Plaque Counts Research Articles

Comparative evaluation of chlorhexidine varnish and fluoride varnish on plaque Streptococcus mutans count – an in vivo study

The aim of the study was to assess and compare the effect of chlorhexidine varnish and fluoride varnish application on Streptococcus mutans counts in plaque of occlusal pits and fissures of permanent mandibular first molars. The study was an in vivo comparative study, conducted among 50 schoolchildren aged 7-8 years under a field setting. The 50 subjects were randomly allocated into two groups. Baseline plaque samples were collected from all the subjects followed by the application of two varnishes, Cervitec and Duraphat. The varnish was applied to pit and fissures of occlusal surface of mandibular first molar. The varnish application was carried out on the first day, fifth day and tenth day after baseline plaque sampling. Subsequent plaque samples were collected at the end of 1 month and at the end of 3 months after the varnish application. The Cervitec varnish has shown a statistically significant reduction at the end of 1 month and at the end of 3 months (P < 0.05). Duraphat varnish did not show a statistically significant difference in reducing the plaque S. mutans count at the end of 1 month and third month (P > 0.05). Cervitec varnish was found to be effective in reducing S. mutans count for a 3-month period, when compared to Duraphat varnish.

Linking protective GAB2 variants, increased cortical GAB2 expression and decreased Alzheimer's disease pathology.

GRB-associated binding protein 2 (GAB2) represents a compelling genome-wide association signal for late-onset Alzheimer’s disease (LOAD) with reported odds ratios (ORs) ranging from 0.75–0.85. We tested eight GAB2 variants in four North American Caucasian case-control series (2,316 LOAD, 2,538 controls) for association with LOAD. Meta-analyses revealed ORs ranging from (0.61–1.20) with no significant association (all p>0.32). Four variants were hetergeneous across the populations (all p<0.02) due to a potentially inflated effect size (OR = 0.61–0.66) only observed in the smallest series (702 LOAD, 209 controls). Despite the lack of association in our series, the previously reported protective association for GAB2 remained after meta-analyses of our data with all available previously published series (11,952-22,253 samples; OR = 0.82–0.88; all p<0.04). Using a freely available database of lymphoblastoid cell lines we found that protective GAB2 variants were associated with increased GAB2 expression (p = 9.5×10−7−9.3×10−6). We next measured GAB2 mRNA levels in 249 brains and found that decreased neurofibrillary tangle (r = −0.34, p = 0.0006) and senile plaque counts (r = −0.32, p = 0.001) were both good predictors of increased GAB2 mRNA levels albeit that sex (r = −0.28, p = 0.005) may have been a contributing factor. In summary, we hypothesise that GAB2 variants that are protective against LOAD in some populations may act functionally to increase GAB2 mRNA levels (in lymphoblastoid cells) and that increased GAB2 mRNA levels are associated with significantly decreased LOAD pathology. These findings support the hypothesis that Gab2 may protect neurons against LOAD but due to significant population heterogeneity, it is still unclear whether this protection is detectable at the genetic level.

A Novel Retro-Inverso Peptide Inhibitor Reduces Amyloid Deposition, Oxidation and Inflammation and Stimulates Neurogenesis in the APPswe/PS1ΔE9 Mouse Model of Alzheimer’s Disease

Previously, we have developed a retro-inverso peptide inhibitor (RI-OR2, rGffvlkGr) that blocks the in vitro formation and toxicity of the Aβ oligomers which are thought to be a cause of neurodegeneration and memory loss in Alzheimer’s disease. We have now attached a retro-inverted version of the HIV protein transduction domain ‘TAT’ to RI-OR2 to target this new inhibitor (RI-OR2-TAT, Ac-rGffvlkGrrrrqrrkkrGy-NH2) into the brain. Following its peripheral injection, a fluorescein-labelled version of RI-OR2-TAT was found to cross the blood brain barrier and bind to the amyloid plaques and activated microglial cells present in the cerebral cortex of 17-months-old APPswe/PS1ΔE9 transgenic mice. Daily intraperitoneal injection of RI-OR2-TAT (at 100 nmol/kg) for 21 days into 10-months-old APPswe/PS1ΔE9 mice resulted in a 25% reduction (p<0.01) in the cerebral cortex of Aβ oligomer levels, a 32% reduction (p<0.0001) of β-amyloid plaque count, a 44% reduction (p<0.0001) in the numbers of activated microglial cells, and a 25% reduction (p<0.0001) in oxidative damage, while the number of young neurons in the dentate gyrus was increased by 210% (p<0.0001), all compared to control APPswe/PS1ΔE9 mice injected with vehicle (saline) alone. Our data suggest that oxidative damage, inflammation, and inhibition of neurogenesis are all a downstream consequence of Aβ aggregation, and identify a novel brain-penetrant retro-inverso peptide inhibitor of Aβ oligomer formation for further testing in humans as a potential disease-modifying treatment for Alzheimer’s disease.

