Salivary Lysozyme Research Articles

Nutritional influences on enzyme activities in saliva of Asian and African elephants

Asian and African elephants show morphological adaptations to their ecological niche including the oral cavity. Variety and preferences of forage plants differ between both herbivorous elephant species. Diet can affect salivary enzymes. Asian elephants were shown to have a higher salivary amylase activity than African elephants. Species-specific differences were presumed to be influenced by feeding during collection procedure. This study aimed to determine the influence of feeding on enzyme activities in saliva of both elephant species to differentiate from species-specific effects. Additionally, season and housing conditions on salivary enzyme activities in non-fed elephants of both species were investigated. Salivary amylase (sAA), lysozyme (sLYS) and peroxidase (sPOD) activity were measured photometrically or fluorometrically. Results of this study reinforce previous observations of higher basic sAA activity in Asian elephants compared to African elephants. Salivary LYS and sPOD activity showed neither species-specific nor housing-specific differences. Independent from season, most elephants of both species revealed a lack of or low sPOD activity. Feeding caused a temporary decrease of sAA, sLYS and sPOD activity in both elephant species kept in four of eight tested zoos. Furthermore, sAA activity in Asian elephants was higher and sLYS activity lower in Spring than in Autumn. This study summarizes that sAA and sLYS are components of Asian and African elephant saliva in an active conformation in contrast to sPOD. Diet varying between season and zoos might influence sAA and sLYS activities primarily in Asian elephants but temporary low effects suggest sufficient buffer capacity of elephant saliva of both species.

Rapid and selective aptamer-based fluorescence detection of salivary lysozyme using plasmonic metal-enhanced fluorescence of ZnSSe alloyed quantum dots-gold nanoparticle nanohybrid

We report on the construction of an aptamer (Apt)-based fluorescent biosensor platform for salivary lysozyme using plasmonic metal-enhanced fluorescence of ZnSSe alloyed quantum dots (QDs)-gold nanoparticle (AuNP) nanohybrid. Salivary lysozyme was used as a model analyte to develop the Apt fluorescence biosensor assay for real-life detection of human saliva. To construct the biosensor, thiolated anti-salivary lysozyme DNA Apt was immobilised on the L-cysteine-ZnSSe QDs surface to form a functionalised Apt-ZnSSe-QDs. Thereafter, citrate-AuNPs was bonded to the Apt-ZnSSe-QDs to form an Apt-ZnSSe-QDs-AuNP fluorescence biosensor probe. Using affinity-based biomolecular binding interaction between the Apt and target lysozyme, the binding effect induced localised surface plasmon resonance (LSPR) signal from AuNPs to amplify the fluorescence signal of the QDs upon lysozyme recognition. Probing the LSPR signal amplifying effects of different plasmonic NPs, our analysis showed that spherical citrate-AuNPs of size 6 nm was most effective in amplifying the fluorescence signal of the biosensor probe. Under optimum experimental reaction conditions, salivary lysozyme was detected quantitatively within 5 min with a detection limit of 22.9 μg/mL and was highly selective against other tested enzymes. Application of the Apt-ZnSSe QDs-AuNP biosensor probe to detect real human saliva samples was successfully achieved.

Combined Chair-Based Exercises Improve Functional Fitness, Mental Well-Being, Salivary Steroid Balance, and Anti-microbial Activity in Pre-frail Older Women.

