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Patients with beta-thalassemia show 5-fold increase in agestandardized lethality due to SARS-CoV-2 infection, representing a high-risk population compared with age- and sex-matched healthy subjects.(1) Vaccination against SARS-CoV-2 is crucial to reduce mortality and morbidity of frail patients.(2) Up to now, limited data have been available on the responses of patients with beta-thalassemia immunized with anti-SARS-CoV-2 mRNA vaccines.(3)

Acyl-CoA-binding protein (ACBP), also known as diazepam binding inhibitor (DBI), is a small phylogenetically conserved protein. This ancestral peptide is multifunctional, performing intracellular activities as ACBP protein or extracellular roles as DBI. Several studies showed its endless facets, including a relevant activity as appetite stimulator and as anabolic factor. High levels of ACBP have been described in erythrocytes, liver, kidney, and gut cells. The aim of this study was to analyze, at immunohistochemical level, the expression of ACBP in fetal human tissues during development, focusing on the developing kidney.Immunohistochemistry for ACBP was performed on 30 human fetal kidneys, from 15 fetuses of gestational age ranging from 13 to 19 weeks. At autopsy, all kidney samples were 10% formalin-fixed, routinely processed and paraffin-embedded. Five micron-thick paraffin sections were stained with Hematoxylin and Eosin and PAS stain for a morphological examination.ACBP was detected in all 30 kidneys analyzed in this study. No significant changes in ACBP expression were observed at different gestational ages. Immunostaining for ACBP was restricted to the epithelium covering the renal pelvis, the papillae, the collecting tubules, and the proximal and distal tubules. On the other hand, medullary regions and in the metanephric mesenchymal stem/progenitor cells did not show any reactivity for ACBP.According to our findings, ACBP should be considered as a new player in the complex field of human nephrogenesis, given that it was detected in all fetal kidneys immunostained. Its preferential localization in the renal structures derived from the Wolf duct, such as pelvis epithelium and collecting ducts, suggests a major role for ACBP in the induction of the metanephric mesenchymal cells toward the differentiation into glomerular structures. ACBP expression in proximal and distal tubules, two structures originating from the metanephric mesenchyme, indicates a further role of this protein in nephron development. In conclusion, ACBP should be added to the multiple molecules involved in human nephrogenesis.

Objective: We hypothesised that prior knowledge of details for a growth modification treatment influences cephalometric measurements and results in a detectable bias. Design: Observational study. Setting: University teaching facility. Methods: Six orthodontic residents assessed 48 lateral cephalograms taken before and after functional appliance treatment from 24 patients. The residents assessed six cephalometric measurements, (Cd-Pog, Cd-Me, Ar-Pog, Ar-Me, Go-Me, SNB) over three separate sessions, in either a random concealed order or as matched pairs with information about treatment and time disclosed. Results: When information was disclosed, five out of the six cephalometric measurements were significantly higher that the corresponding cephalometric measurements taken randomly with undisclosed information. The bias was in the range of 1.6–3.2 mm for linear measurements and was 1.1° for SNB. Conclusion: Disclosing treatment information does introduce systematic errors in cephalometric measurements. Cephalometric analysis in orthodontic clinical research should be carried out by assessors who are blinded to treatment details, to minimise risk of bias.

Paediatric sleep breathing disorders represent an emerging paediatric health concern. Despite risk factors are widely discussed in literature, evidences about protective factors are lacking. The aim of this systematic review was to examine the available evidence about the effect of breastfeeding on snoring and obstructive sleep apnea in childhood, and to methodically describe the underlying mechanism of interaction.The study protocol was registered in advance in PROSPERO (CRD42020212529). Electronic search of the literature was performed up to October 10th, 2020 using four databases: PubMed, ScienceDirect, Medline and Scopus. Two authors independently retrieved potentially relevant articles to meet eligibility criteria. PRISMA guidelines were followed. Prospective, retrospective, case-control, cohort, clinical trial and cross-sectional studies investigating the association between breastfeeding and paediatric sleep-disordered breathing were included and data were extracted. No restrictions on language or date of publication were set. Subsequently, a search in the literature was further done to investigate underlying mechanisms of interaction.This review suggests that breastfeeding can effectively protect children and adolescents from sleep-disordered breathing, especially from habitual snoring, by preventing the associated risk factors. Future prospective studies with more efficient design are required.

