Omega-3 fatty acids as an adjunct for periodontal therapy-a review.

OBJECTIVES: The aim of this article is to present an overview of omega-3 fatty acids, their anti-inflammatory properties and potential use as an adjunct for periodontal therapy. MATERIALS AND METHODS: A general literature search was conducted to provide an overview of omega-3 fatty acids, their metabolism and anti-inflammatory properties. A more specific literature search of PubMed and EMBASE was conducted to identify articles dealing studies investigating the effects of omega-3 fatty acids in the treatment of periodontitis in animals and humans and included cross-sectional, longitudinal and intervention designs. RESULTS: To date, there is good emerging evidence that dietary supplementation with fish oil may be of some benefit and this is enhanced if combined with aspirin. All clinical intervention studies to date have been on small sample sizes, and this indicates there is need for larger and more robust clinical trials to verify these initial findings. CONCLUSIONS: Dietary supplementation with fish oil could be a cost-effective adjunctive therapy to the management of periodontal disease. CLINICAL RELEVANCE: The host modulatory properties of omega-3 fatty acids warrant further assessment of their use as an adjunct in the management of periodontitis.

Azithromycin suppresses P. gingivalis LPS-induced pro-inflammatory cytokine and chemokine production by human gingival fibroblasts in vitro.

OBJECTIVE: Azithromycin is a macrolide antibiotic that appears to have both antibacterial and anti-inflammatory properties. This study aimed to investigate the effect of azithromycin on the production of pro-inflammatory cytokines and chemokines by human gingival fibroblasts (HGF) in vitro. MATERIALS AND METHODS: The effects of azithromycin (0.1 to 10 µg/mL) on the production of interleukin-6 (IL-6), interleukin-8 (IL-8), monocyte chemoattractant protein 1 (MCP-1), and growth-regulated oncogene (GRO) by human gingival fibroblasts cultured in the presence or absence of Porphyromonas gingivalis lipopolysaccharide (LPS) was studied. Cytokine and chemokine protein levels in the culture supernatant were assessed using a Luminex® multiplex immunoassay. RESULTS: P. gingivalis LPS induced cytokine/chemokine (IL-6, IL-8, MCP-1, and GRO) protein production in HGFs, and this effect was suppressed by azithromycin at all concentrations tested. CONCLUSIONS: This study demonstrates that azithromycin suppresses P. gingivalis LPS-induced cytokine/chemokine protein production in HGF, which may explain some of the clinical benefits observed with the adjunctive use of azithromycin in the treatment of periodontitis. CLINICAL RELEVANCE: The current study examines the anti-inflammatory properties of azithromycin which may make it useful as an adjunct treatment to periodontitis. Specifically, we used azithromycin to modulate the production of pro-inflammatory cytokines by gingival fibroblasts known to be important in periodontal inflammation.

Expression of peptidylarginine deiminase-2 and -4, citrullinated proteins and anti-citrullinated protein antibodies in human gingiva.

BACKGROUND AND OBJECTIVE: The presence of citrullinated proteins, and peptidylarginine deiminase types -2 (PAD-2) and -4 (PAD-4) in periodontal tissues, determine the presence of anti-cyclic citrullinated protein antibodies (anti-CCP) in gingival crevicular fluid (GCF) and compare the expression of these proteins between inflamed and non-inflamed sites. MATERIAL AND METHODS: Tissue sections were stained using antibodies against citrullinated proteins, PAD-2 and PAD-4. RT-PCR was performed to investigate PAD-2 and PAD-4 mRNA in inflamed and non-inflamed gingival tissues. Anti-CCP antibodies in gingival crevicular fluid were detected by ELISA. RESULTS: Citrullinated proteins, PAD-2 and PAD-4 were detected in gingiva. There was a correlation between inflammation and expression of these proteins. mRNAs for PAD-2 and PAD-4 were detected in both inflamed and non-inflamed gingival tissues. Antibodies to CCP were found mostly in the GCF of individuals with periodontitis. CONCLUSION: PAD-2 and PAD-4 (protein and mRNA) as well as citrullinated proteins are present in inflamed gingiva, and anti-CCP antibodies can be detected in the GCF of some patients. Tissue expression of citrullinated proteins and PAD increased with the severity of inflammation. The presence of anti-CCP antibodies in GCF was almost exclusive to a subset of patients with periodontitis. Increased expression of these proteins in inflamed gingiva lends support to the notion that periodontal inflammation contributes to the inflammatory burden in a similar way to rheumatoid arthritis.

