Salivary osmolality in individuals with cerebral palsy

Objective

To measure the salivary flow rate, osmolality, electrolyte and total protein concentrations in individuals with cerebral palsy (CP).

Design

Thirty-eight individuals with CP were divided according to the neuromotor abnormality type (total, spastic and dyskinectic) and compared to 22 nondisabled children (control group). Whole saliva was collected under slight suction. The salivary parameters studied were salivary flow rate, osmolality, sodium, potassium, chloride and total protein concentrations.

Results

CP individuals, with both neuromotor abnormality types (spastic and dyskinectic), presented an increase in salivary osmolality, total protein, potassium and chloride concentrations compared to the control group (p < 0.05). Moreover, a reduction in salivary flow rate was verified in spastic individuals (p < 0.05).

Conclusion

The reduction in salivary flow rate and increase in osmolality, total protein and electrolyte concentrations of saliva from cerebral palsy individuals could be caused by hypohydration status.     

Influence of saliva substitute films on the initial adhesion of Candida albicans to dental substrata prior to and after artificial ageing

Objective

The aim of this in vitro study was to investigate whether saliva substitute films influence the adhesion of Candida albicans to different dental substrata prior to and after artificial ageing.

Design

Specimens of a denture base resin (DB) and a veneering composite (VC) were polished and subjected to an artificial ageing protocol (thermal cycling, 2 × 3000 cycles 5/55 °C). Bovine enamel (BE) and glass were used as reference materials. After determination of surface free energy, specimens were rinsed with commercial saliva substitutes (Aldiamed, Saliva natura, Saliva Orthana, Salinum), a positive control (protein mixture), or a negative control (Phosphate Buffered Saline, PBS) for 2 h at 37 °C in a flow chamber. Specimens were then exposed to a C. albicans ATCC 10231 suspension for 4 h at 37 °C. Adherent, viable Candida cells were quantified using a luminometric ATP-based assay. Statistical analysis was performed using 1- and 2-way ANOVA, and post-hocs were analysed using the Tukey-Kramer test (α < .05).

Results

Our data indicated that VC (31.1 mJ/m²) and DB (33.9 mJ/m²) yielded the lowest surface free energy prior to artificial ageing, and BE (43.6 mJ/m²) yielded the highest surface free energy. For C. albicans adhesion, both the materials as well as the saliva substitutes influenced relative luminescence intensities, indicating significant differences in C. albicans adhesion between the various materials and after treatment with the saliva substitutes.

Conclusions

Saliva substitutes may have a decisive influence on C. albicans adhesion, but their impact appears to be dependent on the properties of the underlying substratum material.     

The interactions of epigallocatechin-3-gallate with human whole saliva and parotid saliva

Objective

The aim of the present study was to assess the null hypothesis that the astringency and loss of lubrication in the oral cavity are not related to the properties of the epigallocatechin-3-gallate (EGCG) adlayer, the affinity and the entropy-drive of EGCG binding to saliva.

Methods

The mass, thickness, and viscoelasticity of the EGCG adlayer and the temperature-dependence of EGCG adsorption onto saliva surfaces were determined by quartz crystal microbalance with dissipation (QCM-D). The affinities of EGCG to human whole saliva (WS) and to parotid saliva (PS) were carried out by QCM-D monitoring and fluorescence quenching.

Results

The stiffer and more compact EGCG adlayers were formed on saliva surfaces at various concentrations of EGCG. The affinity for EGCG binding to WS was higher than that to PS. The precipitation of EGCG/saliva was temperature-dependent. The driving force of EGCG binding to saliva is dominated by the hydrogen bond, the hydrophobic reaction, and the entropy-drive, which were confirmed by the FTIR spectra and the measurement of temperature-dependence, respectively.

Conclusion

The viscoelasticity of the EGCG adlayer, the affinity of EGCG to saliva, and the priority of EGCG binding to hydrophobic proteins on the mucosa may account for the oral astringency and loss of lubrication.     

Influence of gender on the metabolism of alcohols in human saliva in vitro

Objective

The aim of this study was to investigate possible gender differences in salivary metabolism of two alcohols, ethanol and 2-propanol. Ethanol and its metabolite acetaldehyde may play important roles in tumour development, especially in the upper digestive tract. 2-Propanol is tested to elucidate our previous findings, where gender-specific differences in salivary acetone levels were seen after exposure to this alcohol.

