Effects of amount of training on the saliva concentrations of cortisol, dehydroepiandrosterone and on the dehydroepiandrosterone: cortisol concentration ratio in women over 16 weeks of training

This study was designed to investigate in the saliva the influence in female athletes of handball or volleyball training on concentrations of cortisol [C], dehydroepiandrosterone [DHEA], and on the [DHEA]:[C] ratio over 16 weeks of training. Data were compared to those of sedentary women. Saliva samples were collected upon waking after an overnight fast during the 1st week (W₁) of the training programme and in the 16th week (W₁₆). The training programme increased the resting concentrations of saliva [DHEA] in all the sportswomen. In contrast, a decrease of [DHEA] was noted in the sedentary group (W₁₆ < W₁; P < 0.05). In none of the women did the [C] at rest change significantly during the study. Between W₁ and W₁₆, the [DHEA]:[C] ratio increased by more than 30% in all the sportswomen. In addition, the athletes with the highest performance levels and greatest amount of training had the lowest [DHEA]:[C] ratio. Negative linear relationships between the amount of training and the [DHEA]:[C] ratio were found both at W₁ (r = −0.53 P < 0.001), and W₁₆ (r=−0.73 P < 0.001), suggesting that the latter could be used as an indicator of the training status of sportswomen. Accepted: 12 May 1998     

Quantifying blood leakage into the oral mucosa and its effects on the measurement of cortisol, dehydroepiandrosterone, and testosterone in saliva

The impact of blood leakage due to microinjury to the oral cavity on the measurement of salivary hormones was examined. Saliva samples were collected before, immediately after, and then every 15 min for 1 h following vigorous tooth brushing. Blood in saliva was quantified by visual inspection of discoloration, Hemastix reagent strips to detect hemoglobin, and an immunoassay for transferrin. The presence of blood in saliva immediately after microinjury was confirmed by all methods. Hemoglobin and transferrin levels remained elevated over baseline for at least 30 min. Levels of salivary testosterone increased over baseline and remained elevated for 30 min in response to microinjury. Microinjury induced change in salivary testosterone was more closely associated with the change in transferrin than hemoglobin levels or discoloration ratings. On average, levels of salivary dehydroepiandrosterone (DHEA) did not increase in response to microinjury. However, individual differences in microinjury induced change in DHEA were associated with discoloration ratings. Salivary cortisol levels, on average, were neither responsive to microinjury, nor were individual differences in cortisol change associated with blood contamination measures. Neither diurnal nor gender-related differences in baseline hormone levels predicted the impact of blood leakage on quantitative salivary measurements. The findings suggest ecologically valid minor-to-moderate level microinjuries to the oral cavity have negligible effects on the measurement of salivary cortisol, but may be important to quantify and control when assessing other hormones especially testosterone.     

Hippocampal damage abolishes the cortisol response to psychosocial stress in humans

The hippocampus (HC) is necessary for learning and memory, but it also plays a role in other behaviors such as those related to stress and anxiety. In support of the latter idea, we show here that bilateral HC damage abolishes the cortisol response to psychosocial stress. We collected salivary cortisol, heart rate, and affective responses to the Trier Social Stress Test (TSST) from 7 participants with bilateral HC lesions, 12 participants with damage outside the HC, and 28 healthy normal comparison participants matched to the HC participants on age and sex. HC participants showed elevated pre-stress cortisol, but no cortisol response to the TSST. Heart rate and affective responses in the HC group were similar to those of the comparison groups. Participants with brain damage outside the HC showed stress responses that were comparable to those of the healthy comparison group. These findings support the idea that the functions of the human HC extend beyond learning and memory, and suggest that the HC is necessary for producing the cortisol response to psychosocial stress.     

Differences in saliva collection location and disparities in baseline and diurnal rhythms of alpha-amylase: a preliminary note of caution

Identified in the early 1980s as a surrogate marker of the sympathetic nervous system component of the stress response, there has been renewed interest in measuring salivary alpha-amylase (sAA) to test biosocial models of stress vulnerability. This brief report presents studies that document that oral fluids from the parotid and submandibular gland areas had higher sAA values than did whole saliva specimens, and sAA values in whole saliva were higher than levels measured in oral fluids from the sublingual gland area. sAA in oral fluids from the parotid and submandibular gland areas showed the highest and more pronounced diurnal variation than levels in whole saliva, and sAA in sublingual saliva showed the lowest and shallowest diurnal variation. When this source of inherent variability in sAA activity levels is not controlled for by collecting oral fluids consistently from specific gland areas, the detection of individual differences, associations between sAA and “behavioral” variables, and intra-individual change in sAA levels may be compromised. Awareness, and management, of this ubiquitous source of measurement error in sAA are essential to ensure the success of future research on the correlates and concomitants of sAA levels, stress-related reactivity and recovery, and diurnal variation.     

