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ObjectiveThe objective of this study was to evaluate the microbiological profile of root canals indicated for endodontic retreatment due to secondary endodontic infections evidenced by the presence of chronic apical periodontitis (G1) or for prosthetic reasons, without clinical and radiographic signs of endodontic reinfection (G2).MethodsMicrobiological samples were collected from thirty teeth (N=30) out of which 15 were indicated for retreatment due to the presence of chronic apical periodontitis (G1) and 15 were indicated exclusively for prosthetic reasons (G2). Samples were collected from root canals before (S1), after chemomechanical preparation (S2), and after 30 days of intracanal medication composed of calcium hydroxide and 2% chlorhexidine gel (S3). The molecular analysis was performed using Nested-PCR for the detection of 17 bacterial species. The efficacy of each stage of the retreatment in reducing the microbial load was verified by counting colony-forming units (CFU). The statistical analysis considered a significance level of 5%.ResultsThe results showed that bacteria were detected in 100% of the cases in S1, in both groups, by Nested PCR. The most frequently found species in S1 in both groups were Enterococcus faecalis, Fusobacterium nucleatum, Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, and Parvimonas micra. The microbial load of G1 was higher than G2 in the initial samples (S1). Endodontic retreatment was effective in reducing the microbial load in G1 and G2. Statistically significant associations were found between specific bacteria and clinical signs and symptoms.ConclusionIt was concluded that the microbial community present in the root canal of teeth indicated for endodontic retreatment is mixed and heterogeneous. G1 and G2 differ in the number of species per case and microbial load.Clinical relevanceEndodontic retreatment was effective in reducing the microbial load. Statistically significant associations were found between specific bacteria and clinical signs and symptoms.

ObjectivesThis study aimed to detect the presence of specific bacteria and to evaluate the levels of lipopolysaccharides (LPS) and lipoteichoic acid (LTA) in symptomatic necrotic root canals associated with acute apical abscess (symptomatic group — GI). It also aimed to compare the findings with those presented by asymptomatic necrotic root canals (asymptomatic group — GII) in the different stages of the endodontic treatment.Materials and methodsMicrobiological samples were collected from 20 root canals, including purulent collection from acute apical abscesses, before and after chemo-mechanical preparation (CMP) preparation (CMP) with chlorhexidine gel 2% and after 30 days of intracanal medication (ICM) with (Ca[OH]2) + chlorhexidine. The culture method was used to evaluate the efficacy of CMP and ICM. Nested PCR was used to detect the presence of 17 specific bacteria. Levels of LPS were measured by using limulus amebocyte lysate, whereas enzyme-linked immunosorbent assay was used for the quantification of LTA.ResultsCMP was effective in reducing the microbial load in both groups (P < 0.05). LPS levels were higher in GI than in GII (P < 0.05). There was a significant reduction in the LPS levels after CMP and ICM (P < 0.05) in GI and GII. LTA levels were significantly reduced in GI after ICM and in GII after CMP and ICM (both P < 0.05). Fusobacterium nucleatum and Enterococcus faecalis were frequently identified in both groups, alone or in combination with each other.ConclusionDifferent species were detected in all stages of the endodontic treatment. CMP was able to reduce bacterial content and the levels of LPS, but not of LTA in the symptomatic group. High levels of LPS were correlated with spontaneous pain and pain to percussion in the symptomatic group.Clinical relevanceThis clinical study showed that chemo-mechanical preparation was able to reduce bacterial load and levels of LPS, but not of LTA in the symptomatic group. Elevated levels of LPS were correlated with the presence of symptomatology.

AimThe objective of this study was to evaluate the presence of microorganisms in the root canals (RC) of teeth submitted to endodontic retreatment due to prosthetic reasons and without evidence of apical periodontitis.Material and methodsTwenty teeth referred to endodontic retreatment due to prosthetic reasons and without evidence of apical periodontitis were included in this study. Gutta-percha (GP) was collected from each third of the RC. After GP removal, microbial samples were also collected from the full extension of RC using paper points and files. The samples were cultivated in Fastidious Anaerobe Agar and the colony-forming units (CFU/mL) were counted. They were also semi-quantitatively analyzed by checkerboard DNA–DNA hybridization using 40 DNA bacterial probes.ResultsMicroorganisms were found in all samples. The coronal third of GP was more contaminated than the apical third (p ≤ 0.05). There was prevalence of Enterococcus hirae and Enterococcus faecalis in all RC thirds and also in the samples collected from the full extension of RCs.ConclusionMicroorganisms were found in all cases referred to endodontic retreatment due to prosthetic reasons and without evidence of apical periodontitis. Enterococcus was the genus most frequently detected.Clinical significanceAn endodontic retreatment should be considered before replacing a prosthesis.

