TRPM7 and MagT1 regulate the proliferation of osteoblast-like SaOS-2 cells through different mechanisms.

A correct magnesium (Mg2+) intake is essential for bone health. In particular, Mg2+ deficiency inhibits the proliferation of osteoblast-like SaOS-2 cells by increasing nitric oxide (NO) production through the upregulation of inducible NO synthase. At the moment, little is known about the expression and the role of TRPM7, a channel/enzyme involved in Mg2+ uptake, and MagT1, a Mg2+ selective transporter, in SaOS-2 cells. Here, we demonstrate that TRPM7 is not modulated by different extracellular concentrations of Mg2+ and its silencing exacerbates growth inhibition exerted by low Mg2+ through the activation of inducible NO synthase and consequent accumulation of NO. Moreover, MagT1 is upregulated in SaOS-2 cultured in high Mg2+ and its silencing inhibits the growth of SaOS-2 cultured in media containing physiological or high Mg2+, without any modulation of NO production. We propose that TRPM7 and MagT1 are both involved in regulating SaOS-2 proliferation through different mechanisms.

Association between pulp and periapical conditions and dental emergency visits involving pain relief: epidemiological profile and risk indicators in private practice in Australia.

AIM: To assess the prevalence of dental emergency visits (DEV) involving pain relief and their relationship with socio-economic and clinical factors in an Australian representative sample in the primary care setting. METHODOLOGY: Data on reason for visit and patient characteristics were collected from a representative random sample of Australian dentists in private practice surveyed in 2009-2010. Information regarding socio-economic (gender, age, health insurance) and clinical factors (number of teeth, number of decayed teeth, diagnosis and reason for visit [DEV, check-up, other reasons not involving pain relief]) were retrieved from compiled questionnaires. Descriptive statistics were reported, and Poisson regression models were used to assess the association between socio-economic and clinical factors and DEV. Prevalence ratio (PR) and 95% confidence interval (CI) were calculated. RESULTS: A total of 1148 dentists responded (67%), resulting in records from 6504 patients. The overall prevalence of DEV was 20.8%. The unadjusted analysis, according to the reason of visit, revealed the following predictors for DEV: male gender (PR = 1.18; 95% CI = 1.08-1.29), age 18-64 years (PR = 2.70; 95% CI = 2.19-3.33) and over 65 years (PR = 2.64, 95% CI = 2.10-3.32), uninsured patients (PR = 1.36; 95% CI = 1.24-1.49), patients with <20 teeth (PR = 1.19; 95% CI = 1.06-1.33), decayed teeth (PR = 1.64; 95% CI = 1.48-1.81). After adjustment for confounding factors (gender, age, insurance status, number of teeth and decayed teeth) apart from 'dental trauma' (PR = 1.37), all remaining diagnoses had lower PR ('other' PR = 0.19, 'decay' PR = 0.34, 'periodontal' PR = 0.51, 'failed restoration' PR = 0.45) compared with 'pulp/periapical disease'. CONCLUSIONS: In the primary care setting, the diagnoses 'pulp/periapical' and 'dental trauma' had a stronger association with DEV compared with visits not involving relief of pain. Both socio-economic (male gender, older age and uninsured individuals) and clinical factors (tooth loss, decayed teeth, endodontic diseases and dental trauma) were identified as independent risk indicators for DEV in this population. Future public health policies should include specific preventive strategies addressing these factors, aiming to reduce the need for DEV.

Real-world efficacy and safety of lenvatinib: data from a compassionate use in the treatment of radioactive iodine-refractory differentiated thyroid cancer patients in Italy.

