Cramoll 1,4 lectin increases ROS production, calcium levels, and cytokine expression in treated spleen cells of rats

This study reports the in vivo stimulatory effects of Cramoll 1,4 on rat spleen lymphocytes as evidenced by an increase in intracellular reactive oxygen species (ROS) production, Ca(2+) levels, and interleukin (IL)-1 beta expression. Cramoll 1,4 extracted from seeds of the Leguminosae Cratylia mollis Mart., is a lectin with antitumor and lymphocyte mitogenic activities. Animals (Nine-week-old male albino Wistar rats, Rattus norvegicus) were treated with intraperitoneal injection of Cramoll 1,4 (235 mu g ml(-1) single dose) and, 7 days later, spleen lymphocytes were isolated and analyzed for intracellular ROS, cytosolic Ca(2+), and IL-6, IL-10, and IL-1 mRNAs. Cell viability was investigated by annexin V-FITC and 7-amino-actinomycin D staining. The data showed that in lymphocytes activated by Cramoll 1,4 the increase in cytosolic and mitochondrial ROS was related to higher cytosolic Ca(2+) levels. Apoptosis and necrosis were not detected in statistically significant values and thus the lectin effector activities did not induce lymphocyte death. In vivo Cramoll 1,4 treatment led to a significant increase in IL-1 beta but IL-6 and -10 levels did not change. Cramoll 1,4 had modulator activities on spleen lymphocytes and stimulated the Th2 response.

The importance of dietary calcium consumption in two species of semi-terrestrial grapsoid crabs

Calcium (Ca) is essential for crustaceans, due to calcium carbonate (CaCO(3)) deposition in the new exoskeleton to harden it. The purpose of this work was to study short term Ca balance in terms of dietary Ca ingestion in two phylogenetically related crabs (Superfamily Grapsoidea) showing different degrees of terrestrial adaptations: Sesarma rectum Randall, 1840 and Neohelice granulata (Dana, 1851). Dietary Ca ingestion was studied using purified diets with different Ca concentrations (0, 2.2 and 6.66% Ca), together with measurements of Ca excretion and Ca hemolymph levels. The results showed that both crabs had the same response to foods containing different levels of Ca, with both species eating more of the high Ca diet. However, S. rectum consumed more per mg body mass at all Ca concentrations (6 mg.g(-1) for S. rectum against 3 mg.g(-1) for N. granulata). Both species excreted/egested Ca differently: S. rectum excreted Ca proportionally to ingestion, whereas N. granulata maintained constant faecal Ca output at all dietary Ca levels. Moreover, Ca hemolymph levels for crabs fed the different diets were independent of dietary Ca. In conclusion, both S. rectum and N. granulata seem to regulate the consumption of diets containing more Ca, which suggests a fine balance for Ca intake.

Mechanism of acetylcholine-induced calcium signaling during neuronal differentiation of P19 embryonal carcinoma cells in vitro

Muscarinic (mAChRs) and nicotinic acetylcholine receptors (nAChRs) are involved in various physiological processes, including neuronal development. We provide evidence for expression of functional nicotinic and muscarinic receptors during differentiation of P19 carcinoma embryonic cells, as an in vitro model of early neurogenesis. We have detected expression and activity alpha(2)-alpha(7), beta(2), beta(4) nAChR and M1-M5 mAChR subtypes during neuronal differentiation. Nicotinic alpha(3) and beta(2) mRNA transcription was induced by addition of retinoic acid to P19 cells. Gene expression Of alpha(2), alpha(4)-alpha(7), beta(4) nAChR subunits decreased during initial differentiation and increased again when P19 cells underwent final maturation. Receptor response in terms of nicotinic agonist-evoked Ca2+, flux was observed in embryonic and neuronal-differentiated cells. Muscarinic receptor response, merely present in undifferentiated P19 cells, increased during neuronal differentiation. The nAChR-induced elevation of intracellular calcium ([Ca2+](i)) response in undifferentiated cells was due to Ca2+ influx. In differentiated P19 neurons the nAChR-induced [Ca2+](i) response was reduced following pretreatment with ryanodine, while the mAChR-induced response was unaffected indicating the contribution of Ca2+ release from ryanodine-sensitive stores to nAChR- but not mAChR-mediated Ca2+ responses. The presence of functional nAChRs in embryonic cells suggests that these receptors are involved in triggering Ca2+ waves during initial neuronal differentiation. (C) 2007 Elsevier Ltd. All rights reserved.

