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Context: Peripheral nerve injuries result in sensory and motor losses in the innervated area and can hinder individuals' daily activities. Objective: The objective was to analyze the influence of applying gallium-aluminum-arsenide (GaAIAs) laser (660Nm) on the functional recovery of the sciatic nerve in rats. Methods: The sciatic nerve of 12 Wistar rats was subjected to injury consisting of neurotmesis and epineural anastomosis. The rats were divided into two groups: control and laser therapy. After the injury, a GaAlAs laser was used (660Nm, 4J/cm2, 26.3mW and 0.63cm2 beam) at three equidistant points on the injury, for 20 days. Footprint impressions were obtained from the animals before and seven, 14 and 21 days after the surgical procedure and the sciatic functional index (SFI) was calculated. Results: Comparison of the SFI did not show any significant difference (p>0.05) between the two groups. Conclusions: The parameters and methods used for the laser therapy did not produce any effect on the SFI over the period evaluated. [ABSTRACT FROM AUTHOR]

Ampelozizyphus amazonicus Ducke, known as "saracura-mirá" in the Amazon rainforest, is valued for its traditional use in malaria prevention and treatment. The plant's roots and bark are employed for these purposes, while the wood is often overlooked. Given the global importance of leishmaniasis, research focused on A. amazonicus anti-Leishmania amazonensis potential. The ethyl acetate extract from the bark (EAEB) exhibited the most effective inhibition of intracellular amastigote growth with IC50 7.0 µg.mL-1 but showed high toxicity (CC50 9.0 µg.mL-1). The wood ethanol (EW) and ethyl acetate (EAEW) extracts demonstrated promising results against L. amazonensis with IC50 15.8 µg.mL-1 and 10.0 µg.mL-1, CC50 50.5 µg.mL-1 and 22.7 µg.mL-1, respectively. High-speed countercurrent chromatography combined with mass spectrometry  off-line detection was employed for the bioassay-guided fractionation of EAEB and EAEW using HEMWat as solvent system. These extracts showed to be rich in saponins and triterpenes, besides minor glycosylated flavonoids. HSCCC was effective in obtaining high purity fractions, leading to the identification of a di-glycosylated triterpene saponin from EAEB, and melaleucic acid from EAEW. These findings highlight A. amazonicus as a potential source for developing new therapeutic alternatives for leishmaniasis and HSCCC as a technique enabling better separation efficiency of complex saponin matrices., (© 2024 Wiley‐VCH GmbH.)

Hepatocellular carcinoma (HCC) represents a global health concern, ranking as the sixth most common malignancy worldwide and the third leading cause of cancer-related mortality. Despite advances in research, the diagnosis and prognosis of such malignancy remain challenging. Alpha-fetoprotein, the current serum biomarker used in the management of HCC, has limited sensitivity and specificity, making early detection and effective management more difficult. Thus, new management approaches in diagnosis and prognosis are needed to improve the outcome and survival of HCC patients. SNHG1 is a long noncoding RNA mainly expressed in the cell and cytoplasm of cells and is consistently upregulated in tissues and cell lines of HCC, where it acts as an important regulator of various processes: modulation of p53 activity, sponging of microRNAs with consequent upregulation of their target mRNAs, regulation of fatty acid, iron and glucose metabolism, and interaction with immune cells. The deregulation of these processes results in abnormal cell division, angiogenesis, and apoptosis, thus promoting various aspects of tumorigenesis, including proliferation, invasion, and migration of cells. Clinically, a higher expression of SNHG1 predicts poorer clinical outcomes by significantly correlating with bigger, less differentiated, and more aggressive tumors, more advanced disease stages, and lower overall survival in HCC patients. This article comprehensively summarizes the current understanding of the multifaceted roles of SNHG1 in the pathogenesis of HCC, while also highlighting its clinicopathological correlations, therefore concluding that it has potential as a biomarker in HCC diagnosis and prognosis.