IntroductionRegular exercise has long been shown to positively impact the immune system responsiveness and improve mental well-being (MWB). However, the putative links between biomarkers of mental health and immune efficiency in exercising subjects have been scarcely investigated. The aim of this study was to verify the effect of a 14-week combined chair-based exercise program (CEP) on salivary steroid hormones and anti-microbial proteins, functional fitness, and MWB indexes in pre-frail older women.MethodsThe participant women (82.8 4.6 years old; n = 32) were randomly divided into the exercising group (CEP, n = 17) and the non-exercising control group (CG, n = 15). The pre/post assessment included: (1) salivary anti-microbial proteins lysozyme; (Lys) and immunoglobulin-A (IgA); (2) salivary steroid hormones of testosterone (TT) and cortisol (COR); (3) functional fitness (gait speed, hand grip strength, and static balance); (4) MWB questionnaires (happiness, depression state, satisfaction with life, and stress).ResultsSignificant differences with large Cohen’s (d) effect sizes were found on increased salivary TT (p < 0.05; d = 0.60) after exercise intervention. The results revealed a decrease in IgA levels after CEP (p < 0.01, d = 0.30). The increase in subjective happiness levels (p < 0.05, d = 0.30) and decrease of stress perception (p < 0.01, d = 2.60) and depressive state (p < 0.05, d = 0.30) were found after intervention in the CEP group. Robust statistical differences in gait speed (p < 0.05; d = 0.60) and balance tests (p < 0.05; d = 0.80) were also found in the CEP group. In control, COR increased moderately (p < 0.05; d = 0.65) while no changes were found for the other indicators. Correlation analyses showed inter-dependence between pre–post variations of MWB, biochemical indexes, and fitness function (e.g., COR inverse correlation with hand grip strength and balance tests).ConclusionThe CEP program was able to improve functional-fitness performance, decrease feelings of stress, and increase happiness. The CEP also induced clinically relevant hormonal and immune responses, which suggests that chair exercises that combine muscular strength, balance, and gait speed training are promising interventions to improve physical and mental health of older pre-frail adults.

Immunodiagnostics, immunotherapy and immunoprophylaxis in the clinic of otolaryngology. Message 2. Indicators of local immunity in assessment of clinical and laboratory status of patients in inflammatory ENT diseases (Analytical summary)

Topicality: It is considered proven that the state of the immune system determines the development and course of many pathological processes, which are based on inflammation. At the same time, taking into account modern trends in assessing the immunological status, a mandatory component is to determine the state of local immunity in the upper respiratory tract. Aim: to conduct an integral analysis of data to determine the state of local immunity in inflammatory diseases of the upper respiratory tract (URT) to select the optimal necessary informative indicators. Materials and methods: A generalization of the data of immunological studies of 168 people with inflammatory pathology of the upper respiratory tract (VDS) was carried out, of whom 42 had chronic tonsillitis (CT), 36 – chronic rhinitis (HR – catarrhal form), 40 – chronic pharyngitis (CP), 28 – secretory mean otitis media (CO), 22 people made up the control group (C) of practically healthy donors. The age of the surveyed was from 14 to 60 years, the duration of the diseases was from 1 to 5 years, female patients predominated (~ 60%). The examination was carried out at the stage of clinical remission. The material for the research was mixed saliva, in which the content of immunolobulins and citrokines was investigated by the ELISA method: enzyme immunoassay, the following factors of immunity and inflammation in the CGS were determined: - secretory and monomeric form of immunoglobulin A (Hema Medica, RF); - immunoglobulins of classes G, M (Hema Medica, RF); - interleukins – 1β; 4; 8; 10 (Cytokine, RF); - interferons α and γ (Cytokine, RF); - pro- and defensins (Nucalt bioteknology, Netherlands); - macrophage inflammatory protein-Mip-1b (Assauro, USA); - salivary lysozyme (Diagnostik Nord, Germany). Statistical processing was carried out using the methods of Student’s t-criterion and angular transformation “φ” according to Fisher. Results: Integral of all methods for determining various types of immunity in diseases of the upper respiratory tract with a chronic course is the insufficiency of factors of both innate and acquired immunity, especially during the period of clinical remission of the disease. A special role in protective reactions belongs to the humoral component, in which the most stable indicator is the level of class A secretory immunoglobulin; it has been shown that almost all the studied components of the immune response had abnormalities in the presence of a chronic inflammatory process in the upper respiratory tract and they can be presented as a line the degree of decrease in the frequency of deviations: secretory IgA, IgG., pro-inflammatory cytokines (IL1β and IL-8), γ-interferon, lysozyme, defensin-β, Mip-1b. Conclusion: The developed criteria and approaches to assessing systemic immunity in patients with inflammatory diseases of the upper respiratory tract can be used in examining patients and assessing the effectiveness of therapy.

