Role of collagen and immunostaining for TGF-ß in the clinical and microscopic findings of pyogenic granuloma and peripheral ossifying fibroma.

BACKGROUND: Collagen is a component of Pyogenic Granuloma (PG) and Peripheral Ossifying Fibroma (POF) and performs different functions in these lesions. The objective of this study is to evaluate the role of collagen and immunostaining for Transforming Growth Factor beta (TGF-ß) in the clinical and microscopic findings of PG and POF. MATERIAL AND METHODS: PG (n=20) and POF (n=20) were selected for clinical evaluation (sex, age, localization, size and evolution time) and microscopic analysis (picrosirius red staining for collagen analysis and immunohistochemistry for TGF-ß) performed in the superficial and deep areas of the two lesions. ANOVA/Bonferroni and t-test, Pearson correlation and χ2 were used to compare the sites and parameters analyzed (p<0.05, GraphPad Prism 5.0). RESULTS: The depth of PG presented the highest amount of collagen (p<0.001), and its surface showed the lowest amount of type 1 collagen (yellow-red strong birefringence). Type 1 collagen gradually increased in depth of PG, surface and depth of POF (p<0.001). The number of TGF-ß+ cells was lower on the surface of PG compared with the depth of PG and the two areas of POF (p<0.001). Sex and localization did not affect these parameters, but the profile of collagen and immunostaining for TGF-ß suffered from modifications by the time of evolution and the size of the lesion. CONCLUSIONS: Although PG and POF are reactive gingival lesions, the expression of TGF-ß and its role in collagen showed different biological behaviors in these lesions, suggesting different biological origins for its components.

Identification of reference genes for studies of quantitative gene expression in male and female quail tissues.

In industrial poultry, quail production has gained increasing prominence over the years. It is known that the intensification of genetic studies has contributed greatly to this growth, through techniques, such as analysis of gene expression by PCR, for example. This study aimed to evaluate stability and recommend reference genes for quantitative real-time PCR in different tissues from male and female broiler quails. The stability of 10 housekeeping genes (GAPDH, RPL5, MRPS27, MRPS30, TFRC, HMBS, EEF1, LDHA, B2M, and UBC) by means Bestkeeper, NormFinder, GeNorm softwares with ΔCq method. The tissues analyzed were: heart, thigh muscle, brain, and spleen, considering that they are tissues commonly used in nutrigenomic, immunological, and poultry performance research. As expected, the reference genes tested showed varying stability depending on the tissue evaluated. According to the present study, the most stable housekeeping genes were MRPS30, TFRC, and HMBS in heart; MRPS30, EEF1, and HMBS in thigh muscle; B2M, GAPDH, and UBC in brain; and EEF1, LDHA, and HMBS in spleen. Therefore, it is recommended to be used as reference genes for gene expression studies of male and female quails.

Key elements in hypersensitivity reactions to chemotherapy: experience with rapid drug desensitization in gynaecological cancer in a Tertiary Hospital.

Summary: Rapid drug desensitization (RDD) is a procedure performed when no alternative drug is considered equally effective. The aim of our study is to describe the experience with RDD to cytostatics in patients being treated for gynaecological cancer in a tertiary hospital, over a period of 5 years. In this paper, we review 22 cases and 107 episodes of RDD; 86.3% of patients had advanced disease and the mortality rate at the time of data collection was 50.0%. RDD was performed on 81.8% patients for platinum, 13.6% for taxanes, and 4.5% for anthracyclines. The reintroduction of antineoplastic drugs in all patients with a previous history of immediate hypersensitivity reaction demonstrated the safety and efficacy of this procedure. There was serious complication (anaphylaxis) in only one case.

Is immunohistochemistry more sensitive than hematoxylin-eosin staining for identifying perineural or lymphovascular invasion in oral squamous cell carcinoma? A systematic review and meta-analysis.

