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Periodontal destruction can be the result of different known and yet-to-be-discovered biological pathways. Recent human genetic association studies have implicated interferon-gamma inducible protein 16 (IFI16) and absent in melanoma 2 (AIM2) with high periodontal interleukin (IL)-1β levels and more destructive disease, but mechanistic evidence is lacking. Here, we sought to experimentally validate these observational associations and better understand IFI16 and AIM2's roles in periodontitis.Periodontitis was induced in Ifi204sup-/-/sup(IFI16 murine homolog) and Aim2sup-/-/supmice using the ligature model. Chimeric mice were created to identify the main source cells of Ifi204 in the periodontium. IFI16-silenced human endothelial cells were treated with periodontal pathogens in vitro. Periodontal tissues from Ifi204sup-/-/supmice were evaluated for alveolar bone (micro-CT), cell inflammatory infiltration (MPO+ staining), Il1b (qRT-PCR), and osteoclast numbers (cathepsin K+ staining).Ifi204-deficient micegt; exhibitedgt;20% higher alveolar bone loss than wild-type (WT) (P lt; 0.05), while no significant difference was found in Aim2sup-/-/supmice. Ifi204's effect on bone loss was primarily mediated by a nonbone marrow source and was independent of Aim2. Ifi204-deficient mice had greater neutrophil/macrophage trafficking into gingival tissues regardless of periodontitis development compared to WT. In human endothelial cells, IFI16 decreased the chemokine response to periodontal pathogens. In murine periodontitis, Ifi204 depletion elevated gingival Il1b and increased osteoclast numbers at diseased sites (P lt; 0.05).These findings support IFI16's role as a novel regulator of inflammatory cell trafficking to the periodontium that protects against bone loss and offers potential targets for the development of new periodontal disease biomarkers and therapeutics.

Aim: This in vivo experimental study investigated bacterial microbiome and metabolome longitudinal changes associated with enamel caries lesion progression and arrest. Methods: We induced natural caries activity in three caries-free volunteers prior to four premolar extractions for orthodontic reasons. The experimental model included placement of a modified orthodontic band on smooth surfaces and a mesh on occlusal surfaces. We applied the caries-inducing protocol for 4- and 6-weeks, and subsequently promoted caries lesion arrest via a 2-week toothbrushing period. Lesions were verified clinically and quantitated via micro-CT enamel density measurements. The biofilm microbial composition was determined via 16S rRNA gene Illumina sequencing and NMR spectrometry was used for metabolomics. Results: Biofilm maturation and caries lesion progression were characterized by an increase in Gram-negative anaerobes, including Veillonella and Prevotella. Streptococcus was associated caries lesion progression, while a more equal distribution of Streptococcus, Bifidobacterium, Atopobium, Prevotella, Veillonella, and Saccharibacteria (TM7) characterized arrest. Lactate, acetate, pyruvate, alanine, valine, and sugars were more abundant in mature biofilms compared to newly formed biofilms. Conclusions: These longitudinal bacterial microbiome and metabolome results provide novel mechanistic insights into the role of the biofilm in caries progression and arrest and offer promising candidate biomarkers for validation in future studies.

The oral cavity plays a critical role as the seeder and gatekeeper of the microbiome that populates the continuum of mucosal surfaces of the gastrointestinal tract, as well as that of the respiratory tract. The mouth has a variety of discrete niches and environmental conditions (microhabitats) that select for and discriminate against a vast array of microorganisms that ultimately determine the microbiome. The oral microbiome is an important contributor to host health and refers specifically to the microorganisms that reside on or in the human oral cavity and its contiguous mucosal surfaces to the distal esophagus. The oral microbiome is composed of approximately 700 species of bacteria, and also includes viruses, fungi, protozoa and archaea associated with the varied microhabitats that define the oral microbial ecosystem. The normal microbiota of the mouth is responsible for maintaining homeostasis of the oral cavity, but is also responsible for two of the most common diseases of bacterial etiology in humans—dental caries and periodontal diseases. Oral diseases have also been linked to systemic chronic diseases including: cardiovascular disease, stroke, abnormal pregnancy outcomes, diabetes, aspiration pneumonia, cancers and Alzheimer’s disease. This Chapter aims to highlight the unique features of the main niches that compose the oral cavity and have influence on its microbiome composition.

The oral cavity is influenced by the dietary characteristics of each individual. It is in the oral cavity that food will cause the first impact within the human body and its microbiome, due to its composition and consistency. On the other hand, the oral microbiome will affect food processing and impact the human gut microbiome, since bacterial biofilm that is processed within saliva forms the food bolus, which will then be swallowed. The mouth is one of the most heavily colonized parts of our bodies and its microbiome consists of microorganisms that live in symbiosis with healthy individuals who have adequate dietary and oral hygiene habits. Nevertheless, perturbations in the microbiome due to certain stress factors, such as high carbohydrate intake and biofilm accumulation, can lead to dysbiosis and the development of oral diseases. The most prevalent diseases in the oral cavity are dental caries and periodontal diseases including gingivitis and periodontitis, but endodontic (pulp) and soft tissue infections are also prevalent. Thus, this chapter will describe the influence of dietary habits on the oral microbiome, the development of prevalent oral diseases, and their relation to the gut microbiome.

An early childhood carie (ECC) is an extremely destructive form of tooth decay. The aim of this study was to investigate the action of ozone (O3), and the association of sodium fluoride (NaF) with chlorhexidine (CHX) on bacteria related to ECC. Overnight culture of the bacteria was performed. On exponential phase the suspension was adjusted (101-108 CFU/mL). A drop (10μL) of each concentration of bacteria was applied on sheep blood agar plates and treated with O3 (2, 20, 200, and 2,000 ppm); after 18 hours, recovery analysis of CFU verified the reduction of bacterial activity. For NaF-CHX, sterile 96-well plates were prepared and divided into groups: G1 (150 µL TSB); G2 (20 µL of bacteria + 25 µL CHX + 25 µL NaF); and G3 (150 µL TSB + 20 µL of bacteria + 50 µL water). The plates were verified by analysis of the optical density (0, 12, 14, 16, and 18 hours). The data from O3 test were submitted to ANOVA and Tukey’s test (p < 0.05). For the data from NaF-CHX, the ANOVA 2-way and Bonferroni’s test (p < 0.05) were used. The number of CFU/mL showed death > 3log10 (99.9%) for all bacteria (ozone ≥ 20ppm), while the combination of NaF-CHX was more effective (p < 0.001) compared to each substance tested alone and the control group. The antimicrobial agents tested were able to inhibit all bacteria tested; O3 seemed to be a good alternative for controlling progression of carious lesions, while the association of NaF-CHX showed to be a good antimicrobial with easy and inexpensive application.

