SOX2 and BCL-2 Expressions in Odontogenic Keratocyst and Ameloblastoma.

BACKGROUND: The purpose of this experimental study was to compare the immunohistochemical expression of SOX2 and BCL-2 in Odontogenic Keratocyst (OKC) and Ameloblastoma (AB) specimens, and to identify a possible correlation in their expression. MATERIAL AND METHODS: Immunohistochemical analysis was performed to evaluate SOX2 and BCL-2 expression in OKC (n = 20) and AB (n = 20). The immunoexpression was analyzed by a quantitative and qualitative scoring system. The comparison between the immunoexpression of SOX 2 and BCL-2 was assessed by the Mann-Whitney U-test. Spearman's correlation coefficient evaluated the correlation between SOX2 and BCL-2 expressions. RESULTS: SOX2 and BCL-2 expression was observed in all specimens of OKC in the full thickness of the epithelium lining. SOX2 immunostaining was higher in OKC, in comparison with AB samples (P<0.05). BCL-2 immunostaining between OKC and AB was not statistically significant. There was no significant correlation between SOX2 and BCL-2 in OKC and AB specimens. CONCLUSIONS: SOX2 and BCL-2 expressions in OKC may suggest their relationship with the biological behavior of this lesion, and the higher expression of SOX2 might be an upstream influence on the Hh signaling pathway.

Oral Lesions in Dogs with Visceral Leishmaniosis.

Canine visceral leishmaniosis (CVL) is an important zoonotic disease, which is endemic in Brazil and several other parts of the world. The disease may affect multiple organs, but lesions in the oral cavity are considered uncommon. Twenty-three cases of oral lesions compatible with CVL were diagnosed from 2015 to 2018 in the Federal District, Brazil. Six cases were confirmed to be CVL based on histopathology, immunohistochemistry and polymerase chain reaction amplification of Leishmania infantum kDNA. Most of the affected dogs were >9 years of age, of mixed breed and were classified as having stage C of the disease. The most important gross findings were ulcerated nodular lesions in the tongue, lip, gingiva and hard palate. Microscopically, there was lymphoplasmacytic to granulomatous inflammation that was diffuse, moderate to severe and associated with ulceration. CVL should be considered in the differential diagnosis of nodular and ulcerative oral lesions in dogs.

Effect of thermally deposited siloxane-methacrylate coating on bonding to glass fibre posts.

AIM: To evaluate the alterations promoted by a thermally deposited siloxane-methacrylate coating on the surface of glass fibre posts and their effect on the bond strength of resin-core materials to the posts. METHODOLOGY: Fibre post surfaces were treated with experimental thermally deposited siloxane-methacrylate coatings or clinically available treatments (i.e. hydrogen peroxide and methylene chloride); nontreated posts were used as controls. The contact angles formed between the post surface and the water/adhesive were measured with a tensiometer. Scanning electron microscopy and electron dispersive spectroscopy were used to examine the topographies and chemical changes in the post surfaces following treatment. Surface roughness was evaluated with laser interferometry. Core resin was bonded to the fibre posts, and microtensile bond strength testing was subsequently performed. The data were individually submitted to anova and Tukey's tests (α = 0.05). RESULTS: The water contact angle was reduced significantly (P < 0.05) by the thermally deposited siloxane-methacrylate coating. All treatments significantly increased the adhesive contact angle (P ≤ 0.016) compared to the control as well as the surface roughness (P ≤ 0.006) and the amount of Si on post surfaces. Greater percentages of Si were observed for the thermally deposited coating. The bond strength to the posts was significantly improved by the thermally deposited coating (P < 0.05), whereas the other treatments did not differ from the control. CONCLUSION: Treating the surface of glass fibre posts with a thermally deposited siloxane-methacrylate coating improved the bond strength to resin-based materials. The coating could be performed by manufacturers of glass fibre posts in order to reduce the number of clinical steps required for luting posts into root canals.

Juçara pulp supplementation improves glucose tolerance in mice.