Background: Although numerous patient specific co-factors have been shown to be associated with worse outcomes in COVID-19, the prognostic value of thalasemic syndromes in COVID-19 patients remains poorly understood. Aims: We studied the outcomes of 137 COVID-19 patients with a history of Transfusion Dependent Thalassemia (TDT) and non-Transfusion Dependent Thalassemia (NTDT) extracted from a large international cohort and compared them with the outcomes from a matched cohort of COVID-19 patients with no history of thalasemia. Results: The mean age of thalassemia patients included in our study was 41±16 years (48.9% male). Almost 81% of these patients suffered from TDT requiring blood transfusions on a regular basis. 38.7% of patients were blood group O. Cardiac iron overload was documented in 6.8% of study patients, whereas liver iron overload was documented in 35% of study patients. 25% of thalassemia patients had a history of splenectomy. 27.7% of study patients required hospitalization due to COVID-19 infection. Amongst the hospitalized patients, one patient died (0.7%) and one patient required intubation. Continuous positive airway pressure (CPAP) was required in almost 5% of study patients. After adjustment for age-, sex- and other known risk factors (cardiac disease, kidney disease and pulmonary disease), the rate of in-hospital complications (supplemental oxygen use, admission to the an intensive care unit for CPAP therapy or intubation) and all-cause mortality was significantly lower in the thalassemia group compared to the matched cohort with no history of thalassemia. Amongst thalassemia patients in general, the NTDT group exhibited a higher rate of hospitalization compared to the TDT group (p=0.001). In addition, the rate of complications such as acute kidney injury and need for supplemental oxygen was significantly higher in the NTDT group compared to the TDT group. In the multivariable logistic regression analysis, age and history of heart or kidney disease were all found to be independent risk factors for increased in-hospital, all-cause mortality, whereas the presence of thalassemia (either TDT or NTDT) was found to be independently associated with reduced all-cause mortality. Conclusions: The presence of thalassemia in COVID-19 patients was independently associated with lower in-hospital, all-cause mortality and few in-hospital complications in in our study. The pathophysiology of this is unclear and needs to be studied in vitro and in animal models. Trial Registration for sources of data extraction: NCT: 04334291, 04746066 Funding Statement: This study was funded by a non-conditioned grant from FUNDACION INTERHOSPITALARIA PARA LA INVESTIGACION CARDIOVASCULAR, FIC. (Madrid, Spain). This nonprofit institution had no role in the study design, the collection, analysis and interpretation of data, the writing of the report, or the decision to submit the paper for publication. Declaration of Interests: The authors declare no conflicts of interest. Ethics Approval Statement: The study was approved by the Ethics Committees in all of the centers involved (Banjarmasin, Bari, Cagliari, Catania, Ferrara, Gela, Genoa, Getafe, Guadalajara, Legan, Madrid, Mannheim, Milan, Monza, Naples, Olbia, Padua, Pavia, Ragusa, Rome, Salerno, Turin, Valladolid and Verona).

