Inhibitory Potential of Different Bilberry (Vaccinium myrtillus L.) Extracts on Human Salivary α-Amylase.

Recently, consumer preferences for bilberries have increased markedly. This fact is probably related to their natural constituents, such as phenolic compounds including anthocyanins and tannins, as well as the vitamins and minerals they contain. Phenolic compounds are known for their numerous beneficial effects on human health. Moreover, bilberry fruits have been shown to inhibit the activity of carbohydrate hydrolyzing enzymes, which can significantly decrease the postprandial increase in blood glucose levels. Thus, the aim of the present study is to investigate the inhibitory effect of Vaccinium myrtillus L. extracts on key enzyme α-amylase, linked to type 2 diabetes. No data have been published on the inhibitory properties of Vaccinium myrtillus L. fruits growing wild in Bulgaria against carbohydrate enzymes. Bilberry extracts were analyzed for total polyphenols, total anthocyanin content, antioxidant activity and their inhibitory properties against α-amylase. The contents of flavonols, anthocyanins and stilbenes were determined by HPLC analysis. The identified flavonols in the analyzed bilberry extracts were mainly represented by quercetin derivatives as rutinoside. The predominant anthocyanins for both aqueous and organic solvents were delphinidin-3-galactoside and malvidin-3-glucoside. The results revealed that bilberry extracts are effective inhibitors of α-amylase, with IC50 values from 20.8 to 194.8 µg GAE/mL. All the samples proved to have antioxidant activity measured by three different in vitro assays (FRAP, CUPRAC and DPPH). The inhibitory properties of V. myrtillus L. extracts may provide a new direction in the development and research of new pharmaceuticals for the suppression of postprandial hyperglycemia in diabetic patients.

Assessment of salivary alpha amylase and mucin-4 before and after non-surgical treatment of peri-implant mucositis.

BACKGROUND: The present study was based on the null hypothesis that there is no difference in clinicoradiographic parameters and whole salivary alpha amylase (AA) and mucin-4 levels before and after non-surgical mechanical debridement (NSMD) of patients with peri-implant mucositis (PM). The aim was to assess whole salivary AA and mucin-4 levels before and after treatment of PM. METHODS: Patients with PM (Group-1) and individuals without peri-implant diseases (Group-2) were included. Demographic data was collected and peri-implant modified plaque and bleeding indices (mPI and mBI, respectively), probing depth (PD) and crestal bone loss were measured at baseline. Levels of AA and mucin-4 were assessed in unstimulated whole saliva samples. All patients underwent full-mouth non-surgical periodontal therapy (NSPT) and NSMD; and clinical parameters and salivary biomarkers were re-assessed after 3 months. Level of significance was set at P < 0.01. RESULTS: Twenty-six and 32 individuals were included in groups 1 and 2, respectively. None of the participants had periodontitis. At baseline clinical periodontal parameters (PI [P < 0.001], GI [P < 0.001], clinical AL [P < 0.001] and PD [P < 0.001]) were significantly high in Group-1 than Group-2. At 3-month follow-up, there was a statistically significant reduction in clinical periodontal and peri-implant parameters (PI [P < 0.01], GI [P < 0.01], and PD [P < 0.01]) in Group-1 compared with their baseline values. At baseline, salivary AA levels were significantly high in Group-1 than Group-2 (P < 0.01). At 3-month follow-up, there was no significant difference in whole salivary AA levels among patients in groups 1 and 2. CONCLUSIONS: The AA and mucin-4 levels are potential biomarkers for evaluation of peri-implant diseases including PM. Mechanical instrumentation continues to be the most predictable treatment option for the management of peri-implant diseases.

Decreased salivary α-amylase activity responding to citric acid stimulation in Myasthenia gravis with malnutrition.