BACKGROUND: This study aimed to analyze whether immunohistochemistry (IHC) is more sensitive than hematoxylin-eosin (H&E) staining for identifying perineural invasion (PNI) or lymphovascular invasion (LVI) in oral squamous cell carcinoma (OSCC). MATERIAL AND METHODS: In this systematic review and meta-analysis (Prospective Register of Systematic Reviews - CRD 42021256515), data were obtained from six databases (PubMed, Scopus, LILACS, Web of Science, EBSCO, LIVIVO, Embase) and the grey literature. Cross-sectional observational studies of the diagnostic sensitivity of IHC for PNI and LVI were included. Studies were selected in two phases: first collection and reference retrieval. The Quality Assessment of Diagnostic Accuracy Studies-2 tool assessed study quality, while the Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) approach assessed evidence quality. The meta-analysis (random effects model) was performed using MedCalc 18.2.1 software (MedCalc®) (p<0.05). RESULTS: Four studies (560 patients with 295 biopsies) were analyzed. The combined sensitivity was 76% (95% confidence interval [CI], 44.30-97.19%) and specificity was 42% (95% CI, 23.40-62.02%). The positive predictive value (PPV) and negative predictive value (NPV) were 61% (95% CI, 49.78-71.53%) and 70% (95% CI, 37.63-94.43%). The overall accuracy was 58% (95% CI, 45.17-70.65%). The risk of bias was low, and GRADE analysis showed a very low certainty of evidence. CONCLUSIONS: Our data suggest that IHC staining to highlight PNI/LVI may be useful in cases in which H&E analysis results in a negative decrease in the prevalence of false-negative cases and underestimated treatment.

Socioeconomic inequalities and temporomandibular joint disorders in adolescents: contributions from a Maranhão cohort.

AIM: To assess the prevalence of temporomandibular disorder (TMD) in adolescents and estimate possible associations with poverty. BASIC RESEARCH DESIGN: A cross-sectional study nested within a prospective birth cohort study conducted in São Luís, Maranhão, Brazil. PARTICIPANTS: 2,412 adolescents aged 18-19 years. MATERIAL AND METHODS: The presence of TMD, classified according to the Fonseca Anamnestic Index, was used as the outcome. The following explanatory variables were assessed: gender, household head, paved/asphalted street, piped water, and socioeconomic background, based on the Brazilian Association of Market Research criteria and the poverty income ratio (PIR). Logistic regression analysis was performed with the estimation of odds ratios (OR) and 95% confidence intervals. RESULTS: TMD was common (51.4%) and was associated with poverty, as it was more frequent among adolescents from social classes D-E (OR=2.60; 95% CI: 1.48-4.55) and C (OR=1.82; 95% CI: 1.12-2.99) compared to A/B, and among poor adolescents using the PIR (OR=1.50; 95% CI: 1.02-2.33). CONCLUSIONS: The prevalence of TMD in socioeconomically disadvantaged adolescents in São Luís is high, and these data allow the early identification of at-risk groups. We recommend carrying out other population-based studies, using diagnostic strategies with greater accuracy.

PMS2: a potential prognostic protein marker in oral squamous cell carcinoma.

BACKGROUND: An increase in oral squamous cell carcinoma (OSCC) cases was observed despite the reduction in exposure to classic risk factors. Although the exact cause of this trend remains unknown, epigenetic factors could be contributing to an increased occurrence of these tumors. This study aims to assess the influence of PMS2 protein immunoexpression on the prognosis of patients with OSCC. MATERIAL AND METHODS: This study comprised 76 cases of OSCC treated between 2011 and 2016. Immunohistochemical staining for PMS2 was performed. For evaluation, 10 fields per histological section were photographed at a 400x magnification and positively-stained cells were counted with Image J. Mann-Whitney and Kruskal-Wallis tests were used to compare the immunolabeling pattern with the clinical-pathological and prognostic characteristics. Survival analysis was performed with Chi-square, Long-Rank Mantel-Cox and Cox regression tests (p<0.05). RESULTS: An overexpression of PMS2 was observed in N0/1 tumors and in oral cancers found in unusual locations. In patients ≤60 years of age, high levels of PMS2 (>60%; p=0.041) were associated with low survival (p=0.029). In multivariate analysis, surgery combined with chemotherapy (p=0.030) and high PMS2 immunoexpression (p=0.042) significantly increased the risk of death for ≤60 years old patients. CONCLUSIONS: The findings of this study indicate that PMS2 can be a potential prognostic protein marker in OSCC patients 60 years of age and younger.

