Development of a nomogram for identifying periodontitis cases in Denmark.

Although self-reported health outcomes are of importance, attempts to validate a clinical applicable instrument (e.g., nomogram) combining sociodemographic and self-reported information on periodontitis have yet to be performed to identify periodontitis cases. Clinical and self-reported periodontitis, along with sociodemographic data, were collected from 197 adults. Akaike information criterion models were developed to identify periodontitis, and nomograms developed based on its regression coefficients. The discriminatory capability was evaluated by receiver-operating characteristic curves. Decision curve analysis was performed. Smoking [OR 3.69 (95%CI 1.89, 7.21)], poor/fair self-rated oral health [OR 6.62 (95%CI 3.23, 13.56)], previous periodontal treatment [OR 9.47 (95%CI 4.02, 22.25)], and tooth loss [OR 4.96 (95%CI 2.47, 9.97)], determined higher probability of having "Moderate/Severe Periodontitis". Age [OR 1.08 (95%CI 1.05, 1.12)], low educational level [OR 1.65 (95%CI 1.34, 2.23)], poor/fair self-rated oral health [OR 3.57 (95%CI 1.82, 6.99)], and previous periodontal treatment [OR 6.66 (95%CI 2.83, 15.68)] determined higher probability for "Any Periodontitis". Both nomograms showed excellent discriminatory capability (AUC of 0.83 (95%CI 0.75, 0.91) and 0.81 (95% CI 0.74, 0.88), good calibration, and slight overestimation of high risk and underestimation of low risk. Hence, our nomograms could help identify periodontitis among adults in Denmark.

Oral conditions are associated with salt taste disability among American adults.

AIM: To explore the association between oral conditions and their interaction with salt taste disability among American adults. METHODS: Data from the 2013-2014 NHANES cycle were used (n = 2373). The exposures were periodontitis, defined by the 2017 EFP-AAP classification, dental caries, missing teeth, and edentulism, as per the DMF-T index, and xerostomia. The outcome was salt taste disability, objectively assessed. Covariates included sex, age, educational level, poverty index, obesity, diabetes, smoking, alcohol consumption, and medications related to mouth dryness. Weighted multivariable logistic regression modeling was used to evaluate the relationship between oral conditions and their interaction and salt taste disability. RESULTS: Participants who reported xerostomia were more likely to have salt taste disability (OR 2.42; 95%CI 1.44-4.07), especially those older than 60 years (OR 3.63; 95%CI 1.72-7.63). Among participants aged 40-59, xerostomia increased the chance of salt taste disability; however, the confidence interval included the null value. The interactions between xerostomia and edentulism increased the chance of salt taste disability. CONCLUSION: Oral conditions seem to influence the ability to taste salt. Dental professionals may help identify individuals with taste alterations and raise their awareness of the risk of systemic diseases that require the reduction of salt intake.

Sugar-sweetened beverage consumption and periodontitis among adults: A population-based cross-sectional study.

AIM: Investigating the association between sugar-sweetened beverages (SSBs) and periodontitis and whether the awareness of diabetes modifies this relationship. MATERIALS AND METHODS: Cross-sectional analysis was conducted using the National Health and Nutrition Examination Survey (NHANES III) data involving US adults aged 30-50. Periodontitis was classified according to the Centers for Disease Control and Prevention and American Academy of Periodontology (CDC-AAP), and SSB consumption as dichotomous (<5 or ≥5, <7 or ≥7 and <14 or ≥14 times/week), ordinal and continuous variables. Confounders included family income poverty ratio, education, race/ethnicity, sex, age, food energy intake, smoking and alcohol. Odds ratios (ORs) were obtained by logistic regressions using inverse probability weighting. Effect modification analysis was performed considering self-reported diabetes. RESULTS: Among 4473 cases analysed, 198 self-reported diabetes. SSBs were associated with periodontitis when individuals consumed ≥5 (OR 1.64; 95% confidence interval [CI] = 1.30-2.06), ≥7 (OR 1.92; 95% CI = 1.50-2.46) and ≥14 (OR 2.19; 95% CI = 1.50-3.18) times/week. The combined effect of consuming SSBs (≥5 and ≥14 times/week) and self-reported diabetes had less impact than the cumulative effect. CONCLUSIONS: SSB consumption was associated with higher odds of periodontitis, and the estimates were reduced among those with awareness of diabetes.