Possible linkage between the ability to change the period (tau) of the prolactin and cortisol rhythms in women and breast cancer risk

The 24 h profiles of plasma hormone concentrations are rhythmic. The circadian period (τ) changes in development, with seasons, and in women with different stages of the menstrual cycle. It is known that the rhythms of prolactin and cortisol are sensitive to environmental time cues, such as changes in day length and phase; however, the importance of these changes is not yet understood. This study investigates whether there is a relation between the ability of a subject to respond to external cues that are associated with seasonal changes causing alteration of the rhythm's periods in cortisol and prolactin and the epidemiologically determined susceptibility to breast cancer. It is shown that the rhythmic output pattern of prolactin and cortisol in vivo is generated by more than one oscillator and structured by more than one rhythmic component. Each cohort of American women, classified on an epidemiologic basis as high risk (HR) or low risk (LR) to develop breast cancer, expresses different rhythmic output patterns of both variables, suggesting that the genetic background as defined by the risk state is related to differences in the circadian time structure, including the ability of the subject to change the rhythm's τ. The LR cohort exhibited a statistically significant change between seasons in the rhythm's τ of both the prolactin and cortisol patterns. In contrast, the HR cohort showed no change in the rhythm's τ between seasons for prolactin and cortisol patterns. These results show that in human beings, the presence of a circannual rhythm in the circadian time structure or the ability to adapt the circadian rhythmic pattern of these variables to external cues, such as seasons, is related to the partly genetically determined risk state to develop breast cancer and may be of importance for human health.     

Influence of CCK and bombesin on ACTH and cortisol secretion in the conscious dog

Bombesin and cholecystokinin (CCK) have a variety of similar actions. Previous investigations have demonstrated that IP injections of bombesin and CCK-33 increased corticosterone secretion in conscious, freely-moving, fed rats. In this study bombesin or CCk-8 was administered by continuous, intravenous infusion to conscious, awake, fasted, mongrel dogs. Following a 30-40 minute control infusion, a progressively-increasing, stepwise infusion of either bombesin (0.1, 1.0 and 2.0 micrograms/kg-hr) or CCK-8 (62.5, 125, and 250 ng/kg-hr) was administered. Each drug dose was infused for 40-45 minutes and blood samples were drawn at 20-22.5 minutes intervals. Bombesin caused significant, dose-dependent increases in plasma cortisol (286 +/- 39% of control) and plasma ACTH (176 +/- 33% of control). CCK-8 had no consistent effect on either cortisol or ACTH secretion. Whether the lack of effect of CCK-8 in dogs, as compared to rats, is due to species variations or to the differing experimental designs is unknown.     

Contact with attractive women affects the release of cortisol in men

Previous studies have shown that situations relevant for human mating can affect the levels of many hormones. This study focused on the hypothalamus-pituitary-adrenal axis by measuring salivary cortisol levels in 84 young men prior to and after a period of short social contact with a woman or man. Results showed that after contact with another man the cortisol levels of the participants declined according to the circadian release pattern of cortisol. However, cortisol levels in men declined less when they had contact with a woman. Furthermore, cortisol levels of men increased when they perceived the woman with whom they had contact as attractive. Our findings provide indirect evidence for the role of the hypothalamus-pituitary-adrenal axis in human courtship. During social contact with attractive women, moderate increases in cortisol levels may reflect apprehension over an opportunity for courtship.     