Accelerometer-based prediction of skeletal mechanical loading during walking in normal weight to severely obese subjects.

There is no objective way to monitor mechanical loading characteristics during exercise for bone health improvement. We developed accelerometry-based equations to predict ground reaction force (GRF) and loading rate (LR) in normal weight to severely obese subjects. Equations developed had a high and moderate accuracy for GRF and LR prediction, respectively, thereby representing an accessible way to determine mechanical loading characteristics in clinical settings. INTRODUCTION: There is no way to objectively prescribe and monitor exercise for bone health improvement in obese patients based on mechanical loading characteristics. We aimed to develop accelerometry-based equations to predict peak ground reaction forces (pGRFs) and peak loading rate (pLR) on normal weight to severely obese subjects. METHODS: Sixty-four subjects (45 females; 84.6 ± 21.7 kg) walked at different speeds (2-6 km·h-1) on a force plate-equipped treadmill while wearing accelerometers at lower back and hip. Regression equations were developed to predict pGRF and pLR from accelerometry data. Leave-one-out cross-validation was used to calculate prediction accuracy and Bland-Altman plots. Actual and predicted values at different speeds were compared by repeated measures ANOVA. RESULTS: Body mass and peak acceleration were included for pGRF prediction and body mass and peak acceleration transient rate for pLR prediction. All pGRF equation coefficients of determination were above 0.89, a good agreement between actual and predicted pGRFs, with a mean absolute percent error (MAPE) below 6.7%. No significant differences were observed between actual and predicted pGRFs at each walking speed. Accuracy indices from our equations were better than previously developed equations for normal weight subjects, namely a MAPE approximately 3 times smaller. All pLR prediction equations presented a lower accuracy compared to those developed to predict pGRF. CONCLUSION: Walking pGRF and pLR in normal weight to severely obese subjects can be predicted with moderate to high accuracy by accelerometry-based equations, representing an easy and accessible way to determine mechanical loading characteristics in clinical settings.

Developmental programming: intrauterine caloric restriction promotes upregulation of mitochondrial sirtuin with mild effects on oxidative parameters in the ovaries and testes of offspring.

According to the developmental origins of health and disease (DOHaD) hypothesis, changes in the maternal environment are known to reprogram the metabolic response of offspring. Known for its redox modulation, caloric restriction extends the lifespan of some species, which contributes to diminished cellular damage. Little is known about the effects of gestational caloric restriction, in terms of antioxidant parameters and molecular mechanisms of action, on the reproductive organs of offspring. This study assessed the effects of moderate (20%) caloric restriction on redox status parameters, molecular expression of sirtuin (SIRT) 1 and SIRT3 and histopathological markers in the ovaries and testes of adult rats that were subjected to gestational caloric restriction. Although enzyme activity was increased, ovaries from female pups contained high levels of oxidants, whereas testes from male pups had decreased antioxidant enzyme defences, as evidenced by diminished glyoxalase I activity and reduced glutathione content. Expression of SIRT3, a deacetylase enzyme related to cellular bioenergetics, was increased in both ovaries and testes. Previous studies have suggested that, in ovaries, diminished antioxidant metabolism can lead to premature ovarian failure. Unfortunately, there is little information regarding the redox profile in the testis. This study is the first to assess the redox network in both ovaries and testes, suggesting that, although intrauterine caloric restriction improves molecular mechanisms, it has a negative effect on the antioxidant network and redox status of reproductive organs of young adult rats.