Geospatial analysis was used to study the epidemiology of Schistosoma mansoni, intestinal parasites and co-infections in an area (Ilha das Flores) in Sergipe, Brazil. We collected individually georeferenced sociodemographic, behavioral and parasitological data from 500 subjects, analyzed them by conventional statistics, and produced risk maps by Kernel estimation. The prevalence rates found were: S. mansoni (24.0%), Trichuris trichiura (54.8%), Ascaris lumbricoides (49.2%), Hookworm (17.6%) and Entamoeba histolytica (7.0%). Only 59/500 (11.8%) individuals did not present any of these infections, whereas 279/500 (55.8%) were simultaneously infected by three or more parasites. We observed associations between S. mansoni infection and various variables such as male gender, being rice farmer or fisherman, low educational level, low income, water contact and drinking untreated water. The Kernel estimator indicated that high-risk areas coincide with the poorest regions of the villages as well as with the part of the villages without an adequate sewage system. We also noted associations between both A. lumbricoides and hookworm infections with low education and low income. A. lumbricoides infection and T. trichiura infection were both associated with drinking untreated water and residential open-air sewage. These findings call for an integrated approach to effectively control multiple parasitic infections.

Femoral vein access is the first choice for percutaneous atrial septal defect closure, and when it cannot be used due to anatomic reasons, the alternative sites should be considered, frequently increasing the complexity of the procedure. Here we report the case of a 3-year-old boy, with situs inversus and dextrocardia, electively referred for percutaneous closure of an ostium secundum atrial septal defect. During the procedure, agenesis of the infra-hepatic segment of the inferior caval vein was diagnosed, and no double inferior caval vein or right superior caval vein were identified by ultrasound or angiography. Therefore, we opted to perform the procedure through the left internal jugular vein, with fluoroscopy and transesophageal echocardiographic guidance. Catheters were navigated through a hydrophilic guidewire, and a Stiff guidewire was positioned in the left ventricle for better support. An Amplatzer septa occluder 19 was successfully deployed without major difficulties and the patient was discharged after 24 hours in good clinical condition. Percutaneous atrial septal defect closure through alternative access sites, especially in the presence of situs inversus, may pose significant challenges to the interventional team. In this case, the left internal jugular vein has shown to be a feasible option, allowing the navigation and manipulation of devices without complications. Provided the expertise of the interventional team, and awareness of the risks involved, alternative access sites can be successfully used for paediatric structural interventions.

Sturge-Weber syndrome (SWS) is a capillary-venous malformation affecting the brain, the eye, and the adjacent trigeminal dermatomes of the skin. This illness is usually diagnosed during the first years of life. If left undiagnosed (and consequently untreated), the condition could develop into severe refractory seizures, ischemic strokes, visual loss, and early cognitive impairment.We report a case of a 23-year-old female patient with a port-wine facial stain, presenting her first convulsive episode in adulthood, associated with a moderate dilation of the deep venous system in the angiography, which raised the diagnostic of Sturge-Weber syndrome.

Regenerative endodontic procedures have become a valuable alternative for the treatment of immature teeth with pulp necrosis. In addition to resolution of periradicular pathosis and promotion of continued root development, positive vitality testing has been observed in some regenerative clinical cases. Importantly, the positive response to electric stimulation of the regenerated tissue requires targeting of periradicular axons into the previously empty root canal space. However, the mechanism by which this process occurs is largely unknown. Since stem cells of the apical papilla (SCAP) have been proposed to populate the root canal following regenerative endodontic procedures, we hypothesized that SCAP regulate neurite outgrowth and axonal targeting. To test this hypothesis, we established primary co-cultures of human SCAP and rat trigeminal neurons, and performed neurite outgrowth assays using ELISA and confocal microscopy to determine the effect of increasing concentration of SCAP on the total neurite outgrowth and axonal targeting. In addition, we evaluated whether SCAP evoked axonal targeting in vivo using a matrigel subcutaneous implant assay. Data were analyzed by ANOVA with Bonferroni's post hoc test, and significance was set at p <0.05. The results demonstrated that SCAP release a soluble factor that regulates neurite outgrowth from cultured trigeminal neurons. Next, we demonstrated that this effect is completely abolished by pretreatment with a neutralizing antibody to brain-derived neurotrophic factor (BDNF), but not by antibodies to other neurotrophins. Further, SCAP release BDNF in a concentration-dependent manner as detected by ELISA, and trigger directed axonal targeting both in vitro and in vivo as demonstrated by microfluidic and matrigel implant experiments, respectively. Collectively, these results suggest that SCAP may be responsible for the chemical signal driving axons to target regenerated tissue via a BDNF-dependent mechanism.