OBJECTIVES: Malnutrition, defined according to Nutritional risk screening (NRS 2002), is commonly observed in patients of Myasthenia gravis (MG), a neuromuscular disorder manifested by varied degrees of skeletal muscle weakness. Because biochemical composition of saliva changes in correspondence to alterations in nutritional status, we tested our hypothesis that a certain saliva component(s) might serve as a biomarker(s) for nutrition status of MG, particularly for those MG patients with high risk of malnutrition. MATERIALS AND METHODS: 60 MG patients and 60 subjects belonging to the healthy control group (HCG) were enrolled in this case-control study. The salivary α-amylase (sAA) activity, salivary flow rate (SFR), pH, total protein density (TPD), and the concentrations of chloride and calcium ions in MG group with or without malnutrition were measured before and after citric acid stimulation. Thereafter, the relationship between sAA activity and BMI was determined in MG and HCG. RESULTS: Compared with HCG, more patients with malnutrition, increased TPD and chloride and calcium concentrations but decreased pH value and SFR both before and after acid stimulation, as well as reduced sAA activity, pH and TPD responses to acid stimulation. MG with malnutrition showed decreased sAA activity and TPD responding to acid stimulation compared with those without malnutrition. Compared with normal BMI, sAA activity response to acid stimulation was reduced in low BMI. There was a significant strong positive correlation between the ratio of sAA activity and BMI in MG. CONCLUSIONS: Salivary biochemical characteristics are abnormally altered in MG with malnutrition. Altered sAA activity responding to acid stimulation was associated with malnutrition. CLINICAL RELEVANCE: Decreased sAA activity responding to acid stimulation can reflect malnutrition state and may be one potential screening marker for MG patients with high risk of malnutrition.

Assessment of salivary alpha-amylase and cortisol as a pain related stress biomarker in dogs pre-and post-operation.

BACKGROUND: The use of salivary biomarkers has garnered attention because the composition of saliva reflects the body's physiological state. Saliva contains a wide range of components, including peptides, nucleic acids, electrolytes, enzymes, and hormones. It has been reported that salivary alpha-amylase and cortisol are biomarkers of stress related biomarker in diseased dogs; however, evaluation of salivary alpha-amylase and cortisol pre- and post- operation has not been studied yet. The aim of this study was to evaluate salivary alpha-amylase and cortisol levels in dogs before and after they underwent surgery and investigate the association between the salivary alpha-amylase and cortisol activity and pain intensity. For this purpose, a total of 35 dogs with disease-related pain undergoing orthopedic and soft tissue surgeries were recruited. Alpha-amylase and cortisol levels in the dogs' saliva and serum were measured for each using a commercially available canine-specific enzyme-linked immunosorbent assay kit, and physical examinations (measurement of heart rate and blood pressure) were performed. In addition, the dogs' pre- and post-operative pain scores determined using the short form of the Glasgow Composite Measure Pain Scale (CMPS-SF) were evaluated. RESULTS: After surgery, there was a significant decrease in the dogs' pain scores (0.4-fold for the CMPS-SF, p < 0.001) and serum cortisol levels (0.73-fold, p < 0.01). Based on their pre-operative CMPS-SF scores, the dogs were included in either a high-pain-score group or a low-pain-score group. After the dogs in the high-pain-score group underwent surgical intervention, there was a significant decrease in their CMPS-SF scores and levels of salivary alpha-amylase, serum alpha-amylase, and serum cortisol. Additionally, there was a positive correlation between salivary alpha-amylase levels and CMPS-SF scores in both the high- and low-pain-score groups. CONCLUSIONS: The measurement of salivary alpha amylase can be considered an important non-invasive tool for the evaluation of pain-related stress in dogs.

Nicotine supplementation enhances simulated game performance of archery athletes.