The localization of Burkholderia cepacia complex (Bcc) bacteria in cystic fibrosis (CF) lungs, alone or during coinfection with Pseudomonas aeruginosa , is poorly understood. We performed immunohistochemistry for Bcc and P. aeruginosa bacteria on 21 coinfected or singly infected CF lungs obtained at transplantation or autopsy. Parallel in vitro experiments examined the growth of two Bcc species, Burkholderia cenocepacia and Burkholderia multivorans , in environments similar to those occupied by P. aeruginosa in the CF lung. Bcc bacteria were predominantly identified in the CF lung as single cells or small clusters within phagocytes and mucus but not as “biofilm-like structures.” In contrast, P. aeruginosa was identified in biofilm-like masses, but densities appeared to be reduced during coinfection with Bcc bacteria. Based on chemical analyses of CF and non-CF respiratory secretions, a test medium was defined to study Bcc growth and interactions with P. aeruginosa in an environment mimicking the CF lung. When test medium was supplemented with alternative electron acceptors under anaerobic conditions, B. cenocepacia and B. multivorans used fermentation rather than anaerobic respiration to gain energy, consistent with the identification of fermentation products by high-performance liquid chromatography (HPLC). Both Bcc species also expressed mucinases that produced carbon sources from mucins for growth. In the presence of P. aeruginosa in vitro , both Bcc species grew anaerobically but not aerobically. We propose that Bcc bacteria (i) invade a P. aeruginosa -infected CF lung when the airway lumen is anaerobic, (ii) inhibit P. aeruginosa biofilm-like growth, and (iii) expand the host bacterial niche from mucus to also include macrophages.

Clinical Relevance All materials tested, especially iBond, have a potential long-term antibacterial effect against the oral bacteria tested.

Background Aerosols and spatter are concerns in health care owing to their potential adverse health effects. The Isolite illuminated isolation system (Isolite Systems) and a saliva ejector were compared for aerosol and spatter reduction during and after ultrasonic scaling. Methods Fifty participants were randomized to control (n = 25, saliva ejector) or test (n = 25, Isolite) groups and received a prophylaxis with an ultrasonic scaler. Aerosols were collected in a petri dish containing transport media, dispersed, and plated to anaerobic blood agar to determine colony-forming units (CFUs). The authors analyzed the data using a t test. Results No significant difference occurred between groups in aerosol and spatter reduction ( P = .25). Mean (standard deviation) of log 10 CFUs per milliliter collected during ultrasonic scaling in the control and test groups were 3.61 (0.95) and 3.30 (0.88), respectively. All samples contained α-hemolytic streptococci, and many samples contained strictly oral anaerobes. Conclusions A significant amount of contamination occurred during ultrasonic scaling in both groups, as indicated by high numbers of CFUs and the identification of strictly oral anaerobes in all plates. Practical Implications Neither device reduced aerosols and spatter effectively, and there was no significant difference in reduction between the 2 devices. Additional measures should be taken with these devices to reduce the likelihood of disease transmission.

Dental caries is the most prevalent disease in humans globally. Efforts to control it have been invigorated by an increasing knowledge of the oral microbiome composition. This study aimed to evaluate the bacterial diversity in occlusal biofilms and its relationship with clinical surface diagnosis and dietary habits. Anamneses were recorded from thirteen 12-year-old children. Biofilm samples collected from occlusal surfaces of 46 permanent second molars were analyzed by 16S rRNA amplicon sequencing combined with the BLASTN-based search algorithm for species identification. The overall mean decayed, missing and filled surfaces modified index [DMFSm Index, including active white spot lesions (AWSL)] value was 8.77±7.47. Biofilm communities were highly polymicrobial collectively, representing 10 bacterial phyla, 25 classes, 29 orders, 58 families, 107 genera, 723 species. Streptococcus sp_Oral_Taxon_065, Corynebacterium matruchotii, Actinomyces viscosus, Actinomyces sp_Oral_Taxon_175, Actinomyces sp_Oral_Taxon_178, Actinomyces sp_Oral_Taxon_877, Prevotella nigrescens, Dialister micraerophilus, Eubacterium_XI G 1 infirmum were more abundant among surfaces with AWSL, and Streptococcus gordonii, Streptococcus sp._Oral_Taxon_058, Enterobacter sp._str._638 Streptococcus australis, Yersinia mollaretii, Enterobacter cloacae, Streptococcus sp._Oral_Taxon_71, Streptococcus sp._Oral_Taxon_F11, Centipeda sp._Oral_Taxon_D18 were more abundant among sound surfaces. Streptococcus mutans was detected on all surfaces in all patients, while Streptococcus sobrinus was detected only in three patients (mean relative abundances 7.1% and 0.6%, respectively). Neither species differentiated healthy from diseased sites. Diets of nine of the subjects were scored as high in fermentable carbohydrates (≧2X/day between meals). A direct association between relative abundances of bacteria and carbohydrate consumption was observed among 18 species. High consumption of fermentable carbohydrates and sound surfaces were associated with a reduction in bacterial diversity. PCoA plots displayed differences in bacterial community profiles between sound and diseased surfaces. Our study showed that, in addition to mutans streptococci, other species may be associated with the initiation of dental caries on occlusal surfaces, and that biofilm diversity of tooth surfaces is influenced by carbohydrate consumption and a surface’s health status.

Coaggregation between Porphyromonas gingivalis and Fusobacterium nucleatum strains was previously studied using either a semi-quantitative macroscopic assay or radioactive tracer assays. A new automated microtiter plate assay is introduced, in which the plate reader (Vmax) was adapted to allow quantitative evaluation of the kinetics of coaggregation. F nucleatum PK 1594 coaggregated with P. gingivalis HG 405 with a maximal coaggregation rate of 1.05 mOD/min, which occurred at a P. gingivalis to F. nucleatum cell ratio of 1 to 2. F. nucleatum PK 1594 failed to do so with P. gingivalis strains A 7436 or ATCC 33277. Galactose inhibition of this coaggregation could be quantitatively measured over a wide range of concentrations to demonstrate its dose-dependent manner. P. gingivalis HG 405 failed to coaggregate with F. nucleatum strains ATCC 25586 and ATCC 49256. The assay used in the present study is a sensitive and efficient quantitative automated tool to study coaggregation and may replace tedious radioactive tracer assays.