Shaping ability of XP-endo Shaper and ProTaper Next in long oval-shaped canals: a micro-computed tomography study.

AIM: To evaluate the shaping ability of the single-file XP-endo Shaper system (XP-S; FKG Dentaire, La Chaux-de-Fonds, Switzerland) employing a different working time and of the multiple-file ProTaper Next system (Dentsply Sirona, Ballaigues, Switzerland) using micro-computed tomography (micro-CT) technology. METHODOLOGY: Twenty long oval-shaped canals in mandibular incisors were matched anatomically and scanned by micro-CT (Skyscan 1172; Bruker micro-CT, Kontich, Belgium). The canals were divided into two groups (n = 10) according to the canal preparation protocol: XP-endo Shaper (XP-S) with an extra 45 s of instrumentation and ProTaper Next (PTN X4). The images recorded before and after preparation were evaluated for morphometric measures of volume, surface area, structure model index and untouched walls. The data were compared statistically (Student's t-test for homogenous variances and Mann-Whitney test) between the two groups (XP-S and PTN X4) at α = 5%. RESULTS: Root canal preparation significantly increased all parameters (volume, surface area, structure model index and untouched walls) tested in each group (P < 0.05). There was no significant difference (P > 0.05) in the percentage increase of volume (107.50%-93.13%), surface area (27.74%-29.68%) or untouched canal wall (13.08%-11.74%) between XP-S and PTN X4, respectively. CONCLUSIONS: The XP-endo Shaper system (single-file) with an extra 45 s of instrumentation and the ProTaper Next system (multiple files) had a similar root canal shaping ability. Neither technique was able to fully prepare the long oval-shaped canals of mandibular incisors.

Clinical-pathological and sociodemographic factors associated with the distant metastasis and overall survival of oral cavity and oropharynx squamous cell carcinoma.

BACKGROUND: The objective of this study was to evaluate the influence of clinical-pathological and sociodemographic factors on the prevalence of distant metastasis (DM) and overall survival in patients with oral cavity and oropharynx squamous cell carcinoma (OOSCC). MATERIAL AND METHODS: Cross-sectional study based on the records of 404 OOSCC patients evaluated for DM, covering the period 2000-2014. We analysed the influence of age, sex, level of schooling, primary tumor subsite, treatment, marital status, family history of cancer, history of smoking and alcohol consumption, type of health care coverage (private vs. public) and overall survival. Findings were submitted to Fisher's exact test, Pearson's chi-squared test, Mantel-Cox log-rank testing and multinomial and Cox regression analysis (SPSS v. 20.0; p<0.05). RESULTS: The prevalence of DM was 5.4% (22/404). The respiratory tract was the most affected DM site (n=9; 40.9%). Male sex (p=0.049), oropharyngeal primary tumor (p=0.008), stage T3-4 (p=0.022), lymph node metastasis (N+) (p<0.001) and palliative treatment (p=0.005) were directly associated with DM. Patients with oral primary tumours (p=0.343) and primary oropharyngeal tumours (p=0.242) did not differ significantly with regard to the prevalence of DM. N+ was an independent risk factor for DM (p=0.017). Five variables independently reduced overall survival: male sex (p=0.035), age >65 years (p=0.046), indigenous/brown racial type (p=0.045), palliative treatment (p=0.035) and DM (p=0.048). CONCLUSIONS: Lymph node metastasis independently increased the prevalence of DM and, along with male sex, older age, brown racial type and palliative treatment, was independently associated with poor prognosis in patients with OOSCC.

Using Genomic Location and Coalescent Simulation to Investigate Gene Tree Discordance in Medicago L.

