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Selected AbstractsQuantitative analysis of MRP-8 in gingival crevicular fluid in periodontal health and disease using microbore HPLCJOURNAL OF CLINICAL PERIODONTOLOGY, Issue 12 2001Fionnuala T. Lundy Abstract Background: The protein components of GCF can be separated by reverse-phase microbore HPLC on a C18 column with detection on the basis of 214 nm absorbance. A single major symmetrical protein peak eluting with a retention time of 26 min (50% acetonitrile) was evident in gingival crevicular fluid (GCF) from periodontitis patients but not in healthy GCF. This protein was identified as human MRP-8 by N-terminal amino acid sequencing and liquid chromotography quadropole mass spectrometry. Aims: To quantify the amount of MRP-8 detectable in GCF from individual healthy, gingivitis and periodontitis affected sites and to study the relationship, if any, between the levels of this responsive protein and periodontal health and disease. Methods: GCF was sampled (30 s) from healthy, gingivitis, and periodontitis sites in peridontitis subjects (n=15) and from controls (n=5) with clinically healthy gingiva and no periodontitis. Purified MRP-8 was sequenced by Edmann degradation and the phenylthiohydantoin (PTH) amino acid yield determined (by comparison of peak area with external PTH amino acid standards). This value was subsequently used to calculate the relative amount of protein in the peak eluting with a retention time of 26.0 min (MRP-8) in individual GCF chromatograms. Results: Higher levels of MRP-8 were detected in inflammatory sites: periodontitis 457.0 (281.0) ng; gingivitis 413.5 (394.5) ng compared with periodontally healthy sites in diseased subjects 14.6 (14.3) ng and in controls 18.6 (18.5) ng, p=0.003. There was at least 20-fold more MRP-8 in the inflammatory compared with the healthy sites studied. Conclusions: The preliminary data indicate that MRP-8 is present in GCF, with significantly greater amounts present at diseased than healthy sites. A systematic study of the relationship of this protein to periodontal disease could prove useful in further clarifying whether MRP-8 could be a reliable GCF biomarker of gingivitis and periodontitis. Zusammenfassung Grundlagen: Der Proteingehalt des GCF, abgetrennt werden mit einer reversphasigen Microbore-HPLC auf einer C18-Säule mit Detektion auf der Basis der Absorption von 214 nm. Ein einziges symmetrisches Protein-Hauptpeak-Eluat, der mit einer Retentionszeit von 26 Minuten eluiert wurde (50% Acetonnitril) war in gingivalen Sulkusfluid (GCF) von Parodontitispatienten deutlich sichtbar, jedoch nicht im GCF von Gesunden. Dieses Protein wurde mitels N-terminaler Aminosären-Sequenzierung und Flüssigkeits-Chromatographie mit Quadropol-Massenspektrometrie als humanes MRP-8 identifiziert. Ziele: Quantifizierung der Menge an MRP-8, die im GCF von gesunden Personen, Patienten mit Gingivitis und mit Parodontitis nachweisbar ist und Studieren der eventuell möglichen Beziehungen zwischen den Titern dieses Reaktionsproteins und parodontaler Gesundheit bzw. Erkrankung. Methoden: Bei Parodontitispatienten (n=15) wurde das GCF von gesunden Bereichen, sowie von Stellen mit Gingivitis und Parodontitis gewonnen (30 Sek.) sowie bei Kontrollpersonen (n=5) mit klinisch gesunder Gingiva und ohne Parodontitis. Das gereinigte MRP-8 wurde mittels Edman-Degradierung Sequenziert und der Phenyl-Thiohydantoin (PTH) Aminosäure-Yield bestimmt (durch Vergleich der Peakbereiche mit externen PTH Aminosäurestandards). Darauffolgend wurde dieser Wert verwendet, um die relative Menge des Proteins im Peak-Eluat mit einer Retentionszeit von 26.0 Min. (MRP-8) in den individuellen Chromatogrammen zu berechnen. Ergebnisse: An entzündeten Stellen wurden höhere Titer von MRP-8 nachgewiesen: Parodontitis 457.0 (281.0) ng; Gingivitis 413.5 (394.5) ng verglichen mit parodontal gesunden Stellen bei erkrankten Patienten 14.6 (14.3) ng und den Kontrollen 18.6 (18.5) ng, p=0.003. An den entzündeten Stellen gab es im Vergleich mit den gesunden Stellen wenigstens 20 mal mehr MRP-8. Schlußfolgerungen: Die vorläufigen Daten zeigen, daß MRP-8 im GCF vorhanden ist und an erkrankten Stellen signifikant höhere Mengen vorhanden sind, als an gesunden Stellen. Eine systematische Studie der Beziehung dieses Proteins zur Parodondalerkrankung könnte sich als nützlich erweisen, um des weiteren zu klären, ob MRP-8 eine verläßlicher Biomarker für Gingivitis und Parodontitis ist. Résumé Origine: Les composants proéiques du fluide gingival peuvent être séparés par HPLC microbore en phase inverse sur une colonne C18 avec une détection sur une base d'absorption de 214 nm. Un unique pic majeur symétrique ayant un temps de rétention de 26 min (50% acetonitrile) était manifeste dans le fluide gingival (GCF) des patients atteints de parodontite, mais pas chez les patients sans. Cette protéine fut identifiée comme étant l'MRP-8 