According to

According to PF-562271 concentration Alasino et al. (2011), SSL helps in maintaining the tearing quality. These authors also verified that the increase of the concentration of SSL produces a beneficial effect on the sensory attributes of bread, including crumb texture score. In general, it can be concluded that breads with added SSL and maltogenic amylase presented an increase in volume and a reduction in firmness on Days 1, 6 and 10 of storage, as well as good acceptance regarding the sensory attributes evaluated. This study presents precise dosage values for practical application in white pan bread. Further research could include the use of combined emulsifier and enzyme in other bakery products, including fiber-enriched

products, cakes, etc., where an increase in shelf-life is technologically and economically important. “
“Theobroma cacao L. (Sterculiaceae) is an important crop of several tropical countries. When ripe, pods are harvested from the trees and opened

to extract the wet beans (∼10% fresh weight of the cacao fruit). After fermentation of surrounding pulp, the beans are dried and bagged, constituting the cocoa of commerce, employed mainly in chocolate manufacturing ( ICCO, 2011a; Kalvatchev, Garzaro, & Cedezo, find more 1998). During the extraction of cocoa beans, pod husks, accounting for approximately 52–76% of the weight of the cacao fruit (Donkoh, Atuahene, Wilson, & Adomako, 1991; Fagbenro, 1988), are thrown away and may cause an environmental problem when dumped around the processing plants. In addition to foul odors due to decomposition, cacao pod husks may be a significant source of disease inocula, such as black pod rot (Barazarte, Sangronis, Methocarbamol & Unai, 2008; Donkoh et al., 1991; Figueira, Janick, & BeMiller, 1993; Kalvatchev

et al., 1998). Because each ton of dry beans produced generates approximately ten tons of cacao pod husks (Figueira et al., 1993; Kalvatchev et al., 1998) and because the world production of dry cocoa beans is projected to rise from approximately 3.6 million tons in 2009/2010 (from October to September) to 3.9 million tons in 2010/2011 (ICCO, 2011b), the burden of cacao pod husk waste continues to increase and represents a serious challenge for waste management. In cocoa producer countries, the processing of this cacao waste may offer economic advantages and decrease the extent of the associated environmental problems. An alternative method of processing cacao pod husks could be their use in pectin production, polysaccharides widely used as gelling and stabilizer agents in a variety of food, cosmetic and pharmaceutical products (Rolin, 1993; Voragen, Pilnik, Thibault, Axelos, & Renard, 1995). Nowadays, commercial pectins come from citrus peel and apple pomace, both by-products of juice production and are generally, extracted with hot, diluted mineral acid (Rolin, 1993; Voragen et al., 1995).

015), higher pain during the muscular palpation of the face (P < 

015), higher pain during the muscular palpation of the face (P < 0.001) and neck (P = 0.002) and more masticatory complaints

(P = 0.002). Pain itself has probably interfered with the mandibular activities, and these findings also support the high frequency of TMD in this sample. Amongst risk factors for TMD, bruxism was commonly observed, but the groups did not statistically differ. Bruxing or clenching the teeth causes an overload on the masticatory muscles and can precipitate TMD. 38 Limitations of this study are the design, which does not allow Bleomycin concentration the investigation of cause–effect associations, and a higher frequency of women in the study group. Chronic pain is more frequent in the female gender,24 and it might have interfered with the results AZD6738 observed. Doses of antidepressants

and anti-hypertensive drugs, which were not investigated, may also have underlain, at least in part, the results as to lower salivary flow in the study group. In conclusion, orofacial pain patients need to be evaluated in regard to their salivary function. They had lower salivary flow and more xerostomia complaints than the controls, which can cause discomfort and effectively contribute to pain. This study was supported by FAPESP (Foundation of Research of the State of Sao Paulo, 2009/00350-6). None declared. This study was approved by the Ethics Committee of the Hospital das Clinicas, Medical School, University of Sao learn more Paulo, Brazil (0901/2008). We would like to acknowledge Raphael Sa, Rodrigo Primiceri da Silva and Maira Caracas for their participation in the study. This study was supported by FAPESP (Foundation of Research of the State of Sao Paulo, 2009/00350-6). “
“The growing obesity epidemic affects millions of people in the modern world and has become a risk factor for the development of many chronic-degenerative diseases such as cardiovascular diseases and diabetes mellitus type II. Several scientific

studies have suggested that obesity contributes effectively to the severity of periodontal disease.1, 2, 3, 4 and 5 Periodontitis is a chronic infectious disease caused predominantly by bacteria that release endotoxins activating pro-inflammatory cytokines (IL-1, TNF-α, amongst others) that affect the supporting tissues of teeth and induce the loss of alveolar bone, cementum and periodontal ligament.6 and 7 The increase in body mass index (BMI) and waist-hip ratio (WHR) are associated with the development of periodontitis.4 Epidemiological data have shown that obese and insulin resistant patients show high plasma concentrations of inflammatory markers. The adipose tissue secretes large quantities of TNF-α and IL-66 and the concentration of these cytokines is proportional to the BMI. The increase in plasma concentration of pro-inflammatory cytokines might explain the relationship between obesity and periodontal disease.

