Community survey of rabies knowledge and exposure to bats in homes - Sumter County, South Carolina, USA.

Subsequent to a human rabies death in Sumter County, South Carolina, we assessed the frequency of exposures to bats in homes and citizens' rabies knowledge. A self-administered survey was mailed to 6033 randomly selected Sumter County addresses. The survey inquired about household exposures to bats and respondents' rabies knowledge. Surveys were returned by mail for descriptive analysis. Of 597 respondents, 3.5% (21/597) reported having bats living in (2.8% or 17/597) or entering their homes (2.5% or 15/597) during 2010-2012. Respondents generally understood that mammals transmit rabies virus through bites, but were less aware of the severity of rabies illness and modern post-exposure vaccine administration. Respondents were unsure about how to exclude bats from homes and ranked highly both healthcare and non-healthcare entities as preferred resources for obtaining assistance with bat-related concerns. We found potential for human exposures to bats in Sumter County households and gaps in citizen knowledge of rabies and bat exclusion. Public health officials should engage non-healthcare partners in assistance disseminating rabies educational materials and for providing appropriate referral for persons potentially exposed to bats.

Effects of autonomic agonists and immunomodulatory cytokines on polymeric immunoglobulin receptor expression by cultured rat and human salivary and colonic cell lines.

OBJECTIVE: Immunoglobulin A (IgA) is transported across glandular epithelial cells by polymeric immunoglobulin receptor (plgR), with each receptor molecule participating in only one round of transcytosis. Nerve-related stimuli rapidly increase salivary secretion of IgA, while concentrations are increased in the autoimmune disease Sjögren's syndrome. Our aim here was to determine whether autonomic agonists and cytokines present in Sjögren's-affected glands can up-regulate salivary cell plgR expression. METHODS: Cultures of rat parotid acinar cells (PAR C5) and human submandibular gland ductal cells (HSG) were exposed to carbachol or adrenaline for 24 h and to interleukin-4 and/or interferon-gamma for 48 h. The human colonic cell line HT-29 served as a positive control for cytokine response. plgR mRNA was quantified by reverse transcription and real-time PCR and protein expression was examined by immunoblotting. RESULTS: Carbachol increased plgR mRNA levels significantly in all cells but adrenaline did so only with PAR cells (P<0.05). HSG and HT-29 cells both up-regulated plgR gene transcription on exposure to interleukin-4 and interferon-gamma either alone or in combination (P<0.05). By contrast, production of plgR mRNA in PAR cells tended to decrease in response to all cytokine treatments. plgR protein levels rose in line with mRNA expression in cytokine-treated HT-29 cultures (P<0.05). CONCLUSIONS: Autonomimetics can up-regulate plgR transcription in transformed and neoplastic salivary and colonic cells, although intracellular coupling mechanisms require further investigation. Immunomodulatory cytokines increased plgR expression in one of the salivary cell lines, but additional work is needed to establish whether this occurs in Sjögren's patients.

Secretory leukocyte protease inhibitor and its potential interactions with elastase and cathepsin B in gingival crevicular fluid and saliva from patients with chronic periodontitis.

BACKGROUND AND OBJECTIVE: Elastase is carried into the oral cavity by gingival crevicular fluid (GCF) from periodontal lesions. Our study investigated the regulation of elastase activity by secretory leukocyte protease inhibitor (SLPI) and the possible action of another GCF protease on this protective salivary component. MATERIAL AND METHODS: Whole-mouth saliva (WMS), parotid saliva (PS) and GCF were obtained from 19 patients with periodontitis. The concentrations of active elastase and cathepsin B were determined using peptide substrates. SLPI and alpha1-proteinase inhibitor (alpha1PI) concentrations were determined using enzyme-linked immunosorbent assays (ELISAs). The molecular forms of SLPI were examined by immunoblotting. RESULTS: The molar concentrations of elastase, cathepsin B and alpha1PI were higher in GCF than in WMS and especially PS (p < 0.0002). The GCF SLPI concentrations were also higher than the WMS SLPI concentrations (p < 0.05). All WMS components increased with GCF content, significantly for elastase and SLPI (p < 0.002). In GCF, the concentration of alpha1PI was higher than the concentration of SLPI (p < 0.0002), while there was no significant difference for WMS. SLPI and elastase levels in GCF and WMS were inversely related (p < 0.005). In SLPI immunoblots, PS contained only the intact 14-kDa molecule of SLPI, while WMS also contained an 8-kDa fragment. For WMS there was a positive correlation between SLPI degradation and cathepsin B (p < 0.002). Incubation of WMS alone or of PS with GCF in the presence of cysteine proteinase activators caused SLPI immunoreactivity to shift to 8 kDa. CONCLUSION: For GCF, serum-derived alpha1PI is the major elastase inhibitor, but in WMS SLPI probably reduces activity. The inflamed gingivae can be an additional source of SLPI in the oral cavity, but here the molecule is apparently cleaved by GCF cysteine proteinases, such as cathepsin B.

