cs.tau.ac.il/expander/overview.html; Tel Aviv University,

Israel). PRIMA achieves www.selleckchem.com/products/gsk2656157.html this by utilizing known models for transcription factor binding sites. Corresponding transcription regulators are considered to be candidate regulators of the corresponding set of genes. We used Ingenuity Pathway Analysis (IPA, http://www.ingenuity.com) as described in detail previously [25] to identify de novo molecular networks that are associated with the NOD altered genes. We compared the whole genome mRNA expression in CD4 T-cells from 2-, 3-, and 4-week old NOD mice to that of two matched control strains, NOR and C57BL/6 (C57). NOR shares ∼88% of its genome with NOD mice, including the diabetogenic H2g7 MHC haplotype and several important

non-MHC T1D susceptibility loci [ [1], [2], [3] and [4], 29]. C57, on the other hand, is a more genetically distantly related strain to NOD. Yet, like C57, NOR is both insulitis- and diabetes-free. We identified 362, 982, and 581 probe sets (genes) with highly significant expression differences between strains (p < 0.005, Benjamini–Hochberg; FDR of 0.5%) at 2, 3, and 4 weeks, respectively. As expected, the majority of these genes had a similar pattern of expression in NOD and NOR compared to C57 ( Fig. 1). The focus of our study was to identify genes in NOR mice whose expression was similar to that in C57 but different from that in NOD, i.e. genes differentially expressed Ribociclib in NOD relative to both NOR and C57, and herein referred to as NOD altered genes. These constitute prospective candidate genes for protection of NOR mice against diabetes.

Thus, we identified a total of 58, 115, and 65 probe sets whose expression was altered in NOD at 2, 3 and 4 weeks, respectively, compared to both controls (clusters of lower or higher expression in NOD are indicated by arrows in Fig. 1). These represented 56, 107, and 60 different genes, respectively, shown in Table 1, Table 2, Table 3 and Table 4. The great majority of these genes were of lower expression in NOD mice compared to controls: ∼70% at 2 weeks; ∼72% at 3 weeks; and ∼87% at 4 weeks. Twenty-five genes (72% of which was of lower expression in NOD mice) were common Pembrolizumab research buy to all 3 ages. Results for qRT-PCR validation of eight genes are shown in Fig. S1 (3 of higher expression in NOD mice: protein tyrosine phosphatase 4a2 (Ptp4a2), biogenesis of organelles complex-1, subunit 6 (Bloc1s6, pallidin) and transmembrane protein 87A (Tmem87a); and 5 of lower expression in NOD mice: tripartite motif-containing 5 (Trim5), tripartite motif-containing 12A (Trim12a), Gatm (glycine amidinotransferase (l-arginine:glycine amidinotransferase)), lymphocyte antigen 6 complex, locus C1 (Ly6c1), and receptor transporter protein 4 (Rtp4) compared to control mice).

The DRG and

TG contain calcium-binding proteins (CaBPs) such as parvalbumin, calretinin, S100 proteins and neurocalcin [13], [14], [15], [16], [17] and [18]. These CaBPs are localized in medium-sized to large neurons with myelinated axons. Previous immunohistochemical studies have demonstrated that encapsulated and unencapsulated corpuscular endings selleck compound in the skin and oral mucosa contain CaBPs [19], [20] and [21]. Thus, CaBP-containing neurons are considered to include low-threshold mechanoreceptors. In addition, large DRG neurons which contain CaBPs send their peripheral axons to muscle spindles [13], [14], [15] and [17]. These findings suggest that CaBPs are also markers for muscular proprioceptors in the DRG. However, cell bodies of proprioceptors innervating masticatory muscles and periodontal ligament are located in the mesencephalic trigeminal tract nucleus (Mes5). Because primary neurons in the Mes5 contain parvalbumin [22], [23] and [24], the CaBP is recognized to be a marker for primary proprioceptors

in the trigeminal nervous system. The development and survival of vertebrate neurons depends on neurotrophic factors such as nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), neurotrophin-3 (NT3) and neurotrophin-4 (NT4) [25], [26], [27], [28], [29], [30], [31], [32], [33] and [34]. These neurotrophins are capable of promoting the survival of specific neuronal populations through their interaction with the tyrosine kinase receptors trkA, trkB and trkC [26], [28], [30], [33] and [34]. NGF binds to trkA. BDNF and LY2109761 purchase NT-4 signal through trkB. NT-3 binds to trkC and, to a lesser degree, trkB. Previous studies have demonstrated that the deletion of genes for these trks results in loss or decrease of various types of sensory neurons in the spinal