BACKGROUND: Nicotine is beneficial to mood, arousal and cognition in humans. Due to the importance of cognitive functioning for archery athletes, we investigated the effects of nicotine supplementation on the cognitive abilities, heart rate variability (HRV), and sport performance of professional archers. METHODS: Eleven college archers were recruited and given 2 mg of nicotine supplementation (NIC group) and placebo (PLA group) in a crossover design. RESULTS: The results showed that at 30 min after the intake of nicotine gum, the "correct rejection" time in the NIC group was significantly lower than that of the PLA group (7.29 ± 0.87 vs. 8.23 ± 0.98 msec, p < 0.05). In addition, the NIC group completed the grooved pegboard test in a shorter time than the PLA group (48.76 ± 3.18 vs. 53.41 ± 4.05 s, p < 0.05), whereas motor reaction times were not different between the two groups. Saliva α-amylase activity was significantly lower after nicotine supplementation (p < 0.01) but increased immediately after the archery test in the NIC group (p < 0.05). In addition, nicotine supplementation significantly decreased HRV and increased the archery score (290.58 ± 10.09 vs. 298.05 ± 8.56, p < 0.01). CONCLUSIONS: Nicotine enhances the performance of archery athletes by increasing cognitive function and stimulating the sympathetic adrenergic system.

Factors contributing to stability and instability in alpha-amylase activity in diluted saliva samples over time.

For the measurement of salivary alpha-amylase (sAA) activity, saliva samples first have to be diluted. There is some evidence for instability, that is, a decline of sAA activity in diluted samples. It is not clear which factors during dilution may contribute to this phenomenon and how quickly this decline of sAA activity occurs. Several experiments were conducted to investigate whether and how the material of the container (polystyrene (PS), polypropylene (PP), glass; experiment 1) and the diluent (saline (NaCl) solution, phosphate buffer saline (PBS), ultra-pure water; experiment 2) may affect sAA stability in diluted samples over a broad time window of up to 5 h. To study the velocity of the phenomenon in a fine-grained temporal resolution, sAA activity during the dilution process was studied (experiment 3). The results suggest that the (in)stability of sAA activity in diluted samples is determined by the interaction of material, diluent, and time. The sAA activity was relatively stable if saliva samples were diluted with a NaCl solution or PBS in glass tubes. However, sAA activity in diluted samples decreased in plastic containers (PS, PP), or if ultra-pure water was used as the diluent. There was a clear time effect on this decline. However, the decline appears to require some time to evolve and may not occur immediately during the dilution process. To conclude, the dilution of saliva samples should preferably be conducted with NaCl solution or PBS in glass containers. If glass containers are not available, PS and PP containers can be used if the dilution is processed quickly (within 25 min) and the measurement is initiated immediately upon dilution.

Salivary alpha-amylase and hormones levels of young adults with different body composition.

The aim of this study was to assess the differences in salivary biomarker levels of young adults classified according to body fat accumulation. One hundred and thirty-four volunteers were evaluated (mean age 21 ± 2 years). Body composition was calculated through skinfold thickness: supra-iliac, biceps, and triceps, sub-scapular. Body fat percentage (BF%) was used to classify subjects according to fat tissue accumulation: normal-weight (n = 37, 19 females, 18 males), overweight (n = 42, 30 females, 12 males), and obese (n = 55, 42 females, 13 males). Saliva samples were collected 30 min after awakening to determine salivary levels of 17-beta-estradiol. For salivary cortisol and alpha-amylase activity (sAA) three samples were obtained, just after awakening, 30 min after awakening and at bedtime. Oral contraceptive intake was considered for the female group. The results showed that overweight and obese females using oral contraceptive presented lower levels of 17-ß-estradiol than normal-weight females. In overweight and obese males, sAA levels were higher 30 min post-awakening when compared with the normal-weight group. The comparison of sAA levels within time showed no difference for males; obese females showed significant higher values at bedtime than 30 min post-awakening. The salivary cortisol concentration showed higher values at morning decreasing significantly at bedtime for all groups. Concluding, differences in 17-ß-estradiol and sAA levels were found in females and males, respectively, according to body fat accumulation, showing the usefulness of salivary biomarkers in the study of systemic conditions. PRACTICAL APPLICATIONS: Saliva is an advantageous biological fluid in innovative methods for diagnosis. Besides being a noninvasive method, salivary steroid measurements have the potential to provide a convenient assessment of serum free steroid concentrations.

Evaluation of adenosine deaminase in saliva and serum, and salivary α-amylase, in canine pyometra at diagnosis and after ovariohysterectomy.

