Postoperative pain after ultrasonically and laser-activated irrigation during root canal treatment: a randomized clinical trial.

AIM: To compare the intensity of postoperative pain after primary root canal treatment of asymptomatic teeth when using ultrasonically (UAI) or laser-activated irrigation (LAI). METHODOLOGY: In this superiority randomized clinical trial (ClinicalTrials.gov ID: NCT03981237) with parallel design, fifty-six patients with an asymptomatic tooth in need of primary root canal treatment were enrolled. After chemo-mechanical canal preparation using rotary instruments and NaOCl irrigation, teeth were randomly assigned to two groups and patients were blinded to the final irrigation protocol. In the UAI group (n = 28), 60s activation with an Irrisafe tip was done per canal. In the LAI group (n = 28), NaOCl was activated with a pulsed Er:YAG (2940 nm) laser, equipped with a conical tip, with settings of 50 µs, 20 mJ at 15 Hz for 60 s. Patients recorded their pain intensity 6, 24, 48 and 72 h after treatment on a 100 mm visual analogue scale (VAS), as well as their analgesic consumption. Pain levels and incidence were compared across groups using the Mann-Whitney U-test and chi-square test. RESULTS: Overall, mean postoperative pain intensity was low, with the majority of patients having no or minimal pain 24 h postoperatively. At 6 h postoperatively, pain intensity and incidence were significantly higher in the UAI group compared to the LAI group (P < 0.05). For the other time intervals, no significant differences in postoperative pain incidence or intensity were found. The frequency of analgesic intake did not differ significantly between the two groups. Neither of the activation methods resulted in any adverse effects. CONCLUSIONS: Ultrasonically and laser-activated irrigation resulted in low and comparable levels of postoperative pain in asymptomatic patients receiving primary root canal treatment.

Knockdown of calreticulin, laccase, and Snf7 Genes Through RNAi Is Not Effective to Control the Asian Citrus Psyllid (Hemiptera: Livideae).

The Asian citrus psyllid, Diaphorina citri Kuwayama, transmits the bacteria Candidatus Liberibacter associated with huanglongbing (HLB), a devastating disease of the citrus industry. The use of genetically modified plants is an alternative to control this vector. Conversely, technology based on RNA interference (RNAi) for silencing specific genes of a target insect could be attempted. This work evaluated the knockdown effect of the target genes calreticulin (DcCRT), laccase (DcLAC), and Snf7 (DcSnf7) by RNAi through feeding D. citri in Murraya paniculata leaves after the uptake of an aqueous solution with dsRNA homologous to each vector target gene. Confocal microscopy revealed the uptake of the fluorescent-labeled dsRNA by detached leaves and the symplastic movement, allowing the ingestion by the feeding insect. A reduction in the survival rate was observed only 144 h after the beginning of feeding with dsRNA targeting DcSnf7; however, no reduction in transcript accumulation. The knockdown of the DcCRT and DcLAC genes was detected only 12 and 96 h after insect feeding, respectively. Additionally, a reduction in amino acid excretion from insects fed with dsRNA targets to DcCRT and DcLAC was observed 120 h after the beginning of feeding. However, the effects of the dsRNAs tested here appear to be minimal, both at the transcriptional and phenotype levels. For most concentrations and time points, no effects were observed. Therefore, the knockdown of genes DcCRT, DcLAC, and DcSnf7 do not appear to have the potential to control of D. citri through RNAi-mediated gene silencing.

Detection and Whole-Genome Analysis of a High-Risk Clone of Klebsiella pneumoniae ST340/CG258 Producing CTX-M-15 in a Companion Animal.

The emergence and dissemination of high-risk clones of Klebsiella pneumoniae producing extended-spectrum ß-lactamases (ESBLs) in animal infections is a critical issue. We report the detection and genomic features of a multidrug-resistant (MDR) ESBL (CTX-M-15)-producing K. pneumoniae infecting a domestic cat. Whole-genome sequencing analysis identified the international ST340 (clonal group CG258), and genes and mutations conferring resistance to ß-lactams, aminoglycosides, macrolides, phenicols, fosfomycin, sulfonamides, tetracycline, trimethoprim, and fluoroquinolones. In addition, the presence of genes encoding resistance to disinfectant and heavy metals hazardous to humans was also confirmed. The MDR profile exhibited by the strain contributed to treatment failure and death of the companion animal. Therefore, active surveillance of critical priority lineages of K. pneumoniae should not only focus on human infections but also on veterinary infections.

