[Show abstract][Hide abstract] ABSTRACT: An important contribution to the variability of any proteome is given by the time dimension that should be carefully considered to define physiological modifications. To this purpose whole saliva proteome was investigated in a wide age range. Whole saliva was collected from 17 preterm newborns with a postconceptional age at birth of 178-217 days. In these subjects sample collection was performed serially starting immediately after birth and within about 1 year follow-up gathering a total of 111 specimens. Furthermore, whole saliva was collected from 182 subjects aged between 0 and 17 years and from 23 adults aged between 27 and 57 years. The naturally occurring intact salivary proteome of the 316 samples was analyzed by low- and high-resolution HPLC-ESI-MS platforms. Proteins peculiar of the adults appeared in saliva with different time courses during human development. Acidic proline-rich proteins encoded by PRH2 locus and glycosylated basic proline-rich proteins encoded by PRB3 locus appeared since 180 days of post-conceptional age, followed at seven months (± two weeks) by histatin 1, statherin and P-B peptide. The other histatins and acidic proline-rich proteins encoded by PRH1 locus appeared in whole saliva of babies from one to three weeks after the normal term of delivery, S-type cystatins appeared at one year (± three months) and basic proline-rich proteins at four years (± one year) of age. All the proteinases involved in the maturation of salivary proteins were more active in preterm than in at-term newborns, on the basis of the truncated forms detected. The activity of the Fam20C kinase, involved in the phosphorylation of various proteins, started around 180 days of post-conceptional age, slowly increased reaching values comparable to adults at about two years (± 6 months) of age. Instead, MAPK14 involved in the phosphorylation of S100A9 was fully active since birth also in preterm newborns.
Full-text · Article · Mar 2015 · Journal of Proteome Research
[Show abstract][Hide abstract] ABSTRACT: Background:
The purpose of this study was to compare the microbiota around natural teeth and dental implants with different restorative platforms. Attention was focused on whether the microbiological environment could change according to the implant platform used i.e. traditional or Platform Switching implants. As the latter show less signs of bone resorption, a correlation with the presence of certain periodontal bacteria was suggested.
Seven partially edentulous patients with dental implants, either traditional or Platform Switching, were included in this study. All the implants were in function at least for 1 year. Gingival crevicular fluid samples were obtained before any periodontal probing from natural teeth and different implant platforms and assayed using DNA extraction and PCR sequences in order to determine quality and quantity of microbiota. Statistical analysis included chi square test were used to establish differences in the microbiological distribution between the two implant platforms.
There were not statistical differences neither regarding the distribution of microbiota around natural teeth and implants nor between the two implant platforms. The presence of B.forsythus was revealed in the majority of the samples (from 90 to 100 percent) while A.actinomycetemcomitans was rarely found (from 0 o 25 percent). As for the other periodontal microbiota, their presence or absence showed a variation according to different sites or patients, without a predictable pattern.
It was not possible to find a link between the colonization of certain types of bacteria and the reduction of bone loss which occurs around Platform Switching implants. Therefore the preservation of bone crest is only due to biomechanical aspects, which are related to the reposition of the implant-abutment interface away from the outer edge of the implant platform and from the bone.
No preview · Article · Nov 2012 · Journal of biological regulators and homeostatic agents
[Show abstract][Hide abstract] ABSTRACT: An HPLC-ESI-MS analysis of adult human whole saliva evidenced three protein masses (M average 11,487±2, 11,301±2 and 22,362±3Da) eluting in the 32.5-35.0min range. Treatment in reducing conditions allowed establishing that they are S-derivatives of N-terminal acetylated cystatin B, namely its S-glutathionyl, S-cysteinyl and S-S dimer. The identification was confirmed by high resolution HPLC-ESI-MS-MS experiments on the intact naturally occurring proteins and their tryptic digests. S-unmodified cystatin B is rarely detectable in whole saliva of healthy adults (5 subjects out of 65) and its percentage does not overcome approximately 20% of total cystatin B (11±9%). In the majority of subjects (60 out of 65) the mean percentages of the S-modified derivatives were S-glutathionyl 53±13%, S-cysteinyl 15±5%, S-S 2-mer 32±13%. Variations of the percentages of these S-modified derivatives of cystatin B could be indicative of oral oxidative stress. As we are aware, this is the first time that S-glutathionylation and S-cysteinylation were described as extensive PTM of a salivary protein and the first time that these PTMs were detected in naturally occurring cystatin B.