Incidence, mortality and survival rates of lip, oral cavity and salivary glands cancers in Singapore: A half-century time trend analysis (1968-2017).

OBJECTIVES: To examine trends in incidence and mortality and evaluate overall survival (OS) of oral cancer in Singapore between 1968 and 2017. METHODS: All diagnosed oral cancers by anatomical sites and population size were extracted from the Singapore Cancer Registry and the Department of Statistics Singapore. The trend of age-standardized incidence rate (ASIR) and mortality rate (ASMR) (per 100 000 person-years) of the lip, oral cavity and salivary gland cancers were evaluated by Prais-Winsten regressions for each ethnicity and gender. Kaplan-Meier curves were performed to evaluate the OS by anatomical sites in each age group by ethnicity and sex. RESULTS: Overall, 49, 3494 and 1066 people were diagnosed, and 28, 2310 and 476 died from lip, oral cavity and salivary gland cancers, respectively. The oral cavity cancer ASIR and ASMR reduced from 3.07 (1968-1972) to 2.01(2008-2012) and from 2.06 (1978-1982) to 1.21 (2013-2017) per 100 000 person-years, respectively, with both highest in Indians throughout the whole period. Male:Female ratio ranged from 3.43 (1973-1977) to 1.75 (2013-2017) and from 3.41 (1978-1982) to 2.40 (2013-2017) for ASIR and ASMR, respectively. However, both salivary gland cancer ASIR and ASMR increased from 0.50 (1968-1972) to 0.80 (2008-2012) and from 0.18 (1968-1982) to 0.42 (1988-1992) per 100 000 person-years, respectively, with both higher in males since 1993. Oral cavity cancer ASIR decreased for males aged ≥60, and Indian females ≥25, but increased among Chinese females aged ≥60. Oral cavity cancer ASMR decreased among Chinese aged 25-59, and among Malay males and Indian females. Salivary gland cancer ASIR increased among Chinese males aged ≥60 and Malay males aged 25-59; while ASMR increased among Chinese males aged ≥60. The median OS for oral cavity, lip and salivary gland cancers were 3.0, 9.3 and 18.1 years, respectively, with females surviving longer than males. CONCLUSIONS: Singapore has experienced a decline in the incidence and mortality of lip, oral cancer, an increase in in the incidence and mortality of salivary gland cancer, with an increase in the median overall survival rate. Monitoring the magnitude of oral cancer burden and the demographic, and temporal variations is necessary for tailoring health planning and setting priorities for future clinical care and research.

Chewing Disability Is Associated With Cognitive Impairment Among Older Adults: A Population-Based Cohort Study.

BACKGROUND: Chewing disability is associated with impaired quality of life, potentially leading to depression, and cognitive impairment. Although the chewing-ability-cognition relationship has been explored, examining whether depression mediates this relationship remains unclear. We investigated the association between chewing disability and cognitive impairment development and a potential mediation via depression among older persons. METHODS: Older persons without cognitive impairment at baseline (n = 973) from the 3 waves of the Panel on Health and Ageing of Singaporean Elderly were investigated. The outcome was incident cognitive impairment by the end of the study, while the exposure was chewing disability over the study period. Time-varying depression was the mediator. Time-fixed confounders included sex, ethnicity, education, marital status, living arrangement, and housing type, and time-varying confounders included age, smoking, cardiovascular diseases, diabetes, number of teeth, and denture wearing. We used marginal structural modeling to evaluate the effect of chewing disability on cognitive impairment development. RESULTS: After 6 years, 11% developed cognitive impairment, and chewing disability was reported by 33%. Chewing disability was associated with higher odds of developing cognitive impairment (OR 1.43, 95% CI: 1.09, 1.87), of which 85.3% was explained by the controlled direct effect of chewing disability, whereas the remaining 14.7% could be eliminated if there was no depression. CONCLUSIONS: Our findings indicate an association between chewing disability and cognitive impairment, while the role of depression could not be fully elucidated. Oral health should be incorporated as part of older persons' care for its potential to assess the risk for other systemic conditions.