BACKGROUND: Lenvatinib is a multi-kinase inhibitor approved for patients with radioactive iodine (RAI)-resistant differentiated thyroid cancer (DTC). Before the drug approval from the Italian National Regulatory Agency, a compassionate use programme has been run in Italy. This retrospective study aimed to analyse data from the first series of patients treated with lenvatinib in Italy. METHODS: The primary aim was to assess the response rate (RR) and progression-free survival (PFS). Secondary end-points include overall survival (OS) and toxicity data. RESULTS: From November 2014 to September 2016, 94 patients were treated in 16 Italian sites. Seventeen percent of patients had one or more comorbidities, hypertension being the most common (60%). Ninety-eight percent of patients were treated by surgery, followed by RAI in 98% of cases. Sixty-four percent of patients received a previous systemic treatment. Lenvatinib was started at 24 mg in 64 subjects. Partial response and stable disease were observed in 36% and in 41% of subjects, respectively; progression was recorded in 14% of patients. Drug-related side-effects were common; the most common were fatigue (13.6%) and hypertension (11.6%). Overall, median PFS and OS were 10.8 months (95% confidence interval [CI], 7.7-12.6) and 23.8 months (95% CI, 19.7-25.0) respectively. CONCLUSION: Lenvatinib is active and safe in unselected, RAI-refractory, progressive DTC patients in real-life setting. RR and PFS seem to be less favourable than those observed in the SELECT trial, likely due to a negative selection that included heavily pretreated patients or with poor performance status.

Histopathological and microbiological findings associated with retrograde peri-implantitis of extra-radicular endodontic origin: a systematic and critical review.

The aim of this study was to systematically review the aetiology, in particular histopathological and microbiological factors, of retrograde peri-implantitis of endodontic origin. The review is registered in the PROSPERO database (CRD42017063898). An electronic search for publications was performed in two databases, from their inception up to October 2018. Subsequently a hand search of the reference lists was conducted. Articles in English and other languages using Latin characters were included. Two independent reviewers selected the studies, extracted and synthesized the data, and assessed the quality. The methodology of the included articles was evaluated using the relevant Joanna Briggs Institute tools. Six studies fulfilled the eligibility criteria and were included in the systematic review. Histopathological examination in the component studies reflected that the presentation of retrograde peri-implantitis involves cyst formation or chronic inflammation. Bacteria found in these lesions included Porphyromonas gingivalis, Corynebacterium, Streptococcus, and Klebsiella pneumoniae. Two studies were judged as having a low possibility of bias and four were judged as having a moderate possibility of bias. This review determined that endodontic complications associated with adjacent teeth, residual infection at the extraction site due to previous apical periodontitis, or refractory apical periodontitis might be considered likely aetiological factors, although the evidence is limited.

Pre-clinical endodontic education outcomes between artificial versus extracted natural teeth: a systematic review.

AIM: To compare the educational outcomes using artificial teeth versus extracted teeth for pre-clinical endodontic training. DATA SOURCES: Literature searches of PubMed, Scopus, Cochrane Library, Trip Database, Web of Science and Open Grey databases were conducted from their inception until November 2018 with no language restriction. Hand searching of most likely relevant journals was performed. The review followed the PRISMA guidelines. STUDY ELIGIBILITY CRITERIA, PARTICIPANTS AND INTERVENTIONS: Studies that compared pre-clinical endodontic training using extracted teeth and artificial teeth were included. STUDY APPRAISAL AND SYNTHESIS METHODS: The quality of included studies was appraised by Joanna Briggs Institute Critical Appraisal tools. The findings were tabulated and summarized according to their outcomes with distinct narrative syntheses. RESULTS: Five studies were included. The component studies included 359 operators in total, mainly consisting of undergraduate students (97%, n = 349) and 10 endodontists (3%). Forty-seven per cent (n = 170) operated on artificial teeth only, whilst 19% (n = 67) worked primarily on extracted teeth, with the final treatment outcome being evaluated by independent observers using objective criteria. Operators in two studies (34%, n = 122) used both artificial teeth and ET and compared their experiences in surveys. Regarding technical outcomes, no significant differences between training with artificial teeth and extracted teeth were found, but the performance tended to be better in artificial teeth than extracted teeth. Operators trained solely on artificial teeth appeared to be adequately educated for subsequent root canal treatment (RCT) in the clinical setting. LIMITATIONS: Due to the scarcity of research on the topic overall, and the methodological variation between the studies, it was not possible to perform a quantitative analysis (meta-analysis). CONCLUSIONS AND IMPLICATIONS OF KEY FINDINGS: Based on the available evidence, the use of artificial teeth for pre-clinical endodontic training achieved similar educational outcomes compared to extracted teeth. However, the experiences reported by the operators diverged. Further studies assessing other artificial teeth available in the market testing other RCT procedures are necessary.