No preview · Article · Oct 2011 · Journal of proteomics
[Show abstract][Hide abstract] ABSTRACT: One of the therapeutic options proposed for reconstruction of the atrophic posterior mandible is inferior alveolar nerve (IAN) mobilization with simultaneous implant placement. However, studies on the functionality of this neurovascular bundle after its mobilization have shown mixed results. This variability can be attributed both to the test methodology, which typically requires subjective answers from patients, and to the surgical procedure itself, which is highly dependent on operator technique. This article reports on a series of 10 cases of IAN mobilization using a device specifically engineered to simplify bone surgery. This device enables the oral surgeon to avoid overstretching the nerve by creating a smaller bone window and using an apicocoronal inclination of instruments to capture the neurovascular bundle. Evaluation by means of neurosurgery function tests over a 36-month period found that all patients had a return to normal sensation after a brief period of neurosensory disturbance. Subjective responses to a patient questionnaire confirmed these findings. The implant success rate was 100%.
No preview · Article · Feb 2010 · The International journal of periodontics & restorative dentistry
[Show abstract][Hide abstract] ABSTRACT: Thymosin beta(4) (Tbeta(4)), its sulfoxide, and thymosin beta(10 )(Tbeta(10)) were detected in human saliva and identified by different strategies based on RP HPLC coupled to electrospray multidimensional IT MS. Tbeta(4 )was almost always detected in whole saliva, its sulfoxide sporadically, Tbeta(10) rarely. Tbeta(4) was undetectable in parotid saliva and less concentrated in submandibular/sublingual saliva than in whole saliva. Analysis of gingival crevicular fluid revealed high relative amounts of Tbeta(4), Tbeta(4) sulfoxide, and Tbeta(10) in all the samples. Tbeta(4) mean concentration was 200 times higher in crevicular fluid (20 micromol/L, N = 9) than in whole saliva (0.1 micromol/L, N = 9). Crevicular fluid concentration of Tbeta(4 )(ca. 5% represented by its sulfoxide) and beta(10 )significantly correlated (r = 0.856; N = 9), and their ratio was about 5. A significant correlation was also observed between Tbeta(4 )concentrations in whole saliva and gingival crevicular fluid (r = 0.738; N = 9). Immunohistochemical analysis of the major salivary glands showed that immunoreactivity for Tbeta(4) is restricted to ductal cells, with minor degree of focal positivity in some acinar cells. On the whole, results indicate that gingival sulcus is a main, although not the sole, source for oral Tbeta(4 )and Tbeta(10).
No preview · Article · Jan 2009 · Journal of Separation Science
[Show abstract][Hide abstract] ABSTRACT: Salivary levels of alpha-defensins 1-4 and histatins 1, 3 and 5 were determined in 11 totally edentulous patients, 11 younger healthy adults with normal gingival mucosa (Control group I) and 8 subjects, age-matched with edentulous patients, having a minimum of 25 teeth (Control group II). Whole saliva was treated with trifluoroacetic acid and the acidic soluble fraction analyzed by High Performance Liquid Chromatography-Mass Spectrometry. The area of the extracted ion current peaks was used for peptide quantification. Levels of alpha-defensins1-4, but not of histatins, were significantly lower in totally edentulous patients with respect to both Control group I and Control group II. The two control groups did not show significant differences. The reduced level of oral alpha-defensins, which are mainly of crevicular origin, is most likely due to the absence of the gingival sulcus in the edentulous subjects. The near absence of alpha- defensins might be in part responsible for the higher vulnerability of the oral cavity to oral pathogen infections observed in totally edentulous patients.
No preview · Article · Oct 2008 · International journal of immunopathology and pharmacology
[Show abstract][Hide abstract] ABSTRACT: To elucidate the localization of post-translational modifications of different classes of human salivary proteins and peptides (acidic and basic proline-rich proteins (PRPs), Histatins, Statherin, P-B peptide, and "S type" Cystatins) a comparative reversed phase HPLC-ESI-MS analysis on intact proteins of enriched granule preparations from parotid and submandibular glands as well as parotid, submandibular/sublingual (Sm/Sl), and whole saliva was performed. The main results of this study indicate the following. (i) Phosphorylation of all salivary peptides, sulfation of Histatin 1, proteolytic cleavages of acidic and precursor basic PRPs occur before granule storage. (ii) In agreement with previous studies, basic PRPs are secreted by the parotid gland only, whereas all isoforms of acidic PRPs (aPRPs) are secreted by both parotid and Sm/Sl glands. (iii) Phosphorylation levels of aPRPs, Histatin 1, and Statherin are higher in the parotid gland, whereas the extent of cleavage of aPRP is higher in Sm/Sl glands. (iv) O-Sulfation of tyrosines of Histatin 1 is a post-translational modification specific for the submandibular gland. (v) The concentration of Histatin 3, Histatin 5, and Histatin 6, but not Histatin 1, is higher in parotid saliva. (vi) Histatin 3 is submitted to the first proteolytic cleavage (generating Histatins 6 and 5) during granule maturation, and it occurs to the same relative extent in both glands. (vii) The proteolytic cleavages of Histatin 5 and 6, generating a cascade of Histatin 3 fragments, take place after granule secretion and are more extensive in parotid secretion. (viii) Basic PRPs are cleaved in the oral cavity by unknown peptidases, generating various small proline-rich peptides. (ix) C-terminal removal from Statherin is more extensive in parotid saliva. (x) P-B peptide is secreted by both glands, and its relative quantity is higher in submandibular/sublingual secretion. (xi) In agreement with previous studies, S type Cystatins are mainly the product of Sm/Sl glands.