This study is primarily aimed at assessing serum changes on a large panel of proteins in patients with chronic back pain following spa therapy, as well as evaluating different spa therapy regimens as a preliminary exploratory clinical study. Sixty-six patients with chronic back pain secondary to osteoarthritis were randomly enrolled and treated with daily mud packs and bicarbonate-alkaline mineral water baths, or a thermal hydrotherapy rehabilitation scheme, the combination of the two regimens or usual medication only (control group), for two weeks. Clinical variables were evaluated at baseline, after 2 and 12 weeks. One thousand serum proteins were tested before and after a two-week mud bath therapy. All spa treatment groups showed clinical benefit as determined by improvements in VAS pain, Roland Morris disability questionnaire and neck disability index at both time points. The following serum proteins were found greatly increased (≥2.5 fold) after spa treatment: inhibin beta A subunit (INHBA), activin A receptor type 2B (ACVR2B), angiopoietin-1 (ANGPT1), beta-2-microglobulin (B2M), growth differentiation factor 10 (GDF10), C-X-C motif chemokine ligand 5 (CXCL5), fibroblast growth factor 2 (FGF2), fibroblast growth factor 12 (FGF12), oxidized low density lipoprotein receptor 1 (OLR1), matrix metallopeptidase 13 (MMP13). Three proteins were found greatly decreased (≤0.65 fold): apolipoprotein C-III (Apoc3), interleukin 23 alpha subunit p19 (IL23A) and syndecan-1 (SDC1). Spa therapy was confirmed as beneficial for chronic back pain and proved to induce changes in proteins involved in functions such as gene expression modulation, differentiation, angiogenesis, tissue repair, acute and chronic inflammatory response.

BACKGROUND The MARTE study investigated the demographic, clinical, and therapeutic characteristics of rheumatoid arthritis (RA) patients ongoing methotrexate (MTX) treatment for longer than 8 years. METHODS This cross-sectional, observational study considered 587 RA patients from 67 Rheumatology Units across Italy. Data collected included demographic, clinical, and therapeutic characteristics, focusing on MTX prescription patterns (route of administration, dosing regimens, treatment duration, and discontinuation). RESULTS As initial therapy, 90.6% of patients received one conventional synthetic Disease Modifying Anti Rheumatic Drug (csDMARD), with treatment started within the first 3 months from diagnosis in half of the patients. MTX was the first csDMARD in 46.2% of patients. The prevalent route of administration at diagnosis was the intramuscular (60.5%), while at study entry (baseline) 57.6% were receiving subcutaneous MTX. Patients who required a higher MTX dose at study entry were those who received a significantly lower starting MTX dose (P

Juvenile Idiopathic Arthritis (JIA) encompasses a wide range of mostly idiopathic autoimmune arthritis which affect growing individuals. Temporomandibular disorders are a diffuse spectrum of diseases which involve temporomandibular joint (TMJ) and associated structures. However, much more need to be defined in order to detect and manage these conditions. Albeit satisfying evidences exist about their impact on the adult population, there is great lack of information in children. The current study has been developed in order to overview what it is known about these disorders and their mutual interactions. A case report as support to the scientific evidence has been further described.A 7-year-old patient affected by an undifferentiated form of JIA developed arthritis to TMJs, complaining pain and functional impairments. After 2 years, the follow-up with combined pharmacological therapies and a modified oral stabilisation appliance shows no objective worsening of the joints' structures. However, the more complex symptomatic management of inflammation highlights the need for further knowledge.The present study shows that both literature and clinical activity highlight a strong relationship between JIA and TMDs, that can affect the quality of life of children and adolescents. Diagnosis and management of these conditions are extremely complex, thus additional studies and evidence are needed. However, the need of an interdisciplinary approach between rheumatologists, paediatricians and dentists has been demonstrated.

Limited information is available on the hematological characterization of the α-thalassemia carrier in pediatric age. The objective of this report was to evaluate the red cell indices according to the α-globin genotype in a cohort of children evaluated in Sardinia. Moreover, we verified the frequency of different α-globin genotypes in this cohort. A total of 453 subjects were investigated for hematological indices and for the most common α-globin defects present in Sardinia. Of them, 352 with HbA2 ≤ 3.2%, and no iron deficiency anemia were taken into consideration to evaluate the red cell indices according to the α-globin genotype in pediatric age. A total of 11 different α-genotypes were detected, confirming the wide heterogeneity of α-thalassemia in Sardinia. Moreover, our results showed that the hematological parameters in normal children may be conditioned by the clinically occult coinheritance of mild α-thalassemia alleles as already described in the adult population while microcytosis and hypocromia in children without iron deficiency should suggest the coexistence of two α-globin defects. We concluded that recognizing the α-globin gene mutations for a particular population with their particular red cell indices may help pediatricians to perform a correct diagnosis distinguishing among physiological and pathological types of microcytosis and hypocromia.