The following information on patient history was obtained: TIA an

The following information on patient history was obtained: TIA and minor strokes we classified into the following categories: retinal TIA, cerebral

TIA or stroke. Documented were the nature of the events such as visual, pure motor, pure sensory, dysarthria, dysphasia, ataxia, apraxia or combination of events. ABCD2 scores were obtained in all patients [6]. MRI findings were classified into cortical infarcts, subcortical infarcts and leucoaraiosis. Infarcts were further subdivided into recent or this website non-recent and left or right sided. The side, severity of the stenosis and presence of plaque ulceration on duplex and CTA were documented as well. Furthermore, blood pressure was documented as well as the current use of anti-thrombotic drugs or anti-coagulants. Documentation of the TCD embolus detection included: the side of insonation, the peak systolic-, mean and end-diastolic velocity, the duration of the measurement and the presence or absence of cerebral embolism by human experts. If experts found cerebral embolism the following parameters LEE011 chemical structure of that embolus were noted: velocity, phase of cardiac cycle (systolic/diastolic) in which the events occurred, intensity, duration and a parameter related to the musical characteristics of the embolus (the zero-crossing index) [7]. Data of stent

procedures and surgery were prospectively documented including the occurrence of neurological or non-neurological complications. The follow-up at three month included a neurological visit at the outpatient clinic. Documented were the TIA and stroke recurrence rate. If complications had occurred in the post-operative phase of angioplasty or surgery they were evaluated including the occurrence of new medical events in the last three months. All data were stored in a downloadable

Internet based electronic management system which allowed online statistical analysis of all included case records. This data management system has been developed by Mediwebdesign© The Netherlands ( A TCD Delica 9 series (Delicate/Shenzen/China) equipped with a 2 MHz TCD transducer and a notebook PC (Acer®, Aspire 1800 Series) were used for this study. A special Delicate headband was used to hold the 2 MHz transducer, which allowed Coproporphyrinogen III oxidase hands-off monitoring. The insonated artery was the middle cerebral artery at its origin, just lateral of the terminal internal carotid artery, on the ipsilateral side of the symptomatic carotid artery territory. Patients were monitored for 30 min. In case of positive embolism the other contra-lateral middle cerebral artery was examined to estimate whether the cerebral embolism was a uni-lateral or bilateral phenomenon. Insonation depth varied between 45 mm and 55 mm. Patients were asked to not speak or move their head during the monitoring session because angular or lateral probe movements may induce false positive embolic events.

The surface water flow through the Sicily Channel is estimated to

The surface water flow through the Sicily Channel is estimated to be approximately 1.4 times the surface water flow through the Gibraltar Strait because: (1) the net evaporation Ion Channel Ligand Library price over the EMB is about three times than the net evaporation over the WMB, (2) deep water convection is more significant in the EMB than the WMB, so the amount of lower-water outflow through the Sicily Channel is more significant than through the Gibraltar Strait. Depending on the two previous

aspects, the amount of inflow water needed to compensate for the loss of water due to net evaporation and outflow is much higher through the Sicily Channel than the Gibraltar Strait. The Sicily Strait is 11 times wider than the Gibraltar Strait, which can explain why the surface flow through

the Sicily Channel is higher than that through the Gibraltar Strait. The calculated SST over the 1958–2010 period followed the reanalysed data with no biases over either studied sub-basin. The surface water Ku-0059436 clinical trial of the EMB was approximately 1.6°C warmer than that of the WMB in the studied period. The Mediterranean Sea surface water displayed a significant warming trend, most pronounced in the 1985–2010 period and over the EMB (Table 5). The modelled sea surface salinity in the 1958–2010 period followed the reanalysed data with a bias of 0.09 and 0.11 g kg−1 for the WMB and EMB, respectively. The surface water of the EMB was approximately 0.87 g kg−1 more saline than that of the WMB. The Mediterranean Sea surface water displayed an insignificant salinity trend (Table 5). In the EMB, this can be explained by a balance between two effects: significant warming (implying increasing salinity) and decreasing freshwater input (implying decreasing salinity). The annual temperature and salinity cycles in the surface and deep layers were realistically simulated using PROBE-MED version 2.0. The calculated evaporation rate and heat balance components agreed well with