Glycogen storage disease type 1a (GSD 1a) is caused by a deficiency in microsomal glucose-6-phosphatase (G6Pase). A variant (GSD 1b) is caused by a defect in the transport of glucose-6-phosphate (G6P) into the microsome and is associated with chronic neutropenia and neutrophil dysfunction. Mutually exclusive mutations in the G6Pase gene and the G6P transport gene establish GSD la and GSD 1b as independent molecular processes and are consistent with a multicomponent translocase catalytic model. A modified translocase/catalytic unit model based on biochemical data in a G6Pase knockout mouse has also been proposed for G6Pase catalysis. This model suggests coupling of G6Pase activity and G6P transport. A 5-mo-old girl with hypoglycemia, hepatomegaly, and lactic acidemia was diagnosed with GSD 1a. She also developed neutropenia, neutrophil dysfunction, and recurrent infections characteristic of GSD 1b. Homozygous G188R mutations of the G6Pase gene were identified, but no mutations in the G6P translocase gene were found. We have subsequently identified a sibling and two unrelated patients with similar genotypic/phenotypic characteristics. The unusual association of neutrophil abnormalities in patients with homozygous G188R mutations in the G6Pase gene supports a modified translocase/catalytic unit model.

The gingival crevicular fluid (GCF) and monocytic secretion of prostaglandin E2 (PGE2) and interleukin 1 beta (IL-1 beta) were measured in a group of 39 insulin-dependent diabetes mellitus (IDDM) patients and 64 systemically healthy individuals. Diabetics were divided into Group A (gingivitis or mild periodontal disease) and Group B (moderate or severe periodontal disease). Diabetics had significantly higher GCF levels of both PGE2 and IL-1 beta as compared to non-diabetic controls who were matched with regard to periodontal disease severity (P0.00001 and P = 0.0005, respectively). Within the diabetic population, the GCF levels of these inflammatory mediators were almost 2-fold higher in Group B as compared to Group A (P = 0.01, P = 0.006, respectively for GCF-PGE2 and IL-1 beta). Furthermore, diabetics as a group had a significantly higher monocytic PGE2 and IL-1 beta production in response to various concentrations of both Escherichia coli and Prophyromonas gingivalis lipopolysaccharide (LPS) as compared to non-diabetic patients with adult periodontitis (P = 0.0001). LPS dose-response curves demonstrated that monocytes from Group B diabetics produced approximately 3 times more PGE2 than Group A monocytes; however, there was no significant difference in monocytic IL-1 beta secretion within the IDDM patients. The levels of GCF or monocytic mediators did not correlate with age, race, or glycosylated hemoglobin (HbA1C) levels. Our data suggest that the high GCF and monocytic secretion of PGE2 and IL-1 beta in IDDM patients may be a consequence of a systemic response trait and that the presence of Gram-negative infections such as periodontal diseases may interact synergistically to yield high local levels of these mediators and a more severe periodontal condition.

The aim of the present study was to identify whether monocytic TNF alpha secretion patterns could serve as a potential phenotypic discriminator for periodontal disease susceptibility within insulin-dependent diabetes mellitus (IDDM) patients. In 32 IDDM individuals the lipopolysaccharide (LPS) stimulated monocytic TNF alpha secretion dose-response characteristics were analyzed and related to two different periodontal status categories. Diabetics were divided into group A (gingivitis or mild periodontal disease) and group B (moderate to severe periodontal disease). In addition, 17 non-diabetic individuals with various degrees of periodontal disease served as control patients. Diabetics as a group had a significantly higher monocytic TNF alpha production in response to increasing Porphyromonas gingivalis A 7436 lipopolysaccharide concentrations (0, 0.003, 0.03, 0.3 and 3.0 micrograms/ml) as compared to non-diabetic patients with gingivitis or adult periodontitis (p < 0.05). A significant difference in the dose response was also noted in the level of TNF alpha secreted as a function of P. gingivalis LPS concentrations between group A and B diabetics, as determined by two-way repeated measurements ANOVA (p < 0.05). Furthermore, there was no significant difference in the mean HbA1C between the two diabetic groups, and the TNF alpha level was not significantly associated with the HbA1C level within diabetic patients. These data suggest that the diabetic state results in an upregulated monocytic TNF alpha secretion phenotype (4.6-fold increase) which, in the presence of Gram-negative bacterial challenge, is associated with a more severe periodontal disease expression. In addition, approximately 40% (10 of 24) IDDM periodontitis patients in group B demonstrated a 62-fold elevation in TNF alpha secretion relative to non-diabetic gingivitis or periodontitis patients and a 13.5-fold increase relative to IDDM group A (gingivitis or mild periodontitis) patients.

Objective. This study compared the leakage resistance of a New Sealer Prototype (NSP) with a traditional sealer (AH 26) in Resilon-filled roots subjected to a bacterial challenge. Study Design. 41 roots were instrumented to ISO size 50 apically. Group 1 ( 𝑛 = 2 0 ) contained Resilon and AH 26 sealer and roots in group 2 ( 𝑛 = 2 1 ) contained Resilon and NSP. Roots were embedded in a dual-chamber model with the upper chamber containing Streptococcus mutans inoculum. Evidence of bacterial penetration was observed for 1 month. Fisher's Test was used to analyze the data. Results. 8 of 20 roots (40%) in the AH 26 group demonstrated leakage whereas 3 of 21 roots (14%) in the NSP group leaked. The difference in leakage rates was not statistically significant ( 𝑃 = 0 . 0 5 3 ). Conclusion. The traditional sealer (AH 26) demonstrated increased leakage rates compared to the New Sealer Prototype (NSP), but the difference did not reach statistical significance in this study.

Interaction of lactoferrin (Lf) with the cell envelope (CE) and outer membrane (OM) of Salmonella typhimurium- type strain ATCC13311 was tested by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western-blot analyses. The peroxidase-labeled bovine Lf (BLf) and human Lf both recognized a heat-modifiable protein with an estimated molecular mass of 38 kD in the OM. Simultaneous immunoblotting with an antiporin monoclonal antibody specific for a conserved porin domain in members of enterobacteriaceae confirmed that the Lf-binding protein is a porin. Such Lf-binding porin proteins (37–39 kD range) were readily detected in nine other common Salmonella species: S. dublin, S. panama, S. rostock, S. abony, S. hartford, S, kentucky, S. pullorum, S. thompson , and S. virchow . The latter six species also demonstrated one to three weak Lf-reactive bands of low molecular weight in their CE. The antibiotic susceptibility of Salmonella in the presence of Lf was examined. A mixture containing sub-minimum inhibitory concentration (MIC) levels of Lf (MIC/4) and cefuroxime (MIC/2) inhibited the bacterial growth. Lf strongly potentiated the action of erythromycin (eightfold), whereas it increased the activity only by two-fold for ampicillin, ciprofloxacin, chloramphenicol, and rifampicin; similarly, these antibiotics also reduced the MIC of BLf by twofold in S. typhimurium . Such antimicrobial potentiation was not observed with BLf mixtures containing cefalexin, gentamycin, or polymyxin B against strain ATCC13311. BLf and cefuroxime also demonstrated potentiation of varying degrees (two to 16-fold) with nine other Salmonella species. These data established the binding of Lf to porins in salmonellae and a potentiation effect of Lf with certain antibiotics .