Graphical Modeling of Gene Expression in Monocytes Suggests Molecular Mechanisms Explaining Increased Atherosclerosis in Smokers

Smoking is a risk factor for atherosclerosis with reported widespread effects on gene expression in circulating blood cells. We hypothesized that a molecular signature mediating the relation between smoking and atherosclerosis may be found in the transcriptome of circulating monocytes. Genome-wide expression profiles and counts of atherosclerotic plaques in carotid arteries were collected in 248 smokers and 688 non-smokers from the general population. Patterns of co-expressed genes were identified by Independent Component Analysis (ICA) and network structure of the pattern-specific gene modules was inferred by the PC-algorithm. A likelihood-based causality test was implemented to select patterns that fit models containing a path “smoking→gene expression→plaques”. Robustness of the causal inference was assessed by bootstrapping. At a FDR ≤0.10, 3,368 genes were associated to smoking or plaques, of which 93% were associated to smoking only. SASH1 showed the strongest association to smoking and PPARG the strongest association to plaques. Twenty-nine gene patterns were identified by ICA. Modules containing SASH1 and PPARG did not show evidence for the “smoking→gene expression→plaques” causality model. Conversely, three modules had good support for causal effects and exhibited a network topology consistent with gene expression mediating the relation between smoking and plaques. The network with the strongest support for causal effects was connected to plaques through SLC39A8, a gene with known association to HDL-cholesterol and cellular uptake of cadmium from tobacco, while smoking was directly connected to GAS6, a gene reported to have anti-inflammatory effects in atherosclerosis and to be up-regulated in the placenta of women smoking during pregnancy. Our analysis of the transcriptome of monocytes recovered genes relevant for association to smoking and atherosclerosis, and connected genes that before, were only studied in separate contexts. Inspection of correlation structure revealed candidates that would be missed by expression-phenotype association analysis alone.

Oral Health Status , Salivary Physical Properties and Salivary Mutans Streptococci among a Group of Mouth Breathing Patients in Comparison to Nose Breathing

Background: Mouth breathing can lead to introduce cold, dry unprepared air that insults the tissue of oral cavity, nasopharynx and lung, leading in turn to pathological changes in oronasal cavity, nasopharyngeal and other respiratory tissue, mouth breathing associated with nasal obstruction may lead to many health problems, in particular oral health problems such as inflammation of gingiva, oral dryness, change in oral environment that may decrease pH, salivary flow rate and increase bacteria and dental caries.Aims of the present study were to assess the oral health condition among mouth breather associated with nasal obstruction, including dental caries, oral cleanliness and gingival health condition as well as to evaluate the changes in salivary physical characteristics and salivary mutans streptococci counts, and their relation to oral variables in comparison to a control group. Materials and Methods: Thirty patients with mouth breathing associated with nasal obstruction (15 females and 15 males) were selected as a study group with an age range (18-22) years old, all subjects were examined by ENT specialist to confirm mouth breathing. A 30 gender and age matched healthy looking subjects without nasal obstruction were selected as control. The diagnosis and recording of dental caries was according to severity of dental caries lesion through the application of D1_4MFS(Manji et al., 1989). Plaque index of (Silness and Loe, 1964) was used for plaque assessment; gingival index of (Loe and Silness, 1963) was used for gingival health condition assessment. Stimulated salivary samples were collected according to (Tenovuo and Lagerlof, 1996) and the following variables were recorded: microbiological analysis included the salivary counts of mutans streptococci, salivary flow rate, salivary pH (potential of hydrogen) and then measurement of salivary viscosity by using Ostwald's viscometer. Results: Results of the present study showed that the mouth breathing group had statistically highly significant, higher plaque and gingival indices than nose breathing group (P<0.01) with a positive highly significant correlation between them in mouth breathing and nose breathing groups (r=0.56, r= 0.64, respectively).The salivary flow rate was lower among mouth breathing with highly significant difference than nose breathing (P<0.01), also salivary pH was lower among mouth breathing but with significant differencecompare to nose breathing (P<0.05); statistically a negative highly significant correlation was recorded among mouth breathing group between salivary flow rate with gingival index (r= -0.56). It has been found that salivary viscosity was not statistically significant difference between mouth breathing group and nose breathing group. The salivary viscosity was found to be inversely significantly correlated with salivary flow rate among mouth breathing group (r= -0.38). While it was positively not significantly correlated with plaque index, gingival index and counts of mutans streptococci among mouth breathing group. Data analysis of the present study showed that salivary mutans streptococci counts among mouth breathing group were higher than that among nose breathing group, difference was statistically highly significant (P<0.01). Conclusion: Mouth breathing associated with nasal obstruction may have an effect on oral health status, leading to an increase in periodontal disease and changes in dental caries.