Several well-documented evolutionary processes are known to cause conflict between species-level phylogenies and gene-level phylogenies. Three of the most challenging processes for species tree inference are incomplete lineage sorting, hybridization and gene duplication, which may result in unwarranted comparisons of paralogous genes. Several existing methods have dealt with these processes but none has yet been able to untangle all three at once. Here, we propose a stepwise method by which these processes can be discerned using information on genomic location coupled with coalescent simulations. In the first step, highly discordant genes within genomic blocks (putative paralogs) are identified and excluded from the data set and, in the second step, blocks of linked genes are grouped according to their hybrid history. Existing multispecies coalescent software can then be applied to recover the principal tree(s) that make up the species tree/network without violating the underlying model. The potential of the approach is evaluated on simulated data derived from a species network composed of nine species, of which one is of hybrid origin, and displaying a single-gene duplication that leads to paralogous comparisons. We apply our method to an empirical set of 12 genes from 7 species sampled in the plant genus Medicago that display phylogenetic discordance. We identify the causes of the discordance and demonstrate that the Medicago orbicularis lineage experienced an episode of ancient hybridization. Our results show promise as a new way to explore phylogenetic sequence data that can significantly improve species tree inference in presence of hybridization and undetected paralogy or other causes leading to extremely discordant gene trees. [Coalescent simulation; gene tree; genomic location; hybridization; incomplete lineage sorting; paralogy; phylogenetic incongruence; principal tree; species tree.].

Effects of rumen-undegradable protein on intake, performance, and mammary gland development in prepubertal and pubertal dairy heifers.

The objective of this study was to evaluate the influence of different amounts of rumen-undegradable protein (RUP) on intake, N balance, performance, mammary gland development, carcass traits, and hormonal status of Holstein heifers at different physiological stages (PS). Sixteen prepubertal (PRE) heifers (initial BW = 106 ± 7.6 kg; age = 4.3 ± 0.46 mo) and 16 pubertal (PUB) heifers (initial BW = 224 ± 7.9 kg; age = 12.6 ± 0.45 mo) were used in an experiment over a period of 84 d. Four diets with increasing RUP contents (38, 44, 51, and 57% of dietary crude protein) and heifers at 2 PS (PRE or PUB) were used in a 4 × 2 factorial arrangement of treatments in a completely randomized design. Throughout the experiment, 2 digestibility trials were performed over 5 consecutive days (starting at d 36 and 78) involving feed and ort sampling and spot collections of feces and urine. At d 0 and 83, body ultrasound images were obtained for real-time carcass trait evaluation. The mammary gland was ultrasonically scanned at d 0 and every 3 wk during the experiment. Blood samples were taken at d 0 and 84 to determine serum concentrations of progesterone, estrogen, insulin-like growth factor I (IGF-I), and insulin. No interaction between PS and the level of RUP was found for any trait. Apparent digestibility of dry matter, organic matter, and neutral detergent fiber corrected for ash and protein was not affected by RUP level but was lower for PRE compared with PUB heifers. Sorting against neutral detergent fiber corrected for ash and protein (tendency only) and for crude protein was greater for PUB than PRE heifers. Pubertal heifers had greater average daily gain (905 vs. 505 g/d) and N retention (25.9 vs. 12.5 g/d) than PRE heifers. In addition, average daily gain and N retention were greatest at 51% RUP of dietary protein. Mammary ultrasonography indicated no effects of RUP amounts on mammary gland composition, whereas PRE heifers had greater pixel values than PUB, indicating higher contents of fat rather than protein in the mammary glands of PRE heifers. Serum progesterone and IGF-I concentration was affected only by PS, and PRE heifers had greater values of progesterone and IGF-I concentrations than PUB heifers. Serum insulin concentration was unaffected by PS but tended to be higher at 51% of RUP. In conclusion, an RUP level of 51% increases body weight, average daily gain, feed efficiency, and N retention in heifers regardless of the PS. In addition, PRE heifers have a lower sorting ability and reduced intake, total-tract digestibility, and N retention. They also have higher amounts of fat in their mammary glands, even at moderate growth rates.