Human β-defensin 2, секреторный иммуноглобулин А и лизоцим в разных биологических средах у детей с атопическим дерматитом

Objective. To evaluate the levels of human beta-defensin 2 (HBD2), secretory immunoglobulin A (sIgA), and lysozyme in various biological specimens from children with atopic dermatitis and to assess their correlation with the severity of clinical manifestations. Patients and methods. This study included 96 children with atopic dermatitis and 31 healthy controls. We measured serum HBD2, salivary and fecal sIgA, and salivary and fecal lysozyme using special ELISA Kits (CUSABIO, Korea). Results. In children with atopic dermatitis, median levels of serum HBD2 (544 pg/mL [range: 288; 719]), salivary sIgA (120 ng/mL [range: 89.5; 214.5]), fecal sIgA (244 ng/mL [range: 196; 367]), and salivary lysozyme (0.59 ng/ml [range: 0.25; 1.82]) were significantly lower than those in controls (1136 pg/mL [range: 741; 1511], 313 ng/mL [range: 208.5; 356.0], 1582 ng/mL [range: 855; 1710], and 1.65 ng/mL [range: 0.18; 2.58], respectively). We found a strong negative correlation between the SCORAD index and serum HBD2 level (ρ = -0.71, p = 0.005), as well as fecal sIgA (ρ = -0.74, p = 0.032). There was also a negative correlation between the SCORAD index and salivary sIgA (ρ = -0.64, p = 0.032) and salivary lysozyme (ρ = -0.58, p = 0.046). Conclusion. Our findings support the hypothesis that children with atopic dermatitis have reduced production of humoral factors of natural resistance and immunity. The level of HBD2, sIgA, and lysozyme can serve as a biomarker of disease severity. Key words: atopic dermatitis, children, defensins, lysozyme, secretory immunoglobulin A

Relationship of oral candidiasis with salivary lysozyme and lactoferrin in HIV-positive patients: a systematic review

Relationship of oral candidiasis with salivary lysozyme and lactoferrin in HIV-positive patients: a systematic review

Dental caries-related primary hypertension in children and adolescents: Cross-sectional study.

Hypertension is a growing clinical problem in pediatric population. Also, the cause of hypertension is usually unknown and it may result from systemic inflammation related to tooth decay. To estimate the potential association in cross-sectional study between tooth decay and hypertension in children and adolescents. Study group-65 children diagnosed with primary arterial hypertension; control subjects-44 normotensive children. Blood pressure, dental examination, measurement of salivary cortisol, alpha-amylase, secretory IgA, and lysozyme concentrations were performed in all of the children. Hyper- and normotensive children had similar peripheral blood morphology and serum biochemical parameters, except of uric acid concentration, which was significantly higher in the study group (p=.047). Salivary evening concentrations of cortisol and alpha-amylase were significantly higher in hypertensive children (p=.002 and p=.004, respectively). Although 24-hr systolic blood pressure (SBP), including daytime and nighttime SBP, correlated with "decay," "microalbuminuria," "BMI," and "glomerular filtration rate" (r>.75, r>.7, r<.68, and r<.43, respectively), in multivariate analysis only "decay" was associated with hypertension both in children and in adolescents (p<.0001). Tooth decay in children/adolescents might be regarded as a potent trigger factor of hypertension in individuals in whom all other causes of secondary arterial hypertension have been excluded.

Effects of exercise in hot and humid conditions and bovine colostrum on salivary immune markers.