An assay for adenosine deaminase (ADA) was validated in serum and saliva in dogs. Changes in ADA and salivary α-amylase activities were analysed in 26 bitches diagnosed with pyometra and compared with activities in 19 healthy bitches. All animals were classified according to the American Society of Anaesthesiologists (ASA) scoring for physical status. In the validation study, the ADA assay had an imprecision<12% and determination coefficients>0.90 in linearity under dilution experiments, with recoveries of 99.2-114.4%. On the day of presentation, salivary ADA activity was significantly higher in dogs with pyometra than in healthy dogs (median values 7.1IU/L vs. 0.8IU/L, respectively; P<0.01). ADA had a moderate positive correlation with leucocyte and band neutrophil counts, haptoglobin, salivary α-amylase and ASA score, and a low positive correlation with C-reactive protein. There were no significant differences in salivary α-amylase activity between dogs with pyometra and healthy dogs (57.3IU/L vs. 27.4IU/L, respectively). Salivary α-amylase had a low correlation with ASA grade, and leucocyte and band neutrophil counts. In 7/26 bitches with pyometra that were sampled 3 and 10days after ovariohysterectomy, there were no significant changes in α-amylase or ADA activities. These results indicate that ADA activity is increased in the saliva of bitches with pyometra, probably related to systemic inflammation.

Stress response and communication in surgeons undergoing training in endoscopic management of major vessel hemorrhage: a mixed methods study.

BACKGROUND: Major vessel hemorrhage in endoscopic, endonasal skull-base surgery is a rare but potentially fatal event. Surgical simulation models have been developed to train surgeons in the techniques required to manage this complication. This mixed-methods study aims to quantify the stress responses the model induces, determine how realistic the experience is, and how it changes the confidence levels of surgeons in their ability to deal with major vascular injury in an endoscopic setting. METHODS: Forty consultant surgeons and surgeons in training underwent training on an endoscopic sheep model of jugular vein and carotid artery injury. Pre-course and post-course questionnaires providing demographics, experience level, confidence, and realism scores were taken, based on a 5-point Likert scale. Objective markers of stress response including blood pressure, heart rate, and salivary alpha-amylase levels were measured. RESULTS: Mean "realism" score assessed posttraining showed the model to be perceived as highly realistic by the participants (score 4.02). Difference in participant self-rated pre-course and post-course confidence levels was significant (p < 0.0001): mean pre-course confidence level 1.66 (95% confidence interval [CI], 1.43 to 1.90); mean post-course confidence level 3.42 (95% CI, 3.19 to 3.65). Differences in subjects' heart rates (HRs) and mean arterial blood pressures (MAPs) were significant between injury models (p = 0.0008, p = 0.0387, respectively). No statistically significant difference in salivary alpha-amylase levels pretraining and posttraining was observed. CONCLUSION: Results from this study indicate that this highly realistic simulation model provides surgeons with an increased level of confidence in their ability to deal with the rare but potentially catastrophic event of major vessel injury in endoscopic skull-base surgery.

The role of week(end)-day and awakening time on cortisol and alpha-amylase awakening responses.

Awakening responses in salivary cortisol (CAR) and alpha-amylase (AAR) constitute proxies of morning activation patterns of the hypothalamic-pituitary-adrenal axis and autonomic nervous system, respectively. Previous studies suggest that the CAR is decreased at weekends and at late awakening. However, it is insufficiently studied (a) whether this also applies to the AAR and (b) whether week(end)-day and awakening time interact with each other. Using an ecological momentary assessment design, 48 healthy young adults (60% women) were investigated over a 7-d period (Study 1), and 27 chronic pain patients diagnosed with fibromyalgia syndrome were examined over a 14-d period (Study 2). For the assessment of the CAR and AAR, participants provided saliva samples each morning (upon awakening, 30 min after awakening). Preprogrammed electronic diary devices were used to track week(end)-day and exact time of saliva sampling (awakening time). In Study 1, CAR was unrelated to weekend, awakening time, or their interaction, whereas only early awakening time was positively associated with AAR. In Study 2, week-days as well as early awakening times on week-days predicted increased CAR. AARs were not predicted by week(end)-day, awakening times, or their interaction. These findings suggest that time-related factors may influence awakening responses, particularly the impact of week(end)-day on the CAR and the impact of awakening times on the AAR. Since week(end)-day and awakening times may negatively affect awakening responses, these potential confounding factors should be assessed and controlled for, particularly in studies assessing both CAR and AAR.