The LABEXTRACT plant extract bank, featuring diverse members of the Myrtaceae family from Brazilian hot spot regions, provides a promising avenue for bioprospection. Given the pivotal roles of the Spike protein and 3CL pro and PL pro proteases in SARS-CoV-2 infection, this study delves into the correlations between the Myrtaceae species from the Atlantic Forest and these targets, as well as an antiviral activity through both in vitro and in silico analyses. The results uncovered notable inhibitory effects, with Eugenia prasina and E. mosenii standing out, while E. mosenii proved to be multitarget, presenting inhibition values above 72% in the three targets analyzed. All extracts inhibited viral replication in Calu-3 cells (EC 50 was lower than 8.3 µg·mL -1 ). Chemometric analyses, through LC-MS/MS, encompassing prediction models and molecular networking, identified potential active compounds, such as myrtucommulones, described in the literature for their antiviral activity. Docking analyses showed that one undescribed myrtucommulone (m/z 841 [M - H] - ) had a higher fitness score when interacting with the targets of this study, including ACE2, Spike, PL pro and 3CL pro of SARS-CoV-2. Also, the study concludes that Myrtaceae extracts, particularly from E. mosenii and E. prasina , exhibit promising inhibitory effects against crucial stages in SARS-CoV-2 infection. Compounds like myrtucommulones emerge as potential anti-SARS-CoV-2 agents, warranting further exploration.

Since the advent of Covid-19, several natural products have been investigated regarding their in silico interactions with SARS-CoV-2 proteases - 3CL pro and PL pro , two of the most important pharmacological targets for antiviral development. Phenylethanoid glycosides (PG) are a class of natural products present in important medicinal plants and a drug containing this group of active ingredients has been successfully used in the treatment of Covid-19 in China. Thus, a dataset with 567 derivatives of this class was built from reviews published between 1994 and 2020, and their interaction against both SARS-CoV-2 proteases was investigated. The virtual screening was performed by filtering the PGs through the evaluation of scores based on the AutoDock Vina, GOLD/ChemPLP, and GOLD/GoldScore evaluation functions. The bRO5 pharmacokinetic parameters of the PGs ranked in the previous step were analyzed and their interaction with key amino acid residues of the 3CL pro and PL pro enzymes was evaluated. Ninety-eight compounds were identified by computational approaches against PL pro and 80 PGs against 3CL pro . Of these, four interacted with key catalytic residues of PL pro , which is an indicative of inhibitory activity, and three compounds interacted with catalytic key residues of 3CL pro . Of these, five PGs occur in plants of the Traditional Chinese Medicine (TCM), while two are components of plants/formulations currently used in the Covid-19 protocols in China. The data presented here show the potential of PGs as selective inhibitors of SARS-CoV-2 3CL pro and PL pro ., Competing Interests: The authors declare no conflict of interest., (© 2023 The Authors. Published by Elsevier B.V. on behalf of Research Network of Computational and Structural Biotechnology.)

Aiming at finding natural sources of antidiabetics agents, 15 extracts from Brazilian medicinal plants of the Atlantic Forest and Amazon region were tested against α-glucosidase enzyme. Plants were selected based on the taxonomic relationships with genera including several species with antidiabetic activity. In this screening, the extracts obtained from the flowers of Hyptis monticola and the leaves of Lantana trifolia and Lippia origanoides resulted endowed with promising anti-α-glucosidase activity. The extracts from H. monticola and from L. origanoides collected in two different areas, were characterised by ultra-high performance liquid chromatography coupled to mass spectrometry. Bioassay-guided fractionation led to the identification of several enzyme inhibiting compounds, among them the mechanism of action of naringenin and pinocembrin was investigated. The two L. origanoides extracts showed differences in bioactivity and in the phytochemical profiles. The fractionation of the extract from H. monticola led to a partial loss of the inhibitory effect.