One or two ligatures inducing periodontitis are sufficient to cause fatty liver.

BACKGROUND: Periodontitis is a chronic disease that due to an intense inflammatory response triggers systemic changes such as hepatic alterations. This study aimed to compare hepatic damage in rats that received experimental periodontitis at one or two periodontal sites with ligatures. MATERIAL AND METHODS: Eighteen rats were separated into three groups: control, without ligature; periodontitis 1, with one ligature; and periodontitis 2, with two ligatures. The following parameters were assessed: gingival bleeding index, probing pocket depth, tooth mobility, alveolar bone loss, malondialdehyde (MDA) and myeloperoxidase (MPO) activity in periodontal tissue; histopathological evaluation of hepatic tissue (steatosis score); glutathione levels (GSH), MDA, MPO, cholesterol and triglycerides in the liver; and serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). RESULTS: Periodontal evaluation data showed that the periodontitis model worked well. The groups with periodontitis did not differ significantly in relation to MPO activity and MDA levels in the gingival samples, but they were significantly different when compared with the control group. Steatosis was observed in the histological analysis of the groups with periodontitis, but between the periodontitis groups, two ligatures did not cause increase in steatosis score. The levels of GSH, MDA, total cholesterol and triglycerides in the hepatic tissue were not altered between groups with periodontitis, but they showed significant differences in comparison with the control group. The activity of MPO in hepatic tissue and serum levels of AST and ALT did not present significant difference among the three groups. CONCLUSION: In conclusion, our results demonstrated that one or two ligatures inducing periodontitis were both sufficient to cause fatty liver. Steatosis caused by two ligatures did not present larger extension and severity than steatosis caused by one ligature.

A cryptic species produced by autopolyploidy and subsequent introgression involving Medicago prostrata (Fabaceae).

Although hybridisation through genome duplication is well known, hybridisation without genome duplication (homoploid hybrid speciation, HHS) is not. Few well-documented cases have been reported. A possible instance of HHS in Medicago prostrata Jacq. was suggested previously, based on only two genes and one individual. We tested whether this species was formed through HHS by sampling eight nuclear loci and 22 individuals, with additional individuals from related species, using gene capture and Illumina sequencing. Phylogenetic inference and coalescent simulations were performed to infer the causes of gene tree incongruence. We found no evidence that phylogenetic differences among M. prostrata individuals were the result of HHS. Instead, an autopolyploid origin of tetraploids with introgression from tetraploids of the M. sativa complex is likely. We argue that tetraploid M. prostrata individuals constitute a new species, characterised by a partially non-overlapping distribution and distinctive alleles (from the M. sativa complex). No gene flow from tetraploid to diploid M. prostrata is apparent, suggesting partial reproductive isolation. Thus, speciation via autopolyploidy appears to have been reinforced by introgression. This raises the intriguing possibility that introgressed alleles may be responsible for the increased range exploited by tetraploid M. prostrata with respect to that of the diploids.

Polarized Raman and Infrared Spectroscopy and ab Initio Calculation of Palmitic and Stearic Acids in the Bm and C Forms.

A complete experimental study on the vibrational properties of palmitic and stearic acids crystallized in the Bm and C forms, both belonging to the monoclinic system with the P21/a (C2h5) space group, through polarized Raman and infrared spectroscopy, is reported in this paper. Density functional theory calculations were also performed to assign the normal modes and to help in the interpretation of the experimental data. The different polarizations were compared and their influence on the spectral profiles, in both the lattice and the internal mode regions, was discussed. In general, the Raman and infrared spectra exhibit accentuated differences among the polymorphic forms, which are associated with the different molecular modifications, defined as gauche and all-trans conformations. Insights about interaction among different groups are also furnished.

Evaluation of the p-AKT, p-JNK and FoxO3a function in oral epithelial dysplasia.