Cortisol y amilasa salival en niñas: variación según la curva diurna, la ingesta de alimentos y la actividad física / Diurnal variation of salivary cortisol and alpha amylase levels in normal girls: effects of meals and physical exercise

Salivary cortisol levels and saliva alpha-amylase enzymatic activity are non-invasive markers of hypothalamic-pituitary-adrenal axis and autonomic nervous system related to stress, which could be associated with excessive energy intake in response to stressors. Aim: To describe the diurnal variation of salivary cortisol levels and á-amylase activity in prepubertal girls and to assess their change after meals and physical activity episodes. Subjects and Methods: Nine normal-weight girls aged 8 to 10 years were monitored for 14 hours, from 6:00 AM to 20:00 h. Three standardized meals were administered across the day with a controlled sport competition performed at the end of the day. Saliva samples were drawn upon awakening, 30 min after awakening, before and after lunch, before and after dinner, and before-after the controlled episode of physical activity. Results: A decreasing salivary cortisol diurnal pattern was confirmed, with an initial increase occurring 30 minutes after waking up. An ascending diurnal pattern was observed for salivary amylase activity. Meals significantly increased cortisol levels, with a non-significant trend to increase amylase activity. The magnitude of physical activity during acute exercise was associated with increased salivary amylase activity (rho = 0.84; P <0.01). Cortisol levels were positively correlated with body mass index z scores (rho = 0.87; P <0.01). Conclusions: We confirmed the existence of a diurnal pattern of salivary cortisol levels and á-amylase activity in saliva that is modulated by diet and exercise. Our preliminary results also show that salivary cortisol might be related with body weight...

Detection of host-specific immunogenic proteins in the saliva of patients with oral squamous cell carcinoma.

The main purpose of this article is to develop a new and reliable saliva-based clinical diagnostic method for the early detection of oral squamous cell carcinoma (OSCC). This study used an immunoproteomic approach which allowed the detection of immunogenic host proteins in patients' samples using pooled human antibodies. In an attempt to investigate potential biomarkers of OSCC, two-dimensional electrophoresis (2-DE) followed by immunoblotting of saliva from patients and controls were compared. The protein spots of interest were analyzed using 2-DE image analyzer and subsequently subjected to MALDI-TOF/TOF and then matched against NCBI database. The result showed that four protein clusters, namely Human Pancreatic Alpha-amylase (HPA), Human Salivary Amylase (sAA), keratin-10 (K-10), and Ga Module Complexed with Human Serum Albumin (GA-HSA), had exhibited immunoreactivity in western blot. The results are suggestive of the potential use of the differentially expressed saliva protein as tumor biomarkers for the detection of OSCC. However, further studies are recommended to validate this finding.

Amylase α-1A (AMY1A): a novel immunohistochemical marker to differentiate chromophobe renal cell carcinoma from benign oncocytoma.