The purpose of this study was to examine the effects of exercise in a hot and humid environment on salivary lactoferrin and lysozyme. A secondary aim was to quantify the effects of 14-day bovine colostrum (BC) supplementation on salivary lactoferrin and lysozyme at rest and following exercise in hot and humid conditions. Using a randomized, double-blind, and counterbalanced design, ten males (20±2 years, VO2max 55.8±3.7mLkg-1min-1, 11.8±2.7% body fat) ran for 46±7.7minat 95% of ventilatory threshold in a 40°C and 50% RH environment after 14-days of supplementation with either BC or placebo. Saliva was collected pre, post, 1-h, and 4-h post exercise, and was analyzed for lactoferrin and lysozyme using ELISA. There was an immediate increase in the concentration and secretion rate of lactoferrin and lysozyme (p<0.05) with exercise, but BC had no effect (p>0.05). Saliva flow rate was not different between conditions [(PLA: pre: 0.54±0.3, post: 0.44±0.3, 1-h: 0.67±0.3, 4-h: 1.0±0.4mLmin-1); (BC: pre: 0.58±0.2, post: 0.37±0.1, 1-h: 0.63±0.2, 4-h: 0.83±0.4mLmin-1)]. There were no differences in thermoregulatory markers (core temperature or physiological strain index) between BC and placebo trials. Interestingly, exercise-induced heat stress did not impair mucosal immune parameters, instead participants showed a transient increase in salivary lactoferrin and lysozyme. Further, 14-day BC supplementation had no effect on mucosal immunity at any time point.

Might Salivary Lysozyme Be An Indicator Of Prolonged Intense Training Load In Athletes? A Preliminary Study In Adolescent Male Gymnasts

Lysozyme is one of the salivary antimicrobial proteins (AMP) which act as a defence at the mucosal surface. While in adult athletes, a decrease in salivary lysozyme (SLys) levels has been reported after prolonged intense training, to our knowledge, no studies have been conducted to study the relationship between SLys levels and long-term physical activity in children or teenagers.The aim of this preliminary study was mainly to evaluate in a group of adolescent male gymnasts undergoing prolonged intense training load whether - in accordance with studies in adult athletes - there will also be a decrease in SLys and if so, whether this phenomenon will be so common that we detect it in a small group of study participants.Twelve adolescent male gymnasts aged from 15.0 ± 1.6 years of national or international performance level were recruited to participate in this study. All participants of the study had their sample of saliva taken: I. Period) after the transitional period (rest), i.e. just before the beginning of the preparatory training period. II. Period) immediately after the end of the preparatory training period that was focused on maximal strength and power development. Preparatory training period lasting 6 weeks consisted of nine 2.5 hour training units (on average) over 6 days in every week. At the same time, three times per week two-a-day training sessions were incorporated. Intensity of the physical exercise was not determined.We found a significant decrease in SLys levels after the preparatory training period (termed as II. period) compared to its level just before the start of the training (i.e. after the rest, termed as I. period).The results of this preliminary study suggest that SLys measurements may be an indicator of prolonged training load in adolescent athletes. Although the intensity of the training load has not been determined, the national and international performance level of the gymnasts enrolled in the study allows at least a rough estimate of its level. However, with respect to some limitations of our study, larger studies on male and female adolescent athletes, applying relevant training load with monitoring of variables such as specific sports performance, physical fitness, nutrition, sleep quality, social and psychological factors, are desirable.

Therapy of recurrent herpetic stomatitis in childrens with bronchial asthma

The work presents the results of a study of the effectiveness of the use of an immunomodulating agent for the treatment of recurrent herpetic stomatitis in children with bronchial asthma. The therapy of recurrent herpetic stomatitis was carried out in 67 children aged 1 to 16 years with bronchial asthma according to the scheme proposed by the authors using an immunomodulator. The immune status of patients before and after the therapy was assessed using the following indicators: the content of immunoglobulin (SIgA) in saliva, salivary lysozyme, an integrated indicator, the balance coefficient (Ksb). The results of the study showed that after the therapy, the incidence rates of recurrent herpetic stomatitis improved, the state of local immunity approached the group of healthy children. It is concluded that the proposed therapy for recurrent herpetic stomatitis in children with bronchial asthma gave a positive result and can be recommended for the treatment of this category of patients.