nervous system [25], [26], [27], [28], [29], [31] and [32]. In addition, the deficiency of intracellular proteins affects the development of nociceptive and proprioceptive neurons in the DRG [35] and [36]. The present review will focus on the developmental dependency of primary sensory neurons on neurotrophins and other proteins second in the trigeminal nervous system. The number of TG neurons is severely decreased in trk-knockout mice as compared to wildtype and heterozygous mice (82%, 39%, and 48% reduction for trkA, trkB and trkC, respectively) [37]. In trkA- or trkC-deficient mice, the number of TG neurons with a variety of cell body sizes is decreased. However, the absence of trkB causes the reduction of medium-sized and large TG neurons. The loss of NGF and trkA results in the disappearance of CGRP-containing neurons in the DRG and their central axons in the superficial laminae of the spinal cord [26] and [29]. In addition, CGRP-containing free nerve endings are absent in the skin of trkA null mutant mice.

After reduction, the highly purified BMP fraction at approximately 30 kDa was

found to consist of several peptides, suggesting that BMP is not a single protein [10], [14], [15] and [16]. Indeed, amino acid selleck chemical sequences of the peptides indicate the presence of distinct but related molecules, including BMP-1, BMP-2/BMP-2A, BMP-3/osteogenin, BMP-4/BMP-2B, BMP-5, BMP-6/Vgr-1 and BMP-7/OP-1 [10], [15], [16] and [17]. Among these compounds, only BMP-1 has a distinct structure and belongs to the metalloproteinase family [14]. The other BMPs are related to each other and can be classified into multiple subclasses of the transforming growth factor-β (TGF-β) family, which contains several other members in both vertebrate and invertebrate species [14], [15], [17] and [18]. Importantly, each of the recombinant proteins, including BMP-2, BMP-4 and BMP-7, has been shown to induce heterotopic bone formation in vivo, confirming the presence of original “BMP” activity [14], [19] and [20]. It was also shown that the bone-inducing activity of the extracts of demineralized bone and dentin could be attributed to several BMPs. Two recombinant Drosophila BMP homologs, Dpp and 60A, also induced heterotopic bone formation in rodents, suggesting that the signal transduction mechanisms

are conserved across vertebrate and invertebrate species [21] and [22]. To date, genes encoding more than 20 members of the BMP and related growth and differentiation factor (GDF) families have been found in the human genome. Several Alectinib solubility dmso BMPs and GDFs Montelukast Sodium have been shown to induce the formation of bone and/or cartilage tissue not only through the implantation of recombinant proteins but also through the transduction of individual cDNAs into sites in vivo using viral vectors or the implantation of cell transduced viral vectors [23] and [24]. TGF-β1 and

TGF-β2 were identified as cartilage-inducing factor-A (CIF-A) and CIF-B, respectively, in bone extracts in a chondrogenesis assay of muscle cells [13] and [25]. However, the implantation of TGF-β1 in vivo failed to induce heterotopic bone formation [26]. Interestingly, a combination of TGF-β1, TGF-β2 or activin with BMP-2 exhibited potent heterotopic bone-inducing activity in vivo [27], [28] and [29]. A small proteoglycan known as osteoglycin has also been shown to enhance the bone-inducing activity of BMP-2 [30] and [31]. The molecular mechanisms underlying the synergistic behavior of BMPs and other molecules remain unclear. Neither the functional receptors nor the downstream effectors of the TGF-β family had been identified when BMPs were first isolated from bone extracts. To examine the details of their intracellular signaling, various cell lines were screened for high responsiveness to BMP-2 in vitro.

, 2009, Kurzrock and Speer, 2001 and Scharnhop and Winterhalter, 2009). The mass errors were lower than 5 ppm confirming the molecular formulas. The results obtained in tandem mass spectrometry