Biomarkers of periodontal inflammation in the Australian adult population.

BACKGROUND: Several inflammatory biomarkers are implicated in the pathogenesis of periodontitis including interleukin-1beta (IL-1beta) and C-reactive protein (CRP). This study investigated the presence of these factors in gingival crevicular fluid (GCF) and their relationship to clinical and social determinants of periodontitis in the Australian population. METHODS: Equal numbers of periodontitis cases and non-cases were sampled during oral epidemiologic examination in the National Survey of Adult Oral Health. GCF was sampled from four sites where probing pocket depth (PPD) and recession were recorded. From these, IL-1beta and CRP were quantified by ELISA and the log amount of GCF IL-1beta (pg) per person and the proportion of adults with detectable CRP was computed. RESULTS: Periodontitis cases (n = 511) had significantly higher levels of IL-1beta and CRP than non-cases (n = 562). PPD, clinical attachment loss, plaque and gingivitis indices were positively associated with elevated levels of both biomarkers. Levels of both were positively associated with age, low socio-economic position and non-Australian birth. CONCLUSIONS: The presence of IL-1beta and CRP in GCF are associated with periodontal disease parameters within the Australian population. The levels of both biomarkers are influenced by age, education and eligibility for public dental care.

Multiple isoforms of the ryanodine receptor are expressed in rat pancreatic acinar cells.

Regulation of cytosolic Ca(2+) is important for a variety of cell functions. The ryanodine receptor (RyR) is a Ca(2+) channel that conducts Ca(2+) from internal pools to the cytoplasm. To demonstrate the presence of the RyR in the pancreatic acinar cell, we performed reverse transcriptase (RT)-PCR, Western blot, immunocytochemistry and microscopic Ca(2+)-release measurements on these cells. RT-PCR showed the presence of mRNA for RyR isoforms 1, 2 and 3 in both rat pancreas and dispersed pancreatic acini. Furthermore, mRNA expression for RyR isoforms 1 and 2 was demonstrated by RT-PCR in individual pancreatic acinar cells selected under the microscope. Western-blot analysis of acinar cell immunoprecipitates, using antibodies against RyR1 and RyR2, showed a high-molecular-mass (>250 kDa) protein band that was much less intense when immunoprecipitated in the presence of RyR peptide. Functionally, permeablized acinar cells stimulated with the RyR activator, palmitoyl-CoA, released Ca(2+) from both basolateral and apical regions. These data show that pancreatic acinar cells express multiple isoforms of the RyR and that there are functional receptors throughout the cell.

Costs and benefits of a subtype-specific surveillance system for identifying Escherichia coli O157:H7 outbreaks.

We assessed the societal costs and benefits of a subtype-specific surveillance system for identifying outbreak-associated Escherichia coli O157:H7 infections. Using data from Colorado, we estimated that if it averted five cases annually, the system would recover all its costs.

Risk for rabies transmission from encounters with bats, Colorado, 1977-1996.

To assess the risk for rabies transmission to humans by bats, we analyzed the prevalence of rabies in bats that encountered humans from 1977 to 1996 and characterized the bat-human encounters. Rabies was diagnosed in 685 (15%) of 4,470 bats tested. The prevalence of rabies in bats that bit humans was 2.1 times higher than in bats that did not bite humans. At least a third of the encounters were preventable.

Diagnosis and management of the autonomously functioning thyroid nodule: the Walter Reed Army Medical Center experience, 1975-1996.