Biofilm model systems for root canal disinfection: a literature review.

The aim of this review was to present an overview of laboratory root canal biofilm model systems described in the endodontic literature and to critically appraise the various factors that constitute these models. The electronic databases MEDLINE, Web of Science and EMBASE were searched up to and including December 2016 to identify laboratory studies using endodontic biofilm models. The following search terms were used in various combinations: biofilm, root canal, in vitro, endodontic, bacteria, root canal infection model, colony-forming unit. Only English papers from journals with an impact factor were selected. The records were screened by two reviewers, and full-text articles were assessed according to pre-defined criteria. The following data were extracted from the included studies: the microbial composition of the biofilm, the substrate, growth conditions, validation and quantification. Seventy-seven articles met the inclusion criteria. In the majority (86%) of the studies, a monospecies biofilm was cultured. In two studies, a dual-species biofilm was grown; others cultivated a multispecies biofilm, containing at least three species. Enterococcus faecalis was the most frequently used test species (in 79% of all studies, 92% of the monospecies studies). Four studies used an inoculum derived directly from the oral cavity. Human dentine was the most frequently used substratum (88% of the studies). Incubation times differed considerably, ranging from one to seventy days. The most common quantification method (in 87% of the studies) was bacterial culturing, followed by microscopy techniques. The variation in laboratory root canal biofilm model systems is notable. Because of substantial variation in experimental parameters, it is difficult to compare results between studies. This demonstrates the need for a more standardized approach and a validated endodontic biofilm model.

Efficacy of sonically, ultrasonically and laser-activated irrigation in removing a biofilm-mimicking hydrogel from an isthmus model.

AIM: To evaluate the efficacy of sonically, ultrasonically and laser-activated irrigation (LAI) in removing a biofilm-mimicking hydrogel from the isthmus in a root canal model. METHODOLOGY: Transparent resin blocks containing two standardized root canals (apical diameter of 0.3 mm, 6% taper, 16 mm long, with a coronal reservoir) connected by an isthmus (0.15 mm wide, 2 mm high) were used as the test model. The isthmus was filled with a hydrogel-containing dentine debris. The canals were filled with irrigant, and the models were randomly assigned to the following activation groups (n = 20): EndoActivator (EA), Eddy, ultrasonically activated irrigation (UAI) with an Irrisafe 25 mm length, size 25 file and LAI with a 2940 nm Er:YAG-laser (20 Hz, 50 µs, 20 mJ, PIPS tip at the canal entrance). All protocols were executed for 3 × 20 s. Needle irrigation (NI) with a 27G needle served as the control. Standardized images of the isthmus were taken before and after irrigation, and the amount of removed hydrogel was determined using image analysis software and compared across groups using Welch anova (P ≤ 0.05). RESULTS: Hydrogel removal was greatest in the LAI group (90.2%) and was significantly greater than that with UAI, EA and NI (P ≤ 0.014), but not significantly different from Eddy (P = 0.498). Hydrogel removal with Eddy (85.9%) was significantly greater than that with NI and EA (P < 0.05), but not significantly different from UAI (P = 0.07). There was no significant difference between the NI and EA groups (P = 1). CONCLUSIONS: Laser-activated irrigation and Eddy resulted in the greatest hydrogel removal and performed better than EA and UAI. The effect of LAI was also not dependent on deep intracanal tip placement.

Laser-induced vapor nanobubbles improve diffusion in biofilms of antimicrobial agents for wound care.