This report describes the effects of two gram-negative bacterial endotoxin (lipopolysaccharide [LPS]) preparations on hamster pregnancy outcome variables. Single intravenous challenges with Escherichia coli and Porphyromonas gingivalis LPS on day 8 of pregnancy produced dose-dependent effects on fetal weight malformation and fetal resorption with E. coli LPS having potent embryolethal effects. Premating maternal exposure to P. gingivalis produced embryolethal effects similar to those of E. coli. These data suggest that maternal exposure to P. gingivalis LPS prior to and during pregnancy can induce deleterious effects on the developing fetus.

The nature of the compartmentalization of catalase in human myeloid cells is an unresolved issue. Using a rabbit polyclonal antibody specific for catalase, indirect immunocytofluorescence of immature leukemic promyelocytes (HL-60 cells) showed a pattern of small, sharp, punctate staining in the cytoplasm of all cells, while mature neutrophils showed a larger diffuse, flocculent pattern of cytoplasmic staining. Differential centrifugation of nitrogen cavitates of HL-60 cells indicated that the putative catalase-containing compartment was relatively fragile compared with the compartment(s) that contained myeloperoxidase (MPO), beta-hexosaminidase, beta-glucuronidase, and lysosomal alpha-mannosidase activities. Parallel studies using dimethylsulfoxide (DMSO)-induced HL-60 cells and mature neutrophils showed that, in the course of differentiation, there was an apparent shift in the localization of catalase from the granule fraction to the cytosolic fraction. Percoll-sucrose density gradient centrifugation of HL-60 cell cavitates showed a catalase-containing compartment with a mean peak density (1.05 g/mL) significantly lower than that of the major myeloperoxidase-containing compartment (1.08 g/mL); in mature neutrophils, catalase activity comigrated with lactate dehydrogenase (LDH) activity. Catalase in isolated fractions was protected from proteolysis in the absence, but not in the presence, of 0.1% Triton X- 100. Digitonin titration experiments confirmed the compartmentalized nature of catalase in immature HL-60 cells and were consistent with a cytosolic localization in mature neutrophils. Ultrastructural localization of catalase by Protein A-gold immunocytochemistry demonstrated four to six catalase-containing compartments in all HL-60 cell profiles. In mature neutrophils, catalase was localized primarily in the cytoplasmic matrix, although in fewer than 2% of the cell profiles, one to two catalase-containing compartments were observed. The changes in catalase localization that occur during myeloid differentiation appear to be similar to the changes that occur during erythroid and megakaryocytic differentiation, and may have potential clinical significance in the classification of acute leukemia and in the development of drug resistance.

The aim of the present study was to determine the antibacterial properties of three resin-based pit and fissure sealant products: Clinpro (3M ESPE), Embrace (Pulpdent), and UltraSeal XT plus (Ultradent).The antibacterial effects of the sealants were tested in both an agar diffusion assay and a planktonic growth inhibition assay using Streptococcus mutans and Lactobacillus acidophilus. The materials were applied to paper and enamel disks in the former and on the side walls of 96-well microtiter plates on the latter.All materials showed either diffusible or contact antibacterial effects in the agar diffusion assays. The effect was diminished when enamel disks were used as substrate. In the planktonic growth inhibition assay, Clinpro had its effect reduced, but retained activity against both bacteria over time. L. acidophilus was more sensitive than S. mutans to UltraSeal. Embrace retained antibacterial activity against both bacteria over time.Within the limitations of this in vitro study it can be concluded that all materials are capable of contact inhibition of L. acidophilus and S. mutans growth. Embrace has the longer lasting antibacterial activity when in solution, especially against S. mutans.

Periodontal disease is a chronic inflammatory disorder that leads to the destruction of tooth-supporting tissue and affects 10–20 million people in the U.S. alone. The oral pathogen Porphyromonas gingivalis causes inflammatory host response leading to periodontal and other secondary inflammatory diseases. To identify molecular components that control host response to P. gingivalis in humans, roles for the NLR (NBD-LRR) protein, NLRP3 (cryopyrin, NALP3), and its adaptor apoptotic speck protein containing a C-terminal caspase recruitment domain (ASC) were studied. P. gingivalis strain A7436 induces cell death in THP1 monocytic cells and in human primary peripheral blood macrophages. This process is ASC and NLRP3 dependent and can be replicated by P. gingivalis LPS and Escherichia coli. P. gingivalis-induced cell death is caspase and IL-1 independent and exhibits morphological features consistent with necrosis including loss of membrane integrity and release of cellular content. Intriguingly, P. gingivalis-induced cell death is accompanied by the formation of ASC aggregation specks, a process not previously described during microbial infection. ASC specks are observed in P. gingivalis-infected primary human mononuclear cells and are dependent on NLRP3. This work shows that P. gingivalis causes ASC- and NLRP3-dependent necrosis, accompanied by ASC speck formation.

The purpose of this study was to subject 2 carrier-based root filling products to a 4-month microbial challenge in a dog model with histologic markers to assess periapical inflammation and bacterial penetration of the 2 filling materials. Histologic evidence of bacterial penetration and periapical inflammation were the outcome parameters used to compare the products.Teeth were aseptically prepared and then filled with carrier-based Resilon (RealSeal 1 [RS-1], n = 25) or with carrier-based gutta-percha (Thermafil, n = 25) and were left exposed for 4 months. The first control group received a coronal seal over either RS-1 or Thermafil root fillings (n = 8). A second control group was instrumented and left completely empty (n = 8).Histologic evidence of periapical inflammation was observed in 29% of the Thermafil group and in 9% of the RS-1 group. This difference was only significant when controlling for a possible tooth position effect on inflammation presence (P.05). Histologic evidence of bacterial penetration was present in 9% of the RS-1 group and in 70% of the Thermafil group. The difference in penetration rates between RS-1 and Thermafil was statistically significant when controlling for any dog or tooth position effects on bacterial penetration (P.001). Furthermore, there was a statistically significant correlation between histologic evidence of inflammation and histologic evidence of infection (P = .002).RS-1 appeared to resist bacterial penetration more effectively than Thermafil under the conditions of this study.