Physical exercise is characterized by an increase in physical and metabolic demand in face of physical stress. It is reported that a single exercise session induces physiological responses through redox signaling to increase cellular function and energy support in diverse organs. However, little is known about the effect of a single bout of exercise on the redox homeostasis and cytoprotective gene expression of white adipose tissue (WAT). Thus, we aimed at evaluating the effects of acute aerobic exercise on WAT redox homeostasis, mitochondrial metabolism, and cytoprotective genic response. Male Wistar rats were submitted to a single moderate-high running session (treadmill) and were divided into five groups: control (CTRL, without exercise), and euthanized immediately (0 h), 30 min, 1 hour, or 2 hours after the end of the exercise session. NADPH oxidase activity was higher in 0 h and 30 min groups when compared to CTRL group. Extramitochondrial ROS production was higher in 0 h group in comparison to CTRL and 2 h groups. Mitochondrial respiration in phosphorylative state increased in 0 h group when compared to CTRL, 30 min, 1, and 2 h groups. On the other hand, mitochondrial ATP production was lower in 0 h in comparison to 30 min group, increasing in 1 and 2 h groups when compared to CTRL and 0 h groups. CAT activity was lower in all exercised groups when compared to CTRL. Regarding oxidative stress biomarkers, we observed a decrease in reduced thiol content in 0 h group compared to CTRL and 2 h groups, and higher levels of protein carbonylation in 0 and 30 min groups in comparison to the other groups. The levels returned to basal condition in 2 h group. Furthermore, aerobic exercise increased NRF2, GPX2, HMOX1, SOD1, and CAT mRNA levels. Taken together, our results suggest that one session of aerobic exercise can induce a transient prooxidative state in WAT, followed by an increase in antioxidant and cytoprotective gene expression., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2021 Leonardo Matta et al.)

Lippia species share various pharmacological activities and are used in traditional cooking and medicine worldwide. Combined chromatographic techniques such as column chromatography, high-performance liquid chromatography, and countercurrent chromatography led to the purification of two new antifungal phenylpropanoid glycosides, lippiarubelloside A (1) and lippiarubelloside B (2), by bioactivity-directed fractionation of an ethanol-soluble extract from Lippia rubella, in addition to the known active related compounds forsythoside A (3), verbascoside (4), isoverbascoside (5), and poliumoside (6). The structures of compounds 1 and 2 were determined by comparison of their NMR spectroscopic data with the prototype active compound 4. Cryptococcus neoformans, which causes opportunistic lung infections, was sensitive to compounds 1-6 in the concentration range of 15-125 μg/mL. A synergistic effect (FIC index = 0.5) between 3 and amphotericin B was demonstrated. The glycosylated flavonoids pectolinarin (7), linarin (8), and siparunoside (9) were also isolated.

Objectives: Evaluate the tissue reaction of periodontium subjacent to furcation perforations in rat molars sealed with Biodentine or mineral trioxide aggregate (MTA)., Materials and Methods: The pulp chamber floor of right upper first molars of 60 rats was perforated and filled with Biodentine, MTA, or cotton pellet (sham); the left first molars were used as control. After 7, 15, 30, and 60 days, maxillary fragments were processed for paraffin-embedding. The periodontal space (PS), volume density of inflammatory cells (VvIC) and fibroblasts (VvFb), number of osteoclasts, and collagen content were obtained. Interleukin-6 (IL-6) and osterix (osteoblast marker) were detected by immunohistochemistry. The data were submitted to ANOVA and Tukey's test (p ≤ 0.05)., Results: At 7 days, high values in VvIC, IL-6-immunolabeled cells, and osteoclasts were accompanied by reduced collagen content in enlarged PS of experimental groups. At all periods, VvIC, number of osteoclasts and IL-6, and PS were higher in sham than in Biodentine and MTA (p < 0.0001). From 7 to 60 days, significant reduction in VvIC, IL-6 immunoexpression, and osteoclasts was accompanied by significant increase in VvFb, osteoblasts, and collagen in Biodentine and MTA groups. At 60 days, significant differences in VvIC, PS, IL-6, osteoclasts, and osteoblasts were not found between Biodentine and MTA. Significant differences in the osteoclast number were not observed among Biodentine, MTA, and control groups while osteoblasts number was higher in Biodentine and MTA groups., Conclusions: Despite the initial inflammatory reaction and bone resorption, the sealing of furcation perforations with Biodentine and MTA favors the repair of periodontal tissues., Clinical Relevance: Biodentine and MTA exhibit potential as repair material in the treatment of furcation perforations.