Collagen degradation by interleukin-1beta-stimulated gingival fibroblasts is accompanied by release and activation of multiple matrix metalloproteinases and cysteine proteinases.

OBJECTIVE: Several collagenolytic matrix metalloproteinases (MMPs) have recently been identified in gingival fibroblasts, while secreted cysteine proteinases could also participate in connective tissue destruction in periodontitis. To clarify their involvement, we examined enzyme release during collagen breakdown by cultured cytokine-stimulated fibroblasts. MATERIALS AND METHODS: Gingival fibroblasts were derived from four chronic periodontitis patients and cultured on collagen gels in serum-free medium for 1-4 days. Collagenolysis was measured by hydroxyproline release into the medium. Proteinases were assessed by electrophoresis and immunoblotting. RESULTS: Adding interleukin-1beta resulted in progressive gel breakdown. This was associated particularly with a shift in MMP-1 band position from proenzyme to active enzyme and the appearance of active as well as proenzyme forms of cathepsin B. There was also partial processing of pro-MMP-13 and increased immunoreactivity for active cathepsin L. In addition, both pro-forms and active forms of MMP-8, membrane-type-1-MMP and MMP-2 were present in control and treated cultures. CONCLUSIONS: Fibroblast MMP-1 was most likely responsible for collagen dissolution in the culture model, while cathepsin B may have been part of an activation pathway. All studied proteinases contribute to extracellular matrix destruction in inflamed gingival tissue, where they probably activate each other in proteolytic cascades.

Collagenase-2 (MMP-8) and collagenase-3 (MMP-13) in adult periodontitis: molecular forms and levels in gingival crevicular fluid and immunolocalisation in gingival tissue.

AIM: To determine the cellular and molecular forms of MMP-8 (collagenase-2) and MMP-13 (collagenase-3) associated with chronic adult periodontitis by examining the species present in gingival crevicular fluid (GCF) and enzyme distribution in gingival tissue. METHODS: 30-s GCF samples were collected directly from the periodontal pockets of 12 untreated patients using filter paper strips. After elution into buffer, the samples were examined by Western immunoblotting with polyclonal antibodies for MMP-8 and MMP-13 and quantification by scanning image analysis. Individual band intensities were expressed as a percentage of total sample absorbance and mean patient values were calculated. Gingival tissue from 6 patients was fixed in formalin and embedded in paraffin wax. MMP-8 and MMP-13 were localised using the same antibodies and an avidin-biotin-peroxidase detecting system. Double staining was performed with a contrasting substrate reaction. RESULTS: The majority of MMP-8 staining in pre-treatment GCF was present in 80, 75 and 60 kD bands corresponding to prepro-, pro- and active forms of PMN-type enzyme. 43 and 38 kD bands evidently represented active, fibroblast-type MMP-8. Immunoreactivities at >100 kD and < or =30 kD were probably enzyme-inhibitor complex and degraded fragments, respectively. MMP-13 was seen mainly as 60 kD proenzyme with some 40 kD active enzyme and a small proportion of >100 kD complex. The percentages of MMP-8 PMN-type enzyme and MMP-13 proenzyme bands correlated significantly with gingival and bleeding indices (p<0.05). Immunohistochemistry demonstrated MMP-8 in PMNs, sulcular epithelial and also plasma cells in inflamed gingival connective tissue. MMP-13 immunoreactivity was detected in the sulcular epithelium and in macrophage-like cells. CONCLUSION: Multiple species and elevated levels of both MMP-8 and MMP-13 from many rather than single cellular sources in the diseased periodontium are identified in untreated periodontitis GCF and active forms contribute to GCF collagenase activity.