Behavioral and metabolic risk factors associated with periodontitis in Brazil, 1990-2019: a multidimensional analysis for the Global Burden of Disease Study 2019.

OBJECTIVES: Periodontitis is a non-communicable disease (NCD) that may be linked to other NCDs through shared risk factors. Accordingly, we analyzed the relationship between periodontitis and behavioral and metabolic risks common to NCDs in Brazilian adults over three decades. METHODS: Indicators of periodontitis, behavioral risks (smoking, alcohol use, sugar-sweetened beverages (SSB), and physical activity), and metabolic risks (overweight/obesity, dyslipidemia, hyperglycemia, and hypertension) in Brazilian adults (25-49 y-old) between 1990 to 2019 were obtained from the Global Burden of Disease Study 2019. Data were adjusted for Gini index. Fixed-effects and Prais-Winsten regressions were performed (p < 0.05). RESULTS: The prevalence of periodontitis has increased among Brazilians since 2005. High-SSB diet, alcohol use, and metabolic risks increased between 1990-2019, whereas smoking decreased. In crude models, periodontitis prevalence increased with alcohol use (2545.1; 95%CI: 2307.9-2782.3), high-SSB diet (365.5; 95%CI: 322.5-408.4), low physical activity (1784.4; 95%CI: 763.7-2805.0), overweight/obesity (172.3; 95%CI: 156.3-188.4), dyslipidemia (734.5; 95%CI: 624.7-844.2), and hyperglycemia (1774.3; 95%CI: 1555.9-1992.7). After adjustment for the Gini index, periodontitis prevalence raised with a high-SBB diet (1416.0; 95%CI: 1120.2-1711.8), overweight/obesity (629.9; 95%CI: 573.1-686.8), dyslipidemia (2035.8; 95%CI: 1728.1-2343.5), and hyperglycemia (8918.1; 95%CI: 7979.8-9856.3). CONCLUSIONS: Periodontitis has increased in Brazil since 2005, despite the smoking reduction. Sugar-sweetened beverage was the behavioral risk that mostly accompanied the periodontal trend. CLINICAL RELEVANCE: Our results support upstream strategies targeting commercial, social, political, and structural determinants to tackle NCDs and reduce oral health inequities.

Obesity, Insulin Resistance, Caries, and Periodontitis: Syndemic Framework.

(1) Background: To investigate the grouping of obesity and insulin resistance with caries and periodontitis from a syndemic perspective through pathways of socioeconomic inequalities, smoking, alcohol, and high sugar consumption in adolescence. (2) Methods: The population-based RPS Cohort study, São Luís, Brazil, in ages 18-19 years (n = 2515) was used. The outcomes were the grouping of pbesity and Insulin Resistance Phenotype (latent variable formed by Triglycerides/HDL ratio, TyG index, and VLDL) and the Chronic Oral Disease Burden (latent variable comprising caries, bleeding on probing, probing depth ≥ 4 mm, clinical attachment level ≥ 3 mm, and visible plaque index ≥ 15%). Socioeconomic Inequalities influencing the Behavioral Risk Factors (latent variable formed by added sugar, smoking, and alcohol) were analyzed using structural equation modeling. (3) Results: Socioeconomic Inequalities were associated with the Chronic Oral Disease Burden [Standardized Coefficient (SC) = 0.222, p < 0.001]. Behavioral Risk Factors were associated with increased Chronic Oral Disease Burden (SC = 0.103; p = 0.013). Obesity was associated with the Insulin Resistance Phenotype (SC = 0.072; p < 0.001) and the Chronic Oral Disease Burden (SC = 0.066; p = 0.005). The Insulin Resistance Phenotype and the Chronic Oral Disease Burden were associated (SC = 0.053; p = 0.032). (4) Conclusion: The grouping of obesity and early events of diabetes with caries and periodontitis call for a syndemic approach in adolescence.