Background: Childhood community-acquired pneumonia is a common and potentially life-threatening illness in developing countries. We assessed the prognostic value of serum procalcitonin level upon admission on clinical response to antibiotic treatment., Methods: Out of 89 patients, the median (IQR) age was 19(12-29) months and 60% were boys. Viral (49.5%), typical bacterial (38%) and atypical bacterial (12.5%) infections as well as probable pneumococcal infections (26%) were diagnosed., Results: Seventy-five (84%) children became afebrile ≤48h after treatment. In 14 children who remained febrile after 48h of treatment, median[IQR] serum procalcitonin (ng/ml) level on admission was higher than in those with rapid recovery (2.1[0.8-3.7] vs 0.6[0.1-2.2]; P=0.025). In the slow-responding children, pneumococcal infections were more common (71% vs 17%; P<0.001). Procalcitonin concentrations on admission were higher in children with pneumococcal pneumonia compared to children with non-pneumococcal pneumonia (2[0.7-4.2] vs 0.5[0.08-2.1]; P=0.002). The ROC curve found that <0.25ng/ml of serum procalcitonin had a high negative predictive value (93%[95%CI:80%-99%]) for pneumococcal infection. All children that remained febrile after 48h of treatment had procalcitonin >0.25ng/ml on admission. The majority of children with pneumonia in a developing country become afebrile within 48h after onset of antibiotic treatment., Conclusions: Serum procalcitonin <0.25ng/ml predicted rapid clinical response and non-pneumococcal etiology., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Aim: To compare the formation of fibrous capsules around Biodentine and MTA Angelus implants as well as the participation of fibroblast growth factor-1 (FGF-1) and mast cells in the tissue response to these endodontic materials., Methodology: Sixty polyethylene tubes filled with Biodentine or MTA, and empty tubes (control group) were implanted into the dorsal subcutaneous tissues of male rats. After 7, 15, 30 and 60 days, the specimens were embedded in paraffin and the number of fibroblasts and mast cells was quantified in the sections stained with Masson's trichrome or Alcian Blue, respectively. FGF-1 and Ki-67 were detected by immunohistochemistry, and the number of immunolabelled cells was computed. The collagen content was estimated in the picrosirius red-stained sections. The data were subjected to two-way ANOVA followed by Tukey's test (P ≤ 0.05)., Results: The capsules were associated with a significant increase (P < 0.0001) in the number of fibroblasts and mast cells, and in the collagen content over time. A significant decrease (P < 0.0001) in the immunoexpression of FGF-1 and Ki-67 was observed in all groups from the 7th-60th day. At 60 days, the number of fibroblasts (P = 0.0226) and the collagen content (P < 0.0001) were significantly greater in MTA than Biodentine specimens, while the greatest number of mast cells and FGF-1-immunolabelled cells was observed in Biodentine specimens (P < 0.0001). A significant difference in Ki-67 immunoexpression was not detected between specimens of Biodentine and MTA., Conclusions: The collagen-rich capsule formed slowly around Biodentine in comparison with MTA. FGF-1 and mast cells participated in capsule remodelling, stimulating fibroblast proliferation and subsequent collagen production, in response to subcutaneous implants., (© 2018 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Novozyme 435, which is a commercial immobilized lipase B from Candida antarctica (CALB), has been proven to be inadequate for the kinetic resolution of rac-indanyl acetate. As it has been previously described that different immobilization protocols may greatly alter lipase features, in this work, CALB was covalently immobilized on epoxy Immobead-350 (IB-350) and on glyoxyl-agarose to ascertain if better kinetic resolution would result. Afterwards, all CALB biocatalysts were utilized in the hydrolytic resolution of rac-indanyl acetate and rac-(chloromethyl)-2-(o-methoxyphenoxy) ethyl acetate. After optimization of the immobilization protocol on IB-350, its loading capacity was 150 mg protein/g dried support. Furthermore, the CALB-IB-350 thermal and solvent stabilities were higher than that of the soluble enzyme (e.g., by a 14-fold factor at pH 5-70°C and by a 11-fold factor in dioxane 30%-65°C) and that of the glyoxyl-agarose-CALB (e.g., by a 12-fold factor at pH 10-50°C and by a 21-fold factor in dioxane 30%-65°C). The CALB-IB-350 preparation (with 98% immobilization yield and activity versus p-nitrophenyl butyrate of 6.26 ± 0.2 U/g) was used in the hydrolysis of rac-indanyl acetate using a biocatalyst/substrate ratio of 2:1 and a pH value of 7.0 at 30°C for 24 h. The conversion obtained was 48% and the enantiomeric excess of the product (e.e. p ) was 97%. These values were much higher than the ones obtained with Novozyme 435, 13% and 26% of conversion and e.e.p, respectively. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:878-889, 2018., (© 2018 American Institute of Chemical Engineers.)