Interaction of biologic therapy with apical periodontitis and periodontitis: a systematic review.

Biologic medications (BMs) are increasingly used for the management of systemic chronic inflammatory diseases. These diseases are often associated with a higher prevalence of apical periodontitis (AP) and periodontitis (P). The purpose of this systematic review was to ascertain the interactions between AP and/or P and BMs. The review was registered in the PROSPERO database (CRD42017054756). Electronic searches were performed on Pubmed Medline, Scopus and The Web of Science from their inception through to 20 March 2018. The references of the articles selected were checked. The keywords were chosen based on a pilot search, which aimed to find the most frequently prescribed BMs. The included studies were appraised qualitatively using appropriate tools. Thirty-five articles met the inclusion criteria, comprising 16 non-randomized clinical studies, 12 in vivo animal studies and 7 case reports. Quality of information was assessed as high in 18 articles, moderate in 16 articles and low in 1 article. BMs in patients suffering from chronic inflammatory diseases seems to inhibit the progression of AP and P, and to enhance the healing response to periodontal and endodontic treatment. A healthier condition of the periodontal tissues seems to be associated with a better response of the patient to BMs therapy.

Methods for measurement of root canal curvature: a systematic and critical review.

The assessment of root canal curvature is essential for clinical and research purposes. This systematic review presents an overview of the published techniques for the measurement of root canal curvature features using imaging and to provide a critique of their clinical application. A database search in PubMed, PubMed Central, Embase, Scopus, EBSCO Dentistry & Oral Sciences Source and Virtual Health Library was conducted, using appropriate key words to identify measurement methods for root canal curvatures. The search strategy retrieved 10594 records in total, and 31 records fulfilled the inclusion criteria. From 2D image acquisitions, eleven studies measured exclusively the angle of curvature, an additional thirteen measured other curvature features (level, height, radius, length and shape). Seven reports described methods from 3D imaging (CBCT, µCT). Root canal curvatures should be measured, for clinical proposes, to facilitate endodontic treatment planning, and in research, to reduce the risk of selection bias. This review has revealed that there are many methods described in the literature; however, no consensus exists on which method should be used. Some of the methodologies have potential clinical translation, whereas others are suitable for research purpose only, as they require a specific software or radiographic exposure in the mesiodistal direction.

Use of a short-term whole blood intracellular staining assay to study the T-cell response in respiratory syncytial virus-infected pediatric patients.

The aim of the present study was the development of a reliable method to evaluate the pattern of the ongoing T-cell response in young infants affected by respiratory infection. To this purpose, we enrolled 44 infants hospitalized with a diagnosis of respiratory syncytial virus bronchiolitis. After a short-term stimulation of whole blood samples, intracellular IFN-g and IL-4 cytokines were measured in CD4+ and CD8+ T-cell subsets by flow cytometry. A stringent staining and gating strategy was used in order to maximize the reduction of background noise and to exclude false positives. The frequencies of cytokine-producing T-cell subsets, albeit low, were easily quantifiable. Cytokine responses were higher in infants sampled > 7 days from the onset of symptoms. The use of a rigorous strategy for cell staining and gating, coupled with a short-term stimulation of whole blood and a careful evaluation of time elapsed from the onset of symptoms constitutes a convincing approach for future clinical studies.

Orthodontic thermoformed retainers: a two-arm laboratory study into post-fabrication outcomes.

BACKGROUND: Retainers are commonly used to maintain post-orthodontic occlusion stability. We aimed to determine post-fabrication thickness and thinning rate of thermoformed retainers. METHODS: Forty-eight retainers were fabricated from polyethylenterepthalat-glycol copolyester or polypropylene blanks, using vacuum- or pressure-thermoforming. Retainer thickness was measured at multiple locations. RESULTS: Thinning rate had a broad range: the mid-labial incisor region of 1 mm polyethylenterepthalat-glycol copolyester pressure-thermoformed mandibular retainers had the greatest thinning rate (68.25 ± 8.26%) and smallest mean post-fabrication thickness (0.32 ± 0.08 mm). Polyethylenterepthalat-glycol copolyester retainers were 0.11 mm thinner than polypropylene (P=0.0222), and polypropylene retainers were 0.21 mm thicker, when pressure-thermoformed (P<0.0001). The interaction of manufacturing method and material used, and tooth type, explained over a third of the variability in the post-fabrication thickness of these retainers. Maxillary retainers made from 1 mm blanks were 0.04 mm thicker in the incisor region compared with the molar region (P=0.0492). CONCLUSIONS: Thermoformed retainers do no thin uniformly against individual teeth and have variable intra- and inter-arch post-fabrication thicknesses. There is no clear benefit in using a specific type of thermoforming machine or material for increasing post-fabrication thickness or reducing thinning rate. Blank thickness and tooth morphology influence these outcomes.