In order to characterize the clinical and laboratory features of autonomously functioning thyroid nodules (AFTNs), and to assess optimal diagnosis and management of patients with this disorder, we performed a retrospective analysis of 49 such patients over a 22-year period encompassing January 1975 to November 1996. The following data were analyzed: thyroid hormone levels, thyroid scintiscan, radioiodine uptake, fine-needle aspiration biopsy, triiodothyronine (T3) suppression testing, thyrotropin-releasing hormone (TRH) stimulation test, and thyroid ultrasound. Clinical outcomes assessed included persistent hyperthyroidism, hypothyroidism, and nodule shrinkage after treatment, or in patients followed without definitive therapy, nodule growth, spontaneous degeneration, and progression to hyperthyroidism. Biochemical hyperthyroidism, often subclinical, was found in 73.5% of patients at presentation and in an additional 24.4% of patients during subsequent follow-up. The introduction of sensitive thyrotropin (TSH) testing during the period of study resulted in a decrease in the use of the T3-suppression test and TRH stimulation test from 100% and 20%, respectively, in the period from 1976-1980, to 4% each in the period from 1991-1996. T3-thyrotoxicosis occurred in 12.2% of patients. Thyrotoxicosis at any time during the course of follow-up was positively correlated with nodule size at diagnosis. Definitive therapy, used in 42.8% of patients, consisted of radioiodine ablation (38.1%) or thyroidectomy (61.9%). No patient had recurrence of thyrotoxicosis after definitive therapy, but 25% became hypothyroid. During follow-up for a mean of 30.9 months, nodules enlarged in 25% of patients overall, or 33% of patients not receiving definitive therapy. Cystic degeneration was documented in 26.5% of patients, although this change rarely reversed subclinical hyperthyroidism. The diagnosis of an AFTN requires a demonstration of TSH-independent nodular hyperfunction. The introduction of sensitive TSH assays has simplified the evaluation of AFTN patients and revealed a high prevalence of subclinical thyroid hyperfunction in this disorder. In view of current increased awareness of adverse consequences associated with subclinical hyperthyroidism and the rarity of spontaneous resolution of hyperthyroidism in AFTN patients (despite a propensity for spontaneous hemorrhage), definitive therapy is recommended. Both radioiodine and hemithyroidectomy have high cure rates and a low posttreatment incidence of hypothyroidism.

Hyaluronan in the exfoliation syndrome.

PURPOSE: To histochemically analyze if the glycoseaminoglycan component of pseudoexfoliation material consists of hyaluronan. METHODS: The hyaluronan binding region of cartilage proteoglycans was utilized in a staining specific for hyaluronan. The technique has been described and employed previously. Human donor eyes with and without the presence of visible pseudoexfoliations were analyzed. RESULTS: Hyaluronan was found to coat the fibrillar exfoliation material on the lens, the zonules, the iris epithelium and the ciliary body. It was also found on a capsular bag following cataract surgery. CONCLUSION: In the anterior chamber hyaluronan constitutes one part of the exfoliation material. Abnormal findings of hyaluronan could be found in the cornea.

Emergency admission of patients to general surgical beds: attitudes of general practitioners, surgical trainees, and consultants in Liverpool, UK.