Porphyromonas gingivalis and Fusobacterium nucleatum are often coisolated from sites of infection, such as suppurative apical periodontitis. The synergistic pathogenicity of mixed infection of P. gingivalis HG 405 with F. nucleatum PK 1594 was studied in the mouse subcutaneous chamber model in groups of seven animals. The minimal dose for P. gingivalis HG 405 that was required to infect 100% of the chambers was reduced by 1,000-fold when animals were inoculated in the same chamber with 1 x 10(9)F. nucleatum PK 1594 (p < 0.001). To benefit from the presence of the fusobacteria, P. gingivalis HG 405 had to be coinoculated; inoculation in separate chambers for the same animal had no such effect (p < 0.001). Subinfective F. nucleatum inocula also benefited from the association with P. gingivalis HG 405 and uniformly established an infection when this partner was present (p < 0.001). These results suggest that the frequent and natural coexistence of P. gingivalis and F. nucleatum in diseased sites may express such a synergism in successful establishment and survival of small inocula.

To evaluate the capability of different adhesive systems to inhibit in vitro caries formation.Four materials were tested: Gluma Comfort Bond (GL; Heraeus Kulzer), Gluma Comfort Bond + Desensitizer (GL+; Heraeus Kulzer), iBond (iB; Heraeus Kulzer), and One-up Bond F (OUB; Tokuyama). Bovine roots were ground to obtain flat mesial and distal dentin surfaces. Nail varnish was applied to the entire root surface except for two 10 mm X 2 mm windows on the flattened surfaces. The adhesives were applied to the exposed areas according to the manufacturers' instructions. The roots were incubated at 37 degrees C for 1 week in a suspension of StreptococcuS mutans and Lactobacillus acidophilus. Control groups either received no root surface treatment or were etched with 37% phosphoric acid (without immersion in the bacterial suspension). Specimens were sectioned perpendicular to the long axis of the roots, stained for 24 hours with Rhodamine B, and analyzed with confocal laser microscopy. The data were analyzed by 1-way ANOVA and Fisher's PLSD test at 95% confidence level.Lesions in the OUB, iB, and GL+ groups were significantly shallower than those in the no-treatment group. Acid etching did not produce any measurable lesions. Mean lesion depth in the GL group was similar to that in the no-treatment group. There was no significant difference between mean lesion depths in the GL+, GL, and iB groups.Fluoride- and glutaraldehyde-containing adhesive systems might be an aid in root caries prevention.

In vitro results are presented for a novel oral drug-delivery system ultimately intended for treatment of oral infections in immunocompromised patients. Test samples of ethylene vinyl acetate copolymer (EVA) containing chlorhexidine diacetate (CDA) showed desirable antimicrobial properties and steady, slow release into aqueous and other media after an initial burst of drug release in the first day of liquid exposure. By washing away this initial burst, the proposed mouthguard device should be capable of sustained delivery of locally effective CDA concentrations far below systemically toxic levels. A prolonged room temperature shelf-life of at least 1 year, and effectivity against a wide range of oral bacteria and Candida species was demonstrated. Drug loaded films showed a top-to-bottom asymmetry in drug release, but good lateral homogeneity, and a linear relationship between initial CDA loading concentration (from 0.63 to 10 wt %) and days 3–14 release rates in a static aqueous environment. The EVA matrix containing CDA appears to possess many suitable properties for localized oral delivery of sustained antimicrobial activity. © 2008 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2008

Purpose: To evaluate the effects of dentin collagen modifications induced by various cross-linkers on the stability of collagen matrix and the inhibition of root caries. Materials and Methods: The following cross-linkers were tested: 5% glutaraldehyde (GA), 0.5% proanthocyanidin (PA), 0.625% genipin (GE). In the first experiment, cross-linker-treated demineralized human root dentin was digested with bacterial collagenase, centrifuged, and the supernatants were subjected to amino acid analysis to determine collagen content. The residues were analyzed by SDS-PAGE and hydroxyproline analysis. In the second experiment, bovine root surfaces were conditioned with phosphoric acid, treated with the cross-linkers, incubated with Streptococcus mutans and Lactobacillus acidophilus for 1 week and the root caries inhibition was evaluated with confocal microscopy. Lastly, the ability of the bacteria to colonize the root surface was evaluated. In this experiment slabs of bovine root were treated with the cross-linkers and incubated in a suspension of S. mutans and L. acidophilus. The slabs were washed, resuspended in water, glucose was added, and the pH measured. Results: While all collagen was digested with collagenase in the control groups, only a small proportion was solubilized in the GA-, PA-, and GE-treated groups. The root caries was significantly inhibited by treatment with PA or GA. Drops in pH in the cross-linker-treated groups were essentially the same as in the untreated group. Conclusion: Naturally occurring cross-linkers, especially PA, could be used to modify root dentin collagen to efficiently stabilize collagen and to increase its resistance against caries.

This study evaluated the clinical efficacy of 2% chlorhexidine (CHX) gel on intracanal bacteria reduction during root canal instrumentation. The additional antibacterial effect of an intracanal dressing (Ca[OH]2 mixed with 2% CHX gel) was also assessed. Forty-three patients with apical periodontitis were recruited. Four patients with irreversible pulpitis were included as negative controls. Teeth were instrumented using rotary instruments and 2% CHX gel as the disinfectant. Bacterial samples were taken upon access (S1), after instrumentation (S2), and after 2 weeks of intracanal dressing (S3). Anaerobic culture was performed. Four samples showed no bacteria growth at S1, which were excluded from further analysis. Of the samples cultured positively at S1, 10.3% (4/39) and 8.3% (4/36) sampled bacteria at S2 and S3, respectively. A significant difference in the percentage of positive culture between S1 and S2 (p 0.001) but not between S2 and S3 (p 0.692) was found. These results suggest that 2% CHX gel is an effective root canal disinfectant and additional intracanal dressing did not significantly improve the bacteria reduction on the sampled root canals. (J Endod 2007;33:1283–1289)

The regulation of neutrophil functions by Type I cGMP-dependent protein kinase (cGKI) was investigated in wild-type (WT) and cGKI-deficient (cGKI−/−) mice. We demonstrate that murine neutrophils expressed cGKIα. Similar to the regulation of Ca2+ by cGKI in other cells, there was a cGMP-dependent decrease in Ca2+ transients in response to C5a in WT, but not cGKI−/− bone marrow neutrophils. In vitro chemotaxis of bone marrow neutrophils to C5a or IL-8 was significantly greater in cGKI−/− than in WT. Enhanced chemotaxis was also observed with cGKI−/− peritoneal exudate neutrophils (PE-N). In vivo chemotaxis with an arachidonic acid-induced inflammatory ear model revealed an increase in both ear weight and myeloperoxidase (MPO) activity in ear punches of cGKI−/− vs WT mice. These changes were attributable to enhanced vascular permeability and increased neutrophil infiltration. The total extractable content of MPO, but not lysozyme, was significantly greater in cGKI−/− than in WT PE-N. Furthermore, the percentage of MPO released in response to fMLP from cGKI−/− (69%) was greater than that from WT PE-N (36%). PMA failed to induce MPO release from PE-N of either genotype. In contrast, fMLP and PMA released equivalent amounts of lysozyme from PE-N. However, the percentage released was less in cGKI−/− (∼60%) than in WT (∼90%) PE-N. Superoxide release (maximum velocity) revealed no genotype differences in responses to PMA or fMLP stimulation. In summary, these results show that cGKIα down-regulates Ca2+ transients and chemotaxis in murine neutrophils. The regulatory influences of cGKIα on the secretagogue responses are complex, depending on the granule subtype.