Association of manual or engine-driven glide path preparation with canal centring and apical transportation: a systematic review.

The role and effect of glide path preparation in root canal treatment remain controversial. This systematic review aims to compare apical transportation and canal centring of different glide path preparation techniques, with or without subsequent engine-driven root canal preparation. A database search in PubMed, PubMed Central, Embase, Scopus, EBSCO Dentistry & Oral Sciences Source and Virtual Health Library was conducted, using appropriate key words to identify the effect of glide path preparation (or its absence) on apical transportation and canal centring. An assessment for the risk of bias in included studies was carried out. Amongst 2146 studies, 18 satisfied the inclusion criteria. Nine studies assessed glide path preparation per se, comparing apical transportation and canal centring of rotary systems and/or manual files; eleven further investigations examined the efficacy of the glide path prior to final canal preparation with different engine-driven systems. Risk of bias and other study design features with potential influence on study outcomes and clinical implications were assessed. Based on the available evidence, and within the limitation of the studies included, preparation of a glide path using rotary sequences performs similarly (in most of the component studies) or significantly better than manual preparation when assessing apical transportation or canal centring. When compared to the absence of a glide path, canal shaping following glide path preparation was of similar, or significantly better quality, in regard to apical transportation or canal centring.

Excessive heat-associated gingival necrosis: a case report.

Heat-delivery devices have been associated with relatively high temperatures. Previous basic research studies suggest that the risk for periodontal tissues is negligible, as the presence of the root filling materials, amongst other factors, reduce the risk of overheating. This report describes the case of a 45 year old male, who was undergoing restorative treatment in a multi-specialty dental practice. As part of his overall treatment planning, orthograde root canal retreatment for his maxillary left incisor teeth was deemed necessary, due to chronic apical periodontitis. Following cold lateral compaction, the coronal portion of the core material was severed using a System B Heat Source (Kerr Dental, Orange, CA, USA), with the patient reporting pain as well as a burning sensation. The following day the patient was seen by a periodontist and gingival necrosis in relation to the central incisor was found. The most likely cause of the complication was considered to be excessive heat during the operative procedure. The patient was followed up for a 2-year period and the soft tissues returned to their original condition without the need for periodontal treatment.

Effect of sonic and ultrasonic activation on organic tissue dissolution from simulated grooves in root canals using sodium hypochlorite and EDTA.

AIM: To compare soft-tissue dissolution by sodium hypochlorite, with an EDTA intermediate rinse, with or without activation with passive ultrasonic activation (PUI) or sonic activation using the Endoactivator (EA) or Eddy tips (ED). METHODOLOGY: The root canals of eighty-three human maxillary central incisors were chemo-mechanically prepared and the teeth split. A standardized longitudinal intracanal groove was created in one of the root halves. Eighty-three porcine palatal mucosa samples were collected, adapted to fit into the grooves and weighed. The re-assembled specimens were randomly divided into four experimental groups (n = 20), based on the final rinse: no activation; EA; PUI; ED, using 2.5% sodium hypochlorite, with an EDTA intermediate rinse. A control group (n = 3) was irrigated with distilled water without activation. The solutions were delivered using a syringe and needle 2 mm from working length. Total irrigation time was 150 s, including 60 s of activation in the specific groups. The study was carried out at 36 ± 2 °C. The porcine palatal mucosa samples were weighed after completion of the assays. Student paired t-test and anova were used to assess the intra- and intergroup weight changes. The multiple comparisons were evaluated using Bonferroni correction (α = 0.05). RESULTS: Weight loss occurred in all experimental groups. Irrigant activation resulted in greater weight loss when compared to the nonactivated group [vs. EA (P = 0.001); vs. PUI (P < 0.001); vs. ED (P < 0.001)]. No significant differences were found amongst the different activation systems. CONCLUSIONS: Activation increased the tissue-dissolving activity of irrigants from artificial grooves in root canals of maxillary central incisors.