OBJECTIVES: To determine (a) whether doctors involved in the process of emergency surgical admission could agree about which patients should be admitted, (b) whether there were consistent differences between doctors in different specialty groups, and (c) whether these opinions were greatly influenced by non-clinical factors. DESIGN: Independent assessment of summarised case histories by three "expert" clinicians (two consultant surgeons and one general practitioner (GP)), by a group of 10 GPs, and by a group of 10 junior and senior surgeons. Experts, but not other observers, scored admissions both independently and as a consensus group. Observers indicated for each patient whether they would admit, would not admit, or were unsure. SETTING: An urban general hospital with teaching status. SUBJECTS: Fifty consecutive patients admitted to the general surgical unit as emergencies during 1995. MAIN OUTCOME MEASURES: Proportion of admissions considered unnecessary or uncertain: agreement between observers on these proportions: effect of social and procedural factors on the admission decision. RESULTS: Between 8 and 34% of admissions were considered unnecessary and 20-38% of unclear necessity. Agreement between the groups of clinicians was not good. GPs and consultant surgeons showed the poorest agreement (kappa = 0.08 to 0.25, 4 comparisons), and the GPs scored a higher percentage of admissions as unnecessary (34 v 8-12%). After discussion, the consensus group achieved good to very good agreement (kappa 0.61-0.84). CONCLUSIONS: Different groups of doctors vary widely in their views about the need for emergency surgical admission. Good agreement can be reached by consensus discussion. GPs are less likely than surgeons to consider emergency surgical admission necessary.

Acyl-coenzyme A causes Ca2+ release in pancreatic acinar cells.

The regulation of cytosolic Ca2+ is important for a variety of cell functions. One non-inositol 1,4,5-trisphosphate (IP3) compound that may regulate Ca2+ is palmitoyl-coenzyme A (CoA), a fatty acid-CoA that is reported to cause Ca2+ release from intracellular stores of oocytes, myocytes, and hepatocytes. To study the role of palmitoyl-CoA in the pancreatic acinar cell, rat pancreatic acini were isolated by collagenase digestion, permeablized with streptolysin O, and the release of Ca2+ from internal stores was measured with fura-2. Palmitoyl-CoA released Ca2+ from internal stores (EC50 = 14 microM). The palmitoyl-CoA-sensitive pool was distinct from, and overlapping with the IP3-sensitive Ca2+ pool. The effects of submaximal doses of IP3 or cyclic ADP-ribose plus palmitoyl-CoA were additive. Fatty acid-CoA derivatives with carbon chain lengths of 16-18 were the most potent and efficacious. Ryanodine and caffeine or elevated resting [Ca2+] sensitized the Ca2+ pool to the actions of palmitoyl-CoA. Fatty acid-CoA levels in pancreatic acini were measured by extraction with 2-propanol/acetonitrile, followed by separation and quantification using reverse phase high performance liquid chromatography, and were found to be 10.17 +/- 0.93 nmol/mg protein. These data suggest the presence of an IP3-insensitive palmitoyl-CoA-sensitive Ca2+ store in pancreatic acinar cells and suggest that palmitoyl-CoA may be needed for Ca2+-induced Ca2+ release.

Mechanisms of cell death after pancreatic duct obstruction in the opossum and the rat.

BACKGROUND & AIMS: Mechanisms of cell death in human and experimental pancreatitis remain poorly understood. The aim of this study was to determine the mechanisms of cell death, apoptosis vs. necrosis, in models of pancreatitis induced by pancreatic duct ligation in the opossum and rat. METHODS: Morphological changes were shown by light and electron microscopy, chromatin condensation and DNA breaks were assessed using Hoechst 33258 staining and DNA nickend labeling, and DNA fragmentation was characterized b gel electrophoresis. RESULTS: Cells with morphology of both necrosis and apoptosis were found in pancreata from duct-ligated animals; however, the apoptosis-necrosis ratio was different for the opossum and the rat. Both apoptotic and necrotic cells stained for breaks in DNA, suggesting that both apoptosis and necrosis are associated with DNA fragmentation and that DNA end labeling cannot specifically detect apoptotic cells. No significant oligonucleosomal DNA fragmentation was found in pancreas from duct-ligated animals, although the endonuclease responsible for oligonucleosomal DNA fragmentation was present in pancreatic nuclei. CONCLUSIONS: Duct ligation resulted predominantly in apoptosis in the rat and necrosis in the opossum. The differences in types of cell death in the two models may result from differences in inflammatory cell infiltration.

Isolation of subcellular agonist-sensitive calcium stores from the pancreatic acinar cell.