studies of m/z 195, 315 and 317 corroborate with the assigned structures in accordance with literature data: caffeine ( Alonso-Salces et al., 2009) (Elab 25 eV: 195 → 138, 110), cafestol ( Scharnhop & Winterhalter, 2009) (Elab 18 eV: 317 → 299, 281, 147, 133) and kahweol ( Scharnhop & Winterhalter, 2009) (Elab 18 eV: 315 → 297, 279, 149, 131). The conditions clearly improve cafestol and kahweol concentrations; methylated fatty acids can be seen at the end of chromatogram in Fig. 3(B), due to methanolysis. Furthermore, the literature highlights the need for anhydrous methanol, which microwave heating showed to be unnecessary, greatly simplifying the methodology and costs ( Bertholet, Selleck Volasertib 1987). Quantification of cafestol and kahweol was accomplished with the external standard method as response factor for the HPLC, obtained by linear regression of known concentrations versus peak area. Linearity was observed for a concentration range of 1–56 μg/mL, with a 5% confidence level and a r correlation coefficient for cafestol and

kahweol higher than 0.99. Coefficients of variation (CV) below 7% were observed for the mixture of free diterpenes. A fast and improved method to obtain a mixture of cafestol (1) and kahweol (2) from green Arabica coffee oil was successfully developed. The microwave-assisted protocol proved to be simple, selleck kinase inhibitor fast, enabled the use of higher reaction amounts and can be carried out at higher temperatures. The rapid

speed of reaction avoided the development of undesired products and increased product yield. In addition, the microwave-assisted method required no clean-up procedure when compared to conventional heating. We thank the Brazilian science foundations FAPERJ, CAPES, CNPq and EMBRAPA CAFÉ for financial assistance. The authors also wish to thank Grão Mestre Café for providing the green coffees. We are grateful to Prof. Paula F. de Aguiar for helping with statistics, Prof. Alberto J. Cavalheiro for the support on HPLC. “
“The edible mushroom Pleurotus ostreatus has a pleasant taste and nutritional properties that are beneficial to health. Daily intake of this mushroom may influence the lipid profile in hypercholesteraemic Teicoplanin patients and improves antioxidant status ( Hossain et al., 2003 and Jayakumar et al., 2007). This mushroom can also be a source of elements, such as iron (Fe), zinc (Zn), selenium (Se), copper (Cu) and molybdenum (Mo), which are involved in many essential biochemical processes ( Zaidman, Yassin, Mahajna, & Wasser, 2005). The bioaccumulation potential of nutrients by fungi enriched with essential elements for human health has been investigated in mycelium and also in mushroom (Munoz et al., 2006, Rabinovich et al., 2007, Silva et al., 2010 and Silva et al., 2012).

dubium seeds were also shown to be highest at 50, 55 and 70 °C, respectively ( Ahmed et al., 2009, Lo Piero et al., 2002 and Teixeira

et al., 2000). Molecular rearrangements in protein structure can lead to increase of enzyme activity ( Purich, 2010). Caseinolytic activity was higher when PP was previously incubated at pH 4.0 and 7.0 (Table 2). A partially purified enzyme from S. dubium seeds also showed proteolytic activity towards azocasein at pH 4.0 but, unlike M. oleifera activity, the enzyme was highly active up to pH 11.0 ( Ahmed et al., 2009). It is known that pH affects the shape, charge Hydroxychloroquine molecular weight properties, the correct positioning of the substrate and the ionisation of side chains of amino acids, in both the active site and in the whole enzyme ( Purich, 2010). Heating of PP from 30 to 40 °C did not interfere in milk-clotting activity, which increased significantly after heating at 50 °C and was neutralised at 70 °C (Table 1). Milk-clotting

enzymes from Bromelia hieronymi, W. coagulans, Solanum esculentum and Solanum macrocarpon are stable proteins, remaining active after heating to 45, 70 and 70 °C, respectively ( Bruno et al., 2010, Guiama et al., 2010 and Naz et al., 2009). A milk-clotting enzyme called religiosin B, purified from Ficus religiosa stem latex, showed highest milk-clotting activity at temperatures of 55 and 60 °C ( Kumari, Sharma, & Jagannadham, 2012). Milk-clotting activity from M. OTX015 molecular weight oleifera flowers was highest after previous incubation of PP at pH 3.0 ( Table 2) and lost of activity was detected when PP was previously incubated at pH values higher than 8.0. Calf rennet showed similar behaviour, acting better in acid PTK6 than in alkaline reaction medium ( Richardson, Nelson, Lubnow, & Schwarberg, 1967). Differently, the milk-clotting enzyme religiosin B showed highest clotting ability at pH 6.0 ( Kumari et al., 2012). High thermal stability and ability to work in a wide pH range are