Clinical and laboratory assessment of the application efficacy of antibacterial combinations in the treatment of chronic generalized periodontitis

Aim. To determine the efficacy of the combination of metronidazole and co-trimoxazole in the treatment of chronic generalized periodontitis.&#x0D; Methods. The studies included patients with slight and moderate chronic generalized periodontitis without concomitant pathology. Patients were divided into two groups with similar age and severity of periodontal diseases. In the first group, metronidazole was administered into the periodontal pocket; in the second group co-trimoxazole and metronidazole combination was used. Oral fluid testing was used to detect ureolytic and glycolytic activity, malondialdehyde (MDA) content, conjugated dienes, catalase activity, Salivary Secretory Immunoglobulin (SIgA) and Lysozyme. Changes in periodontal disease were assessed before and after treatment. The duration of treatment was 12 days.&#x0D; Results. Co-trimoxazole and metronidazole combination more effectively stimulated ureolytic activity and reduced glycolytic effect. It had a more effective antioxidant effect that manifested in: a statistically significant decrease in malondialdehyde levels by 32.8% (p 0.01) and by 36.6% (p 0.01) in patients with slight and moderate periodontitis, respectively; decrease in diene conjugates by 25.0% (p 0.001) and 37.7% (p 0.001); and increased catalase activity 2.6 and 2.9 times (p 0.001). After treatment, it was revealed an increase in lysozyme activity by 11.3% and 17.6% (р 0,001) in patients with slight and moderate periodontitis, respectively, and increase secretory IgA levels by 37.4% (p 0.001) and 53.2% (p 0.001). In the first group, the amount of SIgA in the oral fluid increases by 35.8% (p 0.001), and by 45.6% (p 0.001) compared to the value before treatment in patients with slight and moderate periodontitis, and in the second group by 37.4% (p 0.001) and 53.2% (p 0.001), respectively.&#x0D; Conclusion. The combined treatment with co-trimoxazole + metronidazole leads to the positive changes in the studied parameters causes rapid disappearance of the symptoms characteristic of chronic generalized periodontitis; this 2-drug combination is recommended to be included in the complex therapeutic measures for inflammatory periodontal diseases.

Selected Physicochemical Properties of Saliva in Menopausal Women-A Pilot Study.

The aim of this study was to estimate differences in selected physicochemical properties of saliva between menopausal and premenopausal women. Methods: The study population consisted of 9 menopausal women and 15 women of the control group. Laboratory tests included the determination of pH saliva, salivary flow rate, and concentrations of lactoferrin, lysozyme, immunoglobulin A, and ionized calcium. Results: Among menopausal women, statistically significant differences were observed in values of salivary flow rate and lysozyme and ionized calcium concentrations; however, no statistically significant differences for pH and concentrations of lactoferrin and immunoglobulin A were found. The salivary flow rate in the study group was significantly lower compared to that in premenopausal women. In relation to lysozyme, statistically significant differences were found between control group and menopausal women to the disadvantage of the latter. However, the concentration of ionized calcium in the saliva of menopausal women was distinctly higher than in the saliva of the control group. Conclusion: The saliva of menopausal women appeared significantly different from that of the control group. Differences in physicochemical parameters such as salivary flow rate and lysozyme and ionized calcium concentrations were observed. These differences in saliva properties observed in menopause can potentially affect the oral environment of women in this particular period, possibly increasing the risk of some pathological changes in the oral cavity and consequently indicating the need to take special care of this group of female patients in order to help them maintain proper oral health. Dentists and gynecologists should be aware of the problems associated with menopause and need to provide these women complete health care, including dental care as an integral part.

Effects of Carbohydrate Mouth Rinsing on Salivary Lysozyme, Mood States and Running Performance Among Recreational Runners.