Matrix metalloproteinase-8 levels and elastase activities in gingival crevicular fluid from chronic adult periodontitis patients.

AIM: To make an initial assessment of the periodontal diagnostic potential of immunoreactive matrix metalloproteinase-8 (MMP-8) in gingival crevicular fluid (GCF) by comparison with elastase activity which has previously been associated with disease severity and progression. METHODS: GCF was collected from molar and premolar sites of 16 chronic adult periodontitis patients before treatment and 13 of this group 2 weeks after scaling and root planing. Samples were analysed for MMP-8 by immunofluorometric assay and for elastase activity with a fluorogenic substrate. RESULTS: Mean patient clinical parameters and GCF enzyme totals both decreased significantly after treatment. Total MMP-8 levels and elastase activities generally correlated significantly with gingival and bleeding indices. For GCF concentrations, only MMP-8 showed a significant fall after treatment, and some significant correlations with clinical parameters. Amounts of the 2 enzymes correlated significantly with each other. CONCLUSIONS: Similarities between MMP-8 and elastase probably reflect the fact that both enzymes are associated mainly with neutrophils: MMP-8 levels may have fallen more after treatment because the assay, unlike that for elastase, would most likely not have detected much enzyme bound to alpha-macroglobulin. The immunoassay for MMP-8 is more specific and convenient than functional collagenase assays, and might be suitable for monitoring the periodontal condition.

Differences in bcl-2- and bax-independent function in regulating apoptosis in sensory neuron populations.

Bcl-2 and Bax are cytoplasmic proteins that have antagonistic actions on apoptosis. To investigate the extent to which these proteins function independently in regulating neuronal apoptosis, we studied the in vivo and in vitro development of two populations of sensory neurons of mouse embryos that lack one or both proteins. Absence of Bcl-2 increased neuronal apoptosis and reduced the number of neurons in both the trigeminal and nodose ganglia during the period of naturally occurring neuronal death. Absence of Bax reduced neuronal apoptosis and increased the number of surviving neurons in these ganglia and promoted sustained neuronal survival in neurotrophin-free cultures. In contrast, the elimination of both Bcl-2 and Bax had different consequences for these populations of neurons. In nodose ganglia, apoptosis was suppressed just as effectively in embryos lacking both proteins as in embryos lacking Bax alone, and neurons that lacked both proteins survived just as effectively in neurotrophin-free medium as Bax-deficient neurons. This suggests that for nodose neurons, the suppression of apoptosis by Bcl-2 is entirely dependent on the presence of Bax. In trigeminal ganglia, although neuronal apoptosis was reduced in embryos lacking both proteins compared with wild-type embryos, there were significantly more apoptotic neurons and significantly fewer surviving neurons in embryos lacking both proteins compared with Bax-deficient embryos, and significantly fewer trigeminal neurons from embryos lacking both proteins survived in neurotrophin-free medium compared with trigeminal neurons that lacked Bax alone. This suggests that for trigeminal neurons, Bcl-2 functions partly independently of Bax in regulating survival. Our results therefore suggest that the relative independence of Bcl-2 and Bax in regulating neuronal survival differs from one population of neurons to another.

Advances in periodontal diagnosis. 10. Potential markers of bone resorption.

This paper describes the markers of bone resorption which might serve as potential markers for periodontal disease activity. It firstly describes the bone specific proteins which are involved in bone mineralisation and the role they play in this process. It then explains how they may pass into GCF and reviews those studies which have attempted to relate these factors to periodontal disease severity and activity. It next discusses the difficulties in isolating and detecting these factors in GCF and their possible use as markers for periodontal disease activity. As the final part in the series it lastly discusses the possible uses of predictive diagnostic tests of periodontal disease activity in dental practice.

Advances in periodontal diagnosis. 9. Potential markers of cell death and tissue degradation.