BACKGROUND: The consumption of hyperlipidic and hypercaloric diet is considered a major factor to promote obesity and the consumption of food with antioxidant properties, like Juçara (Euterpe edulis Mart), could be a tool to prevent the deleterious effect of high white adipose deposition. The aim of the present study was to evaluate the effect of administration of juçara pulp in mice fed a high-fat, high-calorie diet on glucose tolerance and adipose tissue inflammatory status. METHODS: Mice were distributed into the following groups: control diet; control diet plus 0.5 % of juçara; control diet plus 2 % of juçara; hypercaloric and hyperlipidic diet; hypercaloric and hyperlipidic diet plus 0.5 % of juçara and hypercaloric and hyperlipidic diet plus 2 % of juçara. Treatments started when mice were 8 weeks old and carried on for a total period of 10 weeks. The serum glucose, triacylglycerol, total cholesterol, insulin, adiponectin, lipopolysaccharides and free fatty acids concentrations were measured. Oral glucose tolerance test was performed. TNF-α, IL-6, and IL-10 protein level were determined by ELISA on mesenteric and epididymal white adipose tissues. Determination of catalase activity was realized in the same tissues. Data were analysed using one-way analysis of variance and post hoc analysis was performed with the Tukey's test. RESULTS: The addition of 0.5 % juçara improved glycemic response in animals that consumed normocaloric as well as hypercaloric and hyperlipidic diets (HC). Supplementation with 0.5 and 2 % did not change the body composition of animals that received the HC diet; however, the animals fed the normocaloric diet with 2 % juçara gained body mass. An intake of 2 % juçara in the HC diet promoted a reduction of catalase activity and IL-10 level in epididymal adipose tissue. CONCLUSIONS: These results suggest that with the administration of 0.5 % juçara, the beneficial effects of polyphenols overcome the deleterious effects of macronutrient composition of juçara, whereas with the administration of 2 % juçara promotes damage by the composition of the fruit and overshadows the beneficial effects of polyphenols on glucose metabolism. On the other hand, higher juçara supplementation improves the inflammatory status targeted by the HC diet.

Etching a fiber post surface with high-concentration bleaching agents.

INTRODUCTION: Commonly, resin composites/cements fail to achieve proper bonding to fiber posts when their surfaces have not been previously etched. This study evaluated the effect of the concentration and application mode of hydrogen peroxide on the surface topography and bond strength of resin composite to glass-fiber posts. METHODS AND MATERIALS: Fiber posts were immersed in 24% or 35% solutions (a high-concentration bleaching agent) of hydrogen peroxide (H2O2), or these solutions were applied over the post surface using a microbrush (n=10). Posts without any treatment were used as a control. After etching, the posts were silanated and an adhesive was applied. The posts were positioned in a mold, and a resin composite was incrementally inserted and light-cured. The post/resin assembly was serially sectioned into several beams that were subjected to a tensile bond strength test. The data were subjected to the two-way analysis of variance and Tukey test (α=0.05). The Dunnet's test was used to compare the experimental conditions to the control. The surface topography was analyzed using scanning electronic microscopy. RESULTS: The non-etched post presented a relatively smooth surface without fiber exposure. Except for the application of 24% H2O2, the other experimental conditions increased the number of exposed fibers and bond strength in relation to the control. Although immersion resulted in higher values for the 24% H2O2 application, the mode of application did not alter bond strength when 35% H2O2 was used. CONCLUSIONS: The effect of the mode of application of H2O2 depended on its level of concentration. A high-concentration bleaching agent improved the bond strength of the resin composite to the post surface, regardless of which mode was used.

Numerical aberrations of chromosome 17 and TP53 in brain metastases derived from breast cancer.

Breast cancer is the second most common origin of brain metastases, after lung cancer, and represents 14-20% of all cases. Abnormalities of chromosome 17 are important molecular genetic events in human breast cancer, and several oncogenes and tumor suppressor genes are located on this chromosome. In about half of all human cancers, the tumor suppressor gene TP53, located at 17p13.1, is either lost or mutated. Loss of p53 protein function influences not only cell cycle checkpoint controls and apoptosis, but also the regulation of other important stages of metastatic progression, such as cell migration and tissue invasion. The aim of our study was to identify numerical aberrations of chromosome 17 and TP53 in 5 subjects with brain metastasis from breast cancer using dual-color fluorescence in situ hybridization experiments. Deletion of TP53 was the most frequent alteration observed, suggesting that if this alteration is present in the primary tumors, breast tumors with loss of TP53 copies have a poorer prognosis and a higher chance for metastasis. If this is true, the analyses of chromosome 17 and TP53 in primary breast cancer could be important for predicting its metastatic potential.