Fresh ginger (Zingiber officinale) has anti-viral activity against human respiratory syncytial virus in human respiratory tract cell lines

Ethnopharmacological relevanceGinger, Zingiber officinale Roscoe, is a common spice and also a widely used medicinal plant in ancient China. Ginger is an ingredient of Ge-Gen-Tang (Kakkon-to; GGT). GGT has been proved to have antiviral activity against human respiratory syncytial virus (HRSV). However, it is unknown whether ginger is effective against HRSV. Aim of the studyTo find a readily available agent to manage HRSV infection, the authors tested the hypothesis that ginger can effectively decrease HRSV-induced plaque formation in respiratory mucosal cell lines. Materials and methodsEffect of hot water extracts of fresh and dried gingers on HRSV was tested by plaque reduction assay in both human upper (HEp-2) and low (A549) respiratory tract cell lines. Ability of ginger to stimulate anti-viral cytokines was evaluated by enzyme-linked immunosorbent assay (ELISA). ResultsFresh ginger dose-dependently inhibited HRSV-induced plaque formation in both HEp-2 and A549 cell lines (p<0.0001). In contrast, dried ginger didn't show any dose-dependent inhibition. 300μg/ml fresh ginger could decrease the plaque counts to 19.7% (A549) and 27.0% (HEp-2) of that of the control group. Fresh ginger was more effective when given before viral inoculation (p<0.0001), particularly on A549 cells. 300μg/ml fresh ginger could decrease the plaque formation to 12.9% when given before viral inoculation. Fresh ginger dose-dependently inhibited viral attachment (p<0.0001) and internalization (p<0.0001). Fresh ginger of high concentration could stimulate mucosal cells to secrete IFN-β that possibly contributed to counteracting viral infection. ConclusionsFresh, but not dried, ginger is effective against HRSV-induced plaque formation on airway epithelium by blocking viral attachment and internalization.

An improved respiratory syncytial virus neutralization assay based on the detection of green fluorescent protein expression and automated plaque counting

BackgroundVirus neutralizing antibodies against respiratory syncytial virus (RSV) are considered important correlates of protection for vaccine evaluation. The established plaque reduction assay is time consuming, labor intensive and highly variable.MethodsHere, a neutralization assay based on a modified RSV strain expressing the green fluorescent protein in combination with automated detection and quantification of plaques is described.ResultsThe fluorescence plaque reduction assay in microplate format requires only two days to complete and is simple and reproducible. A good correlation between visual and automated counting methods to determine RSV neutralizing serum antibody titers was observed.ConclusionsThe developed virus neutralization assay is suitable for high-throughput testing and can be used for both animal studies and (large scale) vaccine clinical trials.

Effectiveness of school‐based dental sealant programs among children from low‐income backgrounds in France: a pragmatic randomized clinical trial