This paper describes the potential markers of cell death and connective tissue degradation which might serve as markers of periodontal disease activity. The first section deals with enzymes released by dead and degenerating cells. Firstly, it describes how these pass from the periodontal tissues into gingival crevicular fluid (GCF) and explains that these enzymes have been used as markers of cell death in medicine for several decades. It then discusses the main enzymes in this group, aspartate amino transferase (AST) and lactate dehydrogenase (LDH) and reviews those studies which have attempted to relate these enzymes to periodontal disease severity and activity. Secondly, it describes the potential markers of connective tissue degradation, fibronectin, hydroxyproline-containing peptides and glycosaminoglycans (GAGs) and explains how these are produced. Finally, it describes the only commercial test kit for markers in this group (GCF-AST).

Advances in periodontal diagnosis. 7. Proteolytic and hydrolytic enzymes link with periodontitis.

Biomarkers of periodontal disease activity may be obtained from potential proteolytic and hydrolytic enzymes of inflammatory cell origin. Studies that have sought to correlate these enzymes with periodontal disease activity are reviewed with special consideration given to collagenases, cysteine, aspartate and serine proteinases, beta-glucuronidase, arylsulphate, alkaline and acid phosphatases, myeloperoxidase, lysozyme and lactoferrin.

Advances in periodontal diagnosis. 8. Commercial diagnostic kits based on GCF proteolytic and hydrolytic enzyme levels.

Biomarkers of periodontal disease activity may be obtained from potential proteolytic and hydrolytic enzymes of inflammatory cell origin. Commercial diagnostic tests and those under development are discussed along with their advantages and disadvantages.

Advances in periodontal diagnosis. 6. Proteolytic and hydrolytic enzymes of inflammatory cell origin.

Potential proteolytic and hydrolytic enzymes of inflammatory cell origin might serve as biomarkers of periodontal disease activity. The role of these enzymes in periodontal pathology, particularly in respect of collagen and proteoglycan degradation, is discussed. The cellular location of these enzymes and their normal control mechanisms by endogenous inhibitors is described.

Advances in periodontal diagnosis. 5. Potential inflammatory and immune markers.

The potential inflammatory and immune markers that might detect periodontal disease severity or activity are examined. The role of inflammatory and immune factors passing from the tissues into the gingival crevicular fluid (GCF) are considered. GCF sampling is a necessary part of all diagnostic tests based on gingival and periodontal tissue factors. Inflammatory and immune factors found in GCF are reviewed with special reference to the humoral immune response, complement, cytokines and prostaglandins. The possible development of diagnostic tests based on these factors are discussed.

Advances in periodontal diagnosis. 3. Assessing potential biomarkers of periodontal disease activity.

The methods for assessing potential biomarkers of periodontal disease activity are considered. This is necessary because a detailed examination of all the relevant research evidence is an essential process in assessing the possible clinical usefulness of a periodontal diagnostic test system based on any one of these markers.

Advances in periodontal diagnosis. 4. Potential microbiological markers.

The potential of bacterial markers for the detection of periodontal disease activity is discussed. Chronic periodontitis has been associated with subgingival bacteria. Techniques are available to detect and distinguish different bacteria. In subgingival plaque, bacterial proteases can be detected. Commercial diagnostic tests based on bacterial factors are described.

Advances in periodontal diagnosis. 2. New clinical methods of diagnosis.

This paper describes the new methods of periodontal diagnosis. An outline of the problems of periodontal probing techniques is followed by a discussion of the advances made by the use of constant pressure electronic probes. We finally highlight the problems of periodontal radiographical techniques, leading on to a description of computer-aided radiographic methods--in particular, digital subtraction radiology and computer-assisted linear radiology.

Cathepsin B, alpha2-macroglobulin and cystatin levels in gingival crevicular fluid from chronic periodontitis patients.