and were strongly correlated with the reanalysed data. This may indicate that the air–sea interaction and turbulent mixing are modelled satisfactorily. Table tetracosactide 5 shows the statistical analysis of net precipitation rates. Calculated net precipitation rates display a positive (negative) trend over the WMB (EMB), most markedly in the 1958–1984 (1985–2010) period. Moreover, the annual average net precipitation rates were −0.88 ± 0.95 and −1.52 ± 1.28 mm day−1 for the WMB and EMB, respectively. This may explain the much more saline surface water in the EMB than the WMB. Different estimation methods are available for calculating net precipitation rates. ERA-Interim reanalysed data indicate that the net precipitation rates over the 1985–2010 period, calculated as long-term means, were −1.4 mm day−1 (trend 0.099 mm day−1 yr−1) and −2.1 mm day−1 (trend −0.139 mm day−1 yr−1) for the WMB and EMB, respectively. Romanou et al.

For the four yield components, PN, SP, SFP, and GW, only SP was n

For the four yield components, PN, SP, SFP, and GW, only SP was not significant between sites. Maximum tiller number per square meter in Nanjing was 313 m‒2 for II You 107 and 335 m‒2 for Xieyou 107, compared with 731 m‒2 for II You 107 and 738 m‒2 for Xieyou 107 in Taoyuan (P < 0.05). Panicle rate was significantly higher at Nanjing than at Taoyuan. The difference of source capacity (LAI at heading stage) and sink capacity between Taoyuan and Nanjing was also significant. The SM at Taoyuan was 1.68-fold higher for II You 107 and

1.63-fold higher for Xieyou PF-562271 in vivo 107 than at Nanjing. Leaf area indexes at heading stage for II You 107 and Xieyou 107 were 1.36 and 1.30-fold higher at Taoyuan than at Nanjing. The CV for SM was larger than that for LAI at heading stage and was identical for the two cultivars. The GD at Taoyuan was 42 d longer for II You 107 and 38 days longer for Xieyou 107 than at Nanjing. The difference in GD between the two sites

was caused mainly by PHP, with averages of 43 days for II You 107 and 39 days for Xieyou 107. No significant difference was found in HM across sites or years. There was a small difference in PH between Taoyuan and Nanjing for both cultivars, and PH was this website stable at approximately 110 cm. Overall, the significant differences between Taoyuan and Nanjing, in descending order, were PHP > GD > PN > MT > SM > LAI > PW > GW > SFP. PH, HM, and SP were relatively stable across locations, and the differences were not significant. The stability of yield-related traits was identical for both cultivars. Compared with the large differences between locations, the differences in the yield-related traits, with the exception of PR, SFP, and GW, between years for both cultivars were not significant. The CV

of 13 yield-related traits was nearly identical for both cultivars. Overall, GD, PH, GW, SFP, and PN were relatively stable across years, with CVs of smaller than 10%. Environment variance (S2) of the two cultivars, II You 107 and Xieyou 107, showed similar stability for GY ( Table 6). However, the stability of PW, GW, and SM of the large-panicle variety, Regorafenib molecular weight II You 107, was higher than that of the heavy-panicle cultivar, Xieyou 107. Among the yield-related traits, independent of large-panicle or heavy-panicle type, HM, PH, SFP, and GW were the most stable with a CV lower than 10%, followed by PW, GD, PHP, LAI, and SP with a moderate CV of 10%–20%. In comparison, MT, PR, and GY were the most unstable traits with the CV above 30%. Grain yield potential is defined as the yield of a cultivar when grown in an environment to which it is adapted, with unlimited nutrients and water and with pests, disease, weeds, lodging, and other stresses effectively controlled [28].