IntroductionCarbohydrate (CHO) mouth rinsing can enhance sports performance through a central action mediated by receptors in the mouth. This study examined the effect of a CHO mouth rinse on salivary lysozyme concentrations, mood states and running performances.MethodsTen males recreational runners were randomised to three running trials with a 1 week recovery period between the trials. Each trial involved running at 75% maximum heart rate (HRmax) for 1 h, followed by a 15 min time trial. The participants used a CHO mouth rinse, placebo (PLA) solution or control (CON, no solution) every 15 min during the exercise. Heart rate (HR), rating of perceived exertion (RPE) and mood states were recorded pre-, during and post-exercise. Saliva samples were collected pre-, post- and 1 h post-exercise.ResultsThere was no significant interaction and time effect (P > 0.05) on the salivary lysozyme concentration and running performance, but it was significant (P < 0.05) for HR and RPE (increase in all trials). However, there was no significant difference (P > 0.05) in salivary lysozyme concentrations, running performances, HR values or RPE between the trials. Mood states were not significantly different (P > 0.05) between the trials, but one of the mood sub-scales showed a significant (P < 0.001) time effect (increase fatigue in all trials).ConclusionCHO mouth rinsing did not affect physiological parameters, salivary lysozyme concentrations, mood states or running performance among recreational runners.

Risk factor of secretory immunoglobulin A and salivary lysozyme level in children aged under 3 years to severe early childhood caries

Severe Early Childhood Caries (SECC) is a significant dental health problem in various countries. The caries pathology process is affected by saliva status. An important antimicrobial component in saliva is sIgA preventing the attachment of bacteria to enamel and lysozyme capability to act as an antibacterial by hydrolyzing bacterial polysaccharide walls. The aim of the study is to compare the sIgAlevel and salivary lysozyme between children with caries free and suffering from SECC, aged under 3 years. This study is an observational analytic with cross sectional design. Saliva samples were collected unstimulated from 68 children. Examination of sIgA level and salivary lysozyme used ELISA sandwich technique. Data were analysed using Mann-Whitney and Kruskal Wallis, while correlation test used Spearman. The results showed,there was no significant differencein sIgAlevel between caries-free children and SECC children (p > 0.05). It has been found that in caries-free children have a higher mean lysozyme level than SECC children. It has been found a medium correlation between deft and lysozyme level (r = -0.492). In conclusion, lysozyme level was a risk factor for the incidence of SECC in children aged under 3 years. The higher deft status, the lower lysozyme level of the child.

Evaluation of lysozyme concentration and S.mutans colonies on children with early childhood caries and caries free after using 0.1% lysozyme toothpaste

The prevalence of Early Childhood Caries (ECC) in various countries is quite high, even at the age of 2 years the incidence is quite apprehensive. One simple and effective ECC alternative prevention is tooth brushing by using lysozyme toothpaste. Lysozyme can kill S.mutans bacteria by hydrolizing bacterial cell walls. This study aims to compare the salivary lysozyme concentration and the number of S.mutans colonies between pre-treatment and after 1 month tooth brushing by using 0.1% lysozyme toothpaste. This research is single masked clinical trial. The sample size of this study is 73 under 3 years old children, consisting of 31 caries-free children and 42 ECC children. The research subjects are obtained from Posyandu in Medan Selayang and Medan Johor District by using purposive sampling. Criteria for ECC is based on the American Association of Pediatric Dentistry (AAPD). Data analysis use Mann-Whitney and Wilcoxon tests because the data are not normally distributed, p < 0.05. The result shows that after tooth brushing by using lysozyme toothpaste, reduction of lysozyme concentration was detected on both groups of caries-free and ECC children compared to the state before tooth brushing, but it is not statistically significant (p > 0.05). Similiarly, reduction in number of S.mutans was detected after tooth brushing by using lysozyme toothpaste, although it is not statistically significant (p > 0.05). As conclusion, lysozyme toothpaste can reduce the growth of S.mutans bacteria, but is only inhibitory or bacteriostatic. Therefore, lysozyme toothpaste can be used as an alternative of caries prevention for early childhood.

Salivary immunity and lower respiratory tract infections in non-elite marathon runners.