A split-mouth randomized clinical trial was carried out to assess the effectiveness of a school-based dental sealant (SBDS) program for French children from low-income backgrounds. The secondary objectives were to determine the effectiveness of the program according to the baseline individual caries risk (ICR) and to assess the sealant retention. The study included 276 pupils from Nice. The sealing was performed in the first or second grades. The permanent first molars were randomized into two groups: one received resin-based sealant and the other no treatment. Active caries on permanent and temporary teeth, visible plaque, Streptococcus mutans (SM) and Lactobacillus counts were recorded at baseline to assess ICR. An intent-to-treat analysis was performed based on the occurrence of new caries at 1 year of follow-up (study outcome). Univariate and multivariable conditional tooth-matched logistic regression analyses (Proc phreg; SAS) were used to assess the effectiveness of the SBDS program. At 1 year of follow-up, 253 children (421 pairs) remained in the analysis. In the adjusted analysis, first permanent molars that received sealants were less at risk of developing new caries at 1 year of follow-up compared with those from the control group (OR = 0.26, 95% CI: 0.14-0.49). Independently of the treatment, SM count, and all other covariates, only the variable 'active caries at baseline' remained related to new caries on first molars at 1 year (OR = 3.11, 95% CI: 1.27-7.62). The effect of the sealants was significant only when the analyses included subjects with active caries (OR = 0.25; 95% CI: 0.12-0.50) or with a high SM count (>10(5) , OR = 0.20; 95% CI: 0.10-0.41) at baseline. At the 1-year follow-up, total retention was recorded in 52.7% (n = 222) of the treated teeth. The 1-year effectiveness of the SBDS program was demonstrated in low socio-economic areas. Selection of schoolchildren according to individual caries risk factors should be considered in a SBDS program.

Pittsburgh compound B and the postmortem diagnosis of alzheimer disease

Deposition of the amyloid-β (Aβ) peptide in neuritic plaques is a requirement for the diagnosis of Alzheimer disease (AD). Although the continued development of in vivo imaging agents such as Pittsburgh compound B (PiB) is promising, the diagnosis of AD is still challenging. This can be partially attributed to our lack of a detailed understanding of the interrelationship between the various pools and species of Aβ and other common indices of AD pathology. We hypothesized that recent advances in our ability to accurately measure Aβ postmortem (for example, using PiB), could form the basis of a simple means to deliver an accurate AD diagnosis. We conducted a comprehensive analysis of the amount of Aβ40 and Aβ42 in increasingly insoluble fractions, oligomeric Aβ, and fibrillar Aβ (as defined by PiB binding), as well as plaques (diffuse and neuritic), and neurofibrillary tangles in autopsy specimens from age-matched, cognitively normal controls (n = 23) and AD (n = 22) cases, across multiple brain regions. Both PiB binding and the amount of sodium dodecyl sulfate (SDS)-soluble Aβ were able to predict disease status; however, SDS-soluble Aβ was a better measure. Oligomeric Aβ was not a predictor of disease status. PiB binding was strongly related to plaque count, although diffuse plaques were a stronger correlate than neuritic plaques. Although postmortem PiB binding was somewhat useful in distinguishing AD from control cases, SDS-soluble Aβ measured by standard immunoassay was substantially better. These findings have important implications for the development of imaging-based biomarkers of AD.

Estimating the Temporal Evolution of Alzheimer's Disease Pathology with Autopsy Data

The temporal growth of Alzheimer's disease (AD) neuropathology cannot be easily determined because autopsy data are available only after death. We combined autopsy data from 471 participants in the Honolulu-Asia Aging Study (HAAS) into latent factor measures of neurofibrillary tangle and neuritic plaque counts. These were associated with intercept and slope parameters from a latent growth curve (LGC) model of 9-year change in cognitive test performance in 3244 autopsied and non-autopsied HAAS participants. Change in cognition fully mediated the association between baseline cognitive performance and AD lesions counts. The mediation effect of cognitive change on both AD lesion models effectively dates them within the period of cognitive surveillance. Additional analyses could lead to an improved understanding of lesion propagation in AD.

Neuropathologically defined subtypes of Alzheimer's disease differ significantly from neurofibrillary tangle-predominant dementia.