Ideally, we would employ a psychosocial mediator such as stress,

Ideally, we would employ a psychosocial mediator such as stress, defined as “the interaction between people and their social Selleckchem Sotrastaurin environment involving psychological processes” (Egan et al.,

2008), but unfortunately such variables were not available in the study. We therefore used the General Health Questionnaire (GHQ-12) to derive a psychological factor for this study. The GHQ-12 comprises 12 self-complete questions describing mood states used to assess psychiatric morbidity, with six of the questions being positively phrased and six negatively phrased (Goldberg and Williams, 1988). Each item of the GHQ-12 has four possible response options and these were scored dichotomously using the GHQ method (all beta-catenin inhibitor items coded 0-0-1-1). Missing items were scored zero. The 12 scores were then summed and a cut-off for mental ill health derived from the mean score. For both waves 1 and 5, mean GHQ scores were approximately 2, setting a cut-off of 3 or more as

a case (‘1’) compared to not being considered a case (‘0’). Data on smoking, alcohol consumption, diet and physical activity were based on self-report. Behavioral variables created for these analyses were based, where possible, on contemporary guidelines, as well as making variables homogeneous between waves. Smoking status at both waves 1 and 5 was defined as current (1) versus ex- or non-smoker (0). Weekly alcohol consumption was used to define respondents as below (‘0’) versus above (‘1’) gender-specific recommended weekly limits (⩽21 versus 22+ units for males; ⩽14 versus 15+ units for females) (Changing Scotland’s Relationship Methocarbamol with Alcohol, 2009) Alcohol strength changed for some drinks during follow-up (Bromley et al., 2003) and we recalculated this variable in wave 5, although this change had no impact

on our results. Physical activity was based on the number of occasions per week that respondents took part in an activity “lasting more than 20 min” that made them “sweat or (be) out of breath”, reflecting guidelines at the time. Respondents were dichotomized into high physical activity (at least 20 min once a week; ‘0’) versus low physical activity (less than once a week; ‘1’). Diet, from food-frequency questionnaires, was based on the number of days per week on which participants reported eating fruit and vegetables. Respondents were classified as having a poor diet (‘1’) if they had at least one day per week with no fruit or vegetables consumed versus not having a poor diet if they consumed fruit and vegetables every day of the week (‘0’) (See Table 1). For each individual measure (e.g. smoking, income, etc.), and for the combined factors, a cumulative measure was generated using data from both waves of survey data such that each mediator could take a value of 0, 1 or 2, with higher scores representing more negative material, psychological or behavioral exposures.

52 mg/L), suggesting that the greater differential gene expressio

52 mg/L), suggesting that the greater differential gene expression potency

and efficacy of SDD decreased as it passed from the duodenum to the jejunum. At day 91, the median rat duodenum and jejunum EC50s were comparable (49 vs. 52 mg/L SDD). Over-represented functions associated with differential gene expression were phenotypically anchored to complementary histopathology FK506 clinical trial and biochemical data (Table 1). The reduction in the GSH/GSSG ratio is suggestive of intestinal epithelium oxidative stress (Thompson et al., 2012). Nrf2 (Nfe2l2) induction (~ 2.6-fold), and subsequent expression of downstream targets (up to 2.7-fold) is consistent with an oxidative stress response. For example, ubiquitination and proteasomal degradation proteins (Vcp, Usp14 and Ube2k), chaperone and stress proteins (Stip1, Cct7, Erp29), and antioxidant proteins (Atf4, Gpx2, and Prdx1) are consistent with oxidative stress. Interestingly, EC50s of 4.2 and 14.2 mg/L SDD for Nrf2 in the duodenum and jejunum at day 8, respectively, provide further evidence that gene expression capability decreased as SDD passed from the duodenum to the jejunum. ToxResponse modeler also calculated EC50 values less than

5.0 mg/L UK-371804 SDD for Nrf2-regulated Usp14, Cct7, and Erp29 at day 8. The Nrf2-mediated oxidative stress response was also observed at day 91 and select QRT-PCR verified genes included the induction of Nrf2, Gclc and Gpx2 ( Fig. 3). Moreover, SDD induced trefoil factor 1 (Tff1), a small secreted protein involved in cell growth that stabilizes the gastrointestinal mucosa and provides a physical barrier against toxic agents. Hydrogen peroxide also induces Tff1 ( Balcer-Kubiczek et al., 2002), while oxidative stress induced by indomethacin and ROS production is reduced by TFF1 ( Chattopadhyay et al., 2006 and Marchbank et al., 1998), further suggesting