Osmolarity and root canal antiseptics.

Antiseptics used in endodontics for disinfection purposes include root canal dressings and irrigants. Osmotic shock is known to cause the alteration of microbial cell viability and might have a role in the mechanism of action of root canal antiseptics. The aim of this review was to determine the role of osmolarity on the performance of antiseptics in root canal treatment. A literature search using the Medline electronic database was conducted up to 30 May 2013 using the following search terms and combinations: 'osmolarity AND root canal or endodontic or antiseptic or irrigation or irrigant or medication or dressing or biofilm; osmolality AND root canal or endodontic or antiseptic or irrigation or irrigant or medication or dressing or biofilm; osmotic AND root canal or endodontic or antiseptic or irrigation or irrigant or medication or dressing or biofilm; osmosis AND root canal or endodontic or antiseptic or irrigation or irrigant or medication or dressing or biofilm; sodium chloride AND root canal or endodontic or antiseptic or irrigation or irrigant or medication or dressing or biofilm'. Publications were included if the effects of osmolarity on the clinical performance of antiseptics in root canal treatment were stated, if preparations with different osmolarities values were compared and if they were published in English. A hand search of articles published online, 'in press' and 'early view', and in the reference list of the included papers was carried out following the same criteria. A total of 3274 publications were identified using the database, and three were included in the review. The evidence available in endodontics suggests a possible role for hyperosmotic root canal medicaments as disinfectants, and that there is no influence of osmolarity on the tissue dissolution capacity of sodium hypochlorite. There are insufficient data to obtain a sound conclusion regarding the role of hypo-osmosis in root canal disinfection, or osmosis in any further desirable ability.

Interaction between chlorhexidine-impregnated gutta-percha points and several chlorine-containing endodontic irrigating solutions.

AIM: To evaluate if the immersion of chlorhexidine-impregnated gutta-percha points in chlorine-containing endodontic irrigants causes colour changes and precipitate formation. METHODOLOGY: Eighty-one size 25 chlorhexidine medicated (Activ Points, Roeko, Langenau, Germany) and 27 size 25 standard gutta-percha points (Roeko) were immersed in microtubes, containing the following solutions: 0.5 and 5.25% NaOCl or Aquatine (Optident, Ilkley, UK). The samples were visually assessed, by two independent observers, at regular intervals over 3 weeks, to detect colour changes and precipitate formation. A score system was used to grade the precipitate intensity. To compare the number of days required for a precipitate to form mean and standard differences were calculated; the means between the groups were compared using a one-way analysis of variance test. Mean intensity scores were calculated and compared between groups using Kruskal-Wallis procedure followed by the Mann-Whitney test. RESULTS: The groups containing 5.25% NaOCl started to produce a visible precipitate after fourteen days (16.48 ± 0.98) for the Activ points; this process was first noticed with the standard gutta-percha group on day 17 (18.33 ± 1.22 days), with a statistically significant difference (P = 0.002). When looking at the intensity scores from day 17 onwards, these two groups differed statistically, with a greater amount for the AP group (P < 0.001). No other test group presented with changes. CONCLUSION: The placement of impregnated gutta-percha points, in 5.25% NaOCl for a 2-week period, caused the formation of a precipitate in the experimental conditions of this study.

Viral infections of the central nervous system in Spain: a prospective study.