A dog model was used to assess and compare in vivo the efficacy of gutta-percha and AH26 sealer versus Resilon with Epiphany primer and sealer [Resilon "Monoblock" System (RMS)] filled roots in preventing apical periodontitis subsequent to coronal inoculation with oral microorganisms. There were 56 vital roots in the premolars of seven adult beagle dogs aseptically instrumented, filled, and temporized. The roots were randomly divided into four experimental groups (Coronal Leakage Model) and one negative control group and filled as follows: group 1-lateral condensation of gutta-percha and AH26 sealer (n=12); group 2-vertical condensation of gutta-percha and AH26 sealer (n=12); group 3-lateral condensation of RMS (n=12); group 4-vertical condensation of RMS (n=10); negative control (n=10)-gutta-percha and AH26 sealer or RMS root fillings using lateral or vertical condensation techniques as in groups 1 to 4. Positive control-57 additional premolar roots were instrumented, infected and not filled (beginning of the Entombment Model experiment). The premolars in groups 1 to 4 were accessed again, inoculated with dental plaque scaled from the dog's teeth, and temporized. This fresh innoculum of microorganisms was repeated on two more occasions at monthly intervals. The teeth in the negative control group were not accessed again and remained undisturbed. On the 14-wk postcoronal inoculation, dogs were euthanized, and jaw blocks prepared for histologic evaluation under a light microcope. Mild inflammation was observed in 82% (18 of 22) of roots filled with gutta-percha and AH26 sealer that was stastistically more than roots filled with RMS (19% or 4 of 21) and roots in the negative control (22% or 2 of 9) (McNemar paired analysis, p < 0.05). The Resilon "Monoblock" System was associated with less apical periodontitis, which may be because of its superior resistance to coronal microleakage.

We have recently found that an extracellular protein, α1proteinase inhibitor (α1PI; α1antitrypsin), is required for in vitro human immunodeficiency virus (HIV) infectivity outcome. We show here in a study of HIV-seropositive patients that decreased viral load is significantly correlated with decreased circulating α1PI. In the asymptomatic category of HIV disease, 100% of patients manifest deficient levels of active α1PI, a condition known to lead to degenerative lung diseases and a dramatically reduced life span. Further, HIV-associated α1PI deficiency is correlated with circulating anti-α1PI immunoglobulin G. These results suggest that preventing HIV-associated α1PI deficiency may provide a strategic target for preventing HIV-associated pathophysiology.

This investigation determined the effects of a distant oral infection (Porphyromonas gingivalis) on a concurrent local enteric infection (Echerichia coli). A modified murine subcutaneous tissue chamber model was used. Subcutaneously implanted chambers with different microbial makeups were monitored for sloughing and their contents assayed for prostaglandin E2. Bilaterally implanted chamber experiments revealed that a distant "chronic" (immunization with heat-killed organism, followed by live challenge) P. gingivalis infection offered protection against a local chronic E. coli infection, as evidenced by delaying the time for 50% of the chambers to reject from day 19 to day 25 and a statistically significant prostaglandin E2 decrease from 529.4 +/- 176.6 ng/ml to 191.5 +/- 100.9 ng/ml (p < 0.01) (Mann-Whitney test). An acute (live challenge only) distant P. gingivalis infection or immunization with the heat-killed organism alone had no effect on "chronic" E. coli infection in this model. These data suggest that the presence of low-grade chronic oral infection may modify the responses to other infectious challenges.

The shifting balance between proteinases and proteinase inhibitors in blood, a function of their relative affinities and concentrations, has long been hypothesized to influence immune competency. The identification of proteinase-activated receptor responses in cells of the mononuclear phagocyte system suggests a potential explanation. The major serum proteinase inhibitor, alpha1proteinase inhibitor (alpha1PI, alpha1-antitrypsin), has been reported to increase in concentration during inflammation. Quantitative determination of serum alpha1PI has traditionally been performed nephelometrically; however, antigenically quantitated levels may not be representative of functional capacity. It has previously been observed that alpha1PI in serum exhibits bimodal behavior as the result of various concentrations of proteinase inhibitors, specifically alpha2macroglobulin (alpha2M) and inter-alpha-trypsin inhibitor, which compete in binding to a panel of serine proteinases. Consequently, it has not previously been possible to assign a numerical value for the specific activity of these competing proteinase inhibitors in serum. By applying known constants representing the association of proteinase inhibitors with porcine pancreatic elastase (PPE), the theoretical relationship between the functional and antigenic values for alpha1PI and alpha2M has been empirically derived allowing, for the first time, the calculation of their specific activities in serum. As predicted, the serum concentration of alpha1PI was found to be highly correlated with residual uninhibited PPE catalytic activity in healthy individuals, but not in individuals exhibiting fragmented or complexed alpha1PI. Using these techniques, both the antigenic and functional levels of alpha1PI were determined in sera from subjects with insulin-dependent diabetes mellitus (IDDM) who had been clinically diagnosed as having either periodontal disease or gingival health. Determination of quantitative levels by antigen-capture suggests that the IDDM subjects with periodontitis manifest dramatically increased levels of fragmented serum alpha1PI compared with their orally healthy counterparts or normal controls. In contrast, functional analysis of serum alpha1PI revealed no differences between the three subject populations. The elevated levels of antigenically determined serum alpha1PI reflect the inflammatory status of periodontal disease. These results support the importance of and provide methodology for determining the functionally active levels of alpha1PI allowing reexamination of changes detected during the acute phase of inflammation, replacement therapy, and longitudinal studies in relevant disease processes including malignancy and diabetes.