Design

Saliva was collected from 25 females and 22 males for in vitro exposure to 2-propanol. In the experiments with ethanol, saliva samples were collected from 17 females and 18 males. The saliva was exposed in vitro to 2-propanol or ethanol. The metabolites acetone, derived from 2-propanol, and acetaldehyde, derived from ethanol, together with the maternal substance were analysed by headspace gas chromatography.

Results

No differences related to gender, age, medication or tobacco intake in the acetone concentration in the saliva samples were found. Gender, age or tobacco intake did not result in difference in the production of acetaldehyde in saliva. However, in the pre-exposure samples the men had a significantly higher concentration of acetaldehyde compared to the women (p = 0.04). Also, there was a tendency (p = 0.05) to higher concentrations of acetaldehyde in the samples (at 1 mM ethanol exposure) from subjects who take medications.

Conclusion

No gender difference in the metabolism of 2-propanol and ethanol in human saliva in vitro were found.     

Calcitonin gene-related peptide- and adrenomedullin-induced facilitation of calcium current in submandibular ganglion

Objective

The control of saliva secretion is mainly under parasympathetic control. The submandibular ganglion (SMG) is a parasympathetic ganglion which receives inputs from preganglionic cholinergic neurons, and innervates the submandibular salivary gland to control saliva secretion. The aim of this study was to investigate if adrenomedullin (ADM) and/or calcitonin gene-related peptide (CGRP) modulate voltage-dependent calcium channel (VDCCs) current (I_Ca) in SMG.

Design

The profile of CGRP and ADM actions in SMG was studied using the whole-cell configuration of the patch-clamp technique.

Results

Both ADM and CGRP facilitated I_Ca. These facilitations were attenuated by intracellular dialysis of the anti-Gα_s-protein and pretreatment of SQ22536 (an adenylate cyclase inhibitor).

Conclusions

ADM and CGRP facilitates VDCCs mediated by Gα_s-protein and adenylate cyclase in SMG.     

The role of salivary factors in persistent oral carriage of Candida in humans

Background

Candida albicans is a commensal oral yeast in approximately one-third to one-half of the healthy population. To date, there are no studies investigating the multiple anti-candidal salivary constituents of healthy individuals with either nil or, consistent oral yeast carriage.

Objective

To compare the composition and anti-candidal activity in stimulated whole saliva of healthy ‘consistent’ oral Candida carriers with Candida-free individuals.

Methods

A sub-sample of 22 consistent, Candida-free individuals and 10 consistent Candida carriers were recruited from a 12 months screening study investigating oral Candida carriage in 97 healthy patients treated by fixed orthodontic appliances. Unstimulated and stimulated saliva samples were collected. The following salivary attributes were measured using standard methodology: the flow rate, pH, lysozyme, lactoferrin and IgA concentration and, the degree of inhibition of blastoconidial growth and blastospore germination.

Results

Saliva from the Candida-free individuals showed 20.2% higher inhibition of blastoconidial growth (p < 0.05) of a reference strain of Candida albicans. No significant differences between the other salivary attributes of the two groups were found.

Conclusion

The fact that saliva of Candida-free individuals significantly inhibited the blastoconidial growth more than Candida-carriers (p < 0.05) suggests that saliva may play a role in modulating oral candidal populations in health. Further studies, with a larger cohort are needed to confirm these findings and determine the factors that confer enhanced salivary anti-candidal activity.     

Antimicrobial activity of super-oxidised water against oral microorganisms

Objective

The radical anion of oxygen (O⁻) is extremely oxidative and shows high reactivity. In this study, the antibacterial activity of water super-oxidised water containing high concentration of O⁻ (O⁻-water) was tested against cultured planktonic cells of cariogenic bacteria, periodontopathic bacteria and Candida albicans.

Methods

O⁻-water was prepared using the AOE-750 (Oxy Japan Corporation, Japan) and its antibacterial activity against pure culture of Streptococcus sobrinus, Porphyromonas gingivalis, Prevotella intermedia, Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum and C. albicans evaluated. Each oral microorganism (10⁴ to 10⁸ CFU/ml) was exposed to three concentrations of O⁻-water at room temperature or 37 °C for 15 s to 24 h.

Results

Exposure to O⁻-water resulted in a bactericidal effect against all cariogenic and periodontopathic bacteria tested. No significant fungicidal effect was observed on C. albicans, however.