A vaccine against S. pyogenes: Design and experimental immune response

Streptococcus pyogenes causes severe invasive infections: the post-streptococcal sequelae of acute rheumatic fever (RF) and rheumatic heart disease (RHD), acute glomerulonephritis, and uncomplicated pharyngitis and pyoderma. Efforts to produce a vaccine against S. pyogenes began several decades ago, and different models have been proposed. Here, we describe the methodology used in the development of a new vaccine model, consisting of both T and B protective epitopes constructed as synthetic peptides and recombinant proteins. Two adjuvants were tested in an experimental inbred mouse model: a classical Freund`s adjuvant and a new adjuvant (AFCo1) that induces mucosal immune responses and is obtained by calcium precipitation of a proteoliposome derived from the outer membrane of Neisseria meningitides B. The StreptInCor vaccine epitope co-administrated with AFCo1 adjuvant induced mucosal (IgA) and systemic (IgG) antibodies as preferential Th1-mediated immune responses. No autoimmune reactions were observed, suggesting that the vaccine epitope is safe. (c) 2009 Elsevier Inc. All rights reserved.

Dosimetric properties of UV irradiated calcium co-doped borate glass-ceramic

The thermoluminescence (TL) response of Dy and Li doped 20CaB(4)O(7)-80CaB(2)O(4) (Wt%) glass-ceramic irradiated with ultraviolet (UV) radiation was studied. In order to act as TL activator ions, the Dy and Li ions were included in the matrix during the melting process to increase its TL efficiency. A single crystalline CaB2O4 phase was present in the glass-ceramic as determined by X-ray diffraction (XRD). The glass-ceramic 20CaB(4)O(7)-80CaB(2)O(4):Dy,Li wt% (named 20CBO7:Dy,Li) is a newly prepared TL material. Its thermoluminescent dosimetric characteristics have shown a linear response under UV radiation exposure and a good TL signal reproducibility, thus proving to be a promising material for using as an ultraviolet radiation dosimeter. (C) 2007 Elsevier B.V. All rights reserved.

Development of impedimetric and optical calcium biosensor by using modified gold electrode with porcine S100A12 protein

We describe the development of a label free method to analyze the interactions between Ca(2+) and the porcine S100A12 protein immobilized on polyvinyl butyral (PVB). The modified gold electrodes were characterized using cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), scanning electron microscopy (SEM) and surface plasmon resonance (SPR) techniques. SEM analyses of PVB and PVB-S100A12 showed a heterogeneous distribution of PVB spherules on gold surface. EIS and CV measurements have shown that redox probe reactions on the modified gold electrodes were partially blocked due the adsorption of PVB-S100A12, and confirm the existence of a positive response of the immobilized S100Al2 to the presence of calcium ions. The biosensor exhibited a wide linear response to Ca(2+) concentrations ranging from 12.5 to 200 mM. The PVB-S100A12 seems to be bound to the gold electrode surface by physical adsorption: we observed an increase of 1184.32 m degrees in the SPR angle after the adsorption of the protein on the PVB surface (in an indication that 9.84 ng of S100A12 are adsorbed per mm(2) of the Au-PVB electrode), followed by a further increase of 581.66 m degrees after attachment of the Ca(2+) ions. In addition, no SPR response is obtained for non-specific ions. These studies might be useful as a platform for the design of new reusable and sensitive biosensing devices that could find use in the clinical applications. (C) 2010 Elsevier B.V. All rights reserved.

Renal- and calcium-dependent vascular effects of Polybia paulista wasp venom

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Signal transducer and activator of transcription 3-regulated sarcoendoplasmic reticulurn Ca2+-ATPase 2 expression by prolactin and glucocorticoids is involved in the adaptation of insulin secretory response during the peripartum period