Intake and serum levels of micronutrients and chronic oral diseases burden.

OBJECTIVE: To investigate pathways from micronutrient intake and serum levels to Chronic Oral Diseases Burden. METHODS: We analyzed cross-sectional data from NHANES III (n = 7936) and NHANES 2011-2014 (n = 4929). The exposure was the intake and serum levels of vitamin D, calcium, and phosphorus. Considering the high correlation of those micronutrients in the diet, they were analyzed as a latent variable dubbed Micronutrient intake. The outcome was the Chronic Oral Diseases Burden, a latent variable formed by probing pocket depth, clinical attachment loss, furcation involvement, caries, and missing teeth. Pathways triggered by gender, age, socioeconomic status, obesity, smoking, and alcohol were also estimated using structural equation modeling. RESULTS: In both NHANES cycles, micronutrient intake (p-value < 0.05) and vitamin D serum (p-value < 0.05) were associated with a lower Chronic Oral Diseases Burden. Micronutrient intake reduced the Chronic Oral Diseases Burden via vitamin D serum (p-value < 0.05). Obesity increased the Chronic Oral Diseases Burden by reducing vitamin D serum (p-value < 0.05). CONCLUSION: Higher micronutrient intake and higher vitamin D serum levels seem to reduce Chronic Oral Diseases Burden. Healthy diet policies may jointly tackle caries, periodontitis, obesity, and other non-communicable diseases.

The Role of Dyslipidemia in Periodontitis.

Studies have suggested an important role of dyslipidemia, a condition with alterations in blood lipid levels, in promoting an additional effect on periodontal breakdown. Thus, this study aimed to explore the theoretical pathways associated with dyslipidemia and periodontitis. We used data from 11,917 US adults with complete periodontal examinations participating in the Third National Health and Nutrition Examination Survey (NHANES III). Our hypothesis was tested using structural equation modelling (SEM). Dyslipidemia was defined according to the National Cholesterol Education Program (NCEP-ATP III) and periodontitis as a latent variable reflecting the shared variance of the number of surfaces with periodontal pocket depth [PPD] = 4 mm, PPD = 5 mm, PPD ≥ 6 mm, clinical attachment level [CAL] = 4 mm, CAL = 5mm, CAL ≥ 6 mm, and furcation involvement. The model also considered distal determinants (age, sex, and socioeconomic status) and proximal determinants (HbA1c, smoking and alcohol consumption, and obesity). The model showed sufficient global fit (Root Mean Squared Error of Approximation = 0.04, 90%CI = 0.04−0.05, Tucker−Lewis Index = 0.93, Comparative Fit Index = 0.95). Age, sex, socioeconomic status, obesity, and smoking were directly associated with periodontitis (p < 0.01). Dyslipidemia revealed a significant direct effect on periodontitis (standardized coefficient [SC] = 0.086, SE 0.027; p < 0.01), also mediated via an indirect pathway through HbA1c (SC = 0.021; SE 0.010; p = 0.02) and obesity (SC = 0.036; SE 0.012; p < 0.01) and resulted in a total effect on periodontitis. Dyslipidemia was associated with periodontitis through a direct pathway and indirectly through HbA1c and obesity in the US population. These results support the need for a multi-professional approach to tackling oral and noncommunicable diseases (NCDs), directed at their common risk factors.