Gingival crevicular fluid (GCF) was collected from 16 molar and premolar sites in each of 20 chronic periodontitis patients before and after periodontal therapy using filter paper strips. These were eluted individually into buffer for determination of cathepsin B and its endogenous inhibitors, alpha2-macroglobulin and cystatin. Cathepsin B activity was assayed with a fluorogenic peptide substrate, alpha2-macroglobulin by enzyme-linked immunosorbent assay and cystatin activity by inhibition of papain. Total amounts of enzyme and inhibitor per GCF sample decreased after treatment and correlated positively with pocket depth and gingival, bleeding and plaque indices. These comparisons were nearly always statistically significant for pooled site data and sometimes so for mean patient values. The amounts of alpha2-macroglobulin and cystatin were greater than those of cathepsin B and, surprisingly, enzyme and inhibitor levels correlated positively with each other. Experiments with purified reagents, however, demonstrated that the cathepsin B: alpha2-macroglobulin complex was still active against the low molecular weight substrate and that cystatin levels in GCF are probably insufficient to inhibit the enzyme substantially These factors may explain why GCF cathepsin B activity reflects the clinical status of periodontal lesions and has been identified in another study as a promising indicator of disease progression.

Advances in periodontal diagnosis. 1. Traditional clinical methods of diagnosis.

This series will consider the limitations of traditional periodontal diagnostic techniques and the recent advances which have sought to overcome them. It will cover improvements in probing and radiographic techniques and also discusses the attempts to find bacterial or tissue-derived markers which will diagnose or predict periodontal disease activity. The first paper describes the traditional methods of periodontal diagnosis.

Outcome after incision and drainage with fistulotomy for ischiorectal abscess.

Concomitant anal fistulotomy (F) and incision and drainage (I&D) of ischiorectal abscesses (IA) are often avoided, for fear of irreversibly impairing anal continence. However, failure to identify and treat the frequently associated trans-sphincteric anal fistula dooms the patient to recurrent anal suppurative disease. We have employed an aggressive approach of performing I&D and F for IA at the time of initial presentation. Adequate drainage is assured by placement of counterincisions and Penrose drains to minimize the time for healing of the perianal wound. Drainage is followed by a careful examination of the anal canal for fistula localization followed by fistulotomy, or less frequently by cutting seton placement. We present our experience with this approach to IA, with special attention paid to the evaluation of recurrence rates and anal continence. This paper represents a retrospective review of 80 patients with IA managed from 1983 to 1996. Operative records and office records were reviewed, and follow-up data were obtained by telephone interview. Internal fistulous openings were identified in 55 (68.8%) patients. Surgeries included: 38 (47.5%) I&D and F, 8 (10%) I&D and seton, and 34 (42.5%) I&D alone. Follow-up data were available on 99 per cent of patients; mean, 44.3 months. Results showed a 44 per cent recurrence rate in those who underwent I&D as compared with 21.1 per cent following I&D and F. 11.8 per cent of patients treated with I&D experienced a change in their level of continence postoperatively as compared to 15.8 per cent treated with I&D and F. The results indicate that an aggressive approach to IA allows identification of a trans-sphincteric fistula in 57.5 per cent of patients with IA. Therefore, optimal surgical management for IA appears to be I&D and F, resulting in a lower recurrence rate and comparable morbidity as compared to I&D alone.

Characterization of protease activities in Capnocytophaga spp., Porphyromonas gingivalis, Prevotella spp., Treponema denticola and Actinobacillus actinomycetemcomitans.

Protease activities in cell sonicates of defined bacterial strains were examined using peptide substrates and class-specific inhibitors. Capnocytophaga spp. all produced serine dipeptidyl peptidase activity and arginine/lysine, elastase- and chymotrypsin-like enzymes with some metalloprotease characteristics. The elastase-like activity was strongest in Capnocytophaga sputigena, but the others were greatest in Capnocytophaga gingivalis. The latter also had a separate arginine-specific enzyme which appeared not to be present in the other two species. Porphyromonas gingivalis showed serine dipeptidyl peptidase activity and very strong arginine and lysine cysteine protease activities. Prevotella spp. had inhibitor-resistant dipeptidyl peptidase activity and arginine cysteine protease activity that was much weaker but biochemically similar to P. gingivalis. Treponema denticola possessed a strong trypsin-like serine protease activity as well as very weak dipeptidyl peptidase and chymotrypsin-like activities that were sensitive to some cysteine protease reagents. Actinobacillus actinomycetemcomitans showed a novel alanine- and lysine-specific activity, but its nature was unclear.