The aim of the study was to determine the incidence of viruses causing aseptic meningitis, meningoencephalitis, and encephalitis in Spain. This was a prospective study, in collaboration with 17 Spanish hospitals, including 581 cases (CSF from all and sera from 280): meningitis (340), meningoencephalitis (91), encephalitis (76), febrile syndrome (7), other neurological disorders (32), and 35 cases without clinical information. CSF were assayed by PCR for enterovirus (EV), herpesvirus (herpes simplex [HSV], varicella-zoster [VZV], cytomegalovirus [CMV], Epstein-Barr [EBV], and human herpes virus-6 [HHV-6]), mumps (MV), Toscana virus (TOSV), adenovirus (HAdV), lymphocytic choriomeningitis virus (LCMV), West Nile virus (WNV), and rabies. Serology was undertaken when methodology was available. Amongst meningitis cases, 57.1% were characterized; EV was the most frequent (76.8%), followed by VZV (10.3%) and HSV (3.1%; HSV-1: 1.6%; HSV-2: 1.0%, HSV non-typed: 0.5%). Cases due to CMV, EBV, HHV-6, MV, TOSV, HAdV, and LCMV were also detected. For meningoencephalitis, 40.7% of cases were diagnosed, HSV-1 (43.2%) and VZV (27.0%) being the most frequent agents, while cases associated with HSV-2, EV, CMV, MV, and LCMV were also detected. For encephalitis, 27.6% of cases were caused by HSV-1 (71.4%), VZV (19.1%), or EV (9.5%). Other positive neurological syndromes included cerebellitis (EV and HAdV), seizures (HSV), demyelinating disease (HSV-1 and HHV-6), myelopathy (VZV), and polyradiculoneuritis (HSV). No rabies or WNV cases were identified. EVs are the most frequent cause of meningitis, as is HSV for meningoencephalitis and encephalitis. A significant number of cases (42.9% meningitis, 59.3% meningoencephalitis, 72.4% encephalitis) still have no etiological diagnosis.

The effect of surface tension reduction on the clinical performance of sodium hypochlorite in endodontics.

Sodium hypochlorite (NaOCl) is recommended as an endodontic irrigant in view of its broad antimicrobial and tissue dissolution capacities. To enhance its penetration into inaccessible areas of root canals and to improve its overall effect, the addition of surface-active agents has been suggested. The aim of this investigation was to review the effect of the reduction of the surface tension on the performance of NaOCl in endodontics. A search was performed in the Medline electronic database (articles published up to 28 July 2012, in English) with the search terms and combinations as follows: 'sodium hypochlorite AND surface tension or interfacial force or interfacial tension or surface-active agent or amphiphilic agent or surface active agent or surfactant or tenside or detergent'. The purpose of this search was to identify publications that compared NaOCl alone and NaOCl modified with the addition of a surface-active agent in endodontics. A hand search of articles published online ('in-press' and 'early view'), and appearing in the reference list of the articles included, was further performed, using the same search criteria as the electronic search. The search identified 302 publications, of which 11 fulfilled the inclusion/exclusion criteria of the review. The evidence available suggests that surface-active agents improve the penetration of NaOCl in the main canal and have no effect on its pulp tissue dissolution ability. There are, however, insufficient data to enable a sound conclusion to be drawn regarding the effect of modifying NaOCl's surface tension on lubrication, antimicrobial and smear layer or debris removal abilities.

Influence of pH changes on chlorine-containing endodontic irrigating solutions.

Chlorine-containing solutions are used for broad disinfection purposes. Water disinfection literature suggests that their disinfectant action depends on pH values as this will influence the available free chlorine forms. Hypochlorous acid (HOCl) has been suggested to have an antimicrobial effect around 80-100 times stronger than the hypochlorite ion. The aim of this paper was to review the influence of pH changes on the efficacy of chlorine-containing endodontic irrigating solutions. An electronic and hand search (articles published through to 2010, including 'in press' articles; English language; search terms 'root canal irrigants AND sodium hypochlorite or hypochlorous acid or superoxidized water or electrochemically activated solution'; 'antimicrobial action AND sodium hypochlorite or hypochlorous acid or superoxidized water or electrochemically activated solution'; 'tissue dissolution AND sodium hypochlorite or hypochlorous acid or superoxidized water or electrochemically activated solution'; 'smear layer AND sodium hypochlorite or hypochlorous acid or superoxidized water or electrochemically activated solution') was performed to identify publications that compared chlorine water solutions with different pH. Of 1304 publications identified, 20 were considered for inclusion in the review. The search resulted in the retrieval of articles studying sodium hypochlorite (NaOCl), superoxidized waters (SOW) and sodium dichloroisocyanurate (NaDCC). Regarding antimicrobial efficacy, the literature suggested that reducing the pH value of NaOCl to between 6 and 7.5 would lead to improved action; SOW was described as having a lower antimicrobial effect. The tissue dissolution activity NaOCl decreased when the pH reached values between 6 and 7.5; NaDCC and SOW had no clinically relevant tissue dissolution capability. Chlorine solutions of different characteristics appeared to have some cleaning efficacy although they should to be used in conjunction with chelating and/or detergent agents.