Conclusion

The results demonstrate that O⁻-water exerts an antibacterial effect on cariogenic and periodontopathic bacteria.     

The effect of sucrose on plaque and saliva urease levels in vivo

Objective

Dietary sugar exposures induce an immediate drop of the plaque pH. Based on in vitro observations, it was hypothesized that oral bacteria may rapidly respond to this environmental change by increasing the activity or expression of alkali-generating pathways, such as the urease pathway. The objective of this exploratory in vivo study was to determine the short-term effect of a brief sucrose exposure on plaque and saliva urease activity and expression, and to relate this effect to caries experience.

Methods

Urease activity levels were measured in plaque and saliva samples collected from 20 children during fasting conditions and 30 min after rinsing with a sucrose solution. Streptococcus salivarius ureC-specific mRNA in saliva was quantified using real-time RT-PCR. The impact of host-related factors, such as age, gender, sugar consumption, salivary mutans streptococci levels and caries status on urease activity was evaluated.

Results

Plaque urease activity under fasting conditions was higher in subjects with low caries and mutans streptococci levels. This difference was not observed after the sucrose exposure. The response of urease to sucrose in vivo did not depend on caries experience or salivary mutans levels. Significant increase in urease activity of plaque and saliva after exposure to sucrose was observed only in the subjects who had low urease levels at baseline.

Conclusions

The findings of this exploratory study suggest that plaque urease activity may have an important long-term influence in caries development but not during a cariogenic challenge.     

Levels of salivary stress markers in patients with anxiety about halitosis

Objective

To investigate the relationship between salivary stress markers and mental stress states in patients complaining of oral malodour. The utility of the salivary stress markers in assessment of mental conditions of those patients was also investigated.

Design

The study population included 74 patients, aged 20-59 years, who complained of oral malodour and were referred to the Breath Odor Clinic at Tokushima University Hospital. Patients were classified into two groups, genuine halitosis (GH) and psychosomatic halitosis (PH), according to the results of organoleptic rating measurement. All patients were subjected to examination by the Cornell Medical Index (CMI) Health Questionnaire. Resting saliva was collected and levels of salivary IgA, cortisol and chromogranin A were determined by ELISA. Twenty-three volunteers not complaining of halitosis were included as the control group. Kruskal-Wallis test and Mann-Whitney's U-test were used for statistical analysis.

Results

A significant increase was observed in the concentrations of salivary cortisol in the PH group as compared with GH and control groups (p < 0.05). Concentrations of IgA and chromogranin A in saliva were not significantly different among the three groups. In addition, higher salivary cortisol concentrations were found in CMI scale III and IV (tendency towards neurosis) than in scale I and II (normal) (p < 0.05). Since salivary cortisol reflects a status of chronic stress condition, psychosomatic halitosis might be closely related to this state of chronic stress.

Conclusions

Determination of cortisol levels in saliva may provide useful information for evaluating the mental status of patients complaining of halitosis.     

Identification of pro- and mature brain-derived neurotrophic factor in human saliva

Objective

Growth factors, including brain-derived neurotrophic factor (BDNF), are polypeptides that are involved in the maintenance, survival, and death of central and peripheral cells. Numerous growth factors have been identified in saliva and are thought to promote wound healing and maintenance of the oral epithelium. The aim of this study was to determine if BDNF is also found in human saliva.

Methods

Whole, unstimulated saliva samples (n = 30) were analyzed by SDS-PAGE and Western blot using an anti-human BDNF antibody. Proteolytic cleavage products were similarly assessed following the incubation of pooled saliva with N-glycanase F and plasmin. Subjects were also genotyped for the BDNF Val66Met single nucleotide polymorphism (SNP).

Results

These experiments revealed the presence of immunoreactive bands at 14, 32 and 34 kDa, corresponding to mature (mBDNF) and proBDNF, as well as a truncated pro-form at 24 kDa. Not every sample contained all forms of BDNF. Treatment with N-glycanase and plasmin reduced the size of the higher molecular weight bands, confirming the glycosylated pro-form of BDNF. mBDNF was detected significantly less often in subjects with the Val66Met SNP, compared to those without the polymorphism (χ² = 4.05; P < 0.05).