Alzheimer's disease (AD) can be classified based on the relative density of neurofibrillary tangles (NFTs) in the hippocampus and association cortices into three subtypes: typical AD, hippocampal-sparing AD (HpSp AD), and limbic-predominant AD (LP AD). AD subtypes not only have pathologic, but also demographic, clinical, and genetic differences. Neurofibrillary tangle-predominant dementia (NFTD), a disorder with NFTs relatively restricted to limbic structures, shares this feature with LP AD raising the possibility that NFTD is a variant of AD. The objective criteria for pathologic diagnosis of NFTD are not available. A goal of this study was to design a mathematical algorithm that could diagnose NFTD from NFT and senile plaque (SP) counts in hippocampus and association cortices, analogous to that used to subtype AD. Moreover, we aimed to compare pathologic, demographic, clinical, and genetic features of NFTD (n = 18) with LP AD (n = 19), as well as the other AD subtypes, typical AD (n = 52) and HpSp AD (n = 17). Using digital microscopy, we confirmed that burden of phospho-tau (CP13) and of an NFT conformational epitope (Ab39) correlated with NFT densities and showed expected patterns across AD subtypes. HpSp AD had the highest and LP AD had the lowest burden of cortical CP13 and Ab39 immunoreactivity. On the other hand, cortical β-amyloid burden did not significantly differ between AD subtypes. Semi-quantitative assessment of SPs in the basal ganglia did show HpSp AD to have significantly more frequent presence of SPs compared to typical AD, which was more frequent than LP AD. Compared to LP AD, NFTD had an older age at disease onset and shorter disease duration, as well as lower Braak NFT stage. NFTs and SPs on thioflavin-S fluorescent microscopy, as well as CP13, Ab39, and Aβ immunoreactivities were very low in the frontal cortex of NFTD, differentiating NFTD from AD subtypes, including LP AD. MAPT H1H1 genotype frequency was high (~70 %) in NFTD and LP AD, and similar to typical AD, while APOE ε4 carrier state was low in NFTD. While it shares clinical similarities with regard to female sex predominance, onset in advanced age, and a slow cognitive decline, NFTD has significant pathologic differences from LP AD, suggesting that it may not merely be a variant of AD.

Clinically concordant variations of Alzheimer pathology in aphasic versus amnestic dementia

Primary progressive aphasia is a neurodegenerative syndrome characterized by gradual dissolution of language but relative sparing of other cognitive domains, especially memory. It is associated with asymmetric atrophy in the language-dominant hemisphere (usually left), and differs from typical Alzheimer-type dementia where amnesia is the primary deficit. Various pathologies have been reported, including the tangles and plaques of Alzheimer's disease. Identification of Alzheimer pathology in these aphasic patients is puzzling since tangles and related neuronal loss in Alzheimer's disease typically emerge in memory-related structures such as entorhinal cortex and spread to language-related neocortex later in the disease. Furthermore, Alzheimer pathology is typically symmetric. How can a predominantly limbic and symmetric pathology cause the primary progressive aphasia phenotype, characterized by relative preservation of memory and asymmetric predilection for the language-dominant hemisphere? Initial investigations into the possibility that Alzheimer pathology displays an atypical distribution in primary progressive aphasia yielded inconclusive results. The current study was based on larger groups of patients with either primary progressive aphasia or a typical amnestic dementia. Alzheimer pathology was the principal diagnosis in all cases. The goal was to determine whether Alzheimer pathology had clinically-concordant, and hence different distributions in these two phenotypes. Stereological counts of tangles and plaques revealed greater leftward asymmetry for tangles in primary progressive aphasia but not in the amnestic Alzheimer-type dementia (P < 0.05). Five of seven aphasics had more leftward tangle asymmetry in all four neocortical regions analysed, whereas this pattern was not seen in any of the predominantly amnestic cases. One aphasic case displayed higher right-hemisphere tangle density despite greater left-hemisphere hypoperfusion and atrophy during life. Although there were more tangles in the memory-related entorhinal cortex than in language-related neocortical areas in both phenotypes (P < 0.0001), the ratio of neocortical-to-entorhinal tangles was significantly higher in the aphasic cases (P = 0.034). Additionally, overall numbers of tangles and plaques were greater in the aphasic than amnestic cases (P < 0.05), especially in neocortical areas. No significant hemispheric asymmetry was found in plaque distribution, reinforcing the conclusion that tangles have greater clinical concordance than plaques in the spectrum of Alzheimer pathologies. The presence of left-sided tangle predominance and higher neocortical-to-entorhinal tangle ratio in primary progressive aphasia establishes clinical concordance of Alzheimer pathology with the aphasic phenotype. The one case with reversed asymmetry, however, suggests that these concordant clinicopathological relationships are not universal and that individual primary progressive aphasia cases with Alzheimer pathology exist where distributions of plaques and tangles do not account for the observed phenotype.