Considering that endodontic sealers release some components which may promote delay in the repair process, our purpose was to evaluate the tissue reaction promoted by MTA Plus and MTA Fillapex in comparison with AH Plus (standard control) and Endofill, which has a long clinical track record. One hundred rats were distributed into five groups: MTA Plus (Avalon Biom Inc., Bradenton, FL, USA), MTA Fillapex (Angelus, Londrina, PR, Brazil), AH Plus (Dentsply DeTrey GmbH, Konstanz, Germany), Endofill (Dentsply, Petrópolis, RJ, Brazil) and CG (control group, empty polyethylene tubes). The polyethylene tubes filled with sealers or empty (CG) were implanted into subcutaneous. After 7, 15, 30 and 60 days, the tubes surrounded by capsules were paraffin-embedded. In HE-stained sections, the volume density of inflammatory cells (VvIC) was estimated in the capsules. The number of interleukin-6-immunolabelled cells (IL-6), a pro-inflammatory cytokine, was also computed in the capsules. The birefringent collagen content was quantified in picrosirius-stained sections. Data were analysed by ANOVA and Tukey tests (p ≤ 0.05). At 7 days, the capsules showed moderate inflammatory reaction. In all groups, VvIC and IL-6-immunostained cells reduced significantly from 7 to 60 days. At 60 days, IL-6 immunoexpression was reduced significantly in MTA Plus and MTA Fillapex in comparison with AH Plus; no difference was found in the VvIC among MTA Plus, MTA Fillapex, AH Plus and CG whereas Endofill exhibited the highest VvIC. The reduction in VvIC was parallel to an increase in the collagen in all the groups, except Endofill. MTA Plus, MTA Fillapex and AH Plus induce a response that culminates in the regression of inflammation and formation of a fibrous capsule over time. The lower IL-6 immunoexpression in the capsules of MTA Plus and MTA Fillapex than AH Plus suggests that the immune response is suppressed more rapidly in the MTA-based sealers.