The function of a trypsin-like enzyme in the saliva of Euplectrus separatae larvae

Larvae of the gregarious ectoparasitoid, Euplectrus separatae, a species that parasitizes Pseudaletia separata, migrate from the dorsal to the ventral side of the host larva for pupation 7 days after parasitization. The parasitized host larvae die after the migration. The body mass of the parasitoid larvae increases while that of the host larva drastically decreases. Most of the tissue in the dead host larvae completely collapses. In this study, we examined the cause of host death and how the tissues collapse. Artificial removal of all parasitoid larvae before their migration on day 7 rescued the host larvae, but removal after parasitoid migration did not rescue the hosts. Tissues of the dead host larvae were completely liquefied. Injection of saliva from day 7 parasitoid larvae into host larvae killed the host larvae. High activity of a trypsin-like enzyme was detected in the saliva of day 7 parasitoids. Though phospholipase B and hyaluronidase were also detected in the saliva, commercial phospholipase B and hyaluronidase did not kill the hosts, whereas an injection of commercial trypsin was lethal. The trypsin-injected hosts showed the same tissue collapse as noted in parasitized and saliva-injected hosts. Leupeptin, a trypsin inhibitor, reduced mortality when injected into day 7 hosts (parasitoids were removal following migration). These observations suggest that the day 7 parasitoid larvae release saliva containing a trypsin-like enzyme to digest the host tissues following migration.     

母亲唾液中2种牙周致病菌对胎儿的影响

目的检测早产低体重儿（pretermlowbirthweight,PLBW）与正常体重儿（normalbirthweight,NBW）母亲的牙周临床状况及唾液中牙龈卟啉单胞菌（porphyromonasgingivalis,Pg）和直形弯曲菌（campylobacterrectus,Cr）检出率,探讨2种牙周病致病菌与PLBW发生的关系。方法选取北京四所医院产后1～1．5年的母亲99人,PLBW（试验组）母亲64人,NBW（对照组）母亲35人,收集母亲唾液样本,行全口牙周检查记录探诊深度（probingdepth,PD）、菌斑指数（plaqueindex,PLI）、出血指数（bleedingindex,BI）及临床附着丧失（clinicalattachmentloss,CAL）。用PCR检测唾液中的P岔和Cr;根据赡、Cr检出与否分为％、Cr阳性组、阴性组。结果（1）PLBW和NBW牙周临床指标及与Pg、cr水平：PLBW和NBW组CAL分别（0．06,0．62）mm（25％,75％）、（0．00,0．18）mm（25％,75％）,差异有统计学意义;PLBW组心检出58人的PD2．49±2.0．37;未检出5人的PD2．03±0．54,差异有统计学意义,％检出率为92．06％;NBW组唯检出28人的PD（2．47±0．35）mm,未检出7人的PD（1．91±0．35）mm,差异有统计学意义,赡检出率为80％。PLBW与NBW的瞻检出率差异无统计学意义。PLBW组cr检出55人检出率为87．30％,检出与未检出之间4项临床指标差异均无统计学意义。NBW组D检出率为91．4％,PLBW与NBW间D检出率差异无统计学意义。（2）P譬、Cr阳性组与阴性组间临床指标及与PLBW关系：Pg阳性组86人,阴性组12人,两组4项临床指标分别为PD（2．48±0．36）mm,（1．96±0．40）mm;PLI1．71±0．46,1．37±0．38;BI（1．82,3．02）（25％,75％）,（1．12,2．29）（25％,75％）;CAL（0．03,0．56）ITIm（25％,75％）,（0．00,0．11）mm（25％,75％）唯阳性组新生儿体重为（2482,78±833．85）g,阴性组为（3172．50±1190．1）g,差异均有统计学意义。cr阳性88人,D阳性11人,两组CAL值分别是（0．36±0．53）mm和（0．11±0．20）mm,差异有统计学意义,PD、PI、BI及新生儿体重差异均无统计学意义。结论珞、D在两组母亲唾液检出水平均高;Pg可能与PLBW发生有关;提示有必要进行孕前预防性牙周干预治疗。     

Acculturation, childhood trauma and the cortisol awakening response in Mexican-American adults