The purpose of the present study was to develop a technique to identify, isolate and partially purify these membrane bound compartments for further characterizations of their Ca2+ transport and storage mechanisms. We 45Ca(2+)-loaded the agonist-sensitive Ca2+ stores in rat pancreatic acini. The loading was accomplished by first depleting the stores with carbachol stimulation followed by the addition of 45Ca2+ and atropine to the extracellular media. After homogenization of the 45Ca(2+)-loaded acini, subcellular fractions were resolved on sucrose and Nycodenz gradients. 45Ca2+ fluxes were minimized during these procedures by inclusion in the media of LaCl3. Five subcellular fractions were identified that specifically accumulated 45Ca2+ after carbachol stimulation. Electron microscopic observations of the fractions demonstrated that three of the fractions consisted of rough membrane vesicles; that one consisted of a mixture of rough and smooth membrane vesicles; and that one consisted of smooth membrane vesicles. All fractions were enriched in glucose-6-phosphatase. All 5 fractions demonstrated ATP dependent 45Ca2+ uptake. By Western blot analysis, all fractions contained calnexin, p58, sarcoplasmic reticulum type Ca(2+)-ATPase, and IP3 receptor. These results demonstrated that the 45Ca(2+)-loading technique can be used to isolate and characterize distinct compartments of the agonist-sensitive Ca2+ store in the pancreatic acinar cell.

Endogenous hyaluronan in corneal disease.

PURPOSE: Hyaluronan (HA) is a disaccharide polymer capable of binding considerable amounts of water. It is present in trace amounts on the cornea endothelium, and it is not normally found in the epithelium or stroma. A specific histochemical stain was used to test for HA in a wide variety of corneal disorders. METHODS. Eighty-six human corneal tissue specimens were examined histochemically for HA. The material consisted of 84 full-thickness corneal buttons, one epithelium scraping, and one pterygium. Cases were analyzed according to the patient's sex, age, diagnosis, and localization of HA staining. RESULTS: The corneal tissue specimens came from 47 women and 39 men, average age 59 years. Fifty-seven percent of the specimens displayed abnormal HA. HA was visualized in Fuch's dystrophy, keratoconus, infections, regrafts, mechanical and chemical trauma, post-excimer ablations, dystrophies, degenerations, pseudophakic bullous keratopathy, congenital opacities, Stevens-Johnson syndrome, and others. Staining was variously seen in the epithelium, stroma, and endothelium, with intensity of staining ranging from trace amounts to extremely heavy. CONCLUSIONS: Endogenous hyaluronan production is seen in virtually the entire spectrum of corneal disorders. The presence of HA was most often associated with dividing, migrating, or fibroblast-like cells and probably represents a nonspecific tissue response to wounding. Its production is biochemically distinct from that of normally present proteoglycans. The abnormal presence of HA may reduce corneal transparency by disrupting the normal spacing between collagen fibrils, creating focal changes in the index of refraction, and altering the normal flow of solutes through the cornea.

Treatment of corneal dystrophies with excimer laser.

Thirty-three eyes with different corneal dystrophies underwent phototherapeutic keratectomy to ablate corneal opacities, surface irregularities and corneal wounds. For most patients the treatment goal was to improve vision but also corneal woundhealing, glare problems and unstable refraction were indications for treatment. In 27 eyes with the treatment goal to improve vision, 23 improved 2 lines or more on the visual chart used. In none of the patients has the treatment led to a worsening of the condition. The mean follow-up time was 9 months. Excimer laser ablation of corneal dystrophies seems to be a good treatment, which can improve visual functions considerably, heal corneal wounds and in this way postpone corneal grafting in many cases.

Nebulae at keratoconus--the result after excimer laser removal.

Ten patients underwent excimer laser ablation due to nebula formation at keratoconus. The nebulae interfered significantly with contact lens fit or wearing time. The mean follow-up time in these patients was 16.5 months. Following surgery all patients could be successfully fitted with a contact lens and thereby obtain good visual acuity. Furthermore, contact lens wearing time was 8 hours or more in all cases. In 2 patients the nebulae recurred but were successfully retreated.