On the quaternary structure of a C-type lectin from Bothrops jararacussu venom--BJ-32 (BjcuL).

BJ-32 (also known as BjcuL) is a C-type lectin from the venom of Bothrops jararacussu with specificity for beta-galactosides and a remarkable ability to agglutinate several species of trypanosomatids. Our objective was to study the oligomerization state of native BJ-32 by using different biophysical and computational methods. Small-angle X-ray light scattering (SAXS) experiments disclosed a compact, globular protein with a radius of gyration of 36.72+/-0.04A and molecular weight calculated as 147.5+/-2.0kDa. From analytical ultracentrifugation analysis, it was determined that the BJ-32 sedimentation profile fits nicely to a decamer model. The analysis of the intrinsic emitted fluorescence spectra for BJ-32 solutions indicated that association of subunits in the decamer is accompanied by changes in the environment of Tryptophan residues. Both ab initio and comparative models of BJ-32 supported the resemblance of the decamer in the crystallographic structure from a close homologue, the rattlesnake venom lectin (RSL) from Crotalus atrox.

Influência do tamanho e da rigidez dos objetos nos ajustes proximais e distais do alcance de lactentes / Influence of object size and rigidity on proximal and distal adjustments to infant reaching

CONTEXTUALIZAÇÃO: Estudos têm identificado que as propriedades dos objetos induzem os ajustes no alcance; no entanto, poucos investigaram a influência específica do tamanho e rigidez dos objetos em lactentes jovens. OBJETIVO: Verificar se lactentes de 4 a 6 meses realizam ajustes proximais e distais ao alcançarem objetos de diferentes tamanhos e rigidez. MÉTODOS: Nove lactentes saudáveis foram posicionados em uma cadeira inclinada a 50º. Quatro objetos foram apresentados, um rígido grande (RG), um rígido pequeno (RP), um maleável grande (MG) e um maleável pequeno (MP), por um período de 1 minuto cada. Em um total de 384 alcances, foram analisados os ajustes proximais (alcance uni e bimanual) e distais (orientação da mão horizontal, vertical e oblíqua; mão aberta, fechada e semi-aberta) e o sucesso do alcance dos objetos. RESULTADOS: Constatou-se ajuste bimanual para o objeto RG e unimanual para os demais. A orientação da mão oblíqua foi predominante no toque dos objetos, enquanto para a preensão dos mesmos, a predominância foi a vertical, principalmente para o objeto RG. A orientação horizontal não foi observada na preensão do objeto RG. A mão semi-aberta foi mais freqüente no início do alcance para todos os objetos, enquanto no toque do objeto RG a mão aberta foi predominante. O sucesso do alcance foi maior para os objetos maleáveis (MG, MP) do que para os rígidos (RG e RP). CONCLUSÃO: Lactentes jovens estudados são capazes de planejar e ajustar seus movimentos baseados na percepção das propriedades físicas dos objetos, o que sugere interação percepção-ação.

BACKGROUND: Studies have identified that object properties lead to adjustments to reaching. However, few have investigated the specific influence of object size and rigidity among young infants. OBJECTIVE: To verify whether four to six-month-old infants make proximal and distal adjustments when reaching for objects of different sizes and rigidity. METHOD: Nine healthy infants were seated on a chair inclined at 50º. Four objects were presented to them: one large rigid (LR), one small rigid (SR), one large malleable (LM) and one small malleable object (SM), each for a one-minute period. A total of 384 reaches were analyzed to verify proximal adjustments (single-hand and two-hand reaching) and distal adjustments (horizontal, vertical and oblique hand orientation; opened, closed and half-open hand), and the success in reaching the objects. RESULTS: The infants exhibited two-hand adjustments for the LR object and single-hand adjustments for the other objects. Oblique orientation was predominant for touching the objects, while vertical orientation was predominant for grasping them, particularly the LR object. Horizontal orientation was not observed for grasping the LR object. At the start of reaching for all objects, the hands were most frequently half-open, while the hands were predominantly open when touching the LR object. Success was greater when reaching for malleable objects (LM, SM) than for rigid objects (LR and SR). CONCLUSION: The young infants studied were capable of planning and adjusting their movements on the basis of their perceptions of the physical properties of the objects, which suggests that perception-action interaction was occurring.