Exposure to chronic and traumatic stress has been associated with the dysregulation of crucial stress response systems. Acculturation has been associated with unique forms of chronic psychosocial stress. The purpose of this study was to examine the effects of exposure to early traumatic stress and acculturation on dysregulation of the cortisol awakening response (CAR) in Mexican-American adults. Salivary cortisol samples were collected at awakening and 30, 45, and 60 min thereafter, on two consecutive weekdays from 59 healthy Mexican-American adult males (26) and females (33), ages 18-38 years. Participants were assessed for level of acculturation and exposure to early trauma. Data were analyzed using a mixed effects regression model with repeated measures at four time points. Mixed effects regression results indicated a significant Early Trauma × Time interaction (p = .0029) and a significant Acculturation × Time interaction (p = .0015), after controlling for age and sex. Subsequent analyses of the interaction of Trauma × Acculturation × Time showed that more than minimal exposure to either risk factor was associated with attenuation of the awakening cortisol response (p = .0002). Higher levels of acculturation with greater Anglo-orientation were associated with attenuation of the CAR in Mexican-American adults. Both moderate and higher levels of exposure to early trauma were associated with an attenuated CAR. However, greater exposure to both risk factors was only incrementally worse than exposure to either one.     

Enrichment and identification of glycoproteins in human saliva using lectin magnetic bead arrays

Aberrant glycosylation of proteins is a hallmark of tumorigenesis and could provide diagnostic value in cancer detection. Human saliva is an ideal source of glycoproteins due to the relatively high proportion of glycosylated proteins in the salivary proteome. Moreover, saliva collection is noninvasive and technically straightforward, and the sample collection and storage is relatively easy. Although differential glycosylation of proteins can be indicative of disease states, identification of differential glycosylation from clinical samples is not trivial. To facilitate salivary glycoprotein biomarker discovery, we optimized a method for differential glycoprotein enrichment from human saliva based on lectin magnetic bead arrays (saLeMBA). Selected lectins from distinct reactivity groups were used in the saLeMBA platform to enrich salivary glycoproteins from healthy volunteer saliva. The technical reproducibility of saLeMBA was analyzed with liquid chromatography–tandem mass spectrometry (LC–MS/MS) to identify the glycosylated proteins enriched by each lectin. Our saLeMBA platform enabled robust glycoprotein enrichment in a glycoprotein- and lectin-specific manner consistent with known protein-specific glycan profiles. We demonstrated that saLeMBA is a reliable method to enrich and detect glycoproteins present in human saliva.     

Oxidative stress and cholinesterase inhibition in saliva and plasma of rats following subchronic exposure to malathion

The aim of this study was to examine whether malathion, a commonly used organophosphate (OP), might induce oxidative stress and cholinesterase (ChE) depression in saliva and plasma in rats following subchronic exposure mimicking human exposure. Malathion was administered orally at doses of 100, 500 and 1500 ppm for 4 weeks. Oxidative stress was determined by measuring the malondialdehyde concentration, the end product of lipid peroxidation, and assessing total antioxidant power. Four weeks oral administration of malathion at doses of 100 ppm, 500 ppm and 1500 ppm depressed plasma ChE activity to 45% (P<0.01), 48% (P<0.01) and 41% (P<0.01) of control, respectively. Malathion at doses of 100 ppm, 500 ppm and 1500 ppm depressed saliva ChE activity to 73% (P<0.01), 75% (P<0.01) and 78% (P<0.01) of control, respectively. Malathion at doses of 100 ppm, 500 ppm and 1500 ppm increased plasma antioxidant power by 33% (P<0.01), 59% (P<0.01) and 118% (P<0.01) of control, respectively. Malathion did not change saliva antioxidant power. Malathion at doses of 100 ppm, 500 ppm and 1500 ppm increased plasma thiobarbituric acid reactive substances (TBARS) by 61% (P<0.01), 69% (P<0.01) and 63% (P<0.01) of control, respectively. Malathion at doses of 500 ppm and 1500 ppm increased saliva TBARS by 19% (P<0.01) and 22% (P<0.01) of control, respectively. Malathion (100 ppm) did not change saliva TBARS level. We concluded that in OP subchronic exposure, depression of ChE is accompanied by induction of oxidative stress that might be beneficial in monitoring OP toxicity.     