Being responsible for delayed wound healing, the presence of biofilms in infected wounds leads to chronic, and difficult to treat infections. One of the reasons why antimicrobial treatment often fails to cure biofilm infections is the reduced penetration rate of antibiotics through dense biofilms. Strategies that have the ability to somehow interfere with the integrity of biofilms and allowing a better penetration of drugs are highly sought after. A promising new approach is the use of laser-induced vapor nanobubbles (VNB), of which it was recently demonstrated that it can substantially enhance the penetration of antibiotics into biofilms, resulting in a marked improvement of the killing efficiency. In this study, we examined if treatment of biofilms with laser-induced vapor nanobubbles (VNB) can enhance the potency of antimicrobials which are commonly used to treat wound infections, including povidone-iodine, chlorhexidine, benzalkonium chloride, cetrimonium bromide and mupirocin. Our investigations were performed on Pseudomonas aeruginosa and Staphylococcus aureus biofilms, which are often implicated in chronic wound infections. Pre-treatment of biofilms with laser-induced VNB did enhance the killing efficiency of those antimicrobials which experience a diffusion barrier in the biofilms, while this was not the case for those compounds for which there is no diffusion barrier. The magnitude of the enhanced potency was in most cases similar to the enhancement that was obtained when the biofilms were completely disrupted by vortexing and sonication. These results show that laser-induced VNB are indeed a very efficient way to enhance drug penetration deep into biofilms, and pave the way towards clinical translation of this novel approach for treatment of wound infections.

Genomic features of a highly virulent, ceftiofur-resistant, CTX-M-8-producing Escherichia coli ST224 causing fatal infection in a domestic cat.

OBJECTIVES: Escherichia coli strains producing extended-spectrum ß-lactamases (ESBLs), especially CTX-M-type, have been largely described in companion animals; however, genomic data are lacking to clarify the clinical impact of ESBL-producing isolates in these hosts. The aim of this study was to present the genomic features of a highly virulent, ceftiofur-resistant, CTX-M-8-producing E. coli isolate from a case of pneumonia in a domestic cat with fatal outcome. METHODS: Genomic DNA was sequenced using an Illumina NextSeq 500 platform and was assembled using CLC Genomic Workbench. Genomic data were analysed using online bioinformatics tools. RESULTS: The genome size was evaluated at 5.1Mb, with 5334 protein-coding sequences. The strain was assigned to sequence type 224 (ST224) and presented genes conferring resistance to ß-lactams (blaCTX-M-8), sulphonamides (sul2), tetracycline (tetA) and trimethoprim (dfrA14) as well as chromosomal point mutations in ParC (S80I), GyrA (S83L) and GyrB (D87N). In addition, the presence of the virulence genes cba, gad, ipfA, iroN, iss, mchF and tsh was detected. CONCLUSION: This draft genome sequence might provide important data for a better understanding of genomic aspects regarding the dissemination of CTX-M-8-producing E. coli in the human-animal-environment interface.

Hepatitis C in Brazil: lessons learned with boceprevir and telaprevir.

In 2012, the first-generation protease inhibitors telaprevir (TVR) and boceprevir (BOC) were introduced in the Brazilian health system for treatment of chronic hepatitis C, after their approval by the National Committee for Health Technology Incorporation (CONITEC). However, these medicines were discontinued in 2015. The short period of use in therapy and their high cost require a discussion about the consequences for patients and for the health system of the early incorporation of new therapies. The article presents a qualitative analysis of the incorporation process of both medications in Brazil and the results of a multicenter study that included patients treated with BOC or TVR between January 2011 and December 2015 in five Brazilian cities. The study included 855 patients (BOC: n=247) and (TVR: n=608). The document analysis showed that CONITEC's decision to incorporate BOC and TVR was based on results of phase III clinical trials that compared sustained virologic response (SVR) rates of patients treated with BOC and TVR with rates of those that received placebo. However, these studies included a low percentage of cirrhotic patients. The SVR rates observed in this multicenter study were worse than clinical trials pointed out (BOC: 45.6%; TVR: 51.8%), but similar to those achieved with previously adopted therapies. The discontinuation rate due to adverse events was (BOC: 15.4%; TVR: 12.7%). Based on these unsatisfactory results, the study brings a discussion that goes beyond the therapy outcomes, exploring the incorporation of these high-cost medicines and the related decision-making process, contributing to future decisions in medicine policies and in the treatment of chronic hepatitis C.

Multidrug-resistant CTX-M-15-producing Klebsiella pneumoniae ST231 associated with infection and persistent colonization of dog.

