESTES, we estimate that approximately half of these no-match ESTs contigs (22%) should represent Apis-specific genes. CONCLUSIONS: The versatile and cost-efficient ORESTES approach produced minilibraries for honey bee life cycle stages. Such information on central gene regions contributes to genome annotation and also lends itself to cross-transcriptome comparisons to reveal evolutionary trends in insect genomes

Salivary nitric oxide and alpha-amylase as indexes of training intensity and load

This study examined the variation in salivary nitric oxide (NO), alpha-amylase (sAA) and serum markers of muscle injury during 21 weeks of training in elite swimmers. Samples of saliva and blood were collected once a month during 5 months from 11 male professional athletes during their regular training season. The variation in each marker throughout the 21 weeks was compared with the dynamics of trainingvolume, intensity and load. Unstimulated whole saliva was assessed for NO and sAA whereas venous blood was assessed for lactate dehydrogenase, creatine kinase, and γ-glutamyltransferase. Nitric oxide and sAA showed a proportional response to the intensity of training. However, whereas the concentration of NO increased across the 21 weeks, the activity of sAA decreased. Similar variations in the concentration of NO and the markers of muscle injury were also observed. The higher concentration of NO might be attributed to changes in haemodynamics and muscle regenerative processes. On the other hand, autonomic regulation towards parasympathetic predominance might have been responsible for the decrease in sAA activity. These findings provide appealing evidence for the utilization of salivary constituents in sports medicine to monitor training programmes

C-reactive protein is inversely and independently associated with maximal oxygen uptake in adolescents with obesity

The purposes of this study were: 1) investigate whether different markers of infl ammation (CRP and TNF-alpha)&nbsp;are correlated to cardiorespiratory fi tness in adolescents with obesity; 2) examine the association of these&nbsp;variables when adjusted by parameters of body composition. Were selected 57 individuals, 34 girls and 23&nbsp;boys, with 16.4±1.56 years and body mass index 36.0±4.3 kg/m2. Anthropometric measurements (weight,&nbsp;height and abdominal circunference) and body composition (BMI, visceral fat, body fat) were assessment.&nbsp;Body composition was estimated by analyzer of tetrapolar bioelectrical impedance. Obesity was defi ned as&nbsp;BMI&gt;95th percentile of the curve proposed by the Center for Diseases Control. Plasma TNF-alpha was measured&nbsp;by a quantitative two-site high-sensitivity enzyme immunoassay and CRP was measured by high-sensitivityimmunoturbidimetric assay. Graded maximal exercise testing was performed to obtain the maximum oxygen&nbsp;consumption (VO2max) and velocity associated with the VO2max (vVO2max). Signifi cant correlations of CRP with&nbsp;VO2max and vVO2max were found (r=-0.40 and r=-0,36, respectively). No correlations were observed between&nbsp;TNF-alpha and VO2max e vVO2max. CRP was associated to the VO2max and vVO2max independently of measurements of&nbsp;body composition. The CRP was independently and inversely associated with direct measurement of VO2max&nbsp;and the indirect variable of cardiorespiratory fitness vVO2max, in obese adolescents, even after adjustments&nbsp;in body composition, a potential confounding factor. No association was found between TNF-alpha and the&nbsp;parameters of cardiorespiratory fitness