Methods of collection for salivary cortisol measurement in dogs

Salivary cortisol has been increasingly used as a measure of stress response in studies of welfare, reaction to stress and human-animal interactions in dogs and other species. While it can be a very useful measure, there are a number of saliva collection issues made evident through studies in the human and animal fields which have not been investigated in the canine species. Collection materials and the volume of saliva that is collected; the use of salivary stimulants; and the effect of food contamination can all dramatically impact cortisol measurement, leading to spurious results. In order to further examine the limitations of the collection method and the effects of collection material and salivary stimulant on salivary cortisol levels, a series of clinical, in vitro and in vivo studies were performed. It was found that there is a large amount of inter- and intra-individual variation in salivary cortisol measurement. Beef flavoring of collection materials leads to unpredictable variability in salivary cortisol concentration. Using salivary stimulants such as citric acid also has the potential to affect cortisol concentration measurement in saliva. Hydrocellulose appears to be a useful collection material for salivary cortisol determination. Recommendations for collection materials and use of salivary stimulants are presented.     

Effects of training for two ball games on the saliva response of adrenocortical hormones to exercise in elite sportswomen

In a group of 20 elite female handball or volleyball players, an evaluation was made of the response to a 4-month training period of cortisol, androstenedione and dehydroepiandrosterone in the saliva. At the beginning of the training programme (W₁) and at the 7th (W₇) and 16th weeks (W₁₆), hormone concentrations were measured on awakening (8 a.m.; resting samples), and also before (t ₀) and at the end (t ₁₂₀) of a 2-h exercise session (simulated match) which took place at 6 p.m. The training programme increased the concentrations of adrenal androgens in the saliva at rest (P<0.05) for both groups of subjects, with no change being noted in cortisol concentrations. In contrast, amongst the volleyball players, stressful stimuli produced by an increase in the amount of training did not affect adrenocortical metabolism during exercise. Indeed, a simulated volleyball match resulted in a decrease in salivary cortisol (P<0.05) and androgen concentrations (P<0.05) without regard for the week chosen for the test investigations. In contrast, a simulated handball match caused an increase in cortisol concentrations at W₁ only (P<0.05), with no change in the adrenal androgens concentration in any week of the training programme. The regulators of these hormones during a period of exercise and during the course of a training programme would seem to be different. Accepted: 30 October 1997     

A note on the kinetics of enzyme action: A decomposition that highlights thermodynamic effects

Michaelis and Menten’s mechanism for enzymatic catalysis is remarkable both in its simplicity and its wide applicability. The extension for reversible processes, as done by Haldane, makes it even more relevant as most enzymes catalyze reactions that are reversible in nature and carry in vivo flux in both directions. Here, we decompose the reversible Michaelis–Menten equation into three terms, each with a clear physical meaning: catalytic capacity, substrate saturation and thermodynamic driving force. This decomposition facilitates a better understanding of enzyme kinetics and highlights the relationship between thermodynamics and kinetics, a relationship which is often neglected. We further demonstrate how our separable rate law can be understood from different points of view, shedding light on factors shaping enzyme catalysis.     

Circadian rhythm characteristics of serum cortisol and dehydroepiandrosterone sulfate in healthy Chinese men aged 30 to 60 years. A cross-sectional study

The relation of adrenal function and aging has been the subject of intense interest in recent years. The circadian variations of plasma cortisol have been described in Caucasians, but little information is available on such hormone variations among the Chinese population, especially its changes with age. This study was, therefore, designed to examine the effects of age on the circadian variations of serum cortisol, dehydroepiandrosterone sulfate (DHEAS) and the molar ratio of cortisol/DHEAS in Chinese men, stratified by 10-year age-groups (i.e. men in their 30-60s, aged from 31 to 63 years old). Circadian variations of serum cortisol and DHEAS were documented at 2-h intervals from 8:00 to 22:00 and hourly from 22:00 to 8:00 in 26 healthy Chinese men. We found that the serum levels of both hormones showed a statistically significant circadian rhythmicity in all age-groups. The circadian pattern of serum cortisol was characterized by peaks (04:00-06:00) and troughs (18:00-24:00) occurring approximately 2h earlier than those usually reported in Caucasians. Aging did not significantly influence serum cortisol concentrations, but serum DHEAS levels declined significantly with age: subjects in their 60s had significantly lower levels, and their cortisol/DHEAS molar ratios were significantly higher than those in the younger age-groups.     

Transformation of ethyl alcohol to ethyl nitrite in acidified saliva: possibility of its occurrence in the stomach

After taking alcoholic beverages, the ethanol is mixed with saliva and then gastric juice. As pH of gastric juice is around 2, the ethanol might be transformed to ethyl nitrite in the stomach by reacting with salivary nitrite. In this study, reactions between ethanol and nitrite in acidified saliva were investigated. The result indicates that nitrite in acidified saliva reacted with ethanol producing ethyl nitrite. It is discussed that ethyl nitrite might be formed in the stomach after drinking alcoholic beverages and that the ethyl nitrite might function as a donor of NO in intestinal and gastric tissues.     