Extended spectrum ß-lactamase (ESBL)-producing bacterial infections in veterinary medicine are a clinical and epidemiological challenge. We report a case of CTX-M-15-producing Klebsiella pneumoniae infection followed by persistent colonization, in a dog presenting with bilateral purulent nasal discharge and dyspnea. In this regard, 5 broad-spectrum cephalosporin-resistant K. pneumoniae isolates were recovered from infection and surveillance cultures, collected during 1 year and eight months study. Genomic analysis of a representative clone of K. pneumoniae (KpPB76) revealed the presence of the human-associated lineage ST231, whereas resistome data confirmed the presence of genes conferring resistance to aminoglycosides, ß-lactams, fluoroquinolones, fosfomycin, phenicols, sulfonamides, tetracyclines and trimethoprim. In the absence of therapeutic options, meropenem therapy was used, contributing to the control of infection during persistent carriage of K. pneumoniae CTX-M-15/ST231. Persistent colonization of companion animals with ESBL-producing bacteria could be result from a variety of situations, including multi introduction from the owner or household family members to pets, or from environmental exposure; whereas colonized animals may serve as an important source for the spread of ESBL-producing strains in the human-animal interface.

Hard tissue debris removal from the mesial root canal system of mandibular molars with ultrasonically and laser-activated irrigation: a micro-computed tomography study.

This study is to investigate the efficacy of different irrigant activation techniques on removal of accumulated hard tissue debris (AHTD) in mesial roots of human mandibular molars. Extracted human mandibular molars with an isthmus between the mesial root canals were selected based on micro-CT (µCT) scans. The mesial canals were instrumented to an apical diameter ISO30 using ProTaper rotary files. Teeth were randomly assigned to three irrigant activation groups (n = 10): ultrasonically activated irrigation (UAI) using a size 20 Irrisafe for 3 × 20 s, laser-activated irrigation (LAI) with an Er:YAG laser (2940 nm) and plain 300 µm fiber tip inside the canal (20 mJ, 20 Hz, 3 × 20 s), and laser-activated irrigation with identical parameters with a 400 µm photon-induced photoacoustic streaming (PIPS) tip held at the canal entrance. All teeth were scanned with µCT before and after instrumentation and after irrigant activation. After reconstruction and image processing, the canal system volume filled with hard tissue debris before and after irrigant activation was calculated. Changes in hard tissue debris volumes were compared between groups using one-way ANOVA. The percentage volume of hard tissue debris (vol%) was significantly lower after irrigant activation in all groups. Although the lowest debris values were observed in the laser groups, no significant differences in the vol% of accumulated hard tissue debris after activation were observed between groups. Accumulated hard tissue debris was reduced significantly in all activation groups. Ultrasonically and laser-activated irrigation regimens performed similarly in this respect. None of the tested methods was able to render the root canal systems free of debris.

Effect of laser-activated irrigation on biofilms in artificial root canals.

AIM: To evaluate the antimicrobial effect of laser-activated irrigation (LAI) on biofilms formed in simulated root canals. METHODOLOGY: A dual-species biofilm of Enterococcus faecalis and Streptococcus mutans was grown in a resin root canal model. Biofilms were formed over 48 h and subsequently subjected to the following treatments, all executed for 20 s: syringe irrigation (SI) with a 27G needle, ultrasonically activated irrigation (UAI) with a size 20 Irrisafe file, and LAI with a 2940 nm Er:YAG laser (20 Hz, 50 µs, 20 or 40 mJ, conical fibre tip at two positions). Tests were performed with both sterile saline as well as NaOCl (2.5%) as the irrigant. Surviving bacteria were harvested and the number of CFU was determined by plate counting and compared across groups (anova, P ≤ 0.05). RESULTS: Using saline as the irrigant, significant reductions in viable counts compared to untreated controls were observed for ultrasonically activated irrigation (0.52 log10 reduction) and for all laser-activated irrigation groups (>1 log10 reduction), but not for syringe irrigation (<0.25 log10 reduction). The reductions in the laser-activated irrigation groups were significantly greater than those of ultrasonically activated irrigation. With NaOCl as the irrigant, significant reductions (>2.2 log10 units) in the number of attached bacteria were observed for all treatment groups with no significant differences between laser-activated and ultrasonically activated irrigation. CONCLUSIONS: Within the limitations of this in vitro set-up, laser-activated irrigation removed more biofilm than ultrasonically activated irrigation when using saline as the irrigant, indicating greater physical biofilm removal. The use of NaOCl resulted in greater biofilm reduction with no significant differences between treatment groups.