[Show abstract][Hide abstract] ABSTRACT: Recent studies describe the potential use of biochemical markers in the evaluation of the severity of periodontitis; moreover, patients suffering from periodontitis frequently complain of halitosis (breath malodour), mainly depending on volatile compounds (e.g. hydrogen sulphide, methyl mercaptan, etc.) produced by anaerobic metabolism of oral bacteria and involving sulphur-containing amino acids. In this study, salivary sulphur compounds, such as cysteine, cysteinylglycine and glutathione and some markers of cellular damage (lactate dehydrogenase and aspartate amino transferase), were measured in periodontitis patients and correlated with the periodontal probing pocket's depth.
Twenty-two periodontitis patients and forty control subjects were studied for the salivary activities of lactate dehydrogenase and aspartate aminotransferase and cysteine, cysteinylglycine and glutathione concentrations. The periodontitis patients were divided into two subgroups based on the severity of periodontal disease, expressed as median periodontal probing pocket depth (> or <5 mm). Enzyme activities were measured by using an automated clinical analyzer; cysteine, cysteinylglycine and glutathione concentrations were measured by HPLC equipped with fluorescence detector.
A statistically significant increase of the salivary parameters level (cysteine, cysteinylglycine, glutathione, aspartate aminotransferase and lactate dehydrogenase) was found in the patient subgroup with periodontal probing pocket depth >5 mm, the salivary cysteine concentrations showing the most significant correlation.
Salivary cysteine, a direct precursor of hydrogen sulphide, could be considered reliable markers for the oral tissue damage severity in periodontitis patients.
No preview · Article · Jun 2007 · Clinical Biochemistry
[Show abstract][Hide abstract] ABSTRACT: Statherin is a multifunctional polypeptide specific of human saliva involved in oral calcium homeostasis, phosphate buffering and formation of protein networks. Salivary P-B peptide is usually included into the basic proline-rich protein family but it shows some similarities with statherin and its specific biological role is still undefined. In this study, various fragments and derivatives of statherin and P-B peptide were consistently detected by RP-HPLC ESI-IT MS in 23 samples of human saliva. They were: statherin mono- and non-phosphorylated, statherin Des-Phe(43) (statherin SV1), statherin Des-Thr(42),Phe(43), statherin Des-Asp(1), statherin Des(6-15) (statherin SV2), statherin Des(1-9), statherin Des(1-10), statherin Des(1-13) and P-B Des(1-5). Statherin SV3 (statherin Des(6-15), Phe(43)) was detected only in one sample. Identity of the fragments was confirmed either by MS/MS experiments or by enzymatic digestion or by Edman sequencing. Detection of the fragments suggests that statherin and P-B peptide are submitted to post-translational proteolytic cleavages that are common to other classes of salivary proteins.
[Show abstract][Hide abstract] ABSTRACT: Saliva is an easily available biological material that is not commonly analysed in clinical chemistry, while it could give useful information especially in several oral diseases.
In this work, the sulphur containing compounds cysteine, cysteinylglycine and glutathione were analysed in saliva of control subjects and periodontopathic subjects by a HPLC method. The detection limit of the method is 0.5, 0.1 and 0.1 micromol/l for cysteine, cysteinylglycine and glutathione, respectively, and it is linear up to 10 mmol/l.
the median values for the control group are 1.2 micromol/l for cysteine and glutathione and 0.4 micromol/l for cysteinylglycine while those of periodontopathic patients are significantly increased (4.4, 2.1 and 11.0 micromol/l for cysteine, cysteinylglycine and glutathione, respectively).
No preview · Article · Jan 2004 · Clinica Chimica Acta