Changes in cortisol release and heart rate and heart rate variability during the initial training of 3-year-old sport horses

Based on cortisol release, a variety of situations to which domestic horses are exposed have been classified as stressors but studies on the stress during equestrian training are limited. In the present study, Warmblood stallions (n = 9) and mares (n = 7) were followed through a 9 respective 12-week initial training program in order to determine potentially stressful training steps. Salivary cortisol concentrations, beat-to-beat (RR) interval and heart rate variability (HRV) were determined. The HRV variables standard deviation of the RR interval (SDRR), RMSSD (root mean square of successive RR differences) and the geometric means standard deviation 1 (SD1) and 2 (SD2) were calculated. Nearly each training unit was associated with an increase in salivary cortisol concentrations (p < 0.01). Cortisol release varied between training units and occasionally was more pronounced in mares than in stallions (p < 0.05). The RR interval decreased slightly in response to lunging before mounting of the rider. A pronounced decrease occurred when the rider was mounting, but before the horse showed physical activity (p < 0.001). The HRV variables SDRR, RMSSD and SD1 decreased in response to training and lowest values were reached during mounting of a rider (p < 0.001). Thereafter RR interval and HRV variables increased again. In contrast, SD2 increased with the beginning of lunging (p < 0.05) and no changes in response to mounting were detectable. In conclusion, initial training is a stressor for horses. The most pronounced reaction occurred in response to mounting by a rider, a situation resembling a potentially lethal threat under natural conditions.     

Proteomic identification of elevated saliva kallikrein levels in the mdx-4cv mouse model of Duchenne muscular dystrophy

Dystrophinopathies are multi-system disorders that affect the skeletal musculature, the cardio-respiratory system and the central nervous system. The systematic screening of suitable biofluids for released or altered proteins promises new insights into the highly complex pathophysiology of X-linked muscular dystrophy. However, standard detection approaches using antibody-based assays often fail to reproducibly detect low-abundance protein isoforms in dilute biological fluids. In contrast, mass spectrometric screening approaches enable the proteome-wide identification of minor protein changes in biofluids. This report describes the findings from the comparative proteomic analysis of whole saliva samples from wild type versus the established mdx-4cv mouse model of highly progressive muscular dystrophy, focusing on the kallikrein protein family. Kallikrein-1 (Klk1) and 13 Klk1-related peptidases were identified in saliva and serum from normal mice. Comparative proteomics revealed elevated saliva levels of the Klk1-related peptidases Klk1-b1, Klk1-b5 and Klk-b22, as well as an increased Klk-1 concentration, which agrees with higher Klk-1 levels in serum from mdx-4cv mice. This indicates altered cellular signaling, extracellular matrix remodeling and an altered immune response in the mdx-4cv mouse, and establishes liquid biopsy procedures as suitable bioanalytical tools for the systematic survey of complex pathobiochemical changes in animal models of muscular dystrophy.     

The effects of a chlorhexidine mouthrinse on culturable microorganisms of the tongue and saliva

A randomized and cross-over design clinical study with normal adult volunteers examined the effects of a 0.12% chlorhexidine (CHX) mouthrinse and a control rinse on a range of microflora including those implicated in halitosis on the tongue and saliva. Saliva and tongue scrapings were collected from 13 subjects prior to treatments with additional samples collected at 3 h post-treatment cultured on media to enumerate anaerobic, Gram-positive and Gram-negative bacteria, odorigenic bacteria producing hydrogen sulphide (H2S) and oral bacteria with proteolytic activity. In comparison to the control, rinsing with CHX demonstrated statistically significant reductions that ranged from 81-90% for tongue microflora with a 89-95% decrease noted on salivary flora (p<0.05). The effects of CHX on anaerobic, Gram-positive and Gram-negative bacteria are in accord with those noted on odorigenic bacteria producing H2S or proteolytic activity. These results help to explain previous clinical results on the malodor efficacy of CHX and highlight microbiological approaches for bacteria implicated in malodor.