Resistência à intoxicação por amorimia septentrionalis em caprinos, induzida pela inoculação ruminal das bactérias pigmentiphaga kullae e ancylobacter dichloromethanicus / Resistance to poisoning by amorimia septentrionalis in goats induced by ruminal inoculation of the bacteria pigmentiphaga kullae and ancylobacter dichloromethanicus

No Brasil, estima-se que as intoxicações por plantas tóxicas que contém monofluoroacetato de sódio (MFA) causam a morte de aproximadamente 500.000 bovinosao ano. A inoculação ruminal de bactérias que degradam MFA tem sido proposta como uma forma de prevenir a intoxicação. O presente trabalho teve como objetivo avaliar, em caprinos, a resistência ao MFA presente em Amorimia septentrionalis, induzida por inoculação ruminal das bactérias Pigmentiphaga kullae e Ancylobacter dichloromethanicus. Doze caprinos, que nunca tiveram contato prévio com plantas que contêm MFA, foram divididos em dois grupos, com seis animais cada. No grupo 1, 60 mL de uma mistura das duas bactérias foi inoculada, diariamente, durante 10 dias em cada caprino. No grupo 2, os caprinos não receberam as bactérias. A partir do 10º dia de inoculação, A. septentrionalis foi administrada, diariamente, na dose de 5g/kg de peso vivo, sendo interrompida em cada animal após a observação dos primeiros sinais clínicos da intoxicação. Os caprinos do grupo 1 apresentaram sinais clínicos 5,83±2,56 dias após a administração da planta o que diferiu significativamente (p=0,037) dos caprinos do grupo 2, que apresentaram sinais clínicos aos 2,67±0,52 dias. A quantidade de planta ingerida pelos caprinos inoculados (28,83±12,97g/kg) e os não inoculados (12,03±3,65g/kg) para desencadear os sinais clínicos foi, também, estatisticamente diferente entre os grupos (p=0,025). Conclui-se que a administração intraruminal de Pigmentiphaga kullae e Ancylobacter dichloromethanicus induz resistência à intoxicação por plantas que contêm MFA.

In Brazil is estimated that poisoning of livestock by sodium monofluoroacetate (MFA) containing plants causes the death of about 500.000 cattle per year. The ruminal inoculation of bacteria that degrade MFA has been proposed as a way to prevent the poisoning. This study aimed to evaluate in goats resistance to the MFA-containing plant Amorimia septentrionalis induced by ruminal inoculation of the bacteria Pigmentiphaga kullae and Ancylobacter dichloromethanicus. Twelve goats, without previous contact with MFA-containing plants, were divided into two groups of six animals each. In group 1, 60ml of a mixture of the two bacteria was inoculated every day for 10 days into each goat. In group 2, the goats did not receive the bacteria. At the 10th day of inoculation, A. septentrionalis began to be administered daily at a dose of 5g/kg body weight to both groups. The administration was interrupted in each goat after first clinical signs of poisoning were observed.. The goats of group 1 showed clinical signs 5.83±2.56 days after the administration of the plant, what differed significantly (p=0.037) from goats of group 2, that showed clinical signs 2.67±0 52 days after the beginning of ingestion. The amount of A. septentrionalis ingested by inoculated goats (28.83±12.97g/kg) to cause clinical sings was significantly greater (p=0.025) than the amount ingested by the non-inoculated (12.03±3.65) goats to cause clinical signs and was also statistically different between the groups. We concluded that the intraruminal administration of Pigmentiphaga kullae and Ancylobacter dichloromethanicus increases the resistance to poisoning by MFA-containing plants.

Endodontic performance by Flemish dentists: have they evolved?