Biofilm elimination is often necessary during antimicrobial therapy or industrial medical manufacturing decontamination. In this context, ultrasound treatment has been frequently described in the literature for its antibiofilm effectiveness, but at the same time, various authors have described ultrasound as a formidable enhancer of bacterial viability. This discrepancy has found no solution in the current literature for around 9 years; some works have shown that every time bacteria are exposed to an ultrasonic field, both destruction and stimulation phenomena co-exist. This co-existence proves to have different final effects based on various factors such as: ultrasound frequency and intensity, the bacterial species involved, the material used for ultrasound diffusion, the presence of cavitation effects and the forms of bacterial planktonic or biofilm. The aim of this work is to analyze current concepts regarding ultrasound effect on prokaryotic cells, and in particular ultrasound activity on bacterial biofilm.

The objective of this study was to assess bone mineral density (BMD) in women on long-term mud-bath therapy (MBT) for osteoarthritis in a Salus per Aquam (SPA) environment. Two hundred and fifty female patients were randomly enrolled in this study in the SPA center of Sardara (Cagliari, Italy) where they were treated with a combination of daily full body mudpacks and bicarbonate-alkaline mineral water baths at cycles of 2 weeks/year. BMD was evaluated by means of calcaneus ultrasonometry (Sahara Hologic Inc., Bedford, MA, USA) and results analyzed according to duration of treatment and clinical variables. In the group of patients undergoing MBT for more than 10 years (group A) and for 3 to 10 years (group B) a reduced frequency of osteopenia and osteoporosis was detected (35.8% and 7.6% group A; 38.4% and 8.5% group B, respectively) compared to controls (group C) (48.9% and 23.4%, P

Obstructive sleep apnoea (OSA) is often treated with autotitrating continuous positive airway pressure (autoCPAP) devices. Clinical and bench tests of these devices have suggested performance limitations. These studies do not indicate whether this is a failure to detect or a failure to respond to airway obstruction. In this randomised, crossover trial, 34 patients with moderate-to-severe OSA underwent polysomnography on two laboratory visits. The autoCPAP device was randomly set to a fixed subtherapeutic pressure (detection assessment) or autotitrating mode (response assessment). Airflow was measured both from the autoCPAP (autoCPAP flow) and directly from the nasal mask, and recorded on polysomnography. Apnoea/hypopnoea indices (AHIs) measured at the two sites and from the autoCPAP download report were compared. Regarding detection, the AHI measured from the nasal mask showed good agreement with the autoCPAP flow AHI, but agreement was lower with the autoCPAP report AHI. In autotitrating mode, there was significant misclassification of those with and without OSA (AHI ≥ 10 events · h(-1)) on the autoCPAP report. Regarding response, residual OSA (AHI ≥ 10 events · h(-1)) was still evident in 24% of patients during autotitration. In some patients, autoCPAP fails to detect and/or respond to sleep apnoea. Clinicians should consider limitations of each device and use caution when using autoCPAP report statistics to verify effective treatment.

To evaluate the association between caffeine exposure and acute kidney injury (AKI) in very low birth weight (VLBW; ≤1500 g) neonates.We retrospectively reviewed a cohort of 140 VLBW neonates consecutively admitted to the University of Virginia's neonatal intensive care unit from March 2011 to June 2012, excluding only those admitted2 days of age or who died at2 days after birth. We separately analyzed a subgroup of 44 neonates who received prolonged invasive respiratory support (mechanical ventilation for first 7 days after birth). The exposure of interest was caffeine exposure in the first week after birth. The primary outcome was AKI within the first 10 days after birth according to the Kidney Disease: Improving Global Outcomes system, modified to include only serum creatinine.Caffeine exposure occurred in 72.1% of all patients and 54.5% of those who received prolonged invasive respiratory support. AKI occurred less frequently in neonates who received caffeine (all patients: 17.8% vs 43.6%; P = .002; prolonged invasive respiratory support: 29.2% vs 75.0%; P = .002). Caffeine exposure was associated with decreased odds for AKI in logistic regression models adjusted for sex, birth weight, gestational age, small for gestational age status, illness severity on admission, and receipt of indomethacin, invasive ventilation, dopamine, aminoglycosides, and vancomycin (all patients: OR 0.22; 95% CI 0.07-0.75, P = .02; prolonged invasive respiratory support subgroup: OR 0.06; 95% CI 0.01-0.57, P = .02).In a cohort of VLBW neonates, those exposed to caffeine were less likely to experience AKI.