humaine après séquençage de l'acide aminé N terminal et spectrométrie de masse quadropole par chromatographie liquide. But: L'objectif est de quantifier la quantité de MRP-8 détectable dans le GCF de site sains, atteints de gingivite ou de parodontite et d'étudier, s'il y en a, la relation entre les niveaux de cette réponse protéique et la santé et la maladie parodontale. Méthodes: Le GCF frut prélevé (30 s) dans des sites sains, atteints de gingivite ou de parodontite, chez des sujets atteints de parodontite (n=15) ou chez des contrôles (n=5), ayant une gencive cliniquement saine sans parodontite. Le MRP-8 purifié fut séquencé par dégradation d'Edmann et le débit d'acide aminé phenylthiohydantoine (PTH) déterminé (par comparaison avec la surface de pic avec des standards d'acide aminé PTH externe). Cette valeur fut ensuite utilisée pour calculer la quantité relative de protéine dans le pic avec un temps de rétention de 26.0 mn (MRP-8) sur des chromatogrammes individuels de GCF. Résultats: De plus hauts niveaux de MRP-8 étaints détectés dans les sites inflammatoires: Parodontite 457.0 (281.0) ng; gingivite 413.5 (394.5) ng par rapport aux sites sains des sujets malades 14.6 (14.3) ng et des sujets contrôles 18.6 (18.5) ng, p=0.003. Il y avait au moins 20× plus de MRP-8 dans les sites inflammatoires par rapport au sites sains. Conclusions: Les données préliminaires indiquent que la MRP-8 est présente dans le GCF, en quantité significativement plus importante dans les sites malades. Une étude systèmatique de la relation entre cette protéine et la maladie parodontale pourrait se révéler utile pour encore plus expliciter si MRP-8 pourrait être un biomarqueur fiable du GCF des gingivites et des parodontites. [source] Plasma lipid and blood glucose levels in patients with destructive periodontal diseaseJOURNAL OF CLINICAL PERIODONTOLOGY, Issue 8 2000Wolfgang Lösche Abstract Hyperlipidaemia and hyperglycaemia are major risk factors for cardiovascular disease. In recent years, some evidence has been presented that periodontal disease is associated with an increased risk of cardiovascular disease. To further elucidate this association, we have studied standard blood chemistry variables known as risk markers for cardiovascular disease in periodontally diseased and healthy subjects. We have measured levels of plasma lipids and fasting blood glucose in 39 subjects with moderate periodontal disease (age 50,60 years) and compared the results with those obtained in 40 age- and sex-matched controls. Both groups were systemically healthy according to their medical history. Total cholesterol, low density lipoprotein cholesterol and triglycerides were significantly higher in periodontally diseased subjects by about 8% (p<0.03), 13% (p<0.003) and 39% (p<0.001), respectively, when compared to controls. Although subjects with diabetes were excluded from the study, we found significantly higher blood glucose levels in the patient than in the control group (85±25 versus 73±17 mg/dl; p<0.02). There was also a significantly higher frequency of pathological plasma lipid profiles in the patient than in the control group. The results indicate that hyperlipaemia and pre-diabetes may be associated with periodontal disease in systemically healthy subjects. These data do not allow us to decide, whether periodontal disease causes an increase in hyperlipaemia and in a prediabetic state or whether periodontal disease and cardiovascular disease share hyperlipidaemia and the prediabetic state as common risk factors. [source] Myeloid-related protein (MRP8/14) expression in gingival crevice fluid in periodontal health and disease and after treatmentJOURNAL OF PERIODONTAL RESEARCH, Issue 4 2010E. Andersen Andersen E, Dessaix IM, Perneger T, Mombelli A. Myeloid-related protein (MRP8/14) expression in gingival crevice fluid in periodontal health and disease and after treatment. J Periodont Res 2010; 45: 458,463. © 2010 John Wiley & Sons A/S Background and Objective:, Myeloid-related protein (MRP8/14) and its subunits are biomarkers of inflammation. The present study evaluated whether gingival crevice fluid levels of these markers discriminate periodontitis from healthy sites in patients with chronic periodontitis or diseased from healthy subjects, and whether these biomarkers detect longitudinal changes after therapy. Material and Methods:, Levels of MRP8/14, MRP14 and total protein were quantified in 19 periodontitis patients before non-surgical periodontal therapy, after 3 and 6 mo of treatment, and were measured once in 11 periodontally healthy subjects. In total, diseased subjects contributed 59 sites with probing depths >4 mm (PP) and 21 sites <4 mm (PH); healthy subjects contributed 91 sites (HH). Results:, Overall, in diseased subjects, MRP8/14, MRP14 and total protein were not significantly different between PP and PH sites. However, at baseline, MRP8/14 and total protein had significantly