During pregnancy, the maternal endocrine pancreas undergoes, as a consequence of placental lactogens and prolactin (PR,L) action, functional changes that are characterized by increased glucose-induced insulin secretion. After delivery, the maternal endocrine pancreas rapidly returns to nonpregnant state, which is mainly attributed to the increased serum levels of glucocorticoids (GCs). Although GCs are known to decrease insulin secretion and counteract PRL action, the mechanisms for these effects are poorly understood. We have previously demonstrated that signal transducer and activator of transcription 3 (STAT3) is increased in islets treated with PRL. In the present study, we show that STAT3 expression and serine phosphorylation are increased in pancreatic islets at the end of pregnancy (P19). STAT3 serine phosphorylation rapidly returned to basal levels 3 days after delivery (U). The expression of the sarcoendoplasmic reticulum Ca2+-ATPase 2 (SERCA2), a crucial protein involved in the regulation of calcium handling in P-cells, was also increased in P19, returning to basal levels at L3. PRL increased SERCA2 and STAT3 expressions and STAT3 serine phosphorylation in RINm5F cells. The upregulation of SERCA2 by PRL was abolished after STAT3 knockdown. Moreover, PRL-induced STAT3 serine phosphorylation and SERCA2 expression were inhibited by dexamethasone (DEX). Insulin secretion from islets of PI 9 rats pre-incubated with thapsigargin and L3 rats showed a dramatic suppression of first phase of insulin release. The present results indicate that PRL regulates SERCA2 expression by a STAT3-dependent mechanism. PRL effect is counteracted by DEX and might contribute to the adaptation of maternal endocrine pancreas during the peripartum period.

Dosimetric properties of UV irradiated calcium co-doped borate glass-ceramic

The thermoluminescence (TL) response of Dy and Li doped 20CaB(4)O(7)-80CaB(2)O(4) (Wt%) glass-ceramic irradiated with ultraviolet (UV) radiation was studied. In order to act as TL activator ions, the Dy and Li ions were included in the matrix during the melting process to increase its TL efficiency. A single crystalline CaB2O4 phase was present in the glass-ceramic as determined by X-ray diffraction (XRD). The glass-ceramic 20CaB(4)O(7)-80CaB(2)O(4):Dy,Li wt% (named 20CBO7:Dy,Li) is a newly prepared TL material. Its thermoluminescent dosimetric characteristics have shown a linear response under UV radiation exposure and a good TL signal reproducibility, thus proving to be a promising material for using as an ultraviolet radiation dosimeter. (C) 2007 Elsevier B.V. All rights reserved.

Photopyroelectric response of PTCa/PEEK composite

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Mechanism of calcium hydroxide-induced neutrophil migration into air-pouch cavity

The aim of this study was to investigate cellular migration induced by calcium hydroxide to air-pouch cavities in mice. The migration was more specific to neutrophil and was dose and time dependent (peaking 96 h after stimulation). This migration was inhibited by pretreatment with thalidomide, indomethacin, MK886, meloxicam, dexamethasone, MK886 associated with indomethacin, and MK886 associated with indomethacin and dexamethasone. The air-pouch exudate from animals stimulated with calcium hydroxide showed an increase of leukotriene-B4 (LTB4), interleukin-1 beta, tumor necrosis factor alpha (TNF-alpha), cytokine-induced neutrophil chemoattractant (KC), and macrophage inflammatory protein 2 (MIP-2) release. Pretreatment with 3% thioglycollate increased the macrophage population in the air pouch but did not change neutrophil migration. Depleting the resident mast cells through chronic pretreatment with compound 48/80 did not alter neutrophil migration in response to calcium hydroxide. It was possible to conclude that calcium hydroxide-induced neutrophil migration to the air-pouch cavity in mice is mediated by LTB4, TNF-alpha, KC, MIP-2, and prostaglandins, but it was not dependent on macrophages or mast cells.

Analysis of the healing process in delayed tooth replantation after root canal filling with calcium hydroxide, Sealapex and Endofill: a microscopic study in rats

The major concern in the therapeutics of tooth replantation refers to the occurrence of root resorption and different approaches have been proposed to prevent or treat these complications. The purpose of this study was to evaluate tissue response to delayed replantation of anterior rat teeth treated endodontically using calcium hydroxide, Sealapex, and Endofill without the placement of gutta-percha cones. Thirty rats had their right upper incisor extracted and maintained in dry storage for 60 min. After removal of the dental papilla, enamel organ, pulp tissue, and periodontal ligament remnants, the teeth were immersed in 2% sodium fluoride phosphate acidulated, pH 5.5, for 10 min. The root canals were dried with absorbent paper points and the teeth were assigned to three groups (n = 10) according to the filling material. Group I - calcium hydroxide and propyleneglycol paste, Group II - Sealapex, and Group III - Endofill. The sockets were irrigated with saline and the teeth were replanted. Replacement resorption, inflammatory resorption and ankylosis were observed in all groups. Although the occurrence of inflammatory resorption was less frequent in Group I, there were no statistically significant differences among the groups. It may be concluded that compared to the paste, filling the root canals with Sealapex and Endofill sealers without the placement of gutta-percha cones did not provide better results.