RationaleRespiratory infections are common after strenuous exercise, when salivary immunity may be altered. We aim to investigate changes in salivary immunity after a marathon and its relationship with lower respiratory tract infections (LRTI) in healthy non-elite marathon runners.MethodsForty seven healthy marathon runners (28 males and 19 females) who completed the 42.195 km of the 2016 Barcelona marathon were studied. Saliva and blood samples were collected the day before the marathon and two days after the end of the race. Salivary IgA, antimicrobial proteins (lactoferrin, lysozyme) and chemokines (Groα, Groβ, MCP-1) were determined using ELISA kits in saliva supernatant. Blood biochemistry and haemogram were analyzed in all participants. The presence of LRTI was considered in those runners who reported infectious lower respiratory tract symptoms during a minimum of 3 consecutive days in the 2 weeks after the race.ResultsEight participants (17%) presented a LRTI during the 2 weeks of follow-up. Higher lysozyme levels were detected after the race in runners with LRTI when compared with those without infection. A decrease in salivary lysozyme, Groα and Groβ levels after the race were observed in those runners who did not develop a LRTI when compared to basal levels. Salivary Groα levels correlated with basophil blood counts, and salivary lysozyme levels correlated with leukocyte blood counts.ConclusionsLRTI are common after a marathon race in non-elite healthy runners. Changes in salivary antimicrobial proteins and chemokines are related to the presence of LRTI and correlate with systemic defense cells, which suggest an important role of salivary immunity in the development of LRTI in non-elite marathon runners.

Effect of storage on long-term stability of salivary α-amylase, lysozyme, lactate dehydrogenase, calcium and phosphorus in dogs

Research or diagnostic conditions may require the storage of salivary samples for long periods before analysis is processed. The aim of this study was to evaluate the stability of canine salivary α-amylase, lysozyme, lactate dehydrogenase (LDH), and total calcium and phosphorus after storage for 1, 3 and 6 months at −20°C. Available saliva samples were 75 immediately after collection (T0) and 46 at 1 month (T1), 31 at 3 months (T3) and 18 at 6 months (T6) of storage, according to the number of aliquots collected from each saliva sample. Compared to T0, LDH declined by 92.3% after 1 month of storage (P<0.001), whereas lysozyme concentration significantly decreased as storage time increased (−29.3%, −43.4% and −59.1% at T1, T3, and T6 respectively, P<0.001). Amylase maintained basically the same concentration for the entire experimental period, whereas total calcium and phosphorus concentration decreased over time (calcium, P<0.001). Assessing the long-term stability of canine salivary analytes stored at −20°C may have important implications in diagnosis and research.

Dynamic Proteome Alteration and Functional Modulation of Human Saliva Induced by Dietary Chemosensory Stimuli.

Saliva flow measurements and SDS-PAGE separation of human whole saliva freshly collected after oral stimulation with citric acid (sour), aspartame (sweet), iso-α-acids (bitter), mono sodium l-glutamate (umami), NaCl (salty), 6-gingerol (pungent), hydroxy-α-sanshool (tingling), and hydroxy-β-sanshool (numbing), followed by tryptic digestion, nano-HPLC-MS/MS, and label-free protein quantitation demonstrated a stimulus- and time-dependent influence of the dietary chemosensates on salivation and the salivary proteome composition. Gene ontology enrichment analysis showed evidence for stimulus-induced alterations of the saliva proteome to boot an efficient molecular defense network of the oral cavity, e.g., 6-gingerol increased salivary lactoperoxidase activity, catalyzing the oxidation of thiocyanate to produce the antimicrobial and antifungal hypothiocyanate, from 0.37 ± 0.02 to 0.91 ± 0.05 mU/mL 45 s after stimulation. In comparison, oral citric acid stimulation induced an increase of myeloperoxidase activity, catalyzing the chloride oxidation to generate antimicrobial hypochloride in saliva, from 0.24 ± 0.04 to 0.70 ± 0.1 mU/mL as well as an increase of salivary levels of lysozyme, exhibiting antimicrobial activity on Gram-positive bacteria, from 6.0-10 to 100-150 μg/mL. Finally, microbial growth experiments clearly demonstrated for the first time that the increase of the salivary lysozyme abundance upon oral citric acid stimulation translates into an enhanced biological function, that is an almost complete growth inhibition of the two lysozyme-sensitive Gram-positive bacteria tested.