Chromophobe renal cell carcinoma (ChRCC) and oncocytoma present with a perplexing overlap of morphologic and immunohistochemical features. ChRCC have deletions in the 1p21.1 region including the amylase α-1A gene (AMY1A). No such deletions are found in oncocytoma. Instead, oncocytomas shared other deletions on chromosome 1: 1p31.3, 1q25.2, and 1q44. We performed AMY1A immunostaining on 75 oncocytomas (57 tissue microarray [TMA] cores, 18 whole slides) and 54 ChRCCs (20 TMA cores, 34 whole slides). Staining was assessed using the H-score method. The intensity was graded as follows: no staining=0, weak=1, moderate=2, and strong=3. The AMY1A immunostain preferentially stained the distal tubules and collecting ducts of normal kidney. All oncocytomas (100%) expressed AMY1A with an H-score that varied from 100 to 300 (mean 205). Mild to moderate heterogeneity in staining intensity was noted within a given oncocytoma. For oncocytomas, 87% (65/75) cases had H-scores of at least 120 with a mean score of 221. Notably, the 13% (10/75) of oncocytoma cases that had an H-score of 100 were derived from the TMA. A total of 87% (47/54) of the ChRCC cases were negative for the AMY1A immunostain. Of the ChRCC cases, 4% (2/54) showed very weak cytoplasmic staining (H-score of 70 each), which was less than the lowest H-score of oncocytoma cases. All 5 cases of ChRCC, which showed an H-score of 100 or more, were referred to as eosinophilic variants of ChRCC. Three of these 5 cases showed a very nondescript, diffuse staining of the cytoplasm. Two of these 5 cases showed an H-score of 130. We think that as the staining pattern of these 2 cases is similar to that of oncocytoma, they should be put in a category of renal oncocytic neoplasms favoring oncocytoma. This result shows that AMY1A staining could be very helpful in further classifying even a subset of the eosinophilic variants of ChRCC. The difference between ChRCC and oncocytoma was statistically significant (χ test, P<0.0001). All cases of clear cell RCC and papillary RCC were negative for AMY1A expression. Overall, sensitivity and specificity of AMY1A staining for oncocytoma was 100% (95% confidence interval, 0.95-1.00) and 96.75% (95% confidence interval, 0.93-0.99), respectively. Similarly, the sensitivity and specificity for distinguishing oncocytoma from ChRCC was 100% (95% confidence interval, 0.95-1.00) and 90.74% (95% confidence interval, 0.80-0.97), respectively. These data show that the novel marker AMY1A can be of great diagnostic utility when trying to differentiate ChRCC (classic and eosinophilic variant) and oncocytoma.

Carbon dioxide insufflation reduces the discomfort due to colonoscopy as objectively analyzed by salivary stress markers.

BACKGROUND: Carbon dioxide (CO2) insufflation has been shown to reduce the procedure-related pain and discomfort during colonoscopy. However, the effects of CO2 insufflation on the improvement of participant's stress had not been objectively analyzed. METHODS: Using a randomized, double-blinded design, 100 consecutive patients undergoing colonoscopy were assigned to have their examination performed with either CO2 or air insufflation. Patients completed a questionnaire to grade their discomfort using a visual analogue scale (VAS). The salivary alpha-amylase (SAA) level was also measured at these times as a marker of stress. RESULTS: The total VAS score did not show any statistically significant differences between the CO2 and air insufflation groups. However, the VAS score for abdominal fullness significantly decreased in the CO2 insufflation group in comparison to the air insufflation group in the patients who had a longer examination. The titer of the maximum SAA was significantly increased by colonoscopy (P <0.01). CO2 insufflation significantly reduced the total SAA after the colonoscopy in comparison with air insufflation (P < 0.05). The examination time and SAA tended to have a positive relationship in the air insuflation group. However, no such relationship was observed in the CO2 insufflation group. CONCLUSIONS: CO2 insufflation significantly reduced the post-examination discomfort, as indicated by a salivary stress marker. The use of CO, rather than air insufflation reduced the patients' stress and may contribute to better acceptance of colonoscopy. The usefulness of CO2 insufflation was more prominent when a longer examination was necessary.

Reduction in salivary α-amylase levels following a mind-body intervention in cancer survivors--an exploratory study.

OBJECTIVE: The main aim of this exploratory study was to assess whether salivary α-amylase (sAA) and salivary cortisol levels would be positively modulated by sleep-focused mind-body interventions in female and male cancer survivors. METHODS: We conducted a randomized controlled trial in which 57 cancer survivors with self-reported sleep disturbance received either a Sleep Hygiene Education (SHE; n=18) control, or one of two experimental mind-body interventions, namely, Mind-Body Bridging (MBB; n=19) or Mindfulness Meditation (MM; n=20). Interventions were three sessions each conducted once per week for three consecutive weeks. Saliva cortisol and sAA were measured at baseline and 1 week after the last session. Participants also completed a sleep scale at the same time points when saliva was collected for biomarker measurement. RESULTS: Our study revealed that at post-intervention assessment, mean sAA levels upon awakening ("Waking" sample) declined in MBB compared with that of SHE. Mean Waking cortisol levels did not differ among treatment groups but declined slightly in SHE. Self-reported sleep improved across the three interventions at Post-assessment, with largest improvements in the MBB intervention. CONCLUSION: In this exploratory study, sleep focused mind-body intervention (MBB) attenuated Waking sAA levels, suggesting positive influences of a mind-body intervention on sympathetic activity in cancer survivors with sleep disturbance.