important criteria for the choice of proteases to be used in industrial processes (Vieille & Zeikus, 1996). In this sense, the milk-clotting enzymes present in PP are promising candidates for application in milk-clotting at an industrial large scale. Additionally, the traditional use of M. oleifera flowers in human diet, being eaten raw or after lightly blanched ( Makkar & Becker, 1996), is an indicative of PP safety for use in cheese production. The evaluation of enzyme activities from M. oleifera flowers in presence of protease inhibitors ( Table 3) showed that the caseinolytic activity on azocasein was not significantly (p > 0.05) altered in presence of PMSF, while milk-clotting activity was significantly (p < 0.05) reduced, by as much as 25%. E-64 significantly (p < 0.05) inhibited only milk-clotting activity (by 30%), while pepstatin A significantly reduced (p < 0.05) caseinolytic and milk-clotting activities, by 25% and 57.5%, respectively.

, 2003). On the other hand, the content of flavonoids found in jambolão fruits in this study was about 7–13 times higher than those previously reported, 13.5 mg/100 g (Luximon-Ramma et al., 2003) and 7 mg/100 g (Benherlal & Arumughan, 2007). This difference can be attributed to the inherent variability of the raw material, as well as to differences

in methodology or standard used. The monomeric anthocyanins content (211 mg/100 g) was in the same range as those found in literature for jambolão fruits, Atezolizumab 134 mg cyd 3-glu/100 g (fresh weight) (Benherlal & Arumughan, 2007) and 230 mg cyd 3-glu/100 g (dry basis) (Veigas et al., 2007). When compared to other fruits from the Myrtaceae family, jambolão showed a high content of monomeric anthocyanins (211 mg/100 g), as compared to those reported for camu–camu (Myrciaria dubia), 30–54 mg/100 g ( Zanatta, Cuevas, Bobbio, Winterhalter, & Mercadante, 2005) and Eugenia myrtifolia, 33 mg/100 g ( Longo, Scardino, Vasapollo, & Blando, 2007). On the other hand, the content of total carotenoids

INCB024360 purchase present in camu–camu, 355–1095 μg/100 g ( Zanatta & Mercadante, 2007) was higher than what was found in jambolão fruits ( Table 1). Table 2 presents the chromatographic, UV–Vis and mass spectrometry characteristics of anthocyanins from jambolão fruit. The chromatogram obtained for these pigments and their structures are shown, respectively, in Fig. S1 and S2 from Supplementary data. The composition of anthocyanins from jambolão was marked by the presence of different aglycones diglucosides. Five of the six aglycones most commonly Etofibrate found in foods were identified by tandem-MS: delphinidin (dpn, m/z 303), cyanidin (cyd, m/z 287), petunidin (ptd, m/z 317), peonidin (pnd, m/z 301) and malvidin (mvd, m/z 331). In all anthocyanins, the hexose was assigned as glucose considering the standards available and that glucose was the only monosaccharide previously found after acid hydrolysis of an anthocyanin extract obtained from fruits of S. cumini ( Veigas et al., 2007). For diglycosilated

anthocyanins (peaks 1, 2, 3, 4a, 5 and 6a), the presence of two glucose unities glycosylated at different positions (probably 3 and 5) rather than a disaccharide at position 3 was assigned considering the presence of two fragments derived from two consecutive losses of 162 u, instead of a fragment resulting from a single loss of 324 u, as reported in previous studies ( De Rosso et al., 2008 and Wu and Prior, 2005). Moreover, the presence of 3,5-diglucosides of dpd, cyd, ptd, pnd, and mvd in jambolão was recently confirmed by nuclear magnetic resonance (NMR) ( Li, Zhang, & Seeram, 2009b). The identification of cyd 3-glucoside (peak 6b), cyd 3,5-diglucoside (peak 2), mvd 3-glucoside (peak 8) and mvd 3,5-diglucoside (peak 5) was confirmed by co-chromatography with the respective standards. Two anthocyanins co-eluted in peak 6.