Simple affinity chromatographic procedure to purify beta-galactoside binding lectins.

Affinity chromatography based on the commercial resin Sepharose CL-6B was used to isolate new C1-beta-type lectins from crude preparations of snake venoms (Bothrops jararaca, Bothrops jararacussu, Bothrops newiedi, Bothrops moojeni, Lachesis muta rhombeata). Most of the C-type lectins could be eluted with almost 100% recovery using the competitor isopropyl-beta-D-thiogalactoside (IPTG) or through Ca2+ sequestration with EDTA. The lectin yield varied considerably among the different snake species, but B. newiedi venom was a particularly rich source of lectin, retaining 2.7 mg of lectin by milliliter of resin in saturating conditions. C1-alpha-lectins from Crotalus durisus terrificus venom, from the jack fruit (jacalin) and from bread fruit seeds extract (frutalin) had no affinity, either with or without Ca2+ added, for Sepharose CL-6B, showing that the resin is specific for C1-beta-type lectins. Sepharose CL-6B used as galactose-affinity chromatography provides a simple and fast method for isolating C-type beta-galactoside binding lectins from crude sample preparations.

TIPUH1 encodes a novel KRAB zinc-finger protein highly expressed in human hepatocellular carcinomas.

To achieve a better understanding of mechanisms that underlie hepatocarcinogenesis and to identify novel target molecules for diagnosis and therapy of hepatocellular carcinoma (HCC), we previously analysed gene-expression profiles of 20 HCC tissues on a cDNA microarray. Among the genes upregulated in the tumor tissues compared with their nontumor counterparts, we focused on a novel gene termed transcription-involved protein upregulated in HCC (TIPUH1) that putatively encoded a 500-amino-acid protein containing 12 zinc-finger domains and a Kruppel-associated box domain. Multiple-tissue northern blot analysis revealed it's testis- and placenta-specific expression in normal tissues. Colony-formation assay in soft agar showed that TIPUH1 conferred anchorage-independent growth to NIH3T3 cells, suggesting its oncogenic activity. Conversely, specific siRNA for TIPUH1 knocked down its expression in HCC cells, which resulted in their growth inhibition. We identified four TIPUH1-interacting proteins including TIF1beta, a transcription-intermediary protein, and three involved in pre-mRNA processing (hnRNPU, hnRNPF, and Nucleolin), suggesting that overexpressed TIPUH1 may play a role in hepatocarcinogenesis by regulating transcription and/or RNA processing of growth control genes. These data may contribute to a better understanding of liver neoplasia, and to the development of novel strategy for treatment of HCCs.

Biochemical and molecular modeling analysis of the ability of two p-aminobenzamidine-based sorbents to selectively purify serine proteases (fibrinogenases) from snake venoms.

Snake venoms contain several trypsin-like enzymes with equivalent physicochemical characteristics and similar inhibition profiles. These are rather difficult to separate by classical purification procedures and therefore constitute a good model for affinity chromatography analysis. Some of these trypsin homologues present fibrinogenase activity, mimicking one or more features of the central mammalian coagulation enzyme, thrombin. It was previously demonstrated that a number of amidine derivatives are able to interact specifically with some of these serine proteases. To understand the enzyme-sorbent interactions we have investigated the ability of two commercially available benzamidine affinity matrices to purify thrombin-like serine proteases (TLSP) with similar biological properties from two snake venoms (Bothrops jararacussu and Lachesis muta rhombeata). Curiously, each sorbent retained a single but distinct TLSP from each venom with high yield. Molecular modeling analysis suggested that hydrophobic interactions within a specific region on the surface of these enzymes could be generated to explain this exquisite specificity. In addition, it was demonstrated that a specific tandem alignment of the two benzamidine sorbents enables the purification of three other enzymes from B. jararacussu venom.