Salivary alpha-amylase during pregnancy: diurnal course and associations with obstetric history, maternal demographics, and mood.

Diurnal patterns of salivary alpha amylase (sAA) in pregnant women have not previously been described. The current study employed ecological momentary assessment to examine the association between the diurnal sAA, obstetric history, maternal demographics, and mood during pregnancy. Saliva was self-collected by 83 pregnant women (89% White, age 25.3-43.0 years; mean gestational age 21.9 weeks, range 6-37 weeks; gravida 1-6) at home over three days. Results indicated that current pregnancy (gestational age and fetal sex) and maternal demographics were not related to diurnal sAA. In contrast, a history of previous miscarriage (Parameter = -.17; SE = .05; p < .05) was associated with an atypical diurnal pattern. Even after accounting for obstetric history, trait anxiety (Parameter = .16; SE = .04; p < .001) was associated with increased sAA over the day while chronic levels of fatigue (Parameter = -.06; SE = .03; p < .05) were associated with decreased sAA. In a separate model, we also tested the time varying covariation of sAA and mood. The effects of momentary mood were in contrast to those for trait mood. Both momentary depression (Parameter = .22; SE = .09; p < .01) and vigour/positive mood (Parameter = .12; SE = .04; p < .001) were associated with momentary increases in sAA while momentary anxiety and fatigue were not related to sAA. The findings suggest that basal sAA during pregnancy is sensitive to emotional arousal. Evaluating diurnal patterns of sAA holds promise for advancing understanding of how emotional arousal during pregnancy may affect fetal development.

Concurrent attenuated reactivity of alpha-amylase and cortisol is related to disruptive behavior in male adolescents.

Attenuated reactivity of salivary alpha-amylase has been proposed as a specific sympathetic marker of disruptive behavior in juveniles and may have additional value to studying other autonomic parameters and hypothalamic-pituitary-adrenal axis activity. Investigating the interrelationships between neurobiological parameters in relation to juvenile disruptive behavior may enhance insight into the complex mechanisms at play. We investigated salivary alpha-amylase, cortisol, heart rate (HR), and heart rate variability (HRV) in response to a standardized public speaking task, and examined interactions between these parameters in relation to disruptive behavior. Participants were 48 delinquent male adolescents (mean age 18.4 years, SD 0.9), with and without a disruptive behavior disorder (resp. DP+, DP-) and 16 matched normal controls (NC). A structured psychiatric interview as well as the Youth Self Report and Child Behavior Checklist were administered to assess disruptive behavior. Alpha-amylase and cortisol reactivity, but not HR or HRV, showed significant inverse associations with dimensional measures of disruptive behavior. Moreover, both cortisol and alpha-amylase reactivity were significantly lower in the DP+ group as compared to the NC group. The mentioned relationships remained present when nicotine use was entered as a covariate. Combining alpha-amylase and cortisol in one model explained a larger part of the variance of disruptive behavior than either single parameter. There were no interactions between alpha-amylase and cortisol or HRV in relation to disruptive behavior. Attenuated alpha-amylase responsivity to stress is a correlate of disruptive behavior in late-adolescent males. Although nicotine use explains a considerable part of the variance of disruptive behavior, both alpha-amylase and cortisol are related to disruptive behavior, over and above the effect of nicotine use. Combining alpha-amylase and cortisol improved insight into neurobiological mechanisms involved with disruptive behavior; concurrent low reactivity of both parameters was related to higher levels of disruptive behavior.

Initial salivary α-amylase activity predicts malignant ventricular arrhythmias and short-term prognosis after ST-segment elevation myocardial infarction.