The importance of legume proteins in cholesterol metabolism has been recognised, but the hindgut contribution is still unclear. The present work was undertaken to address the role of the caecum–colon in cholesterol metabolism in intact (I) and ileo-rectal anastomosed (IRA) pigs fed with casein or extruded soyabean (ES) diets. Four groups of six growing pigs were assigned to the treatments (casein–I, casein–IRA, ES–I, ES–IRA) for 3 weeks. Plasma total cholesterol, LDL- and HDL-cholesterol were not modified by surgery or diet. In the liver, the ES diet significantly depressed non-esterified, esterified and total cholesterol. The treatments did not affect hepatic 3-hydroxy-3-methylglutaryl CoA reductase, cholesterol 7α-hydroxylase or sterol 27-hydroxylase activities. In the gallbladder bile of ES-fed pigs, total cholesterol was depressed while total bile acid concentration was increased. IRA and the ES diet markedly decreased the biliary bile acid microbial metabolites (namely hyodeoxycholic acid) and increased the primary bile acids (mainly hyocholic acid). The concentration of bile hydrophobic acids was decreased only by the ES diet. Faecal neutral sterol output was increased in ES-fed pigs, but the bile acid and the sum of neutral and acidic steroid outputs were not. Microbial transformation of neutral and acidic steroids was markedly reduced by IRA, especially in the ES-fed pigs. Thus, surgery and ES modulated the steroid profile but the caecum–colon did not seem to play a crucial role in determining cholesterolaemia in pigs.

The objectives of this study were to determine the rates of plant litter deposition and decomposition in Marandu pastures (Urochloa brizantha cv. Marandu) under a) three forms of nitrogen (N) supply, b) at different stages of rotational grazing and c) to compare the single-exponential decay constant (‘k’) derived from litterbags with values derived from estimates of deposited and existing litter (DEL technique). The three N supply treatments were: without or with N fertilization (zero or 150 kg N ha-1 yr-1) or with the legume Desmodium ovalifolium. There were no significant differences (p<0.05) between existing litter and rates of litter deposition and decomposition between the three N supply treatments. The litter decomposition rate was estimated using the DEL technique for the 7-day grazing periods and two subsequent 14-day periods in each 35-day grazing cycle. The litter decomposition rate was (P<0.05) higher for the second rest period (days 21 to 35) at 0.089 g g-1 day-1, than for the grazing period (0.038 g g-1 day-1) and for the first rest period (0.040 g g-1 day-1). The mean half-life of the litter was 12 days using the DEL technique while the estimate from the litterbags was 136 days. Results showed that estimates provided by litterbags severely underestimate the decomposition in relation to the DEL technique and predict a long-term accumulation of litter which is not observed. RESUMO: Os objetivos deste estudo foram determinar taxas de deposição e decomposição de serapilheira em pastagens de Marandu (Urochloa brizantha cv. Marandu) sob a) três formas de suprimento de nitrogênio (N), b) em diferentes estágios de pastejo rotativo e c) comparar a constante de decomposição (‘k’) derivada de “litterbags” com os valores derivados das estimativas de serapilheira existente e depositada (técnica SED). Os três tratamentos de suprimento de N foram: sem ou com adubação nitrogenada (zero ou 150 kg N ha-1 ano-1) ou com a leguminosa Desmodium ovalifolium. Não houve diferenças significativas (p<0,05) entre a serapilheira existente e as taxas de deposição e decomposição da serapilheira entre os três tratamentos de N. A taxa de decomposição da serapilheira foi estimada usando a técnica SED para os 7 dias de pastejo e dois períodos subsequentes de 14 dias em cada ciclo de 35 dias. A taxa de decomposição da serapilheira foi (p<0,05) maior no segundo período de descanso (dias 21 a 35) em 0,089 g g-1 dia-1, do que no período de pastejo (0,038 g g-1 dia-1) e no primeiro período de repouso (0,040 g g-1 dia-1). A meia-vida média da serapilheira foi de 12 dias pela técnica SED, enquanto a estimativa dos “litterbags” foi de 136 dias. Os resultados mostram que as estimativas fornecidas pelos “litterbags” subestimam severamente a decomposição em relação à técnica SED, e preveem um acúmulo de serapilheira a longo prazo que não é observado.

Chagas disease remains a serious public health concern with unsatisfactory treatment outcomes due to strain-specific drug resistance and various side effects. To identify new therapeutic drugs against Trypanosoma cruzi, we evaluated both the in vitro and in vivo activity of the organometallic gold(III) complex [Au(Hdamp)(L14)]Cl (L1 = SNS-donating thiosemicarbazone), which was denoted 4-Cl. Our results demonstrated that 4-Cl was more effective than benznidazole (Bz) in eliminating both the extracellular trypomastigote and the intracellular amastigote forms of the parasite without cytotoxic effects on mammalian cells. In very-low-dose in vivo assays, 4-Cl reduced parasitaemia and tissue parasitism in addition to protecting the liver and heart from tissue damage. All these changes resulted in the survival of 100% of the mice treated with 4-Cl during the acute phase. We hypothesised that 4-Cl can act directly on the parasite and may participate in the modulation of IFN- production at the acute stage of the disease. Molecular docking simulations showed that the compound may interact with cruzain, a thiol protease considered a possible antiparasitic drug target, primarily by hydrophobic interactions. These analyses predicted that the Cys25 residue in the cruzain binding site is approximately 3.0 away from the S and Au atoms of the gold compound, which could suggest formation of a possible covalent bond between cruzain and the inhibitor. Overall, we confirmed the potential of 4-Cl as a new candidate for Chagas disease treatment.