AIM: To gather information on aspects of endodontic practice and referral behaviour by Flemish (Dutch-speaking Belgian) dentists and to compare the results with an earlier investigation. METHODOLOGY: A postal questionnaire was distributed to 4468 active Flemish dentists. It consisted of multiple-choice questions on endodontic practice and endodontic referral need. Basic information on age, gender, year of qualification and clinical specialty was also obtained. The questionnaire was free of charge to return. Data were imported in a database, subjected to descriptive and analytical statistics and compared to those of an earlier, comparable survey. RESULTS: The response rate was 18.5%. Of all respondents, 95% were general practitioners. More than half (56%) of the respondents never or seldom used rubber dam during endodontic treatment. The use of sodium hypochlorite as an irrigant was well established (80% of respondents using it), whereas the use of EDTA was limited. Female respondents were more likely to refer to an endodontist than male (70 versus 49%). Referrals for treatment of root canal obstruction (48%), perforations and root resorption (42%) and retreatment (39%) were rated 'very important'. Younger respondents more frequently made use of magnification, rotary instruments and rubber dam. Activation of irrigants was incorporated by a small number of respondents (50% never, 12.6% seldom). Cold lateral condensation was used by most GDPs (35.3% always, 19.5% frequently). CONCLUSIONS: Endodontic practice of general dentists in Flanders did not always comply with quality guidelines. However, when comparing the present data with those of a previous study, an obvious trend towards a more present-day treatment protocol is noticeable, as well as an increased appeal for endodontic referral.

The Trypanosoma cruzi virulence factor oligopeptidase B (OPBTc) assembles into an active and stable dimer.

Oligopeptidase B, a processing enzyme of the prolyl oligopeptidase family, is considered as an important virulence factor in trypanosomiasis. Trypanosoma cruzi oligopeptidase B (OPBTc) is involved in host cell invasion by generating a Ca(2+)-agonist necessary for recruitment and fusion of host lysosomes at the site of parasite attachment. The underlying mechanism remains unknown and further structural and functional characterization of OPBTc may help clarify its physiological function and lead to the development of new therapeutic molecules to treat Chagas disease. In the present work, size exclusion chromatography and analytical ultracentrifugation experiments demonstrate that OPBTc is a dimer in solution, an association salt and pH-resistant and independent of intermolecular disulfide bonds. The enzyme retains its dimeric structure and is fully active up to 42°C. OPBTc is inactivated and its tertiary, but not secondary, structure is disrupted at higher temperatures, as monitored by circular dichroism and fluorescence spectroscopy. It has a highly stable secondary structure over a broad range of pH, undergoes subtle tertiary structure changes at low pH and is less stable under moderate ionic strength conditions. These results bring new insights into the structural properties of OPBTc, contributing to future studies on the rational design of OPBTc inhibitors as a promising strategy for Chagas disease chemotherapy.

Evaluation of Nd:YAG and Er:YAG irradiation, antibacterial photodynamic therapy and sodium hypochlorite treatment on Enterococcus faecalis biofilms.

AIM: To compare the antimicrobial efficacy of two-high power lasers (Nd:YAG and Er:YAG) and two commercial antimicrobial photodynamic therapy (aPDT) systems with that of sodium hypochlorite (NaOCl) action on Enterococcus faecalis biofilms grown on dentine discs. METHODOLOGY: Enterococcus faecalis biofilms were grown on dentine discs in a microtiter plate, incubated for 24 h and subjected to the following treatments: aPDT (Denfotex and Helbo system), Er:YAG laser irradiation (2940 nm, 50 mJ or 100 mJ, 15 Hz, 40 s), Nd:YAG laser irradiation (1064 nm, 2 W, 15 Hz, 40 s) and immersion in 2.5% (w/v) NaOCl for 1, 5, 10 and 30 min. Surviving bacteria were harvested, and the number of CFU per disc was determined by plate counting. RESULTS: Significant reductions (anova, P ≤ 0.05) in viable counts were observed for aPDT (Helbo) (2 log(10) reduction), Er:YAG irradiation using 100 mJ pulses (4.3 log(10) reduction) and all NaOCl treatments (>6 log(10) reduction). NaOCl (2.5%) for 5 min effectively eliminated all bacteria. aPDT (Denfotex), Er:YAG irradiation using 50 mJ pulses and Nd:YAG treatment caused a reduction in the viable counts of <1 log(10) unit; these results were not significantly different from the untreated controls. CONCLUSION: Within the limitations of this particular laboratory set-up, NaOCl was the most effective in E. faecalis biofilm elimination, while Er:YAG laser treatment (100 mJ pulses) also resulted in high reductions in viable counts. The use of both commercial aPDT systems resulted in a weak reduction in the number of E. faecalis cells. Nd:YAG irradiation was the least effective.

