Exploring bacterial diversity of endodontic microbiota by cloning and sequencing 16S rRNA

Introduction

The characterization of microbial communities infecting the endodontic system in each clinical condition may help on the establishment of a correct prognosis and distinct strategies of treatment. The purpose of this study was to determine the bacterial diversity in primary endodontic infections by 16S ribosomal-RNA (rRNA) sequence analysis.

Methods

Samples from root canals of untreated asymptomatic teeth (n = 12) exhibiting periapical lesions were obtained, 16S rRNA bacterial genomic libraries were constructed and sequenced, and bacterial diversity was estimated.

Results

A total of 489 clones were analyzed (mean, 40.7 ± 8.0 clones per sample). Seventy phylotypes were identified of which six were novel phylotypes belonging to the family Ruminococcaceae. The mean number of taxa per canal was 10.0, ranging from 3 to 21 per sample; 65.7% of the cloned sequences represented phylotypes for which no cultivated isolates have been reported. The most prevalent taxa were Atopobium rimae (50.0%), Dialister invisus, Prevotella oris, Pseudoramibacter alactolyticus, and Tannerella forsythia (33.3%).

Conclusions

Although several key species predominate in endodontic samples of asymptomatic cases with periapical lesions, the primary endodontic infection is characterized by a wide bacterial diversity, which is mostly represented by members of the phylum Firmicutes belonging to the class Clostridia followed by the phylum Bacteroidetes.     

Reliability and discriminatory capacity of a clinical scale for assessing abdominal muscle coordination

Objective

This study evaluated the reliability and discriminatory capacity of a novel clinical scale for assessing abdominal muscle coordination. We investigated the interrater reliability of this tool in patients with chronic low back pain (LBP) (reliability section); the ability of this tool to discriminate healthy and LBP subjects (discriminatory section); and the association between the score and measures of pain, disability, and kinesiophobia (correlation section).

Methods

For the reliability section of this study, 14 patients with chronic LBP were included. For the discriminatory section, 10 patients with chronic LBP and 10 pain-free controls were recruited. In the correlation study, data from the 10 chronic LBP patients in the discriminatory section were used. The clinical test was conducted by a blinded examiner while the subjects attempted to independently activate transversus abdominis separate from the rest of the abdominal muscles (hollowing or draw-in maneuver). The intraclass correlation coefficients, receiver operating characteristic curve, and Pearson r correlation coefficients were calculated to assess reliability and validity.

Results

An intraclass correlation coefficient_(2,1) of 0.72 (95% confidence interval, 0.33-0.90) was recorded for interrater reliability. The tool correctly identified the subject condition in 97% of the cases. The score did not correlate substantially with any clinical measures, with Pearson r ranging from 0.122 (P = .737) to 0.493 (P = .148).

Conclusions

This study showed that the proposed scale is a reliable tool and may be useful in discriminating patients with chronic LBP from pain-free controls. The poor correlation between the score and clinical measures may be due to the multidimensional nature of chronic LBP.     

Analogues of orphan nuclear receptor small heterodimer partner ligand and apoptosis inducer (E)-4-[3-(1-adamantyl)-4-hydroxyphenyl]-3-chlorocinnamic acid. 2. Impact of 3-chloro group replacement on inhibition of proliferation and induction of apoptosis of leukemia and cancer cell lines

The parent phenol of adapalene and its (E)-cinnamic acid analogue were found to induce cancer cell apoptosis but cause adverse systemic effects when administered to mice. In contrast, their respective 5-Cl- and 3-Cl-substituted analogues had their adverse effects mitigated without a comparable loss of cancer cell inhibitory activity. As a result, pharmacologic space in this region of the cinnamic phenyl ring scaffold was explored. Various substituents were introduced, and their effects on cancer cell proliferation and viability were evaluated. Cinnamic acids having 3-Br, CN, NO(2), NH(2), OMe, and N(3) groups had activity comparable to that of 4-[3'-(1-adamantyl)-4'-hydroxyphenyl]-3-chlorocinnamic acid. A comparative molecular field analysis study indicated that introduction of an H-bond acceptor at position 3 of the central phenyl ring would favor inhibition of leukemia cell viability, and docking suggested its hydrogen bonding with a polar group in a small heterodimer partner homology model. The 3-CN, NO(2), NH(2), and OH analogues also inhibited MMTV-Wnt1 murine mammary stem cell viability.     