BACKGROUND: Ventricular arrhythmias (VA), including ventricular tachycardia and ventricular fibrillation, are the most common remediable cause of death in patients with acute myocardial infarction. Augmented sympathetic neural activity to the heart and myocardial catecholamine release may be the primary factors in the generation of VA. The aim of this study was to assess the predictive value of salivary α-amylase (sAA) activity, an indicator of sympathetic activity, for malignant VA occurrence and for short-term outcome in patients with ST-segment elevation myocardial infarction (STEMI). METHODS: Patients with STEMI (n=91) were recruited from the Emergency Department during the period 1 December 2008 to 31 April 2010. Correlations of initial sAA activity at presentation with VA, as well as 1-month prognosis were analysed. RESULTS: sAA activity was significantly increased in the VA group (395±173.7; n=9) as compared with the non-VA group (283±89.3; n=82) (p=0.014). The adjusted OR for malignant ventricular arrhythmia occurrence was 1.010 (95% CI 1.001 to 1.018). Eight patients (8.8%) died and 24 (26.4%) had at least one short-term adverse event within the first month after STEMI. Simple logistic regression analysis showed that sAA is an independent predictor for short-term prognosis (p=0.049, OR 1.005, 95% CI 1.000 to 1.009). CONCLUSIONS: Although a prospective study with a large cohort is required, the present results suggest that high initial sAA activity is associated with increased risk of malignant VA and predicts short-term prognosis in patients with STEMI.

Neurobiological and behavioral stress reactivity in children prenatally exposed to tobacco.

This study examined neurobiological and behavioral stress reactivity in children who had been prenatally exposed to tobacco. Neurobiological stress reactivity was measured using salivary cortisol and alpha-amylase levels at five different time points throughout a stressful neuropsychological test session, which involved a competition against a videotaped opponent. Participants (mean age: 10.6 years, SD 1.3) were 14 prenatally exposed (PE) children, 9 children with disruptive behavior problems (DBD), and 15 normal controls (NC). For cortisol responses, no significant differences between the three groups were observed. Normal controls, however, had significantly higher alpha-amylase levels than PE-children throughout the test session, and their alpha-amylase levels also increased throughout the session, whereas these remained low and stable for PE-children. Alpha-amylase levels and trajectory of PE-children were similar to those observed for DBD-children. PE-children also showed significantly increased behavioral stress reactivity compared to NC-children, and neurobiological and behavioral stress reactivity were inversely related in PE-children, again similar to what was observed for DBD-children. These results support the hypothesis that prenatal smoking may lead to long-lasting neurobiological and behavioral changes in exposed offspring.

Measuring salivary analytes from free-ranging monkeys.

Studies of large free-ranging mammals have been revolutionized by non-invasive methods for assessing physiology, which usually involve the measurement of fecal or urinary biomarkers. However, such techniques are limited by numerous factors. To expand the range of physiological variables measurable non-invasively from free-ranging primates, we developed techniques for sampling monkey saliva by offering monkeys ropes with oral swabs sewn on the ends. We evaluated different attractants for encouraging individuals to offer samples, and proportions of individuals in different age/sex categories willing to give samples. We tested the saliva samples we obtained in three commercially available assays: cortisol, salivary alpha amylase, and secretory immunoglobulin A. We show that habituated free-ranging rhesus macaques will give saliva samples voluntarily without training, with 100% of infants, and over 50% of adults willing to chew on collection devices. Our field methods are robust even for analytes that show poor recovery from cotton, and/or that have concentrations dependent on salivary flow rate. We validated the cortisol and SAA assays for use in rhesus macaques by showing aspects of analytical validation, such as that samples dilute linearly and in parallel to assay standards. We also found that values measured correlated with biologically meaningful characteristics of sampled individuals (age and dominance rank). The SIgA assay tested did not react to samples. Given the wide range of analytes measurable in saliva but not in feces or urine, our methods considerably improve our ability to study physiological aspects of the behavior and ecology of free-ranging primates, and are also potentially adaptable to other mammalian taxa.