higher values at sites in periodontally diseased than in healthy subjects. Clinical improvement was associated with a significant decrease of MRP8/14 and MRP14 from baseline to month 6 in PP sites. Interestingly, a similar decrease was observed in PH sites for all three markers. At 6 mo, however, levels of MRP8/14 and protein in PP and PH sites of patients were still significantly higher than in healthy subjects. Conclusion:, Gingival crevice fluid levels of MRP8/14 did not differentiate between clinically diseased and healthy sites in patients with chronic periodontitis. However, this marker was elevated in periodontally diseased compared with healthy subjects, and its values decreased following therapy. MRP8/14 may be used to monitor the response to treatment. [source] An in vitro biofilm model of subgingival plaqueMOLECULAR ORAL MICROBIOLOGY, Issue 3 2007C. Walker Introduction:, Numerous biofilm models have been described for the study of bacteria associated with the supragingival plaque. However, there are fewer models available for the study of subgingival plaque. The purpose of this study was to develop and validate a model that closely mimicked the composition of the subgingival flora. Methods:, The model was developed as follows: calcium hydroxyapatite disks were coated overnight with 10% sterile saliva, placed in flat-bottomed tissue culture plates containing trypticase-soy broth, directly inoculated with a small aliquot of dispersed subgingival plaque, incubated anaerobically, and transferred to fresh medium at 48-h intervals until climax (steady-state) biofilms were formed (,10 days). Results:, The model, based on samples from eight periodontitis patients and eight healthy subjects, yielded a multi-species, heterogeneous biofilm, consisting of both gram-positive and gram-negative species, and comprising 15,20 cultivable species associated with the subgingival flora. The species present and their proportions were reflective of the initial cultivable subgingival flora. Comparisons of the initial plaque samples from healthy subjects and the mature biofilms showed 81% similarity in species and 70% similarity in the proportions present. Biofilms formed from samples obtained from periodontally diseased subjects were 69% similar in species and 57% similar in the proportions present. Conclusions:, The biofilm model described here closely reproduces the composition of the cultivable subgingival plaque both in the species present and in their relative proportions. Differences existed between biofilms grown from diseased and non-diseased sites with the former being characterized by the presence of periodontal pathogens at microbially significant levels. [source] Does ageing have an effect on midbrain premotor nuclei for vertical eye movements?MOVEMENT DISORDERS, Issue 6 2003Craig Henson BSc Abstract Currently, there is debate in the clinical literature as to whether defects in vertical gaze are a consequence of normal ageing or a component of an underlying neurodegenerative disorder. Although pathological changes have been demonstrated in diseased subjects, no study to date has addressed the question of normal ageing effects. In this retrospective study, we examined 23 neurologically and pathologically normal subjects (age 18,91). Using an unbiased, frame-based sampling method, we quantified neuronal and glial cell densities in 10 young (<50) and 13 aged (>65) subjects in the rostral interstitial nucleus of the medial longitudinal fasciculus (riMLF), the key premotor substrate in the vertical gaze pathway. We found no statistically significant difference in neuronal density, glial cell density, or neuron-to-glial cell ratios between the young and the aged. We conclude, therefore, that neuronal loss, neuronal atrophy, or gliosis in the riMLF are not consequences of normal ageing. © 2003 Movement Disorder Society [source] Native Fluorescence Spectroscopy of Blood Plasma in the Characterization of Oral Malignancy,PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 2 2003S. Madhuri ABSTRACT Native fluorescence characteristics of blood plasma were studied in the visible spectral region, at two different excitation wavelengths, 405 and 420 nm, to discriminate patients with different stages of oral malignancy from healthy subjects. The fluorescence spectra of blood plasma of oral malignant subjects exhibit characteristic spectral differences with respect to normal subjects. Different ratios were calculated using the fluorescence intensity values at those emission wavelengths that give characteristic spectral features of each group of experimental subjects studied. These fluorescence intensity ratios were used as input variables for a multiple linear discriminant analysis across different groups. Leave-one out cross-validation was used to check the reliability of each discriminant analysis performed. The discriminant analysis performed across normal and oral