Conclusions

While the function of salivary BDNF still requires elucidation, these findings suggest that it may be possible to use saliva in lieu of blood in future studies of BDNF and the Val66Met polymorphism.     

Inhibition of Streptococcus mutans adherence and biofilm formation using analogues of the SspB peptide

Objective

Streptococcus gordonii is a pioneer colonizer of the enamel salivary pellicle that forms biofilm on the tooth surfaces. Recent reports show the surface protein analogue peptide {400 (T) of SspB 390-402 is substituted to K forming SspB (390-T400K-402)} from S. gordonii interacts strongly with salivary receptors to cariogenic bacteria, Streptococcus mutans. To characterize the analogue peptide biological activities, we investigated its binding and inhibiting effects, and the role of its amino acid moities.

Methods

We measured binding activity of analogue peptides to salivary components using the BIAcore assay; assayed inhibition activities of peptides for bacterial binding and growth on saliva-coated hydroxyapatite beads (s-HA); and describe the peptides interfering with biofilm formation of S. mutans on polystyrene surfaces.

Results

The SspB (390-T400K-402 and -401) peptides significantly bound with salivary components and inhibited the binding of S. mutans and S. gordonii to s-HA without bactericidal activity; but did not inhibit binding of Streptococcus mitis, a beneficial commensal. Further, the lack of D and E-L at position 390 and 401-402 in the peptide, and substituted peptide SspB (D390H- or D390K-T400K-402) did not bind to salivary components or inhibit binding of S. mutans. The SspB (390-T400K-402) peptide inhibited biofilm formation on salivary components-coated polystyrene surfaces in absence of conditioned planktonic cells.

Conclusions

We found constructing the peptide to include positions 390(D), 400(K) and 401(E), two surface positive and negative connective charges, and at least 12 amino acids are required to bind salivary components and inhibit the binding of S. mutans and S. gordonii.     

Availability of saliva for the assessment of alterations in the autonomic nervous system caused by physical exercise training

Objective

The purpose of the study was to assess the alterations of the autonomic nervous system activities caused by physical exercise training under food restriction using a running wheel in mice based on an analysis of saliva.

Methods

Male ICR mice, weighing 39-50 g, at 32 weeks of age were divided into three groups; an exercise with food restriction (EXP), an exercise without food restriction (EXA) and a control (CTL). The EXP group was fed the same amount of diet as the CTL group (pair-feeding). The EXP and EXA groups used a “voluntary running wheel” for exercise. The pilocarpine stimulated whole saliva was collected from the oral cavity by micropipette over 15 min 4, 8 and 12 weeks after the beginning of the experiment. The salivary flow rate, protein concentration and amylase and kallikrein activities were determined, since amylase and kallikrein release have been shown to be evoked by β- and α-adrenergic receptor stimulation, respectively.

Results

There was no significant difference in the sera corticosterone levels among the three experimental groups. The flow rate of saliva per total salivary gland weight in the EXP was significantly lower than that in the CTL and the EXA groups. The total protein secretion and kallikrein activity decreased by 20-30% in the EXP mice between 4 and 12 weeks after the starting of experiments in comparison to the CTL mice, thus suggesting that signal transduction in the α-sympathetic nervous system was downregulated. There was no significant difference in the amylase activity between the EXP and the CTL groups.

Conclusion

Physical exercise under diet control induced alterations in saliva secretion, while exercise training alone did not affect the content of saliva. The results demonstrate the availability of saliva and suggest that the present experimental situation is a suitable experimental model for sports activity performed under diet control.     

Salivary levels of IL-1β, IL-6, IL-8, and TNF-α in patients with burning mouth syndrome

Objective

To compare salivary IL-1β, IL-6, IL-8, and TNF-α levels between patients with burning mouth syndrome (BMS) and controls.

Design

Forty female patients with BMS (mean age: 61.6 ± 10.1 years) and 20 female control subjects (mean age: 65.1 ± 9.0 years) were included in the study. Unstimulated (UWS) and stimulated whole saliva samples (SWS) were collected and their flow rates were determined. Salivary IL-1β, IL-6, IL-8, and TNF-α levels and total protein concentration were also determined. Salivary transferrin level was determined to investigate the level of blood contamination in saliva samples. Gingival index of the subjects was also examined. Student's t-test, Pearson's correlation analysis, and analysis of covariance were used.