oxidative stress protection. Tff1 was induced > 10-fold in rats (EC50 Duodenum and Jejunum Day 8 = 4.2 and 35.3 mg/L SDD), and 53-fold in mice. The induction of Tff1 is consistent with oxidative stress in the rat jejunum 4-Aminobutyrate aminotransferase ( Thompson et al., 2012), and likely represents an adaptive response to SDD. Immune response genes (e.g., Acp5, Anxa5, C3, Ccl24 Cxcl12, Kitlg, Il1rl1, Il33 and C1qa) were also differentially expressed ( Table 1), consistent with the mild to marked histiocytic infiltration at days 8 and 91 ( Thompson et al., 2012). Interestingly, Il1rl1 (5- to 10.9-fold) and Il33 (4.5- to 5.9-fold) exhibited the greatest fold changes with EC50s of 6.8 and 5.4 mg/L SDD, respectively, in the duodenum at day 8. The mouse orthologs were also highly induced immune response genes, although their efficacy of induction was lower with higher EC50s ( Kopec et al., 2012). Several cell cycle, growth and proliferation genes exhibited dose-dependent induction including Myc, Tp53 and their downstream targets.

A large number of osteoprogenitor cells may be implanted at the i

A large number of osteoprogenitor cells may be implanted at the injury site, either alone or combined with a matrix. Autologous bone marrow (BM) is rich in growth factors and osteoprogenitors as MSCs are present in the mononuclear cellular fraction of the bone marrow. Bone marrow MSCs are currently the most appropriate cells for inducing bone repair, as they have a strong osteogenic potential and are easily obtained by culturing iliac crest aspirates. Several MSC-based cell therapy modalities have been developed, i.e., with and without cell culturing, and with or without a matrix. The mononuclear cell fraction of the bone marrow,

which contains the MSCs, can be used directly by percutaneous injection of aspirated BM into the injury site. To increase the number of injected mononuclear cells and consequently of MSCs, it is possible to separate the mononuclear cells by centrifugation and Tanespimycin order concentrate them 3-fold to 6-fold [64] with good results in pseudarthrosis [65]. The healing rate increased in proportion to the injected MSC concentration. Patients whose fractures did

not heal received fewer than 1000 MSCs per mL and fewer than 30,000 MSCs in total, whereas those whose fractures healed received significantly higher MSC concentrations and counts, with a mean of 1500 MSCs per mL and 54,000 MSCs in total, in a volume of 20 mL. Concentrated or unconcentrated mononuclear cells can be mixed in the operating room Axenfeld syndrome with a synthetic or natural osteo-conducting matrix XAV-939 cell line (e.g., allogeneic bone graft or coral) before implantation. Few published studies assessed the combined use of concentrated or unconcentrated BM with a biomaterial [66], [67] and [68]. This method is a valid option for everyday practice, provided that CE-marked (that is, approved for clinical use in Europe) biomaterials are used and concentration (if used) is achieved via an approved procedure. Mononuclear cells may also be cultured in vitro to allow selection and expansion of an adherent fraction corresponding to MSC. This increases the

number of MSC to millions of cells. Expanded MSCs can be extemporaneously mixed with scaffolds during surgery (Fig. 4) so that this composite material is used in the same way as bone grafts. Quarto et al. [69] first reported the use of cultured BM MSCs combined intra-operatively with hydroxyapatite blocks to fill large bone defects (4–7 cm). They successfully treated 3 patients, with defects in the tibia, humerus, and ulna, respectively. A subsequent study confirmed healing of the defects after 6–7 years [70]. The expanded MSCs may also be injected alone percutaneously in the site of fracture or osteotomy with interesting results in two studies [71] and [72]. Tissue engineering combines bone marrow cells or mesenchymal stem cells, synthetic scaffolds and molecular signals (growth or differentiating factors) in order to form hybrid constructs.

Written informed consent was provided by all

subjects Th

Written informed consent was provided by all

subjects. The trial was designed, implemented, and overseen by the LDK378 cost PACTTE Steering Committee. An independent DSMB reviewed the safety data and study progress on an ongoing basis. Outpatient men and women with the following criteria were eligible to enroll: age ≥ 65 years with a hemoglobin concentration of ≥ 9 g/dL and < 11.5 g/dL for women or < 12.7 g/dL for men with unexplained anemia; serum ferritin between 20 and 200 ng/mL (inclusive); ability to walk without the use of a walker or motorized device, or the assistance of another person; lack of significant cognitive impairment defined by a Montreal Cognitive Assessment score of 22 or higher; and ability to understand and speak English (Table 1). The protocol initially included subjects with a serum ferritin between 20 and 100 ng/mL (inclusive) but was modified on March 26, 2012, due to poor recruitment to allow serum ferritin levels between 20 and 200 ng/mL (inclusive). The protocol was additionally modified on