OBJECTIVES: To evaluate the expression of p-AKT, p-JNK, FoxO3a, and Ki-67 in samples of oral squamous cell carcinoma (OSCC) and oral epithelial dysplasias (OEDs) to understand their possible involvement in the malignant transformation process of oral lesions. MATERIALS AND METHODS: Tissue samples of 20 cases of OSCCs, 20 OEDs, and normal oral mucosa were subjected to immunohistochemistry reactions for anti-p-AKT, anti-p-JNK, anti-FoxO3a, and anti-Ki-67 antibodies. It was analyzed using quantitative (number of immunostained cells) and qualitative (immunostaining intensity) parameters in different cell immunostaining sublocations. RESULTS: Nuclear p-AKT was observed significantly greater immunostaining in OSCC (21.2 ± 19.0) than in dysplasias (7.9 ± 8.1) and controls (1.8 ± 4.7) (P = 0.002). Immunostaining of strong nuclear p-JNK was greater in controls (48.3 ± 13.7) than in OEDs (11.0 ± 10.3) and OSCCs (1.1 ± 1.3) (P < 0.001). Strong nuclear immunostaining of FoxO3a proved to be absent in OSCCs (0.0 ± 0.1) with little staining on dysplasias (3.2 ± 5.4) and increased expression in controls (13.5 ± 4.8) (P < 0.001). Immunostaining of strong nuclear Ki-67 was grater in OSCCs (48.1 ± 49.6) than in OED (11.8 ± 10.6) and controls (1.9 ± 2.0) (P < 0.001). CONCLUSIONS: Malignant process of OEDs in this research may involve the same mechanisms of established malignant lesions.

Immune cell profile of dental pulp tissue treated with zoledronic acid.

AIM: To characterize the pulp immune cell profile in the teeth of rats treated with zoledronic acid (ZA). METHODOLOGY: Male Wistar rats (n = 6 per group) received four intravenous infusions of ZA at doses of 0.04, 0.20 or 1.00 mg kg-1 ZA or saline (control). On the 70th experimental day, they were euthanized. The first right molar was examined microscopically and submitted to toluidine blue reaction and immunohistochemical for CD68, tumour necrosis Factor (TNF)-α, interleukin (IL)-1ß, inducible nitric oxide synthase (iNOS), nuclear factor kappa B (NF-kB) and IL-18 binding protein (IL-18 bp). The presence of ectasic/dilated vessels and inflammatory cells was analysed, and mast cells and mononuclear CD68-positive cells were counted along with the intensity of immunostaining (0-3) for inflammatory markers in odontoblasts and nonodontoblasts pulp cells. The Kruskal-Wallis/Dunn's test (scores or quantitative data) and the chi-squared test (categorical data) were used (GraphPad Prism 5.0, P < 0.05). RESULTS: There was no differences in the number of animals exhibiting dilated/ectasic blood vessels (P = 0.242) and inflammatory cells (P = 0.489) or in the number of mast cells (P = 1.000). However, there was an increase in mononuclear CD68-positive cells (P = 0.026), immunostaining of TNF-α (P = 0.020), IL-1ß (P = 0.027) and iNOS (P = 0.001) in odontoblasts, and IL-1ß (P = 0.013) in nonodontoblast pulp cells dose-dependently. NFkB (nucleus and cytoplasm) and IL-18 bp did not differ between groups. CONCLUSION: ZA modified the immune cell profile in the dental pulp, increasing the number of macrophages and expression of pro-inflammatory markers independent of NFkB.

Reliability of the Three-minute All-out Test for Non-motorized Treadmill Tethered Running.