Salivary cytokine expression after non-surgical periodontal therapy in smokers: 12-month follow-up.

BACKGROUND: Diverse smoking trajectories may influence cytokine expression after non-surgical periodontal therapy and supportive periodontal care. Thus, we aimed to describe cytokine profiles in periodontal healing after periodontal therapy in smokers. METHODS: A smoking cessation program and non-surgical periodontal therapy were offered to 80 smokers with periodontitis. Smoking trajectories (quitters/light, moderate, heavy) were observed. The association of salivary interleukin (IL)-1ß, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-13, interferon-gamma (IFN-γ), and tumor necrosis factor-alpha (TNF-α) with smoking trajectories and periodontal outcomes was determined using mixed-effects linear regression. RESULTS: Among quitters/light smokers, IL-1ß was associated with an increase in mean periodontal pocket depth (PPD) and mean clinical attachment level (CAL). IL-6 was associated with a decrease in mean PPD and CAL in heavy smokers, whereas IL-8 was associated with a decrease in PPD among moderate smokers. TNFα was associated with a reduction in mean PPD and CAL among quitters/light smokers, while among moderate smokers, TNFα was associated with an increase in mean PPD and CAL. IL-12 and IL-13 were associated with a decrease in mean PPD in moderate smokers. CONCLUSION: Our findings suggest that distinctive smoking exposures induce differential cytokine expression, which, in turn, seems to influence periodontal repair.

Precursors of insulin resistance underlying periodontitis in adolescents aged 17-18 years.

OBJECTIVE: To investigate the association between insulin resistance markers and periodontitis in adolescents, analyzing confounder variables and the adiposity as a mediator. METHODS: This is population-based study is representative of adolescents aged 17-18 years from public schools in São Luís, Brazil (n = 405). Insulin resistance was assessed using the Model of Assessment of the Homeostasis of the Insulin Resistance Index (HOMA-IR) and its percussor triglycerides/HDL-cholesterol ratio (TG/HDL-c). The outcome was Initial Periodontitis, a latent variable estimated by the common variance shared among bleeding on probing, probing depth ≥ 4 mm, and clinical attachment loss ≥ 4 mm. The association between insulin resistance and Initial Periodontitis was modeled via pathways triggered by socioeconomic status, smoking, alcohol, and Adiposity, using structural equation modeling. RESULTS: Higher TG/HDL-c was directly associated with higher Initial Periodontitis (standardized coefficient [SC] = 0.130, p < 0.001). HOMA-IR was not associated with periodontal outcome (SC = 0.023, p = 0.075), but it was with Adiposity (SC = 0.495, p < 0.001). Higher TG/HDL-c was associated with Adiposity (SC = 0.202, p < 0.001). CONCLUSION: The insulin resistance markers were associated with early signs of periodontal breakdown among adolescents, suggesting a possible relationship between diabetes and periodontitis commences early in life.

Cytokine profiles and the dynamic of gingivitis development in humans.

AIM: To investigate the relationship between cytokine profiles and "fast" and "slow" patterns of gingival inflammation development. MATERIALS AND METHODS: Forty-two adults participated in an experimental gingivitis study, comprising a 2-week hygiene phase (clinical examination and professional cleaning); a 3-week induction phase (absence of oral hygiene); and a 2-week resolution phase (re-establishment of oral hygiene). Plaque and gingival inflammation scores were assessed. Interferon-gamma (IFN-γ), interleukin (IL)-1ß, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-13, and tumour necrosis factor-alpha (TNF-α) from gingival crevicular fluid were collected and measured by multiplex ELISA. Group-based-trajectory-modelling (GBTM) was used to model cytokine profiles over the induction phase. The effect of gingival inflammation on cytokine levels over time was estimated with mixed-effects modelling. RESULTS: GBTM analysis revealed two cytokine profiles, "non-organized response" (IL-4, IL-6, IL-8, IL-12, and IL-13) and "organized response" (IL-2, IL-10, and TNF-α). Among the "slow" responders, neither cytokine profile was associated with gingivitis. In contrast, a "fast" response was associated with a higher "non-organized response" factor (coef. 0.14) and a lower "organized response" factor (coef. -0.03). CONCLUSION: A "fast" gingivitis development was associated with a higher "non-organized response" and a lower "organized response", which may elucidate the role of individual variability in gingivitis susceptibility.