The role of 5-HT₁A receptors in fish oil-mediated increased BDNF expression in the rat hippocampus and cortex: a possible antidepressant mechanism

Epidemiological and dietary studies show that nutritional deficit of omega-3 polyunsaturated fatty acids (ω-3 PUFA) is directly related to the prevalence and severity of depression. Supplementation with docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) during critical periods of development (pregnancy and lactation) is essential for cortical maturation, synaptogenesis and myelination, and may also mitigate the risk for cognitive deficits and psychopathologies in young adults. The present study was performed to evaluate the involvement of serotonin (5-HT) receptors, particularly of 5-HT(1A), and hippocampal brain-derived neurotrophic factor (BDNF) expression in the antidepressant effect of ω-3 PUFA supplementation. In Experiment 1, the antidepressant effects of fish oil were assessed by the modified forced swim test in adult rats. The data indicated a robust antidepressant effect produced by this supplementation and that treatment of the rats with WAY 100135 reversed this effect. In Experiment 2, cortical and hippocampal contents of BDNF, 5-HT, dopamine (DA) and its metabolites, 5-hydroxyindoleacetic acid (5-HIAA), and 3,4-dihydroxyphenylacetic acid (DOPAC), were determined in animals subjected to the same protocol. Increased BDNF expression in the cortex and hippocampus of both age groups was detected. In 90 day-old rats, 5-HT content in the hippocampus was increased, whereas 5-HIAA formation was diminished in the fish oil group. We suggest the occurrence of a reciprocal involvement of 5-HT(1A) receptors activation and the hippocampal BDNF-increased expression mediated by fish oil supplementation. These data corroborate and expand the notion that supplementation with ω-3 PUFA produces antidepressant effects mediated by an increase in serotonergic neurotransmission, particularly in the hippocampus. This article is part of a Special Issue entitled 'Anxiety and Depression'.     

Effect of ganaxolone and THIP on operant and limited-access ethanol self-administration

Recent evidence suggests that GABA(A) receptor ligands may regulate ethanol intake via effects at both synaptic and extrasynaptic receptors. For example, the endogenous neurosteroid, allopregnanolone (ALLO) has a similar pharmacological profile as ethanol, and it alters ethanol intake in rodent models. Additionally, recent evidence suggests that δ-subunit-containing extrasynaptic GABA(A) receptors may confer high sensitivity to both ethanol and neurosteroids. The purpose of the present study was to determine the effects of ganaxolone (GAN; an ALLO analog) and gaboxadol (THIP; a GABA(A) receptor agonist with selectivity for the extrasynaptic δ-subunit) on ethanol intake, drinking patterns, and bout characteristics in operant and limited-access self-administration procedures. In separate studies, the effects of GAN (0-10 mg/kg) and THIP (2-16 mg/kg) were tested in C57BL/6J male mice provided with 2-h access to a two-bottle choice of water or 10% ethanol or trained to respond for 30 min of access to 10% ethanol. GAN had no overall significant effect on operant ethanol self-administration, but tended to decrease the latency to consume the first bout. In the limited-access procedure, GAN dose-dependently decreased ethanol intake. THIP dose-dependently decreased ethanol intake in both paradigms, altering both the consummatory and appetitive processes of operant self-administration as well as shifting the drinking patterns in both procedures. These results add to literature suggesting time-dependent effects of neurosteroids to promote the onset, and to subsequently decrease, ethanol drinking behavior, and they support a role for extrasynaptic GABA(A) receptor activation in ethanol reinforcement.     

Caries inhibition in vital teeth using 9.6-μm CO2-laser irradiation

The aim of this study was to test the hypothesis that in a short-term clinical pilot trial short-pulsed 9.6 μm CO(2)-laser irradiation significantly inhibits demineralization in vivo. Twenty-four subjects scheduled for extraction of bicuspids for orthodontic reasons (age 14.9 ± 2.2 years) were recruited. Orthodontic brackets were placed on bicuspids (Transbond XT, 3M). An area next to the bracket was irradiated with a CO(2)-laser (Pulse System Inc, Los Alamos, New Mexico), wavelength 9.6 μm, pulse duration 20 μs, pulse repetition rate 20 Hz, beam diameter 1100 μm, average fluence 4.1 ± 0.3J∕cm(2), 20 laser pulses per spot. An adjacent nonirradiated area served as control. Bicuspids were extracted after four and twelve weeks, respectively, for a quantitative assessment of demineralization by cross-sectional microhardness testing. For the 4-week arm the mean relative mineral loss ΔZ (vol% × μm) for the laser treated enamel was 402 ± 85 (mean ± SE), while the control showed significantly higher mineral loss (ΔZ 738 ± 131; P = 0.04, t-test). The difference was even larger after twelve weeks (laser arm ΔZ 135 ± 98; control 1067 ± 254; P = 0.002). The laser treatment produced 46% demineralization inhibition for the 4-week and a marked 87% inhibition for the 12-week arm. This study shows, for the first time in vivo, that the short-pulsed 9.6 μm CO(2)-laser irradiation successfully inhibits demineralization of tooth enamel in humans.