Results

No significant differences were found in the salivary levels of IL-1β, IL-6, IL-8, and TNF-α in BMS patients compared with controls. Salivary flow rates and their total protein concentrations did not differ significantly between the groups. The levels of salivary cytokines and total protein concentration correlated significantly with the level of blood contamination in both UWS and SWS.

Conclusion

There were no differences in the salivary levels of IL-1β, IL-6, IL-8, and TNF-α in BMS patients compared with controls. Cytokine levels in whole saliva were affected mainly by the amount of blood contamination.     

Vasodilatory effect of hydroxyethyl methacrylate and triethylene glycol dimethacrylate in rat aorta through calcium antagonistic action

Introduction

Resin-based dental materials contain various diluent monomers that can interfere with vascular function by causing vasodilation. In this study, we evaluated the vasoactive potential of hydroxyethyl methacrylate (HEMA) and triethylene glycol dimethacrylate (TEGDMA) and the possible mechanism of their vascular action on isolated rat aorta.

Methods

Responses of thoracic aorta rings were recorded isometrically by using force displacement transducers. After precontracting aorta rings with phenylephrine, relaxations to HEMA and TEGDMA were recorded in the absence and presence of nitric oxide synthase inhibitor N^ω-nitro-L-arginine methyl ester, cyclooxygenase inhibitor indomethacin, and K⁺ channel inhibitors tetraethylammonium, glibenclamide, and 4-aminopyridine. To investigate the Ca²⁺-channel antagonistic effect of HEMA and TEGDMA in different aorta rings, concentration-response curves to CaCl₂ were obtained in the absence and presence of the test monomers.

Results

Both HEMA and TEGDMA elicited concentration-dependent relaxations. The vasorelaxant effect of HEMA and TEGDMA was not mediated via endothelium-dependent nitric oxide and prostanoid-dependent mechanisms or by K⁺ efflux through K⁺ channels. Both monomers significantly inhibited the contractions induced by CaCl₂.

Conclusions

Our results showed that HEMA and TEGDMA induce vasodilation via Ca²⁺-antagonistic action, whereas nitric oxide and cyclooxgenase pathway and K⁺ channels were not responsible for this vasoactive effect.     

Antibacterial effect of ozone on cariogenic bacterial species

Objective

The aim was to evaluate the antibacterial effect of ozone on cariogenic bacterial species with and without the presence of saliva and a possible effect on the salivary proteins.

Methods

Suspensions of Actinomyces naeslundii (ACTCC 12104^T), Lactobacilli casei (N CTC 151) and Streptococcus mutans (NCTC 10449), in salt buffer or in saliva, were exposed to ozone gas delivered by the ozone generator Healozone™ 2130C. Aliquots of the suspensions were taken after 10, 30 and 60 s ozone exposures and cultivated on agar plates. Initial number of bacteria per ml was 8.0 × 10⁷ (SD 2.2 × 10⁷) (A. naeslundii), 1.0 × 10⁸ (SD 3.1 × 10⁶) (L. casei) and 1.0 × 10⁸ (SD 7.0 × 10⁵) (S. mutans), respectively. The proteins were separated by SDS electrophoresis and visualized by silver staining.

Results

In salt buffer 92%, 73% and 64% of the initial numbers of A. naeslundii, S. mutans and L. casei, respectively, were killed already after 10 s ozone exposure, while approximately 99.9% of the bacteria were dead after a 60 s exposure. After 10 and 30 s, but not after 60 s exposure to ozone, S. mutans and L. casei were less efficiently killed in saliva compared to the salt buffer. Various saliva proteins were degraded by ozone after a 60 s exposure.

Conclusions

The cariogenic species S. mutans, L. casei and A. naeslundii were almost eliminated following 60 s of ozone treatment. This killing was reduced in the presence of saliva although increasing the ozone application time to 60 s overcame these reductants in saliva. Detection of altered salivary proteins indicates that saliva components constitute additional targets for ozone.     

A universal pre-analytic solution for concurrent stabilization of salivary proteins, RNA and DNA at ambient temperature

Objective

Saliva is a biofluid that can be obtained from individuals without supervision by health care providers. To maximize this clinical advantage, it is highly desirable to have a global salivary analyte stabilizer for proteins, RNA and DNA at ambient temperature.