Anticaries effect of dentifrices with calcium citrate and sodium trimetaphosphate

Because of the growing concerns regarding fluoride ingestion by young children and dental fluorosis, it is necessary to develop new dentifrices. Objective: The aim of this study was to evaluate the effect of dentifrices with calcium citrate (Cacit) and sodium trimetaphosphate (TMP) on enamel demineralization. Material and Methods: Enamel blocks (n=70), previously selected through surface hardness analysis, were submitted to daily treatment with dentifrices diluted in artificial saliva and to a pH-cycling model. The fluoride concentration in dentifrices was 0, 250, 450, 550, 1,000 and 1,100 mu g F/g. Crest (TM) was used as a positive control (1,100 mu g F/g). Cacit (0.25%) and TMP (0.25%) were added to dentifrices with 450 and 1,000 mu g F/g. Surface hardness was measured again and integrated loss of subsurface hardness and fluoride concentration in enamel were calculated. Parametric and correlation tests were used to determine difference (p<0.05) and dose-response relationship between treatments. Results: The addition of Cacit and TMP did not provide a higher fluoride concentration in enamel, however it reduced (p<0.05) mineral loss when compared to other dentifrices; the dentifrice with Cacit and TMP and a low fluoride concentration presented similar results when compared to a dentifrice with 1,100 mu g F/g (p>0.05). Conclusions: Dentifrices with 450 and 1,000 mu g F/g, Cacit and TMP were as effective as a gold standard one.

Evaluation of mineral trioxide aggregate and calcium hydroxide cement as pulp-capping agents in human teeth

This study evaluated the histomorphologic response of human dental pulps capped with mineral trioxide aggregate (MTA) and Ca(OH)(2) cement (CH). Pulp exposures were performed on the occlusal floor of 40 human permanent premolars. After that, the pulp was capped either with CH or MTA and restored with composite resin. After 30 and 60 days, teeth were extracted and processed for histologic exam and categorized in a histologic score system. The data were subjected to Kruskal-Wallis and Conover tests (alpha = .05). All groups performed well in terms of hard tissue bridge formation, inflammatory response, and other pulpal findings. However, a lower response of CH30 was observed for the dentin bridge formation, when compared with MTA30 and MTA60 groups. Although the pulp healing with calcium hydroxide was slower than that of MTA, both materials were successful for pulp capping in human teeth.

Short-term evaluation of the pulpo-dentin complex response to a resin-modified glass-ionomer cement and a bonding agent applied in deep cavities

Objectives. To evaluate the response of the pulpo-dentin complex following application of a resin-modified glass-ionomer cement or an adhesive system in deep cavities performed in human teeth.Methods. Deep class V cavities were prepared on the buccal surface of 26 premolars. In Group I the cavity walls (dentin) and enamel were conditioned with 32% phosphoric acid and the dentin adhesive system One Step (Bisco, Inc., Itasca, IL, USA) was applied. In Groups 2 and 3, before total etching and application of bonding agent, the cavity floor was lined with the resin-modified glass-ionomer cement-Vitrebond (3M ESPE Dental Products Division, St. Paul, MN, USA) or the calcium hydroxide cement-Dycal (control group, Dentsply, Mildford, DE, USA), respectively. The cavities were restored using light-cured Z-100 composite resin (3M ESPE). The teeth were extracted between 5 and 30 days and prepared for microscopic assessment. Serial sections were stained with H/E, Masson's trichrome, and Brown and Brenn techniques.Results. In Group 1, the inflammatory response was more evident than in Groups 2 and 3. Diffusion of dental material components across dentinal tubules was observed only in Group 1, in which the intensity of the pulp response increased as the remaining dentin thickness decreased. Bacteria were evidenced in the lateral walls of two samples (Group 2) which exhibited no inflammatory response or tissue disorganization.Conclusions. Based on the experimental conditions, it was concluded total acid etching followed by application of One Step bonding agent cannot be recommended as adequate procedures. In this clinical condition the cavity walls should be lined with a biocompatible dental material, such as Vitrebond or Dycal. 2003 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.