Anti-herpes simplex virus effect of a seed extract from the tropical plant Licania tomentosa (Benth.) Fritsch (Chrysobalanaceae).

Incubation of acyclovir-resistant herpes simplex virus type 1 (ACVr-HSV1), during infection of the HEp-2 cell culture, with an extract prepared from the seeds of Licania tomentosa (Benth.) Fritsch (Chrysobalanaceae) species impaired the productive replication of this virus in a concentration-dependent manner. The extract was able to inhibit extracellular virus (virucidal effect) and also interfered with a very early event of cell infection, at a non-cytotoxic concentration.

Exploring the subsite specificity of Schistosoma mansoni aspartyl hemoglobinase through comparative molecular modelling.

Blood flukes of the genus Schistosoma currently infect millions of people in tropical and subtropical countries. An enzyme playing a major role in hemoglobin (Hb) degradation by Schistosoma mansoni has been cloned and shown to be highly similar to the human cathepsin D aspartyl proteinase, although presenting a distinct substrate specificity from the latter. Investigating the structural features responsible for this difference has a major application in the design of selective anti-schistosomal drugs. In order to achieve this goal a homology model for the S. mansoni aspartyl hemoglobinase was constructed and then used to simulate the complexes formed with two transition state analogues of Hb-derived octapeptide substrates. Comparison with human cathepsin D showed that different pocket volumes and surface electrostatic potentials arise from substitutions in residues comprising the S4, S3, S2 and S3' subsites. Since the primary specificity of the S. mansoni enzyme resembles that of HIV-1 protease, we have discussed the applicability of current retroviral protease inhibitors as leads for the design of new anti-schistosomal drugs.

Paraventricular oxytocin neurons are involved in neural modulation of breathing.

In this study, we determined the projections of oxytocin-containing neurons of the paraventricular nucleus (PVN) to phrenic nuclei and to the rostral ventrolateral medullary (RVLM) region, which is known to be involved in respiratory rhythm generation. Studies were also designed to determine oxytocin-receptor expression within the RVLM and the physiological effects of their activation on respiratory drive and arterial blood pressure. Oxytocin immunohistochemistry combined with cholera toxin B, a retrograde tracer, showed that a subpopulation of oxytocin-containing parvocellular neurons in the dorsal and medial ventral regions of the PVN projects to phrenic nuclei. Similarly, a subpopulation of pseudorabies virus-labeled neurons in the PVN coexpressed oxytocin after injection of pseudorabies virus, a transynaptic retrograde marker, into the costal region of the diaphragm. A subpopulation of oxytocin expressing neurons was also found to project to the RVLM. Activation of this site by microinjection of oxytocin into the RVLM (0.2 nmol/200 nl) significantly increased diaphragm electromyographic activity and frequency discharge (P < 0.05). In addition, oxytocin increased blood pressure and heart rate (P < 0.05). These data indicate that oxytocin participates in the regulation of respiratory and cardiovascular activity, partly via projections to the RVLM and phrenic nuclei.

Acute renal failure due to abdominal compartment syndrome: report on four cases and literature review.

We report on 4 cases of abdominal compartment syndrome complicated by acute renal failure that were promptly reversed by different abdominal decompression methods. Case 1: A 57-year-old obese woman in the post-operative period after giant incisional hernia correction with an intra-abdominal pressure of 24 mm Hg. She was sedated and curarized, and the intra-abdominal pressure fell to 15 mm Hg. Case 2: A 73-year-old woman with acute inflammatory abdomen was undergoing exploratory laparotomy when a hypertensive pneumoperitoneum was noticed. During the surgery, enhancement of urinary output was observed. Case 3: An 18-year-old man who underwent hepatectomy and developed coagulopathy and hepatic bleeding that required abdominal packing, developed oliguria with a transvesical intra-abdominal pressure of 22 mm Hg. During reoperation, the compresses were removed with a prompt improvement in urinary flow. Case 4: A 46-year-old man with hepatic cirrhosis was admitted after incisional hernia repair with intra-abdominal pressure of 16 mm Hg. After paracentesis, the intra-abdominal pressure fell to 11 mm Hg.

Differential cardiorespiratory control elicited by activation of ventral medullary sites in mice.