Gap junction pathology in multiple sclerosis lesions and normal-appearing white matter

Oligodendrocyte gap junctions (GJs) are vital for central nervous system myelination, but their involvement in multiple sclerosis (MS) pathology remains unknown. The aim of this study was to examine alterations of oligodendrocyte and related astrocyte GJs in MS lesions and normal-appearing white matter (NAWM). Post-mortem brain samples from 9 MS and 11 age-matched non-MS control patients were studied. Tissue sections that included both chronic active and inactive lesions were characterized neuropathologically with Luxol Fast Blue staining and immunostaining for myelin oligodendrocyte glycoprotein (MOG) and the microglial marker Iba1. We analyzed the expression of Cx32 and Cx47 in oligodendrocytes and of Cx43, the major astrocytic partner in oligodendrocyte-astrocyte (O/A) GJs by quantitative immunoblot and real-time PCR. Formation of GJ plaques was quantified by immunohistochemistry. Compared to control brains, both Cx32 and Cx47 GJ plaques and protein levels were reduced in and around MS lesions, while Cx43 was increased as part of astrogliosis. In the NAWM, Cx32 was significantly reduced along myelinated fibers whereas Cx47 showed increased expression mainly in oligodendrocyte precursor cells (OPCs). However, OPCs showed only limited connectivity to astrocytes. Cx43 showed modestly increased levels in MS NAWM compared to controls, while GJ plaque counts were unchanged. Our findings indicate that oligodendrocyte GJs are affected not only in chronic MS lesions but also in NAWM, where disruption of Cx32 GJs in myelinated fibers may impair myelin structure and function. Moreover, limited O/A GJ connectivity of recruited OPCs in the setting of persistent inflammation and astrogliosis may prevent differentiation and remyelination.

An evaluation of the impact of MAPT, SNCA and APOE on the burden of Alzheimer's and Lewy body pathology

PurposeThe study investigates the effects of genetic factors on the pathology of Alzheimer's disease (AD) and Lewy body (LB) diseases, including Parkinson's disease and dementia with Lewy bodies.MethodsA multicentre autopsy...

Epifluorescence and atomic force microscopy: Two innovative applications for studying phage–host interactions in Lactobacillus helveticus

Bacteriophages attacking lactic acid bacteria (LAB) still represent a crucial problem in industrial dairy fermentations. The consequences of a phage infection against LAB can lead to fermentation delay, alteration of the product quality and, in most severe cases, the product loss. Phage particles enumeration and phage–host interactions are normally evaluated by conventional plaque count assays, but, in many cases, these methods can be unsuccessful. Bacteriophages of Lactobacillus helveticus, a LAB species widely used as dairy starter or probiotic cultures, are often unable to form lysis plaques, thus impairing their enumeration by plate assay. In this study, we used epifluorescence microscopy to enumerate L. helveticus phage particles from phage-infected cultures and Atomic Force Microscopy (AFM) to visualize both phages and bacteria during the different stages of the lytic cycle. Preliminary, we tested the sensitivity of phage counting by epifluorescence microscopy. To this end, phage particles of ΦAQ113, a lytic phage of L. helveticus isolated from a whey starter culture, were stained by SYBR Green I and enumerated by epifluorescence microscopy. Values obtained by the microscopic method were 10 times higher than plate counts, with a lowest sensitivity limit of ≥ 6 log phage/ml. The interaction of phage ΦAQ113 with its host cell L. helveticus Lh1405 was imaged by AFM after 0, 2 and 5 h from phage–host adsorption. The lytic cycle was followed by epifluorescence microscopy counting and the concomitant cell wall changes were visualized by AFM imaging. Our results showed that these two methods can be combined for a reliable phage enumeration and for studying phage and host morphology during infection processes, thus giving a complete overview of phage–host interactions in L. helveticus strains involved in dairy productions.

Dental health of 6-year-old children in Alpes Maritimes, France

To describe the dental health status of 6-year-old children using the ICDAS-II advanced method and to evaluate the association between the known caries risk factors with the cavitated caries lesion (WHO basic method) or with both non-cavitated and cavitated caries lesion caries (ICDAS II). In this cross-sectional study, a questionnaire was used to evaluate oral health and dietary habits of children. A clinical examination and a Cario analysis test (Pierre Fabre Oral care) were performed. Logistic regression analyses were used to assess the association between caries and daily tooth-brushing, dietary habits, visible plaque and salivary factors. There were 341 children (52% female and 6.25+/-0.46 years of age) in this study. Using the ICDAS-II advanced method, 39% of the children were caries-free. This proportion was larger (67.2%) using the WHO method. In multivariate models, visible dental plaque and Streptococcus mutans count were associated with caries experience registered as ICDAS-II codes 1-6 or codes 3-6. The absence of daily tooth-brushing with fluoridated toothpaste was associated only with caries experience ICDAS-II codes 3-6. The use of WHO or ICDAS-II method changed the proportion of caries-free children but not the clinical caries risk factors associated with caries experience.