Statistical analyses were carried out using the GraphPad Prism software (GraphPad, San Diego, CA, USA) by one-way analysis of variance (ANOVA). Duncan’s multiple range test was employed to test for significant differences between the treatments at p < 0.05 and p < 0.01. The total ginsenoside contents in each tissue of the entire ginseng plant were analyzed. Cultivation of ginseng by hydroponics involves a shorter cultivation period in a greenhouse in which variables such http://www.selleckchem.com/products/s-gsk1349572.html as light, temperature, moisture, and carbon dioxide content can be controlled [30] and [31]. Therefore, we used hydroponically cultured 3-yr-old ginseng

plants (Fig. 1). Fig. 2 shows that ginsenoside accumulations within the aerial parts (leaf and stem) were increased as compared with the control. Total

ginsenoside contents in the leaf were higher than other tissues. In addition, total ginsenoside contents within the underground parts (rhizome, root body, epidermis, KRX-0401 solubility dmso and fine root) were also increased, except in the epidermis. Total ginsenoside contents of the root body in MJ-treated plants increased by approximately twofold compared with that of the control. This result demonstrates that the increase in ginsenoside contents of the root body is the highest among all tested ginseng organs. In rhizome, total ginsenoside accumulation and its composition was significantly increased after MJ treatment. Total ginsenoside content of fine roots was increased by approximately 6 mg/g compared with the control, which is the most increased content observed in underground parts. In the epidermis, total ginsenoside content was only minimally influenced by MJ treatment. Fig. 3 shows the accumulation of individual ginsenosides Pyruvate dehydrogenase in different tissues.

The content of ginsenoside Re in aerial parts (leaf and stem) of the ginseng plant was the highest. In leaf, ginsenoside Re and Rd contents were mainly enhanced. The ratio between PPD-type and PPT-type ginsenosides was significantly changed in the stem. The content of ginsenoside Rd was increased more than other ginsenosides; therefore, the ratio of PPD-type ginsenoside was increased. In rhizome, the ratio of PPD-type ginsenoside was also increased due to accumulated ginsenoside Rd, although the content of ginsenoside Rg1 in the rhizome was the highest. The greatest increase of ginsenoside level was shown in the root body. All individual ginsenoside contents were increased. Levels of ginsenosides Rb1 and Rg1 were doubled as compared with the control. Although the content of ginsenoside Rg1 was the highest, ginsenoside Rd was enhanced fivefold. In addition, ginsenoside Rc and Rb2, which was not detected in the control, accumulated after MJ treatment, showing in the increased ratio of PPD-type ginsenoside. In fine root, all individual ginsenosides were also increased. Fine roots contained mostly ginsenoside Re, but the ratio of ginsenoside Rb1 was enhanced upon MJ treatment.

In contrast, just like in the case of addition or subtraction transformations, they would make no specific prediction as to whether this number word or another number word applies, if one or more individual members of the set are replaced by other individuals – unless the pragmatics of the task leads them to the correct answer. This mTOR phosphorylation explanation in terms

of set identity predicts children’s failure at the one-to-one comparison task, which was left unexplained in Brooks et al.’s (2012) account. Indeed, in both the one-to-one comparison task and the single-set transformation task, children must choose between a previously-heard label and a new label, thus in terms of pragmatics the two tasks are equivalent. In terms of quantities involved, the two tasks are equivalent too. Therefore, if children reason in terms of quantity, they should succeed in

the comparison task when the two sets are equal in number, just as they succeed in the single-set task when no transformation is applied. If however children reason in terms of set identity, then in the one-to-one comparison task there is no reason why information about one set should help them RNA Synthesis inhibitor solve a question about another set. To get a better understanding of this interpretation, think of first names, which are defined in terms of identity. If a set is called “five” and is put in exact one-to-one correspondence with another set, we predict that children are undecided as to whether this second set should be called “five” like the other set. Nevertheless, children should know that if the members of a set called “five” remain in the set, and no new item is added, then the set is still called “five”.5 Interpreting children’s usage of the number words in terms of set identity makes an important prediction. In the published versions of the single-set transformation task

(Brooks et al., 2012 and Sarnecka and Gelman, 2004), the transformation leaving numerosity constant left the identity of the set PIK3C2G unchanged as well. Under our interpretation, subset-knowers should not choose to conserve the initial number word for an identity-changing substitution transformation, even though the cardinal value of the set remains constant in this condition. At 5 years of age, children have clearly overcome the limitations of their understanding of numerical equality, since they know how set transformations impact number words, even for number words that fall beyond their counting range, and even for substitution transformations that keep number constant while altering the identity of a set’s members (Lipton & Spelke, 2006).