Dietary vitamin D and calcium and periodontitis: A population-based study.

Aim: This study aimed to explore the relationship between dietary vitamin D and calcium intake and periodontitis among adults and whether it differs from males to females. Methods: Cross-sectional analysis of a population-based cohort study with adults aged 20 to 60 from Southern Brazil. Intake of vitamin D and calcium were gathered in 2012 using two 24h-dietary recalls. Clinical examination assessed the clinical attachment level and bleeding on probing. Confounders included sex, age, family income, smoking, and obesity. The controlled direct effect of vitamin D and calcium on periodontitis was examined using marginal structural modeling. Analyses were also stratified by sex. Results: Of the 1,066 investigated adults (mean age 35 ± 11.7 years; 49% females), 12.3% (95%CI 10.2;14.7) had periodontitis. Calcium intake had a direct protective effect on periodontitis (risk ratio (RR) 0.61; 95%CI 0.45;0.83), whereas no association between vitamin D and periodontitis was observed (RR 1.13; 95%CI 0.82;1.56). Stratified analyses revealed a null association between both vitamin D and calcium intake and periodontitis among men, but a protective association between calcium and intake and periodontitis among women (RR 0.56; 95%CI 0.38;0.79), while vitamin D remained without any association (RR 1.07; 95%CI 0.72;1.61). Conclusion: Our findings suggest a protective association between dietary calcium intake and periodontitis among women.

Leukocytoclastic Vasculitis as an Extraintestinal Manifestation of Crohn's Disease.

Cutaneous involvement is one of the most common extraintestinal manifestations of inflammatory bowel disease (IBD). More commonly, pyoderma gangrenosum and erythema nodosum are noted, but psoriasis, aphthous stomatitis, Sweet's syndrome, and vasculitis may also occur. Leukocytoclastic vasculitis (LCV) is a rare cutaneous manifestation, characterized by the appearance of palpable purpura, urticaria, and ulcer-necrotic lesions predominantly in the lower extremities that improve with immunosuppressive therapy. In this case, we report a patient with CD and LCV. We also searched the literature on the diagnosis and treatment of LCV in patients with CD. Female, 31, presented with diarrhea containing mucus and blood, abdominal pain, arthralgia, and enanthematous plaques and ulcers with a hematinic background in the lower extremities. The results of the colonoscopy were compatible with CD and skin biopsy showed signs of LCV. Systemic autoimmune disease and primary vasculitis were ruled out. The patient received treatment with a systemic corticosteroid and the skin lesions improved. Outpatient treatment with antitumor necrosis factor therapy was initiated to promote skin healing and IBD clinical remission. As LCV is a rare manifestation of IBD, it is necessary to distinguish this dermatopathy from other systemic vasculitis. The engagement of a multidisciplinary team is essential for the correct diagnosis and management.

Immunomodulatory effect of short-term supplementation with Bacillus toyonensis BCT-7112T and Saccharomyces boulardii CNCM I-745 in sheep vaccinated with Clostridium chauvoei.