cancerous subjects classified 94.7% of the original grouped cases and 93.7% of the cross-validated grouped cases. A classification algorithm was developed on the basis of the score of the discriminant functions (discriminant score) resulted in the analyses. The diagnostic potentiality of the present technique was also estimated in the discrimination of malignant subjects from normal and nonmalignant diseased subjects such as liver diseases. In the discriminant analysis performed across the three groups, normal, oral malignancy (including early and advanced stages) and liver diseases, 99% of the original grouped cases and 95.9% of the cross-validated grouped cases were correctly classified. Similar analysis performed across normal, early stage of oral malignancy, advanced oral malignancy and liver diseases correctly classified 94.9% of the original grouped cases and 91.8% of the cross-validated grouped cases. [source] Genetic Variation in the Indoleamine 2,3-Dioxygenase Gene in Pre-eclampsiaAMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY, Issue 1 2010Haruki Nishizawa Citation Nishizawa H, Kato T, Ota S, Nishiyama S, Pryor-Koishi K, Suzuki M, Tsutsumi M, Inagaki H, Kurahashi H, Udagawa Y. Genetic variation in the indoleamine 2,3-Dioxygenase gene in pre-eclampsia. Am J Reprod Immunol 2010; 64: 68,76 Problem, To investigate the contribution of genomic variations in the indoleamine 2,3-dioxygenase (IDO) gene to the onset of pre-eclampsia. Method of study, We examined sequence variations in the IDO1 gene using placental genomic DNA from 35 pre-eclamptic patients and 32 normotensive pregnant women. Results, A case,control study revealed that none of the common variants influences the risk of disease. Sequencing of each IDO1 exon in diseased subjects revealed rare variants. This variation, c.-147_150delGAAA, was located within the 5,-untranslated region of the IDO1 gene, and its homozygote was identified only in pre-eclamptic subjects. However, despite the low levels of IDO expression and enzyme activity in the c.-147_150delGAAA homozygote, reporter assays indicated that this variation does not affect gene expression. Conclusion, Our findings indicate that genetic alteration of fetal IDO gene does not appear to be a primary cause of pre-eclampsia. [source] The effect of screening for cardio-renal risk factors on drug use in the general populationBRITISH JOURNAL OF CLINICAL PHARMACOLOGY, Issue 6 2007Jarir Atthobari What is already know about this subject ,,Screening of the population may result in medicalization. ,,There is no report about the effect of a health screening programme on drug prescribing. What this study adds ,,Screening of the general population for cardiovascular risk factors does not lead to more drug prescribing, for either screening-related or screening-unrelated drugs. ,,The incidence of drug use increases in screened subjects with high risk, but only for drugs related to the purpose of screening. ,,For screening to be successful, i.e. increased drug use in the detected diseased subjects, it has to be performed in a population expected to be at increased risk. Aim To evaluate the effect of a cardio-renal screening programme on desired and undue drug use. Methods Data from the PREVEND cohort (Prevention of REnal and Vascular ENd-stage Disease) were used. The drug use of screened (randomly) selected subjects (n = 2650) was compared with unscreened subjects, matched for age and sex (n = 10 434). Drug use in the overall PREVEND cohort, enriched for albuminuria (n = 6751), was also studied. Screening-related drugs (antihypertensive, antilipidaemic, antidiabetic and antithrombotic) were selected, as well as screening-unrelated drugs (benzodiazepines, drugs for acid-related disorders and painkillers). Time to first prescription after screening is presented as Kaplan,Meier curves. Results After 6.5 years of follow-up, the incidence of drug use was not significantly different between the screened, randomly selected and unscreened cohorts. Antihypertensives were used by 21.5 and 20.8%, respectively; antilipidaemic 12.8 and 10.2%, antidiabetics 4.0 and 3.9%, and antithrombotic 11.4 and 12.0%. Screening-unrelated drugs were used at comparable frequencies. Compared with the unscreened cohort, screening-related drugs were prescribed more frequently for subjects in the enriched cohort (25.8, 15.5, 5.5 and 13.5% for antihypertensive, antilipidaemic, antidiabetic and antithrombotic, respectively), whereas screening-unrelated drugs were used at comparable frequencies. Conclusions The incidence of drug use did not differ between the screened, randomly selected and unscreened cohorts. Screening does not lead to more drug prescription, thus arguing against the fear of undue medicalization after screening. The data also show that, for screening to be successful, it should be performed in a targeted population, such as one enriched for albuminuria. [source] |