The 3-min all-out test was developed and validated on a cycle ergometer using a modification of a linear mathematical equation (1/time vs. power) obtained from the original critical power model. The purpose of this development was to obtain, in a single test, the aerobic and anaerobic capacity parameters and identify the exercise transition moment from heavy to severe intensity. The aim of this study was to propose an adaptation of the all-out 3-min cycle ergometer to a non-motorized treadmill with tethered running. In addition, we tested the reproducibility of this adapted protocol, highlighting the need for mechanical power evaluation using a specific ergometer. Consequently, 10 physically active individuals visited the laboratory 4 times for testing and data collection. The results suggested that the protocol adaptation for the 3-min all-out test for non-motorized treadmill with tethered running was reproducible and feasible. It was also possible to show that the AO3 application in this ergometer ensures the specificity of the sports that involve the running exercise, from assessment of both aerobic and anaerobic parameters, accomplished in a single day of application.

Immune cellular profile of bisphosphonate-related osteonecrosis of the jaw.

OBJECTIVES: Characterize the cell profile and immunostaining of proinflammatory markers in an experimental model of bisphosphonate-related osteonecrosis of the jaw (BRONJ). MATERIALS AND METHODS: Male Wistar rats (n = 6-7) were treated chronically with saline solution or zoledronic acid (ZA) at 0.04, 0.20, and 1.00 mg kg(-1) (1.4 × 10(-7) , 6.9 × 10(-6) , and 3.4 × 10(-5)  mol kg(-1) ), and subsequently, the first left inferior molar was extracted. Were performed counting of viable and empty osteocyte lacunae, viable and apoptotic osteoclasts, polymorphonuclear neutrophil, mast cells (toluidine blue), and the positive presence cells for CD68, tumor necrosis factor-alpha (TNF-α), IL (interleukin)-1ß, inducible nitric oxide synthase (iNOS), nuclear factor-kappa B (NF-kB) and IL-18 binding protein (IL-18 bp). RESULTS: BRONJ was showed in ZA treated with 0.20 and 1.00 mg kg(-1) . There is a dose dependent increase in percentage of empty osteocyte lacunae (P < 0.001) and apoptotic osteoclasts (P < 0.001), counting of total osteoclasts (P = 0.003), polymorphonuclear neutrophil cells (P = 0.009), cytoplasmic-positive cells of CD68 (P < 0.001), TNF-α (P = 0.001), IL-1ß (P = 0.001), iNOS (P < 0.001), NF-kB (P = 0.006), and nuclear-positive cells of NF-kB (P = 0.011). Consequently, there is no difference in mast cells (P = 0.957), and IL-18 bp immunostaining decreases dose dependently (P = 0.005). CONCLUSIONS: BRONJ is characterized by increases in immunostaining for proinflammatory markers and NF-kB and inversely associated with cells exhibiting IL-18 bp.

Functional screening for cellulolytic activity in a metagenomic fosmid library of microorganisms associated with coral.

Cellulases are enzymes that degrade cellulosic materials. Cellulose is the most abundant renewable carbon resource on Earth, and cellulases are used in various industrial sectors. Although cellulases are obtained from a variety of sources, this is the first description of cellulolytic activity isolated from a coral metagenomic library. A metagenomic fosmid library of microorganisms associated with the coral Siderastrea stellata, comprising 3552 clones, was screened for cellulolytic activity; this allows access to non-cultivable microorganisms by exploiting the full biotechnological potential. Clones were grown on LB agar plates supplemented with 0.5% carboxymethylcellulose and cellulase positive clones revealed by staining with Congo red. Using this approach, six positive clones with cellulolytic activity were identified. The enzymatic index (EI) of the positive clones was calculated by the ratio between the hydrolysis zone diameter and colony diameter. All positive clones had an EI greater than 1.5. Digestion of the DNA isolated from the six positive clones, using the HindIII restriction endonuclease, revealed different restriction patterns in each clone, indicating that the DNA of each clone is different. There is a growing interest for new cellulolytic enzymes in various industry sectors. Here, we present the initial selection of potential clones for cellulose degradation that could be targets for future studies of enzymatic characterization.