The bacterium Clostridium chauvoei is the causative agent of blackleg in livestock, and vaccination is the most effective means of prevention. The aim of this study was to assess the effect of short-term supplementation with Bacillus toyonensis and Saccharomyces boulardii on the immune response to a C. chauvoei vaccine in sheep. Sheep were vaccinated subcutaneously on day 0 and received a booster dose on day 21, with 2 mL of a commercial vaccine formulated with inactivated C. chauvoei bacterin adsorbed on aluminum hydroxide. Probiotics were orally administered B. toyonensis (3 × 108 cfu) and S. boulardii (3 × 108 cfu) over five days prior to the first and second doses of the vaccine. Sheep supplemented with B. toyonensis and S. boulardii showed significantly higher specific IgG, IgG1, and IgG2 titers (P<0.05), with approximately 24- and 14-fold increases in total IgG levels, respectively, than the nonsupplemented group. Peripheral blood mononuclear cells from the supplemented group had increased mRNA transcription levels of the IFN-γ, IL2, and Bcl6 genes. These results demonstrate an adjuvant effect of short-term supplementation with B. toyonensis and S. boulardii on the immune response against the C. chauvoei vaccine in sheep.

Influence of Bacterial Profiles in Cytokine and Clinical Features of Endodontic Disease.

INTRODUCTION: We verified the association between selected bacterial profiles and levels of cytokines, chemokines, and the expression of signs and symptoms of primary endodontic infection with apical periodontitis. METHODS: Samples were collected from 21 root canals, and macrophages were stimulated for 24 hours. Tumor necrosis factor alpha (TNF-α), interleukin (IL)-6, IL-10, IL-12p70, interferon gamma, and chemokine (C-C motif) ligand 2 (CCL2) were measured using cytometric bead array. We investigated the overlapping networks between cytokines and chemokines with regression analysis. Checkerboard DNA-DNA hybridization was used to assess 40 target bacteria species. Using factor analysis, bacterial species aggregated in 2 factors. The association of bacteria species-based factors on cytokine and chemokine levels and clinical features was estimated with regression analysis. RESULTS: A negative relationship between IL-10 (anti-inflammatory cytokine) and CCL2, TNF-α, and IFN-γ (proinflammatory cytokines) (all P < .05) was observed. CCL2 was positively correlated with TNF-α (P < .01). Thirty-eight bacteria species were detected in primary endodontic infection with apical periodontitis. The first bacteria species-based factor was associated with the size of the radiolucent area (coefficient = 15.42) and tenderness to percussion/pain on palpation (coefficient = 20.79). The second factor was associated with CCL2 levels (coefficient = 1.28). CONCLUSIONS: Different bacterial profiles can be differentially related to the expression of inflammatory proteins and the experience of clinical features.

Expression cassette and plasmid construction for Yeast Surface Display in Saccharomyces cerevisiae.

OBJECTIVES: Develop a Cell Surface Display system in Saccharomyces cerevisiae, based on the construction of an expression cassette for pYES2 plasmid. RESULTS: The construction of an expression cassette containing the α-factor signal peptide and the C-terminal portion of the α-agglutinin protein was made and its sequence inserted into a plasmid named pYES2/gDαAgglutinin. The construction allows surface display of bovine herpesvirus type 5 (BoHV-5) glycoprotein D (gD) on S. cerevisiae BY4741 strain. Recombinant protein expression was confirmed by dot blot, and indirect immunofluorescence using monoclonal anti-histidine antibodies and polyclonal antibodies from mice experimentally vaccinated with a recombinant gD. CONCLUSIONS: These results demonstrate that the approach and plasmid used represent not only an effective system for immobilizing proteins on the yeast cell surface, as well as a platform for immunobiologicals development.

Recombinant bovine IL17A acts as an adjuvant for bovine herpesvirus vaccine.