Background Anchorage onto permanent dentition is a common procedure in rapid maxillary expansion. However, replacing first permanent molars with the second deciduous molars seems to be an option to reduce some negative side effects during orthodontic treatment. The purpose of this study was to evaluate the dental effect of rapid maxillary expansion with anchorage exclusively onto deciduous teeth performed in the first period of transition. Methods Twenty patients with a lateral cross-bite treated exclusively by a Haas expander in early mixed dentition were retrospectively analyzed before treatment, at appliance removal, and at 21 months out of retention. The sagittal and transverse dimensions, together with the inter-canine arch and irregularity index, were digitally measured on scanned images of dental casts. The patients were compared with three balanced control groups (in total, 60 individuals) matched for gender. Two control groups had the same canine dental class as the treated group at T1, were in the inter-transitional period, and either had or lacked a lateral cross-bite. The last control group was comprised of adolescents in permanent dentition with a dental class I. The statistical analysis was performed by means of repeated-measures ANOVA for paired data and one-way ANOVA, the Kruskal-Wallis test, and the Mann-Whitney test for independent measures (α-level p

Periodontal probing aims at detecting the presence, type and gravity of periodontal diseases influencing distance between gingival margin and connective ligament. Measurements in vivo are affected by substantial uncertainty, owing, for example, to probe features, anatomic variations and operator’s skill. Inadequate reproducibility in periodontal probing may lead to diagnostic mistakes and inappropriate therapeutic decisions. In vitro evaluation of reproducibility of measurements of periodontal pockets was aimed at while developing a Periodontal Calibration Box designed to calibrate periodontal probe operators, catering also for simulating contraction observed in vivo. Probe type and clinical experience of operators were found to affect substantially probing errors.

A reversed-phase high-performance liquid chromatography (HPLC) method with diode-array detection for the quantification of several human salivary peptides is described. Sample pretreatment consisted of the acidification of whole saliva by phosphate buffer. This treatment produced precipitation of mucins, alpha-amylases and other high-molecular-mass salivary proteins, simultaneous inhibition of intrinsic protease activities and reduction of sample viscosity. Direct HPLC analysis by diode-array detection of the resulting acidic sample allowed one to quantify histatin 1, histatin 3, histatin 5, statherin, as well as uric acid, in normal subjects. Moreover, the groups of peaks pertaining to proline-rich proteins and cystatins were tentatively identified. The method can be useful in assessing the concentration of salivary peptides from normal subjects and from patients suffering oral and/or periodontal diseases.

To assess the reliability and simplicity of a method chosen for selecting subjects to be treated in the orthodontic system and for the acceptance of the method by the community.490 orthodontic patients received full dental examination. For each patient a table was prepared for the detection of malocclusion in which the occlusal characteristics according to the Norwegian Orthodontic Treatment Index (NOTI), the molar class and the teeth present were recorded. The parents of the examined children completed a questionnaire aimed at identifying their social ranking. In the same questionnaire, how many siblings the patient had was also asked. Each of the three parameters were rated alphanumerically and a score of 1 was given to each child; the sum of three scores gave the final result for the subject. Acceptance of the procedure in the general community was evaluated by the number of complaints received by the Public Relations Office (URP).The social classes most represented in the sample are "blue collar class" (42.2%) and "white collar class" (35.6%). There was an average of 2.1 children per family (SD=0.6). The distribution of the type and grading of the treatment need was similar throughout the different social classes.The chosen method proved reliable for two reasons: no complaints with the local URP and ease of application of the selected criteria.