Aim: To evaluate the influence of the addition of microparticulate (micro) and nanoparticulate (nano) zirconium oxide (ZrO 2 ) and niobium pentoxide (Nb 2 O 5 ) to a calcium silicate-based cement (CS) on the subcutaneous healing process in rats compared with MTA Angelus™., Methodology: In each rat, two polyethylene tubes filled with the following materials: (i) MTA; (ii) CS + ZrO 2 micro; (iii) CS + ZrO 2 nano; (iv) CS + Nb 2 O 5 micro or (v) CS + Nb 2 O 5 nano were implanted subcutaneously; empty polyethylene tubes were used in the Control group. After 7, 15, 30 and 60 days, the specimens (n = 5 per group in each period) were fixed and embedded in paraffin. Masson's trichrome sections were used to obtain the volume density of the inflammatory cells (VvIC) and fibroblasts (VvFb). The sections were also stained with Picrosirius-red to calculate the birefringent collagen content. Fibroblast growth factor-1 (FGF-1) was detected by immunohistochemistry, and the number of immunolabelled cells was obtained. The data were subjected to two-way anova followed by Tukey's test (P ≤ 0.05)., Results: At all periods, the VvIC was significantly lower (P < 0.001) in all the CS and Control groups than in the MTA group. At all periods, the VvFb was reduced significantly (P = 0.023) in the MTA group in comparison with the other groups. In addition, the number of immunolabelled cells in the capsules of the CS groups was significantly higher (P < 0.001) than in the MTA group at all time-points., Conclusions: The experimental materials (CS + ZrO 2 and CS + Nb 2 O 5 ) induced fibroblast proliferation and accelerated the regression of the inflammatory reaction. However, the addition of nanoparticulate radiopacifiers did not improve the biological properties of a calcium silicate-based cement when compared to microparticulate agents., (© 2017 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Purpose: Several studies have suggested worse outcomes for patients requiring medical care at night or on weekends. However, whether or not children should be extubated only during in-hours has not been studied yet. We sought to compare outcomes and complications of in-hours versus off-hours extubated patients., Methods: We prospectively included all children receiving invasive mechanical ventilation (MV) in a pediatric intensive care unit. Off-hours extubations included patients who were extubated at nighttime (8:00 pm-7:59 am) plus weekends/holidays whereas the in-hours extubations included regular daytime weekdays (Monday to Friday: 8:00 am-7:59 pm)., Results: Of the 480 patients, 346 (72%) were extubated during in-hours period and 134 (28%) were extubated during off-hours. In-hours patients spent a longer time to have planned extubation and had a longer MV duration and pediatric intensive care unit stay compared to those extubated at off-hours. Kaplan-Meier curve showed that in-hours patients were more likely to have a longer time until the first extubation (log-rank test: P=.006, HR: 5.05)., Conclusion: Patients extubated at off-hours had more favorable outcomes with similar complications rate compared with those extubated at in-hours. These results provide no support for delaying extubations until in-hours period. Further studies are required to confirm these findings., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Background and Aims: Although iatrogenic withdrawal syndrome (IWS) has been recognized in patients exposed to opioids and benzodiazepines, very few studies have used a validated tool for diagnosis and assessment of IWS in critically ill children. We sought to determine IWS rate, risk factors, and outcomes of IWS patients., Methods: Prospective observational study conducted in a pediatric intensive care unit. A total of 137 patients (31 with IWS and 106 with no IWS) received a continuous infusion of fentanyl and midazolam for 3 or more days. The Sophia Observation withdrawal Symptoms scale was repeatedly applied when children were weaned off sedation/analgesia., Results: The overall incidence of IWS was 22.6%. Of the 31 IWS patients, 6 showed IWS with less than 5 days sedation or analgesia. Logistic regression showed that the median peak dose of midazolam was associated with IWS development (odds ratio 1.4). Receiver-operating curve showed a cut-off value of 0.35 mg/kg/h for midazolam peak dose (sensitivity 96.7%, specificity 51%, positive predictive value 36.6%, and negative predictive value 98.2%), with area under the curve of 0.80. IWS patients had a longer time on mechanical ventilation, prolonged pediatric intensive care unit, and hospital stays, and required prolonged period to have drugs discontinued., Conclusions: Although length of sedation/analgesia for at least5 days has been widely proposed for monitoring IWS, our data suggest that initiating monitoring after 3 sedation days is highly recommended. In addition, patients requiring infusion rates of midazolam above 0.35 mg/kg/h should be considered at high risk for IWS.