Pulpal regeneration and root development after subcutaneous transplantation of cryopreserved immature teeth in rats.

The purpose of this in vivo study was to investigate revascularization and root growth after autotransplantation of cryopreserved immature teeth. Immature molar teeth were extracted in 4-week-old Wistar rats. In the test group, teeth were cryopreserved for 1 week and transplanted subcutaneously to the abdomen. In the control group, teeth were transplanted subcutaneously immediately after extraction. Material was collected in test and control animals at intervals of 1, 2, 4 and 10 weeks post-transplantation and histological and microradiographical examination was performed. Results showed that during the first weeks after transplantation, pulpal repair was similar in both groups although degenerated pulpal tissue was replaced slower in cryopreserved teeth and some differences in types of hard tissue formation were found between test and control teeth. After 10 weeks, the differences in the regenerated pulpal tissue between cryopreserved and control teeth observed during the first weeks were no longer detectable. No root growth was detected microradiographically 10 weeks after transplantation in any of the transplanted teeth. The presence of dentin-like tissue in the pulp cavity of some autotransplanted cryopreserved teeth, suggests survival of pulpal tissue after cryopreservation.

The major leucyl aminopeptidase of Trypanosoma cruzi (LAPTc) assembles into a homohexamer and belongs to the M17 family of metallopeptidases.

BACKGROUND: Pathogens depend on peptidase activities to accomplish many physiological processes, including interaction with their hosts, highlighting parasitic peptidases as potential drug targets. In this study, a major leucyl aminopeptidolytic activity was identified in Trypanosoma cruzi, the aetiological agent of Chagas disease. RESULTS: The enzyme was isolated from epimastigote forms of the parasite by a two-step chromatographic procedure and associated with a single 330-kDa homohexameric protein as determined by sedimentation velocity and light scattering experiments. Peptide mass fingerprinting identified the enzyme as the predicted T. cruzi aminopeptidase EAN97960. Molecular and enzymatic analysis indicated that this leucyl aminopeptidase of T. cruzi (LAPTc) belongs to the peptidase family M17 or leucyl aminopeptidase family. LAPTc has a strong dependence on neutral pH, is mesophilic and retains its oligomeric form up to 80°C. Conversely, its recombinant form is thermophilic and requires alkaline pH. CONCLUSIONS: LAPTc is a 330-kDa homohexameric metalloaminopeptidase expressed by all T. cruzi forms and mediates the major parasite leucyl aminopeptidolytic activity. Since biosynthetic pathways for essential amino acids, including leucine, are lacking in T. cruzi, LAPTc could have a function in nutritional supply.

[Lasers: principles, characteristics and tissue interactions]. / Lasers: principes, caractéristiques et interaction tissulaire.

Since their appearance in 1960, lasers have been considered useful light sources for medical applications. Laser light is monochromatic, the bundle is parallel and can be directed and focussed, as a result of which very high energy densities can be achieved. Several applications in dentistry have been investigated over the past decades. This article describes the physics behind lasers, the characteristics of the laser beam and overviews the laser wavelengths currently used in dentistry. The different interactions between the laser beam and the target are explained.

[Cavity preparation using an Er:YAG laser in the adult dentition]. / La préparation de cavités à l'aide du laser Er:YAG dans la denture adulte.

Many lasers (i.e., different wavelengths) are available today for clinical applications. Not all lasers are to be used for cavity preparation and removal of carious tissues. Erbiumlasers (Er:YAG and Er,Cr:YSGG) are suitable for these purposes (the wavelengths coincide with the highest absorption peaks of water and hydroxyapatite). The advantages during cavity preparation and caries removal are smear layer free cavity walls, selective and localised removal of tooth substance, a restricted need or the absence of anaesthesia, and cavity walls with a higher acid resistance resulting in a better protection against secondary decay. Possible side effects of erbium lasers with water cooling and used with respect for correct power setting are minimal and can be compared with those of rotary instruments. Erbium laser tooth preparation is now one of the options in the minimal invasive approach. The needs for acid etching has been debated for long. At present it has become clear that acid etching is mandatory to obtain a good adhesion and retention with resin composites.