Microbial neonatal infections are responsible for considerable morbidity and mortality and for this reason there is a growing interest for new approaches in the clinical government of this human affection. Using an integrated statistical model, this work investigated the role of the C-reactive protein (CRP) in the diagnosis of sepsis and therapy assessment in newborns admitted in neonatal intensive care unit.386 newborns admitted in neonatal intensive care unit were enrolled in this work. Different clinical-laboratory parameters, such as: CRP level, blood culture, complete blood cell count, urine and other blood tests were assessed for the first 7 days after birth. Several statistical methods have been used to estimate the correlation CRP-septicaemia, using Chi-squared, Pearson, analysis of the variance and Poisson distribution.a statistical positive correlation (CRP value vs. septicaemia status) was observed to integrate the analysis of the variance and Poisson distribution methods, especially in the first days after birth.A correct statistical evaluation of CRP values could be significant for risk prediction and subsequent prompt therapy in neonatal sepsis.

The purpose of this study is to compare, by means of mechanical tests, the resistance of two types of dental restoration in the uncomplicated crown fracture of human permanent maxillary central incisor: rebonding of the fractured tooth fragment and application of ceramic veneers.Thirty human maxillary central incisors were divided into three groups of ten teeth each. The teeth of the first group (Group A) were kept intact and used as controls, while the remaining teeth were sectioned, in order to simulate a crown fracture, and restored respectively with either the rebonding of the fragment (Group B) or with the use of ceramic veneers (Group C). All samples underwent mechanical tests by means of a universal test machine.The mean resistance to fracture recorded is respectively 425.2N for Group A, 233.3N for Group B, and 347.3N for Group C. One-way ANOVA analysis demonstrates statistical significance for at least one couple of values (Group B compared to Group A). The group of restorations with rebonding of fractured fragments has a mean value of resistance to fracture that is lower than both intact teeth and teeth restored with ceramic veneers. The resistance of teeth with rebonded restorations is 54.9% of that of, intact teeth, and resistance increases for teeth restored with ceramic veneers (81.8% of that of intact teeth).Ceramic veneers showed a higher resistance than rebonding and resulted to be more reliable in case of further trauma.

Tooth eruption is defined as the movement of a tooth from its site of development within the alveolar process to its functional position in the oral cavity [Massler et al., 1941]. The process of tooth eruption can be divided into different phases: pre-eruptive bone stage, alveolar bone stage, mucosal stage, preocclusal stage, occlusal stage and maturation stage [Andreasen et al., 1997]. Any disturbance in these phases can lead to eruptive anomalies. The incidence of unerupted teeth is usually higher among permanent teeth than among deciduous ones [Walker et al., 2004; Otsuka et al., 2001; Amir et al., 1982; Broadway, 1976; Pinborg et al., 1970]. Of the primary teeth reported as unerupted, second primary molars are the teeth most frequently involved [Walker et al., 2004; Otsuka et al., 2001; Bianchi et al., 1991; Ranta et al., 1988; Tsukamoto et al., 1986; Amir et al., 1982], followed by primary central incisors [Otsuka et al., 2001].This paper presents a case of inversion of the intraosseous position of a second unerupted deciduous molar and the succedaneous second premolar.

The acidic soluble fraction of whole saliva of type 1 diabetic children was analyzed by reversed phase (RP)(1)-HPLC-ESI-MS and compared with that of sex- and age-matched control subjects. Salivary acidic proline-rich phosphoproteins (aPRP), histatins, α-defensins, salivary cystatins, statherin, proline-rich peptide P-B (P-B), beta-thymosins, S100A8 and S100A9*(S100A9* corresponds to S100A9 vairant lacking the first four amino acids), as well some naturally occurring peptides derived from salivary acidic proline-rich phosphoproteins, histatins, statherin, and P-B peptide, were detected and quantified on the basis of the extracted ion current peak area. The level of phosphorylation of salivary acidic proline-rich phosphoproteins, histatin-1 (Hst-1), statherin and S100A9* and the percentage of truncated forms of salivary acidic proline-rich phosphoproteins was also determined in the two groups. The study revealed that statherin, proline-rich peptide P-B, P-C peptide, and histatins, were significantly less concentrated in saliva of diabetic subjects than in controls, while concentration of α-defensins 1, 2 and 4 and S100A9* was higher. The low concentration of P-C peptide was paralleled by high levels of some of its fragments. On the whole, the study highlighted the severe impairment of the repertoire of peptides involved in the safeguard of the oral cavity in children who have diabetes, as well as an higher concentration of the proinflammatory mediator S100A9* with respect to healthy children.