Harm Avoidance and Risk of Alzheimer's Disease

To test the hypothesis that harm avoidance, a trait associated with behavioral inhibition, is associated with the risk of developing Alzheimer's disease (AD). A total of 791 adults 55 years and older without dementia completed a standard self-report measure of harm avoidance. They then underwent annual evaluations that included detailed cognitive testing and clinical classification of mild cognitive impairment (MCI), dementia, and AD. In a uniform neuropathologic examination of those who died, counts of neuritic plaques, diffuse plaques, and neurofibrillary tangles were standardized and combined to yield a pathologic measure of disease. The relation of harm avoidance to incidence of AD and related outcomes was estimated in analyses adjusted for age, sex, and education. During a mean of 3.5 years of annual observation, 98 people (12.4%) developed incident AD. A high level of harm avoidance (90th percentile) was associated with a more than two-fold increase in risk of AD compared with a low score (10th percentile). Higher harm avoidance was also associated with increased incidence of MCI and more rapid decline in episodic memory, working memory, and perceptual speed (but not semantic memory or visuospatial ability). In 116 participants who died and underwent brain autopsy, harm avoidance was not related to a composite measure of plaques and tangles. A high level of the harm avoidance trait, indicating a tendency toward behavioral inhibition, is related to the risk of developing AD and its precursor, MCI.

Rapid and accurate determination of the potency of varicella vaccine by quantitative polymerase chain reaction

The potency of varicella vaccines is currently determined by a plaque assay technique, which usually takes seven days and is laborious and has considerable inter- and intra-assay variability. Here, we report a new potency assay for varicella vaccine based on quantitative polymerase chain reaction in conjunction with a much more efficient virus infection step. Potency results can be obtained within 24h of infection and demonstrates acceptable accuracy and reproducibility when compared with the plaque assay, which relies on manual counting of plaques formed one week after viral infection. Using multiple vaccine lots from 7 manufacturers, we found no significant difference in infectivity determined between the new assay and plaque assay. The optimized conditions for viral infection and polymerase chain reaction are of significant value for the potency determination of the vaccine due to its rapidity, accuracy and the high throughput capacity of the assay.

Development and evaluation of an automatable focus reduction neutralisation test for the detection of measles virus antibodies using imaging analysis

A plaque reduction neutralisation test (PRNT) is still regarded as the gold standard for the investigation of anti-measles immunity. In this study, an alternative simplified automatable focus reduction neutralisation test (AFRNT) based on the classical PRNT was developed. The AFRNT uses the conventional Edmonston strain of measles, immunoperoxidase staining with monoclonal antibodies, and automated plaque counts performed with AID ViruSpot software. The assay is performed in 96-well plates, requires 2 days, and is fully automatable. The AFRNT was evaluated in comparison with PRNT and Enzygnost anti-measles enzyme immunoassay (EIA). A total of 130 samples, which included two available WHO international anti-measles standards, sera from 90 patients, and 38 different lots of immunoglobulin products, were tested. Overall, good agreement was observed between EIA and both neutralisation tests; however, the EIA values for the immunoglobulin products and international standards were slightly but significantly higher than those of the neutralisation tests. The Bland–Altman analysis showed excellent agreement between AFRNT and PRNT. AFRNT is a fully automatable high-throughput neutralisation assay, which can be performed with measles and other types of viruses, including wild-type strains. It is perfectly suited for epidemiological and vaccine studies.

New Diagnostic Criteria for Alzheimer‘s Disease

More than 100 years ago, Alois Alzheimer described the clinical presentation and neu‐ ropathological changes, including neuronal loss, extracellular plaques and intraneuronal tangles, in a single patient with the disease that now bears his name [1]. Over the years, it became clear that the pathological character‐ istics of Alzheimer’s disease (AD) were com‐ mon in individuals who died demented. The finding of a correlation between plaque counts and dementia severity put great focus on the involvement of plaques in the disease process [2]. Similar but even stronger correlations were later seen between dementia severity and the number of tangles in the cerebral cortex [3]. However, it took almost 80 years from Alzheimer’s case presentation to reveal the molecular compo‐ sition of plaques and tangles (i.e., aggregated amyloid‐b [Ab] [4,5] and hyperphosphorylated tau proteins [6,7]).