Effects of prolonged running in the heat and cool environments on selected physiological parameters and salivary lysozyme responses

IntroductionLysozyme is one of the salivary antimicrobial proteins which act as the “first line of defence” at the mucosal surface. The effects of prolonged exercise in the hot and cool environments among recreational athletes on salivary lysozyme responses are very limited in the literature, especially in the Asian countries. ObjectiveTo determine the effects of prolonged running in the hot and cool environments on selected physiological parameters and salivary lysozyme responses among recreational athletes. MethodsRandomised and cross-over study design. Thirteen male recreational athletes (age: 20.9 ± 1.3 years old) from Universiti Sains Malaysia participated in this study. They performed two separate running trials; 90 min running at 60% of their respective maximum oxygen uptake (V˙O2max) One running trial was performed in the hot (31ºC) while the other was in the cool (18ºC) environment and this sequence was randomised. Each running trial was started with a 5 min warm-up at 50% of participant's respective V˙O2max Recovery period between these two trials was one week. In the both trials, saliva samples, blood samples, heart rate, ratings of perceived exertion, skin and tympanic temperatures, oxygen consumption, nude body weight, room temperature, and relative humidity were collected. ResultsParticipants' skin temperature, tympanic temperature, body weight changes, heart rate, ratings of perceived exertion, and plasma volume changes were significantly higher (p < 0.05) in the hot trial compared to the cool trial. Saliva flow rate was not significantly (p = 0.949) different between the hot (0.32 ± 0.08 ml/min) and cool (0.27 ± 0.05 ml/min) trials. However, in each trial, it significantly decreased (p < 0.05) at post-exercise as compared to pre-exercise but it returned to baseline value at 1 h post-exercise. In addition, there were no significant differences between and within hot and cool trials in salivary lysozyme concentration (p = 0.925; 4.79 ± 1.37 and 4.44 ± 1.11 μg/ml respectively) and secretion rate (p = 0.843; 1.67 ± 1.1 and 1.17 ± 1.0 μg/min respectively). ConclusionThis study found similar lysozyme responses between both hot and cool trials. Thus, room/ambient temperature did not affect lysozyme responses among recreational athletes. Nevertheless, the selected physiological parameters were significantly affected by room temperature.

Effects of probiotics supplementation and circuit training on immune responses among sedentary young males.

Growing evidence suggests that probiotics may have positive benefits on immune responses following endurance exercise. However, little attention has been given to its possible beneficial effects on immune responses following resistance exercise. Forty-one healthy sedentary males were recruited and randomised into four groups: sedentary control with placebo (C), probiotics (P), circuit training with placebo (Ex), and circuit training with probiotics (PEx) groups. Participants in the Ex and PEx groups performed a progressive load of circuit training at 3 times/week for 12 weeks. Each circuit comprised 10 exercises with work to rest ratio of 1:2. Participants consumed either multi-strain probiotics or placebo twice daily for 12 weeks. Body height and weight, blood pressure, resting heart rate, saliva and blood samples were collected at pre- and post-tests. Saliva flow rate and salivary IgA, α-amylase, lactoferrin and lysozyme responses were not significantly different (P>0.05) between groups and also between pre- and post-test within each group. Similarly, total leukocytes, total lymphocytes, T lymphocytes, T-helper, T-cytotoxic, B lymphocytes, and natural killer cells counts were not significantly affected (P>0.05) by the probiotics and/or circuit training. However, circuit training significantly increased (P<0.05) immune cells count at post-test as compared to pre-test. Yet, a combination of circuit training and probiotics showed no significant (P>0.05) effects on immune cells count. This study did not provide enough support for the positive effects of probiotics on immune responses among sedentary young males following resistance exercise. However, 12 weeks of circuit training enhanced immune cells count.