Objectives: Fibromyalgia (FM) is a syndrome characterized by chronic, diffuse musculoskeletal pain and by a low pain threshold at specific anatomical points (tender points). Numerous other conditions (Irritable bowel syndrome, tension-type headache, migraine headaches, etc.) may overlap with FM. Aim of this study was to evaluate the quality of life and associated clinical distress in patients with FM. Methods: 53 females affected by primary fibromyalgia and 40 healthy females were examined were examined by an experienced rheumatologist and interviewed using the Fibromyalgia Impact Questionnaire (FIQ). Clinical monitoring included Visual Analogue Scale for pain and pain pressure threshold measurements. Results: Mean FIQ scores were 66.39±14.94 in FM patients and 13.15±5.37 in control subjects and the difference was statistically significant. Among associated clinical distress higher frequencies have been found for paraesthesia (87%), sleep disturbance (72%), tension type headache (70%), oto-vestibule syndrome (72%) and irritable colon (60%). An R.O.C. bend was developed in the presence of paraesthesias and oto-vestibule syndromes at the same time. This allowed us to identify a FIQ cut off value of 66.85 so FM patients were divided into 2 groups according to their FIQ scores: severe degree and mild or slight degree. Conclusions: Based on our data, it would appear possible to use a FIQ value equal to or higher than 66.85 for the clinical picture of FM to be classified as severe.

Human saliva from a healthy donor was subjected to fractionation by gel chromatography and six pools were collected and analysed by MALDI-TOF-MS and HPLC-ESI-MS. Three peptides, corresponding to 888.3, 687.3, and 524.1 amu and SNYLYDN, YLYDN, and LYDN sequences (determined by automated Edman sequencing), were isolated from pool 4. YLYDN was not previously described in human saliva. The peptides show the common C-terminal sequence of histatin 3 and histatin 1. To exclude the possibility that the three peptides were an artifact of the purification procedure, nine samples of human saliva were collected from healthy donors, immediately acidified with 0.2% TFA, and analysed by RP-HPLC-ESI-MS. The three peptides were detected in all the analyzed samples. SNYLYDN was always found at a concentration higher than that of YLYDN and LYDN. A correlation analysis performed on quantitative data indicated that the three peptides derive only from histatin 3. Other already known histatins also were searched for in the chromatogram. Histatins 1, 2, 3, 5, 6, 7, 8, and 10 were observed, although not in all samples analyzed, whereas other minor histatins were not detected.

Human salivary cystatins, five major (S, S1, S2, SA, SN) and two minor (C and D), are multifunctional proteins playing a different role in the oral environment. Salivary cystatin SN is able to effectively inhibit lysosomal cathepsins B, C, H and L and cystatin SA inhibits cathepsins C and L in vitro. These activities suggest, particularly for cystatin SN, an important role in the control of proteolytic events in vivo. Differently, cystatins S are involved, together with statherin, in the mineral balance of the tooth. Due to their distinct role, a reliable method for identification and quantification of the different cystatins, as well as of possible truncated and derived forms, could be helpful for the assessment of the status of the oral cavity. To this purpose high-performance liquid chromatography electrospray ionization mass spectrometry (HPLC-ESI MS) was applied to the analysis of human saliva obtained from healthy subjects. All known salivary cystatins, with the exception of cystatin C, were detected. Strong evidence was also obtained for the presence in saliva of post-translational modified isoforms of cystatins, which may be related to donor habits. Cystatin SN and cystatins S, S1 and S2 were well separated by HPLC-ESI MS coupling from other components and thus this approach can be successfully applied to their quantification.