Steroid treatment of myopic regression: acute refractive and topographic changes in excimer photorefractive keratectomy patients.

We documented acute changes in corneal topography and refraction after readministration of topical steroids in five patients with myopic regression after photorefractive keratectomy (PRK). One was a high myope (-10.00 sphere); the others were moderate myopes (preoperative spherical equivalents -4.25 to -6). All patients had been off steroids for 3 to 8 months before reinstitution. The changes in refractive spherical equivalents ranged from +0.75 to +3.00 diopters, and were observed from 6 days to 4 weeks later. Refractive hyperopic shifts were accompanied by pronounced corneal flattening as measured by computer-assisted corneal topographic analysis. Two to seven months later, three patients have become even less myopic, and two patients have again become slightly more myopic. Topical steroid use can modulate refractive changes after excimer laser treatments in some patients.

Excimer laser photorefractive keratectomy for myopia. Clinical results in sighted eyes.

PURPOSE: To evaluate the refractive results of excimer laser photorefractive keratectomy (PRK) performed on normal, sighted myopic eyes; to assess the role of postoperative topical steroid treatment in patients with PRK; and to study the regression of effect. METHODS: An argon fluoride 193-nm excimer laser was used. Photorefractive keratectomy was performed on 420 eyes with preoperative refraction ranging from -1.25 to -7.50 diopters (D). Minimum follow-up time was 12 months, and 194 of the eyes were followed for 15 months. Postoperative treatment generally consisted of topical dexamethasone for 3 months, but in a sub-study, some eyes were treated for only 5 weeks. RESULTS: Mean refraction (spherical equivalent +/- standard deviation) at 12 months was -0.04 +/- 0.84 D and at 15 months -0.22 +/- 0.78 D. At 12 months postoperatively, 86% of the eyes were within 1.00 D of emmetropia, at 15 months 87%. At 12 months, 91% of the eyes had an uncorrected visual acuity of at least 20/40, at 15 months 87%. Eyes treated with dexamethasone for 3 months regressed significantly less than those treated for only 5 weeks (P < 0.01). Dexamethasone also was effective in reversing regression later in the postoperative course. Eyes with preoperative myopia up to 4.90 D had significantly better refractive results at 12 months than eyes with myopia ranging from 5.00 to 7.50 D (P < 0.01). CONCLUSION: These data show that excimer laser PRK can correct myopia with good predictability. Results at 12 and 15 months tend to suggest stability of postoperative refraction. Regression of effect was more common in higher myopes. Topical steroids postoperatively seem to play a crucial role for the refractive result.

Phototherapeutic keratectomy: long-term results in 166 eyes.

One hundred sixty-six eyes with corneal disease were treated with the excimer laser in order to ablate opacities, irregularities, or to improve wound healing. Postoperative follow up time was 15 months (range 6 to 28 months). An individual goal was set for each patient and 84% achieved the goal. The visual acuity change is given for each subgroup. Phototherapeutic keratectomy (PTK) was shown to be an advantageous surgical method, where the success is in the careful preoperative evaluation. One problem with the technique is the induced hyperopia.

Hyaluronic acid in the rabbit cornea after excimer laser superficial keratectomy.

Hyaluronic acid (HA) is not normally found in the corneal stroma. Rabbit corneas were examined for the presence of stromal HA after excimer laser treatment. One eye in each of 28 rabbits received a 60 microns deep superficial keratectomy with the excimer laser. After 1, 8, 21, and 60 days, the corneas were analyzed by quantitative and histochemical methods specific for HA. A statistically significant increase in the HA concentration compared to the baseline amount in the untreated fellow eye was seen at 8, 21, and 60 days. HA was visualized histochemically in the anterior stroma of the excimer-treated eyes at all times tested. The presence of HA after excimer surgery may influence the hydration, thickness, and transparency of the cornea. The reactive production of HA in the stroma may represent a nonspecific corneal tissue response to injury.