The Bovine herpes virus type 5 glycoprotein D (gD) is essential for viral penetration into host permissive cells. The Herpes virus gD glycoprotein has been used for bovine immunization, being efficient in reduction of viral replication, shedding and clinical signs, however sterilizing immunity is still not achieved. Recombinant subunit vaccines are, in general, poorly immunogenic requiring additional adjuvant components. Interleukin 17A (IL17A) is a pro-inflammatory cytokine produced by T helper 17 cells that mediate mucosal immunity. IL17 production during vaccine-induced immunity is a requirement for mucosal protection to several agents. In this study, we investigated the potential of a recombinant IL17A to act as an adjuvant for a recombinant BoHV-5 glycoprotein D vaccine in cattle. Three cattle groups were divided as: group 1) rgD5 + alumen + rIL-17A; 2) rgD5 + alumen; and 3) PBS + alumen. The cattle (3 per group) received two doses of their respective vaccines at an interval of 21 days. The group that received rIL17 in its vaccine formulation at the 7th day after the prime immunization had significant higher levels of specific rgD-IgG than the alumen group. Addition of rIL17 also led to a significant fold increase in specific anti-rgD IgG and neutralizing antibodies to the virus, respectively, when compared with the alumen group. Cells stimulated with rIL17A responded with IL17 transcription, as well IL2, IL4, IL10, IL15, Bcl6 and CXCR5. Our findings suggest that the rIL17A has adjuvant potential for use in vaccines against BoHV-5 as well as potentially other pathogens of cattle.

RA92A recombinant protein as immunogen to protect cattle against tick challenge in Brazil and Uganda

In this study, the recombinant gut protein rRa92A produced in Pichia pastoris yeast cells was used to immunize cattle in two experiments, one in Brazil and the other in Uganda. In both experiments, the animals were intramuscularly (IM) injected with 200 µg of recombinant protein in Brazil on days 0, 30 and 51 and in Uganda on days 0, 30. Blood samples for sera separation were collected from different days in both experiments. These samples were analyzed by ELISAs. In Brazil, ticks collected from the animals during the experimental period were analyzed for biological parameters. At Uganda, blood was collected to assess blood parameters, clinical signs were recorded and adult tick (Rhipicephalus appendiculatus) counts were performed. All animals of the vaccinated groups were shown to produce antibodies, and it was not possible to detect an effect on Rhipicephalus microplus. All the clinical parameters were considered within the normal ranges for both the experimental and control groups in Uganda. Antibody absorbance was elevated after each immunization and remained high until the end of the experiments, remaining low in the control animals. The results of stall test carried out in Brazil using R. microplus tick showed efficacy of 21.95%. The rRa92A immunization trial experiments in Uganda showing a decrease of 55.2% in the number of engorged adult ticks, which was statistically significant (p<0.05). Assessment of the immunogenicity of Ra92A produced in the P. pastoris expression system in bovines is reported for the first time, and the protein acted as a concealed antigen.

Biological and mechanical characterization of commercial and experimental periodontal surgical dressings

Abstract The objective of this study was to evaluate the biocompatibility and mechanical properties of two commercially available and one experimental periodontal dressing materials. The cytotoxicity of Periobond ® , Barricaid ® and one experimental periodontal dressing based on Exothane ® 8 monomer was tested on 3T3/NIH mouse fibroblast. Genotoxicity was assessed by micronuclei formation, and cell alterations were analyzed using light microscopy. Both biological assays were performed using the eluate obtained from specimens after 24, 72, or 168 hours of incubation. Mechanical characterization was assessed through the ultimate tensile strength and the water sorption and solubility tests. The significance level of α = 0.05 was used for all statistical analyses. All the materials promoted a cell viability lower than 60% in all evaluated times. In general, the cell viability was significantly reduced after 72 and 168h of specimens' incubation. Considering the factor material, there were not statistical differences in the cell viability (p = 0.156). The genotoxicity was not statistically significant among the groups in the different periods of time (p > 0.05). Differences in the ultimate tensile strength values were not statistically significant different among the groups (p = 0.125). Periobond ® showed the higher water sorption values (p < 0.001). Regarding solubility, there were no statistical differences between the groups (p = 0.098). All the periodontal dressing materials evaluated in this study exerted a cytotoxic effect against mouse fibroblasts, and their toxicity became more evident over time. Among the materials evaluated, the experimental light-cure type has shown overall similar properties to the commercial references.