“
“Leaf area as photosynthetically active area is one of

the main drivers for tree growth and thus an important tree characteristic for tree growth studies. For silvicultural purposes trees have to be considered as parts of stands, and individual tree growth has to be investigated in relation to stand structure. Thus, O’Hara (1988) selleck screening library used the area for individual trees as a measure of site occupancy. Leaf area in relation to stand parameters, e.g., ground area potentially available (APA), which could be named as individual tree leaf area index, but also leaf area in relation to stemwood increment which is described as growth efficiency (Waring, 1983) are important research issues. However, leaf area is hard to determine precisely and non-destructively. For leaf area index determination of stands various optical instruments like LAI-2000 (Li-Cor) or SunScan (Delta-T) are available. But these instruments are limited by the complexity of the canopy structure and improvement in accuracy is still needed (Moser et al., 1995, Chen et al., 1997, Pokorny and Marek, 2000 and Pokorny et al., 2004). Another

way to determine stand leaf area index is to use the individual tree leaf area. Hence, different approaches to estimate individual tree leaf area in an indirect way were and are investigated. Such investigations aim at strong relations of leaf area to other tree characteristics. Based on the pipe model of Shinozaki et al. (1964), Luminespib which supposed that a given leaf area is supplied Rebamipide with water from a respective quantity of conducting pipes, mainly sapwood area (e.g., Waring et al., 1982, Bancalari et al., 1987 and Meadows and Hodges, 2002), early sapwood area (Eckmüllner and Sterba, 2000), and diameter at breast height (e.g., Gholz et al., 1979 and Baldwin, 1989) are used as estimators for leaf area or leaf biomass. A few studies deal with estimating leaf area with allometric functions based on different other

tree characteristics (e.g., Pereira et al., 1997 and Kenefic and Seymore, 1999). The majority of studies dealing with indirect leaf area estimation describe sapwood area as the most accurate estimator for leaf area (e.g., Long et al., 1981, O’Hara and Valappil, 1995 and Meadows and Hodges, 2002). But to get continuously information about leaf area and related characteristics, e.g., growth efficiency, and their development over time, the determination via sapwood area is not feasible, because from the same trees cores cannot be taken every 5 or 10 years over a long term. Additionally, it is well known that the relationship between leaf area and sapwood area, even within species, is not constant. Differences could be shown between sites, crown classes, stand density, and age (Long et al.

If this approach works similarly in the clinical setting including to the point of being able to circumvent the

need for an interappointment intracanal medication still needs to be shown by clinical trials. Although studies have revealed that PUI may enhance cleaning of root canal irregularities, many of these studies also showed that along with other tested irrigation approaches, PUI was not able to completely remove debris in the apical part of the root canal 21 and 22. As for disinfection, in vitro findings about the effectiveness Vemurafenib price of PUI in reducing bacterial populations have been somewhat inconclusive. One study showed that it was superior to syringe irrigation (30), and another one found no significant difference between the two techniques (31). PUI was not superior than syringe irrigation or passive sonic activation, all using 5.25% NaOCl, in eliminating E. faecalis from root canals of extracted teeth (32). The present findings with PUI alone corroborate those from studies showing no significant additional antibacterial effects. However, when combined with a final rinse with CHX, the whole approach was significantly

effective. A variation in PUI with the irrigant being pumped under a high flow rate through a needle attached to an ultrasonic handpiece has been proposed 19 and 33 and shown to improve cleaning (19) and disinfection 33 and 34. The antibacterial effects of the PUI approach with constant irrigation remain to be evaluated N-acetylglucosamine-1-phosphate transferase in oval-shaped canals. In conclusion, the present in vitro study showed that PUI followed by CHX rinsing significantly reduced the PS-341 supplier bacterial counts and the incidence of positive

cultures after chemomechanical preparation of oval-shaped root canals. Therefore, there seems to be a benefit of using this combined approach as supplementary steps in the treatment of infected root canals. Further clinical studies are required to confirm these results. Also, the search for effective alternative or supplementary measures to predictably disinfect oval-shaped canals should be encouraged. The authors thank Fernando A. Magalhães for his excellent technical support. The authors deny any conflicts of interest related to this study. “
“Fracture of nickel-titanium (NiTi) endodontic instruments is not an uncommon incident during root canal treatment (1). Fatigue and shear failure are cited as the main reasons for fracture, and the failure mode of the instrument is related to the canal preparation technique (2). Recent clinical studies document that the prognosis for endodontic treatment is not significantly affected by the fracture and retention of a fractured instrument 3 and 4. However, the prognosis is lower when a fractured instrument compromises the effective disinfection of a root canal associated with periapical pathology 3, 4 and 5. Therefore, the management of a case with a broken instrument might involve an orthograde or a surgical approach (6).