Aim: To evaluate the inflammatory process induced by Biodentine and mineral trioxide aggregate (MTA) in rat subcutaneous tissues., Methodology: A polyethylene tube filled with Biodentine (n = 20) or MTA (n = 20) was placed into the dorsal subcutaneous of forty male rats; in the control group (CG; n = 20), empty tubes were implanted. After 7, 15, 30 and 60 days, the polyethylene tubes surrounded by connective tissue were fixed and embedded in paraffin. The number of inflammatory cells was estimated in HE-stained sections; numerical density of interleukin-6 (IL-6)-immunolabelled cells was also performed. The differences amongst the groups were analysed statistically by Tukey's test (P ≤ 0.05)., Results: A high number of inflammatory cells and IL-6-positive cells were observed at 7 days, in all groups; however, in the Biodentine group, the number of inflammatory cells and IL-6-immunolabelled cells was significantly higher (P ≤ 0.05) in comparison with the other groups at 7 and 15 days. In the capsules of animals from all groups, a gradual and significant reduction (P ≤ 0.05) of these parameters was seen over time. At 60 days, the capsules exhibited numerous fibroblasts and bundles of collagen fibres; in addition, the number of IL-6-positive cells was not significantly different amongst Biodentine, MTA and control groups., Conclusions: There was a significant regression in the inflammatory reaction in the capsules indicating, therefore, that Biodentine is a biocompatible material., (© 2015 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Representatives of the genus Trypanosoma have been traditionally found in epimastigote, espheromastigote and trypomastigote flagellated forms in axenic cultures. Trypanosoma caninum is a trypanosomatid that has recently been reported infecting dogs in endemic areas of canine leishmaniasis in Brazil. It presents specific biological characteristics and it is found exclusively on healthy skin. Here, we describe the evolutive forms of this parasite showing not only the forms commonly found in culture, but also epimastigote forms with no free flagellum. The study was conducted using scanning and transmission electron microscopy and, we demonstrate that typical flagellated epimastigotes originate from forms without flagellum, although the latter may remain without differentiation in the culture. Two hypotheses are considered and discussed in this paper: (i) the aflagellated epimastigotes are a typical developmental forms of T. caninum and (ii) the emergence of these aflagellated forms could be resultant from a disturbed process during cell division caused by interfering specific proteins, which leads to inability to form and regulate the flagellum length. In any case, considering that T. caninum is a parasite that is still little studied, the information brought by our study adds data which may be useful to clarify aspects on the cell cycle of this intriguing parasite that has been found in different regions of Brazil., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Aim: The aim of the following study is to evaluate the cleaning capacity of a hybrid instrumentation technique using Reamer with Alternating Cutting Edges (RaCe) system files in the apical third of mesial roots of mandibular molars., Materials and Methods: Twenty teeth were selected and separated into two groups (n = 20) according to instrumentation technique as follows: BioRaCe - chemomechanical preparation with K-type files #10 and #15; and files BioRaCe BR0, BR1, BR2, BR3, and BR4; HybTec - hybrid instrumentation technique with K-type files #10 and #15 in the working length, #20 at 2 mm, #25 at 3 mm, cervical preparation with Largo burs #1 and #2; apical preparation with K-type files #15, #20, and #25 and RaCe files #25.04 and #30.04. The root canals were irrigated with 1 ml of 2.5% sodium hypochlorite at each change of instrument. The specimens were histologically processed and photographed under light optical microscope. The images were inserted onto an integration grid to count the amount of debris present in the root canal., Results: BioRaCe presented the highest percentage of debris in the apical third, however, with no statistically significant difference for HybTec (P > 0.05)., Conclusions: The hybrid technique presented similar cleaning capacity as the technique recommended by the manufacturer.

An unknown Trypanosoma species was isolated from an axenic culture of intact skin from a domestic dog captured in Rio de Janeiro, Brazil, which was co-infected with Leishmania (Viannia) braziliensis. Giemsa-stained smears of cultures grown in different media revealed the presence of epimastigotes, trypomastigotes, spheromastigotes, transitional stages, and dividing forms (epimastigotes or spheromastigotes). The highest frequency of trypomastigotes was observed in RPMI (15.2%) and DMEM (9.2%) media containing 5% FCS, with a mean length of these forms of 43.0 and 36.0 mum, respectively. Molecular analysis by sequential application of PCR assays indicated that this trypanosome differs from Trypanosoma cruzi and T. rangeli when specific primers were applied. On the other hand, a PCR strategy targeted to the D7 domain of 24salpha rDNA, using primers D75/D76, amplified products of about 250 bp in that isolate (stock A-27), different from the amplification products obtained with T. cruzi and T. rangeli. This organism differs from T. cruzi mainly by the size of its trypomastigote forms and kinetoplasts and the absence of infectivity for macrophages and triatomine bugs. It is also morphologically distinct from salivarian trypanosomes reported in Brazil. Isoenzyme analysis at 8 loci demonstrated a very peculiar banding pattern clearly distinct from those of T. rangeli and T. cruzi. We conclude that this isolate is a new Trypanosoma species. The name T. caninum is suggested.

This study evaluated the temperature rise on external root surfaces by simulating the removal of metallic posts by applying ultrasound, with or without refrigeration and at different times. The crowns of 30 healthy maxillary central incisors were removed and the roots were treated endodontically. The prosthetic spaces were prepared and impressions were taken with self-curing acrylic resin. After casting with a copper-aluminum alloy, the posts were blasted with aluminum oxide and cemented with resin cement. The specimens were randomly assigned to three groups according to the time of ultrasonic vibration (30, 60, or 120 seconds) and were subdivided into two groups, those cooled with and without water refrigeration. A thermocouple connected to a digital thermometer was positioned on the root external surface and the maximum temperature was recorded. Data were submitted to ANOVA and Tukey's test. Analysis of the results revealed statistical difference among groups (p < 0.01). Ultrasound (with and without water spray) increased root temperature significantly, reaching values of more than 50 degrees C. The exception was the group that used 30 seconds of ultrasound with refrigeration; root temperature reached 35.4 degrees C, making it the only tested condition that remained below the critical value (50 degrees C).