We studied the respiratory and blood pressure responses to chemical stimulation of two regions of the ventral brainstem in mice: the rostral and caudal ventrolateral medulla (RVLM and CVLM, respectively). Stimulation of the RVLM by microinjections of the excitatory amino acid L-glutamate induced increases in diaphragm activity and breathing frequency, elevation of blood pressure (BP), and a slight increase in heart rate (HR). However, activation of the CVLM induced a decrease in breathing frequency, mainly due to prolongation of expiratory time (TE), and hypotension associated with a slight slowing of HR. Because adrenergic mechanisms are known to participate in the control of respiratory timing, we examined the role of alpha(2)-adrenergic receptors in the RVLM region in mediating these inhibitory effects. The findings demonstrated that blockade of the alpha(2)-adrenergic receptors within the RVLM by prior microinjection of SKF-86466 (an alpha(2)-adrenergic receptor blocker) significantly reduced changes in TE induced by CVLM stimulation but had little effect on BP responses. These results indicate that, in mice, activation of the RVLM increases respiratory drive associated with an elevation of BP, but stimulation of CVLM induces prolongation of TE via an alpha(2)-adrenergic signal transduction pathway.

alpha(2)-adrenergic receptors are not required for central anti-hypertensive action of moxonidine in mice.

In the mouse medulla oblongata, we characterized binding properties and functional responses of two recognition sites for imidazoline compounds: I(1)-imidazoline and alpha(2)-adrenergic receptors. The mouse medulla expresses a higher density of I(1)-receptors than in the rat, whereas alpha(2)-receptor densities were similar between the two species. In anesthetized, ventilated and paralyzed mice, we tested the hypotensive actions of the I(1)/alpha(2) agonist moxonidine, determined its central site of its actions, and the relative roles of I(1) and alpha(2)-receptors. Experiments were performed in C(57)Bl(6) wild type and alpha(2A)-adrenergic receptor deficient mice. In both types of mice, neuronal activation within the rostral ventrolateral medulla (RVLM) region by glutamate microinjection elicited increases in arterial pressure. Moxonidine (0.5 nmol/site/10 nl) microinjected bilaterally into this vasopressor region decreased arterial pressure by 30% and heart rate by 11% in wild type mice. Efaroxan, the I(1)/alpha(2) antagonist (0.4 nmol) when microinjected into the RVLM elevated blood pressure itself and abolished the action of moxonidine, whereas alpha(2)-blockade with SK&F 86466 had no significant effect on blood pressure and did not attenuate moxonidine's effect. To more definitively test the role of alpha(2)-adrenergic receptors in the action of moxonidine, moxonidine was microinjected into the RVLM of alpha(2A)-adrenergic deficient mice. The decreases in arterial pressure were nearly identical to those of wild type mice, whereas bradycardia was attenuated. Thus, in the mouse moxonidine acts within the RVLM region to lower arterial pressure mainly through the I(1)-imidazoline receptor independent of alpha(2)-adrenergic receptors.

Respiratory effects of kynurenic acid microinjected into the ventromedullary surface of the rat

Several studies demonstrate that, within the ventral medullary surface (VMS), excitatory amino acids are necessary components of the neural circuits involved in the tonic and reflex control of respiration and circulation. In the present study we investigated the cardiorespiratory effects of unilateral microinjections of the broad spectrum glutamate antagonist kynurenic acid (2 nmol/200 nl) along the VMS of urethane-anesthetized rats. Within the VMS only one region was responsive to this drug. This area includes most of the intermediate respiratory area, partially overlapping the rostral ventrolateral medulla (IA/RVL). When microinjected into the IA/RVL, kynurenic acid produced a respiratory depression, without changes in mean arterial pressure or heart rate. The respiratory depression observed was characterized by a decrease in ventilation, tidal volume and mean inspiratory flow and an increase in respiratory frequency. Therefore, the observed respiratory depression was entirely due to a reduction in the inspiratory drive. Microinjections of vehicle (200 nl of saline) into this area produced no significant changes in breathing pattern, blood pressure or heart rate. Respiratory depression in response to the blockade of glutamatergic receptors inside the rostral VMS suggests that neurons at this site have an endogenous glutamatergic input controlling the respiratory cycle duration and the inspiratory drive transmission.