Some factors influencing the stability of Sterilox(®), a super-oxidised water.

Super-oxidised waters, particularly Sterilox(®), have been suggested for the disinfection of dental unit water lines and dental impression materials owing to their antimicrobial efficacy. One of the previously suggested characteristics is their short shelf life. The purpose of this investigation was to understand the effect of storage conditions on Sterilox(®)'s stability. Eight bottles (four completely full, four half-full) of freshly prepared solution were divided into four groups and subsequently stored by being either exposed to or protected from sunlight. The chlorine concentration was monitored using chlorine test strips until the concentration reached zero, or until the thirteenth week. Statistically significant differences between the groups exposed to sunlight and the non-exposed groups (p <0.001) were found. The mean loss of chlorine per day for the non-exposed samples was 1.01 mg/L, whilst the mean for the exposed samples was 2.42 mg/L. The presence of air did not affect the chlorine decomposition in the bottles. The results of this investigation indicate that when the solution is exposed to sunlight, the decrease of chlorine starts at day 4, whilst for the groups sheltered from sunlight, the process started after day 14. Therefore, Sterilox(®) solutions appear to be more stable than previously surmised.

A preliminary study investigating the survival of tetracycline resistant Enterococcus faecalis after root canal irrigation with high concentrations of tetracycline.

AIM: To compare the ability of two Enterococcus faecalis strains to survive exposure to an irrigation solution containing a high concentration of tetracycline in the root canals of bovine teeth. METHODOLOGY: The root canals of twelve bovine incisor root sections were chemo-mechanically prepared using commercially available drills, sodium hypochlorite and ethylenediamine tetra-acetic acid. The root sections were divided into two groups and inoculated with either a tetracycline sensitive or resistant strain of E. faecalis. The strains are isogenic, however one contains a conjugative transposon related to Tn916 which confers resistance to tetracycline, and the other strain is sensitive to the antibiotic. After 26 days of incubation the root canals were irrigated using one of three solutions (sterile distilled water, 50% ethanol or tetracycline at a concentration of 30 mg mL(-1)). The roots were sampled by grinding dentine and canal contents and the debris collected were incubated in broth to assess growth. RESULTS: Irrigation with sterile distilled water or 50% ethanol did not remove all of the cells present. The tetracycline containing solution was efficient in preventing any growth of sensitive E. faecalis, however the resistant strain was able to survive a 5 min exposure at 30 mg mL(-1). CONCLUSIONS: The presence of the Tn916-like conjugative transposon containing the tetracycline resistance gene tet(M) allowed an E. faecalis strain to survive irrigation using a solution containing an extremely high concentration of tetracycline in a root canal model.

Detection and identification of orthopoxviruses using a generic nested PCR followed by sequencing.

Some orthopoxviruses are considered to be potential biological weapons. After the smallpox eradication campaign ended, routine vaccination was stopped around the world. Consequently, a significant portion of the population is now completely unprotected from infection by variola virus and related orthopoxviruses. Some of the symptoms associated with non-variola infections can be similar to smallpox, causing alert and panic situations. These infections should be considered as real public health concerns, so suitable tools for their differential diagnosis are needed. This study aims to devise a simple and easy-to-perform method that is able to detect and identify any orthopoxvirus that might cause infection in humans. In addition, the similarity of the different genes in the genomes of several species of orthopoxviruses is investigated, and orthopoxvirus-universal primer pairs in the tumour necrosis factor receptor II homologue gene are designed, taking full account of nucleotide similarity. A strategy is devised for their sensitive, rapid and cost-effective detection and identification, based on a nested PCR followed by sequencing. The efficacy of the method is tested with samples sent by the European Network of Imported Viral Diseases as part of two external quality control assays. All human orthopoxviruses assayed were detected and identified.