August 20, 2012, at sites with Spanish-speaking study staff to include subjects who were able to speak and understand Spanish. Unexplained anemia was defined, similar to published criteria [13] and [14], as not meeting criteria for any known etiology of anemia, including vitamin B12, folate, or iron deficiency (defined as serum ferritin < 20 ng/mL); renal insufficiency (defined as glomerular filtration rate of less than 30 [16] using the four-variable Modification of Diet in Renal Disease equation [17]); thyroid dysfunction; myelodysplastic PCI32765 syndrome; anemia of inflammation; plasma cell

dyscrasia; thalassemia trait; alcohol overuse; any prior history of hematologic malignancy; unexplained splenomegaly or lymphadenopathy; or the presence of any condition reasonably assumed to be causing anemia and not corrected for 3 months (Table 2). Subjects were excluded if they had received a red blood cell transfusion, intravenous iron, or an erythropoiesis stimulating agent within 3 months prior to enrollment; had unstable angina, a myocardial infarction, a stroke, or a transient ischemic attack within 3 months prior to enrollment; had uncontrolled hypertension; had a positive fecal occult blood test during the screening period; had significant impairment in liver else function; had a documented history of anaphylactic reaction to iron sucrose infusion; had recently initiated oral iron supplementation; or if the distance walked on the 6-minute walk test (6MWT) was above the median for age and sex, to avoid a ceiling effect (Table 1; Appendix A). Subjects were randomized to start IVIS either immediately (immediate intervention group) or after a 12-week wait list period (wait list control group) at a 1:1 ratio via an interactive voice and web response system. The randomization sequence was computer-generated with random block sizes. Neither subjects nor investigators were blinded.


H&E slides were reviewed


H&E slides were reviewed GSK2118436 chemical structure to confirm the diagnosis. The tissues removed were classified as cysts whenever a partial or total epithelium lining was present. The diagnosis of cysts was based mainly on radiographic and histopathologic examination. DC intensely inflamed and cysts with inadequate tissue samples were excluded. A total of 40 cysts were selected for the study (20 RC and 20 DC). Clinical and radiographic information, including age, gender, and anatomic site, were obtained from biopsy forms submitted by the clinicians. For immunohistochemical analysis, 3 μm thick paraffin embedded tissue sections were placed on 3-aminopropyltriethoxy-silane coated glass slides (Sigma Chemical Co., St Louis, MO, USA). The samples were deparaffinised

with xylene, rehydrated in graded alcohols, and washed in deionised water and phosphate-buffered saline (PBS). Samples were then incubated with 3% hydrogen peroxide and immersed in a citrate buffer, pH 6.0 for 20 min. Sections were then blocked by incubation with 3% normal goat serum at room temperature for 20 min, and slides were incubated at 4 °C, overnight, in a humidified chamber with the following primary rabbit polyclonal antibodies: anti-OPG (N-20; Santa Cruz Biotechnology, Santa Cruz, CA) diluted 1:200; anti-RANK (C-20; Santa Cruz Biotechnology, Santa Cruz, CA) diluted 1:200; and anti-RANKL (N-19; Santa Cruz Biotechnology, Santa Cruz, CA) diluted 1:200. After washing FG-4592 molecular weight in TBS (tris-buffered saline), the sections were treated with a labelled streptavidin-biotin kit (LSAB; Dako, Glostrup, Denmark). Peroxidase activity was visualised by immersing tissue sections in 3,3′-diaminobenzidine (D5637; Sigma Chemical, St. Louis, MO) and counterstained with Mayer’s haematoxylin. A central giant cell granuloma was used as positive control.9 Negative controls were obtained by the omission of primary antibodies and substitution of primary antibodies by nonimmune rabbit serum (X0902; Dako). Immunoexpression of RANK, RANKL and

OPG was evaluated in lining epithelium and fibrous capsule. The epithelial immunoexpression was semiquantitatively evaluated by Baf-A1 molecular weight two observers, using 400× magnification and classified according to the scores: 0 or no staining (<10% of positive immunostaining cells), 1 or weak (11–25%), 2 or moderate (26–75%) and 3 or strong (>76%).23 In fibrous capsule, the analysis was quantitative and the number of positive cells was counted in 10 representative and consecutive microscopic high-power fields (1000×) over totally counted cells,12 irrespectively of cell type. Digital images were loaded on the software IMAGE J® (National Institutes of Health, Bethesda, Maryland, USA) 24 to count the number of immunostained cells. Results are expressed as the mean percentage of observations per field, with the following modifications.