This study evaluated in vitro the retention of intracanal cast posts cemented with dual-cure resin varying the application method of the primer/adhesive solution and luting agent in the prosthetic space prepared to receive the posts. Sixty endodontically treated maxillary canines had their crowns discarded, and their roots were embedded in acrylic resin. The prosthetic spaces were prepared with Largo burs mounted on a low-speed handpiece coupled to a parallelometer in order to maintain length and diameter of intraradicular posts constant and to guarantee that the preparations were parallel after casting. Two groups (n = 30) were randomly formed according to the device used to apply the adhesive system: microbrush or standard bristle brush (control). Each group was divided into 3 subgroups (n = 10) according to the technique used to place the luting agent into the root canal: using only a lentulo spiral before setting the post, applying it onto the post surface, or combining both methods. After 72 hours, the tensile force required to dislodge each post was determined by a universal testing machine (Instron 4444) set at a speed of 1 mm/min. The results indicated that the use of the microbrush yielded higher bond strength values (0.1740 +/- 0.04 kN) than those recorded for the bristle brush tip (0.1369 +/- 0.04 kN, p < 0.001). Bonferroni's test demonstrated a higher retention (p < 0.001) in radicular post cemented with the technique that combined both methods (lentulo + post: 0.1787 +/- 0.03 kN) than that obtained with lentulo (0.1461 +/- 0.065 kN) or post (0.1416 +/- 0.03 kN) alone. The interactions between the adhesive system and luting agent application techniques presented statistical difference (p < 0.001). It was concluded that the best performance in terms of tensile strength among the tested conditions was obtained when the adhesive system was applied with a microbrush and the luting agent was taken into the root canal with lentulo spirals alone (0.1961 +/- 0.04 kN) and combining both methods (lentulo + post: 0.1911 +/- 0.02 kN).

The efficacy of ultrasound in removing cast metal posts was evaluated in this in vitro study using one or two ultrasound units and ultrasonic vibration for various lengths of time. The crowns of 30 healthy maxillary canines were removed, the roots were embedded in acrylic resin blocks, and the root canals were treated endodontically. The canals were prepared and their impressions were taken with self-curing acrylic resin. After casting with copper-aluminum alloy, the posts were blasted with aluminum oxide and cemented with Panavia F resin cement. The specimens were divided into five groups. In groups I and II, only one ultrasound unit was used for 30 and 60 s, respectively; in groups III and IV, two ultrasound units were used simultaneously for 30 and 60 s, respectively; in group V, ultrasound was not used (control). Ultrasonic vibrations were applied with an Enac OE-5 ultrasound unit and an ST-09 tip. All samples were subjected to traction on an Instron machine (model 4444) at a cross-head speed of 1 mm/min. Analysis of the results revealed a statistically significant difference between the groups (ANOVA, P < 0.01); however the difference between groups II and IV was not statistically significant. The efficacy of ultrasound in removing intraradicular posts was confirmed, and the most effective technique was the use of two ultrasound units, independent of the length of time ultrasound was applied.

The efficacy of ultrasound, with and without water spray, was evaluated in vitro on the amount of force necessary to dislodge posts cemented with resin or zinc phosphate. Forty-two samples were divided into six groups: groups 1, 2, and 3, posts cemented with zinc phosphate; groups 4, 5, and 6, posts cemented with resin (Panavia F); groups 1 and 4 (controls), no ultrasound; groups 2 and 5, ultrasound without water spray; and groups 3 and 6, ultrasound with water spray. The Instron testing machine was used. Ultrasound without water spray significantly reduced (71%) the force necessary to displace posts cemented with Panavia F (p < 0.01); however, this value was similar to the efficacy of ultrasound with water spray for posts cemented with zinc phosphate (reduction of 75%). We conclude that cooling with ultrasound interferes with the force necessary for post removal, depending on the type of cement used.