The antimicrobial spectrum of lactoferrin (Lf) includes statsis, cidal, cationic, phagocytic, and colonization/decolonization effects on susceptible microorganisms. The selective interaction of Lf with microbial surface seems to play an essential role in regulating many of these events. Lf bound to specific outer membrane (OM) pore-forming proteins (porins) of various enteric bacteria. The magnitude of Lf-microbe interaction showed a direct relationship with the bacterial susceptibility to Lf. Although most bacteria expressed porins, certain strains showed resistance to Lf effects and did not demonstrate specific Lf binding. This Lf “resistance” was attributed to the shielding of porin accessibility by the carbohydrate O-antigenic chains of lipopolysaccharide (LPS). Mutants with progressive deletions in O-side-chain and core polysaccharide demonstrated increased Lf binding and progressive susceptibility to Lf effects. Lf interaction with bacterial surface affected the porin channels in the OM and potentiated the diffusion of antibiotics. Lf also affected the intestinal colonization of Escherichia coli. Lf inhibited the expression of various bacterial fimbriae and also blocked bacterial interaction with host subepithelial matrix components, such as fibronectin, fibrinogen, various collagens, and laminin. Our studies suggested that specific interaction with cell target involved the anion-binding ligands of Lf, followed by coordinate binding and oxidation of Fe2+ to Fe3+ on bacterial surface. This metal transition in the presence of suitable oxygen species seems to fulfill the necessary components of Haber-Weiss reaction and promote a targeted radical generation, with iron (Fe) serving as the Lf prosthetic group on the bacterial surface.

While it is clear that CD4 is the receptor for the gp120 envelope protein of HIV-1, substantial evidence suggests that other host cell proteins are required for successful membrane fusion. Studies were initiated to examine the potential for a protein receptor which has an elastase-like character to participate in fusion of HIV-1 with permissive host cells. A synthetic elastase inhibitor was shown to significantly reduce HIV-1 infectivity when present during, but not after, the initial contact between virus and cells. A human T cell elastase-like membrane component was purified and shown to be lipid-associated. By competitive inhibition, the purified protein was shown to bind gp160 within the HIV-1 fusion domain. The binding parameters of whole T cell membrane extract, with a hydrophobic pentapeptide representative of the fusion domain, suggested an elastase-like protein is the single, secondary T cell receptor for HIV-1 (K = 1 x 10(3) M-1). The pentapeptide interacted with porcine and human (epithelial and polymorphonuclear leukocyte), but not murine, elastase isoforms, suggesting its participation in the permissiveness of host cells to infection.

The pathogenesis of periodontal tissue destruction involves the orchestrated sequential activation of several components of the host response, each of which have the potential for providing diagnostic information regarding the health or disease status of the periodontium. These events include the bacterial triggering of serum components, the release of vasoactive compounds, the recruitment of inflammatory cells, the activation of phagocytes, the local secretion of immunoglobulins and inflammatory mediators, as well as connective tissue remodeling. For purposes of diagnosis, much attention has been directed toward the measurement of local levels of specific byproducts of the pathogenic process either within the tissues or in the adjacent crevicular fluid. These are site-directed assessments of the local concentration of these byproducts, and with the important exception of antibody levels, very few byproducts of the periodontal pathogenic process have been studied on a systemic level. However, since the risk associated with periodontal disease progression is primarily patient-based and secondarily site-based, there is an increasing need to identify patients at risk for intervention strategies. Furthermore, there are substantial data that periodontal diseases are specific infections that occur in an appropriately susceptible host. The expression of periodontal disease has a strong genetic component, which presumably defines the host's response thereby affecting susceptibility. For this reason, there is increased interest in defining those elements of the host response which result in susceptibility to disease. The ultimate objective of a diagnostic strategy is to identify patients who are potentially at risk for acquiring disease to enable preventive measures.

This report describes the clinical and microbiological features of 30 refractory patients and their response to a combined local and systemic therapy at 6 weeks and 3 years following treatment. The refractory treatment protocol (RefTx) consisted of a 2-week regimen of amoxicillin/clavulanate potassium in conjunction with professional, intrasulcular delivery of povidone iodine, and chlorhexide mouthwash rinses b.i.d. Eighty-seven percent of the patients had favorable clinical responses to the RefTx and could be divided into 3 groups (A, B, C) based upon initial flora patterns and the shifts that occurred following treatment. Pretreatment prevalence of Porphyromonas gingivalis (P.g.) was 36.7%. The RefTx was effective in reducing P.g. below detection levels in 10 of the 11 positive patients at P0.01; each of these 10 patients (Group A) demonstrated significant gain in attachment. Other black-pigmented Bacteroides species (OBP) were isolated from 70% of the patients at baseline. Nine of these patients did not harbor P.g., showed clinical improvement upon treatment, and were OBP negative following treatment (Group B). Group C patients (7) demonstrated clinical improvement with therapy and did not fit into either Group A or B based upon microbial patterns. Group D patients (4) did not show clinical improvement with 3 patients harboring either P.g. or OBP after treatment. The RefTx was effective at reducing probing pocket depth with a 56% decrease in the number of pockets greater than 6 mm at 6 weeks. This was accompanied by an overall gain ofor = 1 mm of probable attachment in 45% of all sites. The clinical effects of the RefTx were shown to persist at 34.3 months with an apparent attachment gain ofor = 1 mm in 41.2% of sites. These data suggest that P.g. and OBP are important pathogens in refractory periodontitis and that the RefTx protocol is an acceptable, non-invasive alternative for the management of these patients.

Activated polymorphonuclear leukocytes have been associated with neoplasia, atherogenesis and reperfusion injury. Since some of these conditions are also correlated with dietary fat, we examined the functional characteristics of leukocytes isolated from subjects before and after consumption of a lipid-rich meal. There was up to 2-fold greater superoxide generation in response to agonists in leukocytes obtained post-prandially; the maximum increase was observed about 4 h after eating and followed the peak (2–4 h) in serum triglycerides. Neutrophils isolated post-prandially also exhibited impaired chemotaxis and defective bacterial killing, but normal phagocytosis. These findings provide a new variable that should be considered in studies of leukocytes.

The objective of this study was to evaluate the polymorphonuclear leukocyte (PMN) function in a poorly controlled adult insulin-dependent diabetic patient (IDDM) with severe recurrent periodontitis, while describing the microbiological and clinical findings. Chemotaxis, superoxide production, and phagocytosis and killing of Porphyromonas (Bacteroides) gingivalis by the IDDM PMN were evaluated 1 week before treatment relative to a healthy, matched control. These analyses revealed a significant (P less than .05) depression in the number of IDDM PMNs migrating along an FMLP gradient (Boyden chamber assay). In addition, a significant (P less than .05) enhancement of IDDM PMN superoxide production in response to opsonized zymosan (cytochrome C reduction) was observed. Phagocytosis and killing (fluorochrome phagocytosis assay) by IDDM PMN of two P. gingivalis strains was also impaired significantly (P less than .05). The subgingival microflora contained significant levels of P. gingivalis, Actinobacillus actinomycetemcomitans, Eikenella corrodens, and Peptostreptococcus micros. Periodontal treatment consisted of extraction of hopeless teeth, scaling and root planing and 3 weeks of Augmentin therapy. The antibiotic therapy resulted in unrecoverable numbers of the putative pathogens and a reduction in both gingival inflammation and disease progression. The IDDM healing response to previous surgical treatment and extractions was poor, presumably due to a marked thrombocytopenia (91 x 10(3) platelets/mm3).