Design

Whole saliva, saliva supernatant and saliva filtrate (5.0 μm) were treated with RPS at room temperature (RT) for up to 6 days and then subjected to SDS-PAGE. Immunoblotting of β-actin and cystatin C were used to evaluate protein stability. For salivary DNA/RNA, whole saliva was incubated with RPS at RT for up to 10 weeks. After extracting total DNA/RNA in samples at week 0, 2, 6 and 10, DNA stability was assayed by chromosome 18 DNA qPCR and RNA stability by β-actin mRNA RT-qPCR.

Results

β-actin completely degraded in all types of saliva samples after 6-day incubation at RT. However, 24.0%, 91.4% and 89.3% of β-actin remained intact with RPS for whole saliva, saliva supernatant and filtrate, respectively. Similarly, 70.3% of cystatin C in supernatant remained intact in the presence of RPS. For salivary DNA/RNA, the cycle threshold (Ct) values showed no significant change for chromosome 18 DNA and β-actin mRNA in RPS-incubated saliva during the 10-week time course while significant increase in Ct values were observed in controls without RPS for both β-actin mRNA and DNA.

Conclusions

RPS provided effective concurrent stabilization to salivary DNA/RNA in whole saliva for up to 10 weeks and proteins in saliva filtrate for 6 days at RT. We also achieved separation of saliva supernatant from cellular elements by a simple filtration step (bypassing the need for centrifugation).     

Characteristics of neurokinin A-induced salivary fluid secretion in perfused rat submandibular gland

Tachykinins such as neurokinin A (NKA) and substance P have been demonstrated to induce salivary fluid secretion in vivo. However, characteristics of salivary fluid secretion induced by tachykinins in salivary glands have not been well elucidated. In this study, the effects of the tachykinin NKA on salivary fluid secretion were investigated in isolated, perfused rat submandibular gland. NKA provoked salivary fluid secretion, which consisted of transient and sustained phases, in a dose-dependent manner. In fura-2-loaded dispersed cells of the rat submandibular gland, the doses of NKA in which induced salivary fluid secretion caused an increase in intracellular Ca²⁺ concentration. When Ca²⁺ was removed from the perfusate to examine the effect of Ca²⁺ mobilization on NKA-induced fluid secretion, only the transient salivary fluid secretion occurred. When the gland was perfused with the Ca²⁺-free perfusate containing the intracellular Ca²⁺ chelator BAPTA-AM, NKA failed to induce salivary fluid secretion. NKA also induced an increase in oxygen consumption, but which was reduced by the removal of Ca²⁺ from perfusate. Salivary fluid is secreted via transcellular and paracellular pathways in acinar cells of salivary glands. To examine the contribution of paracellular pathway to NKA-induced salivary fluid secretion, the glands were perfused with a perfusate containing Lucifer yellow (LY), a cellular impermeable substance, and then were stimulated with NKA, which provoked secretion of LY in the saliva. These results suggest that the NKA-induced salivary fluid secretion is Ca²⁺-dependent and that the paracellular pathway contributes to the secretion.     

Transaminases in the acquired pellicle

Objective

Transaminases (AST, aspartate amino transferase; ALT, alanine amino transferase) are relevant enzymes in physiology and pathology of the human organism. The aim of the present in situ study was to demonstrate the presence of these enzymes in the enamel pellicle.

Methods

Bovine enamel slabs were fixed on buccal sites of individual upper jaw splints and worn for 3, 30 and 120 min by 5 subjects to allow pellicle formation. The in situ pellicles were tested for AST and ALT. Enzyme activities were measured photometrically via determination of the products pyruvate and oxalacetate using lactate-dehydrogenase and malate-dehydrogenase, respectively.

Results

Enzymatic AST- as well as ALT-activities are present in the acquired pellicle within 3 min. The enzyme activities exposed at the pellicles’ surfaces increased slightly with the pellicle formation time (ANOVA, AST: n.s., ALT: p = 0.021). However, the two enzymes show considerable intraindividual and interindividual variability. The mean AST-activity of the pellicle amounted to 1.07 ± 0.81 mU/cm² (ALT 1.18 ± 0.52 mU/cm²). The ALT-activity of the centrifuged saliva was 26.62 ± 11.09 mU/ml (AST 35.98 ± 29.35 mU/ml).

Conclusions

AST as well as ALT are present in the in situ pellicle layer and may contribute to the intrinsic maturation of pellicle proteins.