Lactoferrin has been previously shown to be bactericidal for Legionella pneumophila. The current study showed that CaCl2, Mg(NO3)2, and MgCl2, but not NaCl, blocked killing. Activity was pH dependent with the greatest activity at 5.0. Sensitivity of the organism was dramatically affected by the growth conditions. Log phase 12 h, broth-grown cells were most sensitive, with older cultures becoming more resistant. Plate-grown cells were completely resistant. Lactoferrin binding, as detected by immunofluorescence microscopy, was temperature dependent (no binding at 4 °C), but was independent of killing.Key words: Legionella pneumophila, lactoferrin, mucosal proteins.

A group of 25 subjects with selective IgA deficiency formed part of a study to investigate the relationship between secretory IgA and the incidence of dental caries. Eight of these subjects were found to have significant levels of IgM in their whole, parotid, and submandibular-sublingual saliva. Parotid saliva and serum samples were assayed for the presence of naturally induced antibodies to five serotypes of Streptococcus mutans. Agglutinin titers to strains AHT, BHT, 10449, LM-7, and 6715 were equivalent in control groups and IgA-deficient subjects with elevated salivary IgM. Fractionation of parotid saliva on Sephadex G-200 and immunofluorescent and serological procedures indicated that the anti-S. mutans antibody activity was associated exclusively with IgM. In contrast, in the control population the immunoglobulin responsible for antibody activity was principally IgA. No antibody could be detected in the Ig-deficient saliva samples. Immunochemical analyses indicated that the IgM antibody was, at least in part, associated with secretory component. The IgM-compensating subjects as a group had markedly lower caries experience compared to subjects with Ig-deficient saliva. These data indicate a biological activity for secretory IgM which may function to compensate for the absence of s-IgA, and this antibody may be an important determinant in the oral, as well as overall, health of selective IgA-deficient patients.

A lectin isolated from the tropical jackfruit, jacalin, previously reported to precipitate human immunoglobulin A (IgA), and conjugated to agarose was used to separate the two subclasses of IgA from secretions. Jacalin-agarose binds specifically to the D-galactose moiety of IgA1 but not to IgA2 which has a different carbohydrate content and structure. IgA2 passed through the jacalin-agarose column and was collected in the void volume. IgA1 was eluted from the lectin by 0.8 M galactose. Of a representative diluted anti-alpha chain-purified colostral IgA preparation containing 50.2 micrograms IgA1 and 55.8 micrograms IgA2, 40.3 micrograms IgA1 (80.3% of the original) and 49.6 micrograms IgA2 (88.9%) was collected following jacalin-agarose chromatography. The jacalin-purified IgA1 fraction contained 8.0% IgA2 and the IgA2 fraction contained no IgA1. In addition, the IgA1 and IgA2 fractions had naturally occurring antibody activity to a normal oral bacterium. The method is easy, reproducible and specific and has many applications to mucosal immunological investigations.

Ingestion of capsules which contained killed Streptococcus mutans by four healthy human subjects led to the appearance of specific antibodies in external secretions. Salivary and lacrymal antibodies were detected within 1 wk of ingestion and continued to increase throughout a 14-day immunization period, with a gradual decline during the 2 ensuing months. A second period of immunization resulted in a pronounced increase of specific antibody levels which occurred earlier than in the primary immunization period and reached peak levels by day 10. No change was detected in serum antibody levels throughout either immunization period. The antibody activity in all secretions was associated with the immunoglobulin A class, as determined by immunochemical analyses. These data indicate that ingestion of bacterial antigens selectively stimulates the immune response in secretions.

Patients with localized juvenile periodontitis (LJP) exhibit defective neutrophil functions to a variety of environmental and host stimuli. It is not clear, however, how many of the measurable functions are defective and whether individual patients exhibit single or multiple dysfunctions. The purpose of this study was to evaluate chemotaxis, phagocytosis, specific granule release and superoxide production in a group of 23 previously unreported LJP patients. Our results indicate that all 23 of these LJP patients exhibited chemotaxis depression to N-formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP) and endotoxin-activated serum (EAS). Smaller groups from the 23 chemotactically defective LJP group were used to test other function due to inability to obtain sufficient quantities of blood. Fourteen of 14 LJP patients tested exhibited defective phagocytosis. Ten LJP patients were evaluated for specific granule release, and 14 LJP patients were evaluated for superoxide production. Both granule release and superoxide production were found to be normal in chemotactically defective LJP patients. Since both defective and normal responses noted in the same neutrophil populations are mediated by the same receptor, it is hypothesized that the cellular defect lies in a post receptor pathway.

Streptococcus mutans and Vibrio cholerae, but not Escherichia coli, were killed by incubation with purified human apolactoferrin. Concentrations of lactoferrin below that necessary for total inhibition resulted in a marked reduction in viable colony-forming units. This bactericidal effect was contingent upon the metal-chelating properties of the lactoferrin molecule.

A major problem that has received considerable attention concerns a method for effectively stimulating a selective IgA-associ-ated immune response on mucous surfaces. Earlier immunization attempts indicated that an antigen applied to the mucous membrane penetrates these membranes and encounters B-lymphocytes that have the potential of differentiating into IgA-producing plasma cells of a given specificity. The IgA produced is then actively transported through epithelial cells on the surface of the membrane where the original antigen stimulation occurred. Numerous examples clearly demonstrate the efficiency of this pathway of immunization. For example, the stimulation through the conjunctival sac of one eye led to the appearance of antibodies in tears from the immunized site but not in secretions from the other eye (1). Similarly, the application of viral antigen into the large intestine of individuals who have had double-barrelled colostomies induced s-IgA antibodies restricted to the site of antigen application but failed to induce antibodies in other parts of the intestine, in glandular secretions or in nasal secretions (2). These are only two examples of experiments that are suggestive of an IgA-associated immune response that results from topical antigen application.

This study was undertaken in order to correlate the immune status of persons with various types of immune dysfunctions with the incidence of dental caries. Preliminary data on caries experience and the microbial composition of plaque in these individuals are presented. The findings suggest that persons with immunoglobulin dysfunctions have a greater susceptibility to dental caries and have a greater frequency of harboring S mutans than do normal persons.